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Sample records for clinical isolates bovine

  1. Isolation of nuc mutant isolates of Staphylococcus aureus from bovine clinical mastitis.

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    Zastempowska, E; Orczykowska-Kotyna, M; Lassa, H

    2014-06-01

    Isolates of Staphylococcus aureus with a mutation in the nuclease (nuc) gene were recovered from cases of bovine mastitis in Poland. Three S. aureus isolates from cows in one herd had a 42 base pair duplication in the nuc gene. These isolates belonged to sequence type 97 (ST97) and clonal complex 97 (CC97). They had a different spa type and multiple-locus variable-number tandem-repeat fingerprinting (MLVF) subtype than a S. aureus isolate without the nuc mutation from the same herd. Isolation of nuc mutant S. aureus strains from cases of bovine mastitis may confound diagnostic PCRs based on detection of the nuc gene.

  2. Differences in Virulence Between Bovine-Derived Clinical Isolates of Pasteurella multocida Serotype A from the UK and the USA in a Model of Bovine Pneumonic Pasteurellosis.

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    Dagleish, M P; Bayne, C W; Moon, G G; Finlayson, J; Sales, J; Williams, J; Hodgson, J C

    2016-07-01

    The time of onset and subsequent degree and progression of clinical signs, bacterial colonization and tissue pathology during experimental disease induced by intratracheal inoculation of either a UK or USA isolate of Pasteurella multocida serotype A recovered from clinical cases of bovine pneumonia were determined. Calves aged 8 weeks were challenged with 300 ml phosphate buffered saline (PBS) alone (group 1, n = 3, negative control) or containing 7.1 × 10(8) colony forming units (cfu) of UK isolate (group 2, n = 8) or 5.8 × 10(8) cfu of USA isolate (group 3, n = 8). Bronchoalveolar lavage (BAL) at 0, 1 and 4 days post challenge (dpc) and at the time of necropsy examination (7-8 dpc) showed no significant differences between groups 2 and 3 in bacterial numbers recovered. No P. multocida were recovered from group 1 animals. No clinical disease was present in group 1 calves and in group 3 was limited to scour in 1 calf at 1 dpc. All calves in group 2 had reduced food intake at 4-5 dpc, five had periods of dullness, three a mild nasal discharge at 1 dpc, four had mild to substantial respiratory stridor and one was killed at 6 dpc for humane reasons. Rectal temperatures remained about 39°C in group 1 calves, but increased in P. multocida-challenged calves to 40-41°C within 8-12 h of challenge. Significantly (P = 0.01) greater percentages of lung surface area were consolidated in group 2 (mean ± SD, 21 ± 10.1) compared with group 3 (7 ± 8.6) calves. Significantly more extensive and severe histological lesions were present in the lung lobes (P = 0.006) and lymph nodes (P = 0.02) of group 2 compared with group 3 calves. Pleurisy was present in group 2 calves only and no pathology was present in group 1. Pulsed-field gel electrophoresis (PFGE) produced 11 (group 2, UK isolate) or 10 (group 3, USA isolate) bands with differences in banding patterns. Results overall showed that two isolates, distinct geographically and genetically (by PFGE

  3. Comparative molecular analysis of ovine and bovine Streptococcus uberis isolates.

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    Gilchrist, T L; Smith, D G E; Fitzpatrick, J L; Zadoks, R N; Fontaine, M C

    2013-02-01

    Streptococcus uberis causes clinical and subclinical mastitis in cattle and sheep, but it is unknown whether the composition of Strep. uberis populations differs between host species. To address this, we characterized a collection of bovine and ovine Strep. uberis isolates with shared geographical and temporal origins by means of an expanded multilocus sequence typing scheme. Among 14 ovine and 35 bovine isolates, 35 allelic profiles were detected. Each allelic profile was associated with a single host species and all but one were new to the multilocus sequence typing database. The median number of new alleles per isolate was higher for ovine isolates than for bovine isolates. None of the ovine isolates belonged to the global clonal complexes 5 or 143, which are commonly associated with bovine mastitis and which have a wide geographical distribution. Ovine isolates also differed from bovine isolates in carriage of plasminogen activator genes, with significantly higher prevalence of pauB in ovine isolates. Isolates that were negative for yqiL, one of the targets of multilocus sequence typing, were found among ovine and bovine isolates and were not associated with a specific sequence type or global clonal complex. One bovine isolate carried a gapC allele that was probably acquired through lateral gene transfer, most likely from Streptococcus salivarius. We conclude that ovine isolates are distinct from bovine isolates of Strep. uberis, and that recombination between isolates from different host species or bacterial species could contribute to changes in virulence gene profiles with relevance for vaccine development. PMID:23200465

  4. Observations on macrolide resistance and susceptibility testing performance in field isolates collected from clinical bovine respiratory disease cases.

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    DeDonder, Keith D; Harhay, Dayna M; Apley, Michael D; Lubbers, Brian V; Clawson, Michael L; Schuller, Gennie; Harhay, Gregory P; White, Brad J; Larson, Robert L; Capik, Sarah F; Riviere, Jim E; Kalbfleisch, Ted; Tessman, Ronald K

    2016-08-30

    The objectives of this study were; first, to describe gamithromycin susceptibility of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni isolated from cattle diagnosed with bovine respiratory disease (BRD) and previously treated with either gamithromycin for control of BRD (mass medication=MM) or sham-saline injected (control=CON); second, to describe the macrolide resistance genes present in genetically typed M. haemolytica isolates; third, use whole-genome sequencing (WGS) to correlate the phenotypic resistance and genetic determinants for resistance among M. haemolytica isolates. M. haemolytica (n=276), P. multocida (n=253), and H. somni (n=78) were isolated from feedlot cattle diagnosed with BRD. Gamithromycin susceptibility was determined by broth microdilution. Whole-genome sequencing was utilized to determine the presence/absence of macrolide resistance genes and to genetically type M. haemolytica. Generalized linear mixed models were built for analysis. There was not a significant difference between MM and CON groups in regards to the likelihood of culturing a resistant isolate of M. haemolytica or P. multocida. The likelihood of culturing a resistant isolate of M. haemolytica differed significantly by state of origin in this study. A single M. haemolytica genetic subtype was associated with an over whelming majority of the observed resistance. H. somni isolation counts were low and statistical models would not converge. Phenotypic resistance was predicted with high sensitivity and specificity by WGS. Additional studies to elucidate the relationships between phenotypic expression of resistance/genetic determinants for resistance and clinical response to antimicrobials are necessary to inform judicious use of antimicrobials in the context of relieving animal disease and suffering. PMID:27527782

  5. Prevalence and Characterization of Oxacillin Susceptible mecA-Positive Clinical Isolates of Staphylococcus aureus Causing Bovine Mastitis in India

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    Mistry, Hiral; Sharma, Paresh; Mahato, Sudipta; Saravanan, R.; Kumar, P. Anand; Bhandari, Vasundhra

    2016-01-01

    Bovine mastitis caused by multidrug resistant Staphylococcus aureus is a huge problem reported worldwide, resulting in prolonged antibiotic treatment and death of livestock. The current study is focused on surveillance of antibiotic susceptibility along with genotypic and phenotypic characterization of the pathogenic S. aureus strains causing mastitis in India. One hundred and sixty seven milk samples were collected from mastitis-affected cows from different farms in India resulting in thirty nine isolated S. aureus strains. Antibiotic sensitivity profiling revealed the majority of the strains (n = 24) to be multidrug resistant and eleven strains showed reduced susceptibility to vancomycin (MICs = 2μg/ml). All strains were oxacillin sensitive, but 19 strains were positive for the mecA gene, which revealed the occurrence of oxacillin susceptible mecA positive strains (OS-MRSA) for the first time from India. Additionally, 32 strains were positive for the pvl gene, a virulence determinant; of these 17 were also OS-MRSA strains. Molecular characterization based on multilocus sequence typing (MLST), spa typing, agr typing and SCCmec classification revealed strains belonging to different groups. Moreover, strains showed spa types (t2526, t9602) and MLST sequence types, ST-72, ST-88 and ST-239 which have been earlier reported in human infections. The prevalence of OS-MRSA strains indicates the importance of including both the genetic and phenotypic tests in characterizing S. aureus strains. Increased genotypic variability with strain related to human infections and pvl positive isolates indicates a worrisome situation with the possibility of bilateral transfer. PMID:27603123

  6. STAPHYLOCOCCAL MASTITIS IN BOVINES AND SOME PROPERTIES OF STAPHYLOCOCCAL ISOLATES

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    M. ARSHAD, G. MUHAMMAD1, M. SIDDIQUE, M. ASHRAF AND H.A. KHAN

    2006-01-01

    Full Text Available Milk samples of 100 clinically mastitic bovines (50 each from buffaloes and cows were processed for bacterial isolation. A total of 90 bacterial isolates, including 33 of staphylococci and 57 of other bacterial species were recovered. Of the 33 staphylococcal isolates, 23 were identified as Staphylococcus aureus and 10 as Staphylococcus epidermidis. All Staphylococcus aureus isolates were positive for deoxyribonuclease and mannitol fermentation test and produced ß-haemolysis on blood agar. All Staphylococcus epidermidis isolates were negative to these tests and failed to produce ß-haemolysis on blood agar containing 5% sheep erythrocytes. Penicillin resistance was detected in 13(56.5% isolates of Staphylococcus aureus and 8(80% isolates of Staphylococcus epidermidis. Penicillinase was produced by 4 of the 13 penicillin resistant isolates of Staphylococcus aureus and 2 of the 8 penicillin resistant Staphylococcus epidermidis isolates.

  7. Puesta en evidencia del virus diarrea viral bovina en bovinos clínicamente afectados Isolation of the bovine viral diarrhoea virus from tissue of clinically affected cattle

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    M.O CELEDON

    1997-01-01

    Full Text Available Para conocer la presencia del virus diarrea viral bovina (VDVB en animales sospechosos de estar cursando un cuadro clínico provocado por este virus, se trabajó con un total de 33 animales, correspondiendo a 23 fetos abortados, 2 mortinatos, un nonato, 3 vacas: una madre de mortinato, una madre de aborto y una muerta, 2 novillos muertos y 2 terneros muertos. Muestras de órganos se inocularon en cultivos primarios de pulmón fetal bovino (PFB y en la línea MDBK. Después del primer pasaje en células de PFB, se detectó la presencia de antígenos del VDVB por la prueba de inmunoperoxidasa indirecta (IPI. Todas las muestras con reacción positiva a IPI se inocularon por segunda y tercera vez en células de PFB, aplicándose la prueba de IPI en el tercer pasaje. Sobre un cuarto pasaje se aplicó la prueba de inmunofluorescencia direccta (IFD. Todas las muestras, positivas y negativas a IPI, se inocularon en 3 pasajes seriados en las células MDBK. En 23 de los 33 animales se aisló VDVB cepas no citopatogénicas (NCP, correspondiendo a 14 fetos abortados, un nonato, un mortinato, 3 vacas, 2 novillos y 2 terneros. En 6 fetos abortados, independiente de los infectados con el VDVB, se aisló el virus de la rinotraqueítis infecciosa bovina (RIB. Se concluye que la presencia del VDVB es de alta frecuencia en muestras clínicas de ganado bovino con patologías asociables al VDVB, desconociéndose el rol patógeno del virus en estos aislados.Cattle infected with the bovine viral diarrhoea (BVD virus can present a variety of clinical signs. This research studied the presence of BVD virus in cattle by virus isolation in primary cell cultures of bovine embryo lungs. Virus identification was done using the immunoperoxidase staining assay and the direct fluorescent antibody staining. As a result, 23 out of 33 animals were identified as positive to BVD virus: 14 foetal abortions, 2 stillbirths, 3 dams, 2 steers and 2 calves. No cytopathogenic isolates were

  8. A peptide derived from human bactericidal/permeability-increasing protein (BPI) exerts bactericidal activity against Gram-negative bacterial isolates obtained from clinical cases of bovine mastitis

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    Gram-negative bacteria are responsible for approximately one-third of the clinical cases of bovine mastitis and can elicit a life-threatening, systemic inflammatory response. Lipopolysaccharide (LPS) is a membrane component of all Gram-negative bacteria and is largely responsible for evoking the de...

  9. Clinical applications of bovine colostrum therapy

    DEFF Research Database (Denmark)

    Rathe, Mathias; Müller, Klaus; Sangild, Per Torp;

    2014-01-01

    Bovine colostrum, the first milk that cows produce after parturition, contains high levels of growth factors and immunomodulatory components. Some healthy and diseased individuals may gain health benefits by consuming bovine colostrum as a food supplement. This review provides a systematic...... to populations, outcomes, and methodological quality, as judged by the Jadad assessment tool. Many studies used surrogate markers to study the effects of bovine colostrum. Studies suggesting clinical benefits of colostrum supplementation were generally of poor methodological quality, and results could...... not be confirmed by other investigators. Bovine colostrum may provide gastrointestinal and immunological benefits, but further studies are required before recommendations can be made for clinical application. Animal models may help researchers to better understand the mechanisms of bovine colostrum supplementation...

  10. Antibacterial Effect of Copper on Microorganisms Isolated from Bovine Mastitis.

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    Reyes-Jara, Angelica; Cordero, Ninoska; Aguirre, Juan; Troncoso, Miriam; Figueroa, Guillermo

    2016-01-01

    The antimicrobial properties of copper have been recognized for several years; applying these properties to the prevention of diseases such as bovine mastitis is a new area of research. The aim of the present study was to evaluate in vitro the antimicrobial activity of copper on bacteria isolated from subclinical and clinical mastitis milk samples from two regions in Chile. A total of 327 microorganisms were recovered between March and September 2013, with different prevalence by sample origin (25 and 75% from the central and southern regions of Chile, respectively). In the central region, Escherichia coli and coagulase negative Staphylococci (CNS) were the most frequently detected in clinical mastitis cases (33%), while in the southern region S. uberis, S. aureus, and CNS were detected with frequencies of 22, 21, and 18%, respectively. Antibiotic susceptibility studies revealed that 34% of isolates were resistant to one or more antibiotics and the resistance profile was different between bacterial species and origins of isolation of the bacteria. The minimum inhibitory concentration of copper (MIC-Cu) was evaluated in all the isolates; results revealed that a concentration as low as 250 ppm copper was able to inhibit the great majority of microorganisms analyzed (65% of isolates). The remaining isolates showed a MIC-Cu between 375 and 700 ppm copper, and no growth was observed at 1000 ppm. A linear relationship was found between the logarithm of viable bacteria number and time of contact with copper. With the application of the same concentration of copper (250 ppm), CNS showed the highest tolerance to copper, followed by S. uberis and S. aureus; the least resistant was E. coli. Based on these in vitro results, copper preparations could represent a good alternative to dipping solutions, aimed at preventing the presence and multiplication of potentially pathogenic microorganisms involved in bovine mastitis disease. PMID:27199953

  11. Antibacterial Effect of Copper on Microorganisms Isolated from Bovine Mastitis

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    Reyes-Jara, Angelica; Cordero, Ninoska; Aguirre, Juan; Troncoso, Miriam; Figueroa, Guillermo

    2016-01-01

    The antimicrobial properties of copper have been recognized for several years; applying these properties to the prevention of diseases such as bovine mastitis is a new area of research. The aim of the present study was to evaluate in vitro the antimicrobial activity of copper on bacteria isolated from subclinical and clinical mastitis milk samples from two regions in Chile. A total of 327 microorganisms were recovered between March and September 2013, with different prevalence by sample origin (25 and 75% from the central and southern regions of Chile, respectively). In the central region, Escherichia coli and coagulase negative Staphylococci (CNS) were the most frequently detected in clinical mastitis cases (33%), while in the southern region S. uberis, S. aureus, and CNS were detected with frequencies of 22, 21, and 18%, respectively. Antibiotic susceptibility studies revealed that 34% of isolates were resistant to one or more antibiotics and the resistance profile was different between bacterial species and origins of isolation of the bacteria. The minimum inhibitory concentration of copper (MIC-Cu) was evaluated in all the isolates; results revealed that a concentration as low as 250 ppm copper was able to inhibit the great majority of microorganisms analyzed (65% of isolates). The remaining isolates showed a MIC-Cu between 375 and 700 ppm copper, and no growth was observed at 1000 ppm. A linear relationship was found between the logarithm of viable bacteria number and time of contact with copper. With the application of the same concentration of copper (250 ppm), CNS showed the highest tolerance to copper, followed by S. uberis and S. aureus; the least resistant was E. coli. Based on these in vitro results, copper preparations could represent a good alternative to dipping solutions, aimed at preventing the presence and multiplication of potentially pathogenic microorganisms involved in bovine mastitis disease. PMID:27199953

  12. Clinical manifestations of bovine leukosis

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    Sorensen, D.K.

    1979-01-01

    The diagnosis of animals infected with BLV can be accurately identified with the available serologic tests. Diagnosis of animals in the incipient stage of leukosis is extremely difficult and can only be diagnosed by a positive tissue biopsy. Animals with frank tumor involvement can be suspected and diagnosed on a tentative clinical basis on the signs reported. Positive diagnosis must be made on the basis of a biopsy of the tumor or in some cases on a hemotological examination.

  13. Genetic characterization of a noncytopathic bovine viral diarrhea virus 2b isolated from cattle in China.

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    Wang, Wei; Shi, Xinchuan; Chen, Chaoyang; Wu, Hua

    2014-10-01

    In January 2013, several clinical signs of cattle with diarrhea, cough, nasal discharge, and fever were reported in Jilin province, China. One virus named SD1301 was isolated and identified. Complete genome of the virus is 12258nt in length and contains a 5'UTR, one open reading frame encoding a polyprotein of 3,897 amino acids and a 3'UTR. Phylogenetic analysis of 5'UTR, N(pro), E1 and E2 gene demonstrated the virus belonged to BVDV 2b, and genetically related to the BVDV strain Hokudai-Lab/09 from Japan in 2010. This bovine viral diarrhea virus displays a unique genetic signature with 27-nucleotide deletion in the 5'UTR, which is similar to the bovine viral diarrhea virus C413 (AF002227). This was the first confirmed isolation of ncp BVDV2b circulating in bovine herd of China.

  14. Prevalence of bacterial agents isolated from clinical cases of bovine mastitis in the dry period and the determination of their antibiotic sensitivity in Tabriz, Iran

    Institute of Scientific and Technical Information of China (English)

    Samad Mosaferi; Reza Ghabouli Mehrabani; Mansoor Khakpoor; Nader Ghabouli Mehrabani; Amir Maleksabet; Faezeh Hamidi

    2015-01-01

    Objective: To determine the prevalence of mastitis-causing bacteria in the dry period and its antibiotic sensitivity. Methods: In this study, 852 dry cows were examined. A total of 30 cows with clinical mastitis symptoms were detected and their milk samples were collected. In order to purify the bacteria, brain heart infusion and blood agar media were applied and single colonies were used for Gram staining, oxidase and catalase testing, cultivating in O-F medium to determine the genus and species of bacteria. Then, antimicrobial susceptibility was tested by the agar disk diffusion method. Results: The prevalence of isolated bacteria was 2.46%, in which coagulase positive Staphylococcus, coagulase negative Staphylococcus, Streptococcus dysgalactiae, Streptococcus faecalis, Escherichia coli, Pseudomonas, Bacillus and yeast were (9/99)%, (6/66)%, (13/32)%, (3/33%), (6/66)%, (13/32)%, (9/99)% and (6/66)%, respectively. After tests of antibiotic susceptibility, the most and the least sensitivity were reported to enrofloxacin and ampicillin respectively. Conclusions: This study indicated that Streptococcus dysgalactiae is the most commonly isolated bacteria with the greatest sensitivity to enrofloxacin and tetracycline which can be used to treat mastitis in the dry period in Tabriz.

  15. Prevalence of bacterial agents isolated from clinical cases of bovine mastitis in the dry period and the determination of their antibiotic sensitivity in Tabriz, Iran

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    Samad Mosaferi

    2015-09-01

    Full Text Available Objective: To determine the prevalence of mastitis-causing bacteria in the dry period and its antibiotic sensitivity. Methods: In this study, 852 dry cows were examined. A total of 30 cows with clinical mastitis symptoms were detected and their milk samples were collected. In order to purify the bacteria, brain heart infusion and blood agar media were applied and single colonies were used for Gram staining, oxidase and catalase testing, cultivating in O-F medium to determine the genus and species of bacteria. Then, antimicrobial susceptibility was tested by the agar disk diffusion method. Results: The prevalence of isolated bacteria was 2.46%, in which coagulase positive Staphylococcus, coagulase negative Staphylococcus, Streptococcus dysgalactiae, Streptococcus faecalis, Escherichia coli, Pseudomonas, Bacillus and yeast were (9/99%, (6/66%, (13/32%, (3/33%, (6/66%, (13/32%, (9/99% and (6/66%, respectively. After tests of antibiotic susceptibility, the most and the least sensitivity were reported to enrofloxacin and ampicillin respectively. Conclusions: This study indicated that Streptococcus dysgalactiae is the most commonly isolated bacteria with the greatest sensitivity to enrofloxacin and tetracycline which can be used to treat mastitis in the dry period in Tabriz.

  16. Diversity and antimicrobial susceptibility profiling of staphylococci isolated from bovine mastitis cases and close human contacts.

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    Schmidt, T; Kock, M M; Ehlers, M M

    2015-09-01

    The objectives of this study were to examine the diversity of Staphylococcus spp. recovered from bovine intramammary infections and humans working in close contact with the animals and to evaluate the susceptibility of the staphylococcal isolates to different antimicrobials. A total of 3,387 milk samples and 79 human nasal swabs were collected from 13 sampling sites in the KwaZulu-Natal province of South Africa. In total, 146 Staph. aureus isolates and 102 coagulase-negative staphylococci (CNS) were recovered from clinical and subclinical milk samples. Staphylococcusaureus was isolated from 12 (15.2%) of the human nasal swabs and 95 representative CNS were recovered for further characterization. The CNS were identified using multiplex-PCR assays, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and tuf gene sequencing. Seven Staphylococcus spp. were identified among the CNS of bovine origin, with Staph.chromogenes (78.4%) predominating. The predominant CNS species recovered from the human nasal swabs was Staph.epidermidis (80%) followed by Staph.chromogenes (6.3%). The antimicrobial susceptibility of all staphylococcal isolates was evaluated using disk diffusion and was supplemented by screening for specific antimicrobial resistance genes. Ninety-eight (67.1%) Staph.aureus isolates of bovine origin were pansusceptible; 39 (26.7%) isolates were resistant to a single class, and 7 (4.8%) isolates were resistant to 2 classes of antimicrobials. Two Staph. aureus (1.4%) isolates were multidrug-resistant. Resistance to penicillin was common, with 28.8% of the bovine and 75% of the human Staph. aureus isolates exhibiting resistance. A similar observation was made with the CNS, where 37.3% of the bovine and 89.5% of the human isolates were resistant to penicillin. Multidrug-resistance was common among the human CNS, with 39% of the isolates exhibiting resistance to 3 or more classes of antimicrobials. The antimicrobial

  17. Bovine immunoglobulin protein isolates for the nutritional management of enteropathy.

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    Petschow, Bryon W; Blikslager, Anthony T; Weaver, Eric M; Campbell, Joy M; Polo, Javier; Shaw, Audrey L; Burnett, Bruce P; Klein, Gerald L; Rhoads, J Marc

    2014-09-01

    The gastrointestinal tract is responsible for a multitude of digestive and immune functions which depend upon the balanced interaction of the intestinal microbiota, diet, gut barrier function, and mucosal immune response. Disruptions in one or more of these factors can lead to intestinal disorders or enteropathies which are characterized by intestinal inflammation, increased gut permeability, and reduced capacity to absorb nutrients. Enteropathy is frequently associated with human immunodeficiency virus (HIV) infection, inflammatory bowel disease, autoimmune enteropathy, radiation enteritis, and irritable bowel syndrome (IBS), where pathologic changes in the intestinal tract lead to abdominal discomfort, bloating, abnormal bowel function (e.g., diarrhea, urgency, constipation and malabsorption). Unfortunately, effective therapies for the management of enteropathy and restoring intestinal health are still not available. An accumulating body of preclinical studies has demonstrated that oral administration of plasma- or serum-derived protein concentrates containing high levels of immunoglobulins can improve weight, normalize gut barrier function, and reduce the severity of enteropathy in animal models. Recent studies in humans, using serum-derived bovine immunoglobulin/protein isolate, demonstrate that such protein preparations are safe and improve symptoms, nutritional status, and various biomarkers associated with enteropathy. Benefits have been shown in patients with HIV infection or diarrhea-predominant IBS. This review summarizes preclinical and clinical studies with plasma/serum protein concentrates and describes the effects on host nutrition, intestinal function, and markers of intestinal inflammation. It supports the concept that immunoglobulin-containing protein preparations may offer a new strategy for restoring functional homeostasis in the intestinal tract of patients with enteropathy. PMID:25206275

  18. Genetic Features Differentiating Bovine, Food, and Human Isolates of Shiga Toxin-Producing Escherichia coli O157 in The Netherlands

    NARCIS (Netherlands)

    Franz, E.; Hoek, van A.H.A.M.; Wal, van der F.J.; Boer, de A.G.; Zwartkruis-Nahuis, A.; Zwaluw, van der K.; Aarts, H.J.M.; Heuvelink, A.E.

    2012-01-01

    The frequency of Escherichia coli O157 genotypes among bovine, food, and human clinical isolates from The Netherlands was studied. Genotyping included the lineage-specific polymorphism assay (LSPA6), the Shiga-toxin-encoding bacteriophage insertion site assay (SBI), and PCR detection and/or subtypin

  19. Clinical applications of bovine colostrum therapy: a systematic review.

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    Rathe, Mathias; Müller, Klaus; Sangild, Per Torp; Husby, Steffen

    2014-04-01

    Bovine colostrum, the first milk that cows produce after parturition, contains high levels of growth factors and immunomodulatory components. Some healthy and diseased individuals may gain health benefits by consuming bovine colostrum as a food supplement. This review provides a systematic, critical evaluation of the current state of knowledge in this area. Fifty-one eligible studies were identified from the following databases: Medline, Embase, Global Health, the Cochrane Library, and the Cumulative Index to Nursing and Allied Health Literature. Studies were heterogeneous with regard to populations, outcomes, and methodological quality, as judged by the Jadad assessment tool. Many studies used surrogate markers to study the effects of bovine colostrum. Studies suggesting clinical benefits of colostrum supplementation were generally of poor methodological quality, and results could not be confirmed by other investigators. Bovine colostrum may provide gastrointestinal and immunological benefits, but further studies are required before recommendations can be made for clinical application. Animal models may help researchers to better understand the mechanisms of bovine colostrum supplementation, the dosage regimens required to obtain clinical benefits, and the optimal methods for testing these effects in humans. PMID:24571383

  20. Isolation, identification, and complete genome sequence of a bovine adenovirus type 3 from cattle in China

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    Zhu Yuan-Mao

    2011-12-01

    Full Text Available Abstract Background Bovine adenovirus type 3 (BAV-3 belongs to the Mastadenovirus genus of the family Adenoviridae and is involved in respiratory and enteric infections of calves. The isolation of BAV-3 has not been reported prior to this study in China. In 2009, there were many cases in cattle showing similar clinical signs to BAV-3 infection and a virus strain, showing cytopathic effect in Madin-Darby bovine kidney cells, was isolated from a bovine nasal swab collected from feedlot cattle in Heilongjiang Province, China. The isolate was confirmed as a bovine adenovirus type 3 by PCR and immunofluorescence assay, and named as HLJ0955. So far only the complete genome sequence of prototype of BAV-3 WBR-1 strain has been reported. In order to further characterize the Chinese isolate HLJ0955, the complete genome sequence of HLJ0955 was determined. Results The size of the genome of the Chinese isolate HLJ0955 is 34,132 nucleotides in length with a G+C content of 53.6%. The coding sequences for gene regions of HLJ0955 isolate were similar to the prototype of BAV-3 WBR-1 strain, with 80.0-98.6% nucleotide and 87.5-98.8% amino acid identities. The genome of HLJ0955 strain contains 16 regions and four deletions in inverted terminal repeats, E1B region and E4 region, respectively. The complete genome and DNA binding protein gene based phylogenetic analysis with other adenoviruses were performed and the results showed that HLJ0955 isolate belonged to BAV-3 and clustered within the Mastadenovirus genus of the family Adenoviridae. Conclusions This is the first study to report the isolation and molecular characterization of BAV-3 from cattle in China. The phylogenetic analysis performed in this study supported the use of the DNA binding protein gene of adenovirus as an appropriate subgenomic target for the classification of different genuses of the family Adenoviridae on the molecular basis. Meanwhile, a large-scale pathogen and serological epidemiological

  1. Infection of human keratinocytes by Streptococcus dysgalactiae subspecies dysgalactiae isolated from milk of the bovine udder.

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    Roma-Rodrigues, Catarina; Alves-Barroco, Cynthia; Raposo, Luís R; Costa, Mafalda N; Fortunato, Elvira; Baptista, Pedro Viana; Fernandes, Alexandra R; Santos-Sanches, Ilda

    2016-04-01

    Streptococcus dysgalactiae subsp. dysgalactiae (SDSD) are considered exclusive animal pathogens; however, a putative zoonotic upper limb cellulitis, a prosthetic joint infection and an infective endocarditis were described in humans. To unravel if bovine SDSD isolates are able to infect human cells, the adherence and internalization to human primary keratinocytes of two bovine SDSD strains isolated from milk collected from udder were analyzed. Bacterial adhesion assays and confocal microscopy indicate a high adherence and internalization of SDSD isolates to human cells, suggesting for the first time the ability of bovine isolates to infect human cells. PMID:26655883

  2. Assembly and structure of neurofilaments isolated from bovine spinal cord

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    佟向军; 陈建国; 刘洁; 庞世瑾; 翟中和

    1999-01-01

    Neurofilaments (NFs) are neuron-specific intermediate filaments. The NFs were isolated from bovine spinal cord by differential centrifugation. The NFs were detected with electron microscopy and scanning tunneling microscopy (STM). Under STM, two kinds of sidearm of NFs were revealed: one was short, the other was long. They were arrayed along the 10-nm width core filaments one by one. The intervals between two adjacent long sidearms or two short sidearms were 20—22 nm, while those between two adjacent long and short sidearms were 10—11 nm. It was proposed that the rod domain of NF triplet prnteins was 3/4-staggered. The assembly properties of NF triplet proteins were also studied. Immuno-colloidal-gold labeling assay showed that NF-M and NF-H are able to co-assemble into long filaments with NF-L. NF-M and NF-H can also co-constitute into winding filaments.

  3. Phage types and antimicrobial resistance among Danish bovine Staphylococcus aureus isolates since the 1950s

    DEFF Research Database (Denmark)

    Vintov, Jan; Aarestrup, Frank Møller; Zinn, C. E.;

    2003-01-01

    A total of 292 bovine Staphylococcus aureus isolates obtained from the 1950s (86 isolates), 1992 (107 isolates), and 2000 (99 isolates) were examined for antimicrobial susceptibility and phage typing. The same types of S. aureus (80, 52, 3A, 3A/3C, 42E, 77) were found among the isolates from all...... three time periods, representing 43.3% of the typeable isolates. This indicates that the Danish S. aureus population related to bovine mastitis has remained relatively unchanged over the last 50 years. The occurrence of antimicrobial resistance has remained low in Denmark in comparison to other...

  4. Clinically isolated laryngeal sarcoidosis

    DEFF Research Database (Denmark)

    Plaschke, Christina Caroline; Owen, Hanne Hoejris; Rasmussen, Niels

    2011-01-01

    to a combination of CO(2)-laser excision of supraglottic tissue and closure of the incision with sutures. All serological tests were negative or normal, including angiotensin 1 converting enzyme. The clinical expression was uniform with pale, smooth swellings of the supraglottic structures. Surgery proved...

  5. Viral infections and bovine mastitis: a review

    NARCIS (Netherlands)

    Wellenberg, G.J.; Poel, van der W.H.M.; Oirschot, van J.T.

    2002-01-01

    This review deals with the role of viruses in the aetiology of bovine mastitis. Bovine herpesvirus 1, bovine herpesvirus 4, foot-and-mouth disease virus, and parainfluenza 3 virus have been isolated from milk from cows with clinical mastitis. Intramammary inoculations of bovine herpesvirus 1 or para

  6. Genomic and Evolutionary Analysis of Two Salmonella enterica Serovar Kentucky Sequence Types Isolated from Bovine and Poultry Sources in North America

    Science.gov (United States)

    Haley, Bradd J.; Kim, Seon Woo; Pettengill, James; Luo, Yan; Karns, Jeffrey S.; Van Kessel, Jo Ann S.

    2016-01-01

    Salmonella enterica subsp. enterica serovar Kentucky is frequently isolated from healthy poultry and dairy cows and is occasionally isolated from people with clinical disease. A genomic analysis of 119 isolates collected in the United States from dairy cows, ground beef, poultry and poultry products, and human clinical cases was conducted. Results of the analysis demonstrated that the majority of poultry and bovine-associated S. Kentucky were sequence type (ST) 152. Several bovine-associated (n = 3) and food product isolates (n = 3) collected from the United States and the majority of human clinical isolates were ST198, a sequence type that is frequently isolated from poultry and occasionally from human clinical cases in Northern Africa, Europe and Southeast Asia. A phylogenetic analysis indicated that both STs are more closely related to other Salmonella serovars than they are to each other. Additionally, there was strong evidence of an evolutionary divergence between the poultry-associated and bovine-associated ST152 isolates that was due to polymorphisms in four core genome genes. The ST198 isolates recovered from dairy farms in the United States were phylogenetically distinct from those collected from human clinical cases with 66 core genome SNPs differentiating the two groups, but more isolates are needed to determine the significance of this distinction. Identification of S. Kentucky ST198 from dairy animals in the United States suggests that the presence of this pathogen should be monitored in food-producing animals. PMID:27695032

  7. Extensive sequence divergence among bovine respiratory syncytial viruses isolated during recurrent outbreaks in closed herds

    DEFF Research Database (Denmark)

    Larsen, Lars Erik; Tjørnehøj, Kirsten; Viuff, B.

    2000-01-01

    The nucleotides coding for the extracellular part of the G glycoprotein and the full SH protein of bovine respiratory syncytial virus (BRSV) were sequenced from viruses isolated from numerous outbreaks of BRSV infection. The isolates included viruses isolated from the same herd (closed dairy farms...

  8. Antimicrobial susceptibilities and bacteriological characteristics of bovine Pseudomonas aeruginosa and Serratia marcescens isolates from mastitis.

    Science.gov (United States)

    Ohnishi, Mamoru; Sawada, Takuo; Hirose, Kazuhiko; Sato, Reiichiro; Hayashimoto, Mizuki; Hata, Eiji; Yonezawa, Chizuko; Kato, Hajime

    2011-12-29

    The presence of metallo-β-lactamase (MBL)-producing and multidrug-resistant Pseudomonas aeruginosa (MDRP) strains among bovine isolates of Gram-negative bacilli, and O-serotypes of bovine Serratia marcescens and P. aeruginosa isolates have been reported rarely. The aims of this study were to (1) elucidate antimicrobial susceptibilities and O-serotypes of P. aeruginosa and S. marcescens isolates from bovine mastitis and the presence of MBL-producers and MDRP strains among them and (2) evaluate their relationships to human isolates. We investigated the MICs of 24 antimicrobials and O-serotypes for 116 P. aeruginosa and 55 S. marcescens isolates in Japan, primarily in 2006. A total of 171 isolates exhibited high antimicrobial susceptibilities with the exception of a partial drug. P. aeruginosa isolates exhibited high susceptibilities of ≥ 95.7% to ciprofloxacin, imipenem, meropenem, piperacillin, ceftazidime, cefepime, cefoperazone/sulbactam, amikacin, tobramycin, and gentamicin; however, they exhibited a susceptibility of only 69.8% to aztreonam. They exhibited substantial resistances to ceftriaxone, enrofloxacin, cefotaxime, and moxalactam. S. marcescens isolates exhibited high susceptibilities of ≥ 90.9% to kanamycin, ceftiofur, sulfamethoxazole-trimethoprim, and the 15 aforementioned drugs, but exhibited resistance to minocycline. Neither MBL-producers nor MDRP strains were detected among the 171 strains. The dominant serotypes of P. aeruginosa isolates were OG, OA, OB, OI, OF, OE, and OK; those of S. marcescens isolates were O6 and O5. Every S. marcescens isolate was pigmented. These findings suggest that bovine P. aeruginosa and S. marcescens isolates differ from human isolates from both antibiogram and phenotypic perspectives, and could help to evaluate differences in bacteriological characteristics between bovine and human isolates.

  9. Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR.

    Science.gov (United States)

    Ali, Shahzad; Ali, Qurban; Melzer, Falk; Khan, Iahtasham; Akhter, Shamim; Neubauer, Heinrich; Jamal, Syed M

    2014-01-01

    Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell's serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n = 5), aborted fetuses (n = 13), and vaginal swabs (n = 12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.

  10. Pathogenicity of local isolate virus BHV-1 as the aetiological agent of Infectious Bovine Rhinotracheitis in Bali Cattle

    OpenAIRE

    Rini I Damayanti; Sudarisman

    2005-01-01

    Infectious Bovine Rhinotracheitis is a disease of cattle characterised by clinical signs of the upper respiratory tract, reproductive tract and nervous system. A study to define the pathogenicity of four BHV-1 local isolates has been conducted. Fourteen Bali cattle that were free of BHV-1 has been selected and divided into four treatment groups. Each group of three was infected with virus isolate I, II, III and IV respectively with approximately a dose of 108TCID50 /10 ml and two cattle were ...

  11. Antigenic variability in bovine viral diarrhea virus (BVDV) isolates from alpaca (Vicugna pacos), llama (Lama glama) and bovines in Chile.

    Science.gov (United States)

    Aguirre, I M; Quezada, M P; Celedón, M O

    2014-01-31

    Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities where they have been introduced worldwide. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus and mainly to bovine viral diarrhea virus (BVDV). Seventeen Chilean BVDV isolates were analyzed by serum cross neutralization with samples obtained from five llama, six alpacas, three bovines, plus three reference strains belonging to different subgroups and genotypes. The objective was to describe antigenic differences and similarities among them. Antigenic comparison showed significant differences between different subgroups. Consequently, antigenic similarities were observed among isolates belonging to the same subgroup and also between isolates from different animal species belonging the same subgroup. Among the analyzed samples, one pair of 1b subgroup isolates showed significant antigenic differences. On the other hand, one pair of isolates from different subgroups (1b and 1j) shared antigenic similarities indicating antigenic relatedness. This study shows for the first time the presence of antigenic differences within BVDV 1b subgroup and antigenic similarities within 1j subgroup isolates, demonstrating that genetic differences within BVDV subgroups do not necessary corresponds to differences on antigenicity.

  12. Isolation and Genetic Analysis of Bovine Viral Diarrhea Virus from Infected Cattle in Indiana

    Directory of Open Access Journals (Sweden)

    Roman M. Pogranichniy

    2011-01-01

    Full Text Available Species and biotype distribution was determined in 44 bovine viral diarrhea virus- (BVDV- positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL in Indiana during 2006–2008. BVDV RNA was detected in the 5′-untranslated region and Npro region using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory distress, reproductive, persistent infection (PI, and mucosal disease (MD. Of 44 BVDV-positive samples, 33 were noncytopathic (ncp, 10 were cytopathic (cp, and one presented both ncp and cp biotypes. Sequencing analysis demonstrated that all samples belonged to BVDV-1a, BVDV-1b, or BVDV-2. The most common isolate was ncp BVDV-1b, (44% followed by ncp BVDV-2a (24%. Among the six categories, respiratory clinical signs were the most common (36% followed by PI (25% and MD (16%.

  13. The molecular structure and lubricating activity of lubricin isolated from bovine and human synovial fluids.

    Science.gov (United States)

    Swann, D A; Silver, F H; Slayter, H S; Stafford, W; Shore, E

    1985-01-01

    Lubricin was isolated from bovine ankle, metacarpophalangeal and knee and human knee synovial fluids. The lubricins isolated from the bovine joint fluids had the same amino acid and carbohydrate compositions, but differences were observed in the relative molecular masses. The Mr values of bovine metacarpophalangeal and ankle lubricin determined by light-scattering measurements were about 200 000, whereas values of 132 000 and 143 000 were obtained for the bovine knee lubricin. The human knee lubricin had a similar carbohydrate composition to bovine knee lubricin except for the higher glucosamine content, and the amino acid composition differed slightly. The human sample had a lower glutamic acid content and a leucine/isoleucine ratio of 2:1 compared with 1:1 in the bovine. The Mr value of the human knee lubricin (166 000) was also lower than that of the bovine metacarpophalangeal and ankle samples. The Mr value of the bovine knee lubricin determined by sedimentation-equilibrium measurements was 171 000. The length measurements determined by electron microscopy and also the sedimentation measurements showed considerable polydispersity and indicate that the degree of extension of lubricin molecules can vary. Friction measurements showed that the human knee synovial-fluid lubricin had equivalent lubricating ability in a test system in vitro to that observed for lubricin isolated from normal bovine synovial fluids. The lubricating ability of lubricin was concentration-dependent, and each lubricin sample was able to act as a lubricant in vitro in an equivalent manner to whole synovial fluid at concentrations that are thought to occur in vivo. PMID:3977823

  14. Distribution of Serotypes and Antimicrobial Resistance Genes among Streptococcus agalactiae Isolates from Bovine and Human Hosts

    OpenAIRE

    Dogan, Belgin; Schukken, Y. H.; de Santisteban, C.; Boor, Kathryn J.

    2005-01-01

    To better understand the emergence and transmission of antibiotic-resistant Streptococcus agalactiae, we compared phenotypic and genotypic characteristics of 52 human and 83 bovine S. agalactiae isolates. Serotypes found among isolates from human hosts included V (48.1%), III (19.2%), Ia and Ib (13.5% each), and II (5.8%). Among isolates from bovine hosts, molecular serotypes III and II were predominant (53 and 14.5%, respectively). Four and 21 different ribotypes were found among human and b...

  15. Characterization of Staphylococcus simulans strains isolated from cases of bovine mastitis

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Larsen, H. D.; Jensen, N. E.

    1999-01-01

    This study was conducted to characterize Staphylococcus simulans isolated from cases of bovine mastitis. A total of 134 isolates of S. simulans selected from 80 quarters from 61 cows or heifers in 37 different herds were characterized by EcoRI ribotyping. From 22 quarters two to seven consecutive......, where 76 paired or multiple isolates were at disposal, the same ribotype was constantly found in the same quarter. This study showed that S. simulans causing bovine mastitis could be divided into relatively large number of different types, but that two types predominated. More than one type could...

  16. Species distribution and resistance profiles of coagulase-negative staphylococci isolated from bovine mastitis in Switzerland

    OpenAIRE

    Moser, A.; Stephan, R.; DE ZIEGLER, D.; Johler, S.

    2013-01-01

    Coagulase-negative staphylococci (CNS) are the predominant cause of bovine intra-mammary infections. They can lead to chronic infections and were reported to significantly increase milk somatic cell counts. The goal of our study was to determine the species distribution and antimicrobial susceptibility profiles of CNS in bovine mastitis milk samples in Switzerland. Between March 2011 and February 2012, a total of 120 CNS were isolated from mastitis milk samples from 117 different animals at 7...

  17. Large genomic differences between Moraxella bovoculi isolates acquired from the eyes of cattle with infectious bovine keratoconjunctivitis versus the deep nasopharynx of asymptomatic cattle.

    Science.gov (United States)

    Dickey, Aaron M; Loy, John D; Bono, James L; Smith, Timothy P L; Apley, Mike D; Lubbers, Brian V; DeDonder, Keith D; Capik, Sarah F; Larson, Robert L; White, Brad J; Blom, Jochen; Chitko-McKown, Carol G; Clawson, Michael L

    2016-02-13

    Moraxella bovoculi is a recently described bacterium that is associated with infectious bovine keratoconjunctivitis (IBK) or "pinkeye" in cattle. In this study, closed circularized genomes were generated for seven M. bovoculi isolates: three that originated from the eyes of clinical IBK bovine cases and four from the deep nasopharynx of asymptomatic cattle. Isolates that originated from the eyes of IBK cases profoundly differed from those that originated from the nasopharynx of asymptomatic cattle in genome structure, gene content and polymorphism diversity and consequently placed into two distinct phylogenetic groups. These results suggest that there are genetically distinct strains of M. bovoculi that may not associate with IBK.

  18. Multilocus sequence types of Finnish bovine Campylobacter jejuni isolates and their attribution to human infections

    Directory of Open Access Journals (Sweden)

    Corander Jukka

    2010-07-01

    Full Text Available Abstract Background Campylobacter jejuni is the most common bacterial cause of human gastroenteritis worldwide. Due to the sporadic nature of infection, sources often remain unknown. Multilocus sequence typing (MLST has been successfully applied to population genetics of Campylobacter jejuni and mathematical modelling can be applied to the sequence data. Here, we analysed the population structure of a total of 250 Finnish C. jejuni isolates from bovines, poultry meat and humans collected in 2003 using a combination of Bayesian clustering (BAPS software and phylogenetic analysis. Results In the first phase we analysed sequence types (STs of 102 Finnish bovine C. jejuni isolates by MLST and found a high diversity totalling 50 STs of which nearly half were novel. In the second phase we included MLST data from domestic human isolates as well as poultry C. jejuni isolates from the same time period. Between the human and bovine isolates we found an overlap of 72.2%, while 69% of the human isolates were overlapping with the chicken isolates. In the BAPS analysis 44.3% of the human isolates were found in bovine-associated BAPS clusters and 45.4% of the human isolates were found in the poultry-associated BAPS cluster. BAPS reflected the phylogeny of our data very well. Conclusions These findings suggest that bovines and poultry were equally important as reservoirs for human C. jejuni infections in Finland in 2003. Our results differ from those obtained in other countries where poultry has been identified as the most important source for human infections. The low prevalence of C. jejuni in poultry flocks in Finland could explain the lower attribution of human infection to poultry. Of the human isolates 10.3% were found in clusters not associated with any host which warrants further investigation, with particular focus on waterborne transmission routes and companion animals.

  19. Isolation and identification of a bovine viral diarrhea virus from sika deer in china

    Directory of Open Access Journals (Sweden)

    Wang Nan

    2011-02-01

    Full Text Available Abstract Background Bovine viral diarrhea virus (BVDV infections continue to cause significantly losses in the deer population. Better isolation and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from sika deer is seldom reported in literature. In this study, we collected some samples according to clinical sign of BVDV to isolation and identification of BVDV from sika deer. Results we isolated a suspected BVDV strain from livers of an aborted fetus from sika deer in Changchun (China using MDBK cell lines, named as CCSYD strain, and identified it by cytopathic effect (CPE, indirect immunoperoxidase test (IPX and electron microscopy(EM. The results indicated that this virus was BVDV by a series of identification. The structural proteins E0 gene was cloned and sequenced. The obtained E0 gene sequence has been submitted to GenBank with the accession number: FJ555203. Alignment with other 9 strains of BVDV, 7 strains of classical swine fever virus (CSFV and 3 strains of border disease virus(BDV in the world, showed that the homology were 98.6%-84.8%, 76.0%-74.7%, 76.6%-77.0% for nucleotide sequence, respectively. The phylogenetic analysis indicated that new isolation and identification CCSYD strain belonged to BVDV1b. Conclusion To the best of our knowledge, this is the first report that BVDV was isolated and identified in sika deer. This current research contributes development new BVDV vaccine to prevent and control of BVD in sika deer.

  20. Research on Isolation and Clone of Embryonic Stem Cell-Like in Bovine

    Institute of Scientific and Technical Information of China (English)

    AN Li-long; YANG Qi; XIAO Mei; FENG Xiu-Liang; YANG Chun-rong; LEI An-min; GAO Zhi-min; DOU Zhong-ying; QIU Huai

    2002-01-01

    Bovine embryonic stem cell would be invaluable for researching the aspect of animal cloning, production transgenic animal and discussion of gene function in vitro. With the object of establishing an effective culture system for isolation and clone of bovine pluripotent stem cell, we cultured bovine embryos and mouse embryos including morula blastula and hatached blastula and obtained animal ICM on Primary marine embryonic fibroblast (Primary murine embryonic fibroblast, PMEF) feeder layer with tissue medium(DMEM supplemented with 15ml/100ml NBS ,0.1μmol/L Na2SeO3, 0. 1mmol/L β-mercaptoethanol, 1 000ng/ml LIF,10 ng/ml IGF, 1mmol/L necessary amino acid and 1mmol/L L-glutamine), then, we obtained mouse ICM and bovine ICM. Moreover, we isolated and cloned the 6 passage bovine ES like cells(12 cell lines) and 9 passage marine ES like cells (52 cell lines) deriving from bovine ICM and murine ICM respectively on the feeder layer of PMEF by disaggregating ICM and ES cell clones of bovine and murine into smaller clumps through digesting with 0. 125g/100ml trypsin and 0.02g/100ml EDTA and scattering with a glass needle. The pluripotency of both murine and bovine ES like cells was identified with morphological character, histochemistry identification, karyotype analysis and differentiation of ES cells in vitro or in vivo. This result showed that bovine embryonic stem cell and murine embryonic stem cell had developmental pluripotency.

  1. Detection of multiple virulence-associated genes in Listeria monocytogenes isolated from bovine mastitis cases.

    Science.gov (United States)

    Rawool, D B; Malik, S V S; Shakuntala, I; Sahare, A M; Barbuddhe, S B

    2007-01-25

    Clinical samples (n=725) were collected from bovines (n=243) which were positive for mastitis using the California mastitis test (CMT) and somatic cell count (SCC). The clinical samples comprising blood (n=239), milk (n=243), and faecal swabs (n=243) were examined for the presence of pathogenic Listeria spp. Isolation of the pathogen was done using selective enrichment in University of Vermont Medium and plating onto Dominguez-Rodriguez isolation agar. Confirmation of the isolates was based on biochemical tests and Christie, Atkins, Munch-Petersen (CAMP) test followed by pathogenicity testing. Pathogenicity of the isolates was tested by phosphatidylinositol-specific phospholipase C (PI-PLC) assay as well as in vivo tests namely, chick embryo and mice inoculation tests. The isolates were subjected to PCR assay for five virulence-associated genes, plcA, prfA, hlyA, actA and iap. Listeria spp. were isolated from 12 (1.66%) samples. Of these 4 (0.55%) and 1 (0.14%) were confirmed as Listeria monocytogenes and Listeria ivanovii, respectively. L. monocytogenes and L. ivanovii were recovered from milk samples (2) and faecal (3) of mastitic cattle (3) and buffaloes (2). L. monocytogenes recovered from the milk of mastitic cattle and L. ivanovii from the faecal swab of buffalo turned out to be pathogenic. However, the remaining three hemolytic isolates exhibiting positive CAMP test turned out to be negative in PI-PLC assay, chick embryo and mice inoculation. L. monocytogenes and L. ivanovii isolates characterized as pathogenic by PI-PLC assay and in vivo pathogenicity tests were found to possess all the five virulence-associated genes and three genes, plcA, prfA and actA respectively. The remaining three hemolytic but non-pathogenic L. monocytogenes isolates were negative for plcA by PCR. It seems that the plcA gene and its expression (in the PI-PLC assay) have an important role as virulence determinants in pathogenic Listeria spp. In conclusion, the PI-PLC assay and

  2. The Comparison of Streptococcus agalactiae Isolated from Fish and Bovine using Multilocus Sequence Typing

    Directory of Open Access Journals (Sweden)

    Angela Mariana Lusiastuti

    2013-12-01

    Full Text Available Multilocus sequence typing (MLST has greater utility for determining the recent ancestral lineage and the relatedness of individual strains. Group B streptococci (GBS is one of the major causes of subclinical mastitis of dairy cattle in several countries. GBS also sporadically causes epizootic infections in fish. The aim of this study was to compare the evolutionary lineage of fish and bovine isolates in relation to the S. agalactiae global population as a whole by comparing the MLST profiles. Twenty S. agalactiae isolates were obtained from dairy cattle and fish. PCR products were amplified with seven different oligonucleotide primer pairs designed from the NEM316 GBS genome sequence. Clone complexes demonstrated that bovine and fish isolates were separate populations. These findings lead us to conclude that fish S. agalactiae is not a zoonotic agent for bovine. MLST could help clarify the emergence of pathogenic clones and to decide whether the host acts as a reservoir for another pathogenic lineage.

  3. Isolation of bovine serum albumin from whey using affinity chromatography

    NARCIS (Netherlands)

    Besselink, T.; Janssen, A.E.M.; Boom, R.M.

    2015-01-01

    The adsorption of bovine serum albumin (BSA) to a chromatography resin with immobilised llama antibody fragments as affinity ligands was investigated. The maximum adsorption capacity of the affinity resin was 21.6 mg mL-1 with a Langmuir equilibrium constant of 20.4 mg mg-1. Using packed bed chromat

  4. Activity of bacteriocins synthesized by Bacillus thuringiensis against Staphylococcus aureus isolates associated to bovine mastitis.

    Science.gov (United States)

    Barboza-Corona, José Eleazar; de la Fuente-Salcido, Norma; Alva-Murillo, Nayeli; Ochoa-Zarzosa, Alejandra; López-Meza, Joel E

    2009-07-01

    Antimicrobial therapy is a useful tool to control bovine mastitis caused by Staphylococcus aureus, as consequence an increase in staphylococci resistant cases has been registered. Alternative strategies are desirable and bacteriocins represent attractive control agents to prevent bovine mastitis. The aim of this work was to evaluate the activity of five bacteriocins synthesized by Bacillus thuringiensis against S. aureus isolates associated to bovine mastitis. Fifty S. aureus isolates were recovered from milk composite samples of 26 Holstein lactating cows from one herd during September 2007 to February 2008 in México and susceptibility of those isolates to 12 antibiotics and 5 bacteriocins from B. thuringiensis was evaluated. S. aureus isolates were mainly resistant to penicillin (92%), dicloxacillin (86%), ampicillin (74%) and erythromycin (74%); whereas susceptibility to gentamicin, trimethoprim and tetracycline was detected at, respectively, 92%, 88%, and 72%. All S. aureus isolates showed susceptibility to the five bacteriocins synthesized by B. thuringiensis, mainly to morricin 269 and kurstacin 287 followed by kenyacin 404, entomocin 420 and tolworthcin 524. Our results showed that S. aureus isolates had differences in the antimicrobial resistance patterns and were susceptible to bacteriocins produced by B. thuringiensis, which could be useful as an alternative method to control bovine mastitis. PMID:19359107

  5. Genotyping and study of the pauA and sua genes of Streptococcus uberis isolates from bovine mastitis.

    Science.gov (United States)

    Perrig, Melina S; Ambroggio, María B; Buzzola, Fernanda R; Marcipar, Iván S; Calvinho, Luis F; Veaute, Carolina M; Barbagelata, María Sol

    2015-01-01

    This study aimed to determine the clonal relationship among 137 Streptococcus uberis isolates from bovine milk with subclinical or clinical mastitis in Argentina and to assess the prevalence and conservation of pauA and sua genes. This information is critical for the rational design of a vaccine for the prevention of bovine mastitis caused by S. uberis. The isolates were typed by random amplified polymorphic DNA (RAPD) analysis and by pulsed-field gel electrophoresis (PFGE). The 137 isolates exhibited 61 different PFGE types and 25 distinct RAPD profiles. Simpson's diversity index was calculated both for PFGE (0.983) and for RAPD (0.941), showing a high discriminatory power in both techniques. The analysis of the relationship between pairs of isolates showed 92.6% concordance between both techniques indicating that any given pair of isolates distinguished by one method tended to be distinguished by the other. The prevalence of the sua and pauA genes was 97.8% (134/137) and 94.9% (130/137), respectively. Nucleotide and amino acid sequences of the sua and pauA genes from 20 S. uberis selected isolates, based on their PFGE and RAPD types and geographical origin, showed an identity between 95% and 100% with respect to all reference sequences registered in GenBank. These results demonstrate that, in spite of S. uberis clonal diversity, the sua and pauA genes are prevalent and highly conserved, showing their importance to be included in future vaccine studies to prevent S. uberis bovine mastitis. PMID:26507633

  6. Isolation and characterization of a virulent bacteriophage SPW specific for Staphylococcus aureus isolated from bovine mastitis of lactating dairy cattle.

    Science.gov (United States)

    Li, Longping; Zhang, Zhiying

    2014-09-01

    Mastitis in dairy cattle continues to be an economically important disease. However, control is complicated by a high prevalence of resistance to antibiotics. Phage therapy, therefore, is considered as an alternative way of controlling bacterial infections and contaminations. In this study, we have described isolation and characterization of a highly virulent phage SPW from wastewater of dairy farm, which possesses a strong lytic capability against mastitis-associated Staphylococcus aureus, the most important pathogen in bovine clinical and subclinical mastitis. The phage SPW produced large, round and clear plaques on bacterial culture plates. TEM showed phage SPW has an icosahedral head 62.5 nm in diameter and long tail of 106 nm, head and tail were held together by a connector of 18 ± 1.5 nm long and can be classified as a member of the Myoviridae family. Restriction analysis indicated that phage SPW was a dsDNA virus with an approximate genome size of 65-69 kb. One-step growth kinetics showed a short latency period of about 10-15 min and a rise period of 50 min and a relatively small burst size was 44 ± 3 phages particles/infected cell. Moreover, adsorption rates were not influenced by calcium ions and phage SPW was relatively stable in a wide range of temperature and pH values, and resistant to chloroform and isopropanol. The optimal multiplicity of infection (MOI) was 0.01. When phage SPW was used to infect five other clinically isolated pathogenic isolates, it showed relatively wide spectrum host range. Phage SPW was capable of eliciting efficient lysis of S. aureus, revealing it potentially as an effective approach to prophylaxis or treatment of S. aureus-associated mastitis in dairy cows.

  7. Molecular and in vitro Characterization of Field Isolates of Bovine Herpesvirus-1

    Institute of Scientific and Technical Information of China (English)

    Julian Ruiz-Saenz; Jairo Jaime; Gloria Ramirez; Victor Vera

    2012-01-01

    Bovine Herpesvirus-1 (BoHV-1) is distributed worldwide and is a major pathogen in cattle,being the causal agent of a variety of clinical syndromes.The aim of this study was to isolate and to characterize (molecular and biological characterization) BoHV- 1 from 29 immunosuppressed animals.It was possible to obtain 18 isolates,each from a different animal,such as from the respiratory and reproductive tracts.In some cases the cytopathic effect was visible 12 hours post-inoculation,and became characteristic after 36-48 hours.Biological characteristics were evaluated and compared with Iowa and Colorado-1 reference strains,and differences were found in plaque size,virus titer measured by TCID50 and PFU/mL,and one step virus curves.These results showed that some isolates had a highly virulent-like behavior in vitro,compared to the reference strains,with shorter eclipse periods,faster release of virus into the supematants,and higher burst size and viral titer.There were no differences in glycoprotein expression of BoHV-1 isolates,measured by Western blot on monolayers.Moreover,using restriction endonucleases analysis,most of the viruses were confirmed as BoHV-1.1 and just one of them was confirmed as BoHV-1.2a subtype.These findings suggest that some wild-type BoHV-1 isolates could be useful as seeds to develop new monovalent vaccines.

  8. VTEC O157 subtypes associated with the most severe clinical symptoms in humans constitute a minor part of VTEC 0157 isolates from Danish Cattle

    DEFF Research Database (Denmark)

    Roldgaard, Bemt Bjørn; Scheutz, Flemming; Boel, Jeppe;

    2004-01-01

    -positive VTEC 0 157 isolates (63 of bovine origin and 86 from human clinical cases) isolated between 1987 and 2001. All were analysed by vtx-PCR-RFLP and phage typing. The vtx-PCR-RFLP showed that isolates carrying the vtx2 gene was more than four times as prevalent among the human clinical isolates (55...

  9. Association between phage types and antimicrobial resistance among bovine isolates of Staphylococcus aureus in 10 countries

    DEFF Research Database (Denmark)

    Vintov, J.; Aarestrup, Frank Møller; Zinn, C. E.;

    2003-01-01

    associated with penicillin resistance in contrast to phage group I (P = 0.0023) and phage complex-80 (P = 0.0066). This study confirms that a large number of phage types of S. aureus cause bovine mastitis, but that some types predominate. In addition, these findings could indicate that the use of penicillin...... in the bovine environment has selected for specific types of S. aureus in countries with a high frequency of resistance. (C) 2003 Elsevier B.V. All rights reserved.......This study was conducted to investigate the diversity of phage types and associations between penicillin resistance and phage types among 815 Staphylococcus aureus isolates from bovine mastitis in nine European countries and USA. All isolates were examined for susceptibility to antimicrobial agents...

  10. Complete Genome Sequence of a Bovine Viral Diarrhea Virus Strain Isolated in Southern China

    OpenAIRE

    Xie, Zhixun; Fan, Qing; Xie, Zhiqin; Liu, Jiabo; Pang, Yaoshan; Deng, Xianwen; Xie, Liji; Luo, Sisi; Khan, M. I.

    2014-01-01

    We report here the full-length RNA genomic sequence of the bovine viral diarrhea virus (BVDV) strain GX4, isolated from a cow in southern China. Studies indicate that BVDV GX4 belongs to the BVDV-1b subtype. This report will help in understanding the epidemiology and molecular characteristics of BVDV in southern China cattle.

  11. Social isolation may influence responsiveness to infection with bovine herpesvirus 1 in veal calves

    NARCIS (Netherlands)

    Reenen, van C.G.; Mars, M.H.; Leushuis, I.E.; Rijsewijk, F.A.M.; Oirschot, van J.T.; Blokhuis, H.J.

    2000-01-01

    An experiment was performed to develop a model to study the impact of stress on responsiveness to infection with bovine herpesvirus 1 (BHV1) in veal calves. Social isolation after previous group-housing was used as a putatively stressful treatment. Group-housed specific pathogen-free veal calves (n=

  12. A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

    DEFF Research Database (Denmark)

    Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars;

    components. Here we present a novel strategy for a short, gentle and non-denaturing isolation of skim milk-derived membrane vesicles. Methods: Untreated fresh bovine milk was defatted to remove milk fat globules. The resulting skim milk was subjected to ultracentrifugation. The resulting ochre-coloured...

  13. Production of cattle immunotolerant to bovine viral diarrhea virus.

    OpenAIRE

    McClurkin, A W.; Littledike, E T; Cutlip, R C; Frank, G H; Coria, M F; Bolin, S R

    1984-01-01

    Inoculation of bovine virus diarrhea virus into 58 to 125 day old fetuses of bovine virus diarrhea virus seropositive pregnant cows, or inoculation of bovine virus diarrhea virus into seronegative cows 42 to 114 days pregnant, may produce clinically normal calves which are persistently infected with the specific isolate of bovine virus diarrhea virus yet seronegative to the homologous and heterologous isolates. Reinoculation of these persistently infected cattle with their homologous isolate ...

  14. A clinical comparison of purified bovine and purified porcine insulins.

    OpenAIRE

    Olczak, S A; Greenwood, R H

    1985-01-01

    Twenty four patients with established insulin dependent diabetes treated with twice daily soluble and isophane bovine insulins were changed to equivalent doses of either purified bovine Neusulin and Neuphane (Wellcome) or purified porcine Actrapid and Monotard (Novo) insulins. After 6 months treatment the porcine group showed a 35% fall in insulin binding antibodies and a 14% reduction in insulin dosage. The group changed to purified bovine insulins showed no significant change in insulin bin...

  15. Isolation of Leptospira spp from dogs, bovine and swine naturally infected

    Directory of Open Access Journals (Sweden)

    Freitas Julio Cesar de

    2004-01-01

    Full Text Available Leptospira isolation allows definitive diagnosis of the infection. Contamination by microorganisms is one of the inconveniences of the culture. The objective of this study was to describe the isolation of Leptospira from dogs, bovine and swine naturally infected. Urine samples from 14 dogs and three bovines, and kidney, liver, ovary, and uterus body samples from 36 slaughtered sows with unknown health history, were used. The urine and organ samples were cultured in culture medium. Modified Ellinghausen-McCullough-Johnson-Harris medium (EMJH culture medium was used with addition of 5-fluorouracil, chloramphenicol, vancomycin, nalidixic acid and neomycin. Incubation was performed at 28oC for 24 hours, followed by subculture in modified EMJH without antibiotics. The cultures were assessed weekly for up to eight weeks for the dog and swine samples and for up to 16 weeks for the bovine samples. With this methodology, Leptospira spp could be isolated from 11 dogs, two bovines and liver fragments from two sows.

  16. Pathogenicity of local isolate virus BHV-1 as the aetiological agent of Infectious Bovine Rhinotracheitis in Bali Cattle

    Directory of Open Access Journals (Sweden)

    Rini I Damayanti

    2005-10-01

    Full Text Available Infectious Bovine Rhinotracheitis is a disease of cattle characterised by clinical signs of the upper respiratory tract, reproductive tract and nervous system. A study to define the pathogenicity of four BHV-1 local isolates has been conducted. Fourteen Bali cattle that were free of BHV-1 has been selected and divided into four treatment groups. Each group of three was infected with virus isolate I, II, III and IV respectively with approximately a dose of 108TCID50 /10 ml and two cattle were used as control animals. Isolate I and III were originated from semen from IBR positive bulls number G 867 and G 148 respectively whereas isolate II was collected from vaginal mucosa and isolate IV was from nasal mucosa of IBR positive cattle treated with dexamethasone. Clinical response, gross-pathological and histopathological changes were observed. Immunohistochemical staining was applied to detect the antigen in tissue section. The results show that the BHV-1 local isolates could produce IBR syndrome namely fever and changes in the respiratory and reproductive tracts even though the clinical responses seemed to be disappeared by 21 days PI. Grossly there were hyperaemic nasal and vaginal mucosa and pneumonia whereas histologically there were non suppurative rhinitis, tracheitis, pneumonia and vulvovaginitis. Immunohistochemically the antigen was detected in the nasal concha and trachea. Dexamethasone treatment at 60-64 days PI could produce less severe clinical features and the second necroppsy at 69 days PI also results in less severe pathological responses. The findings also suggest that the pathogenicity of BHV-1 local isolates were as follows: isolates I, II, IV and III.

  17. Pheno- and genotyping of Staphylococcus epidermidis isolated from bovine milk and human skin

    DEFF Research Database (Denmark)

    Thorberg, B. M.; Kuhn, I.; Aarestrup, Frank Møller;

    2006-01-01

    The purpose of this study was to improve our knowledge concerning the epidemiology and strain diversity of Staphylococcus epidermidis isolated from bovine milk in commercial dairy herds. A total of 341 S. epidermidis isolates obtained from cows' milk (317), farmers (17) and patients (7) were...... different S. epidermidis types exist in milk samples. Antibiotic resistance patterns matched the SmaI profiles closely in the two herds, but poorly in the routinely collected milk samples. Isolates from herd I showed one to five patterns, depending on the typing method used. Isolates from the milker's skin...... showed one pattern, which was identical to the most common pattern found in the milk isolates. Isolates from herd 2 showed three to four patterns, two of these being identical to skin isolates from the milker. As dairy cows are not a natural host for S. epidermidis the results suggest a human source...

  18. Isolation and Purification of Bovine Colostrum sIgA and IgG

    Institute of Scientific and Technical Information of China (English)

    YAO Yonghao; GAO Xuejun; LIU Xiaofei

    2008-01-01

    To further utilize bioactive substance such as bovine colostrum sIgA and lgG, slgA and IgG were isolated and purified simultaneously by salting out,ultrafiltration and gel chromatography,etc.The analysis of results were showed quantitatively by nonhydrogenized SDS-PAGE,and quanlities of sIgA and IgG were respectively detected by Western Blot. The results showed that the purity and yield of bovine colostrum slgA were 85.3% and 42.8%,respectively,while the purity and yield of bovine colostrum IgG were respectively 97.2% and 64.4%.This preparative method provides theoretical and experimental foundation for sIgA industrial production.

  19. Whole-Genome Sequences of Mycobacterium bovis Strain MbURU-001, Isolated from Fresh Bovine Infected Samples

    Science.gov (United States)

    Lasserre, Moira; Berná, Luisa; Greif, Gonzalo; Díaz-Viraqué, Florencia; Naya, Hugo; Castro-Ramos, Miguel; Juambeltz, Arturo

    2015-01-01

    Bovine tuberculosis in cattle has a high incidence in Uruguay, where it is considered a disease of national importance. We present the genome sequence of Mycobacterium bovis strain MbURU-001, isolated from pectoral lymph nodes of a bovine host from a cattle farm. PMID:26543108

  20. Genome Sequences of Four Staphylococcus aureus Strains Isolated from Bovine Mastitis

    OpenAIRE

    Kant, Ravi; Taponen, Suvi; Koort, Joanna; Paulin, Lars; Åvall-Jääskeläinen, Silja; Palva, Airi

    2015-01-01

    Staphylococcus aureus is a major causative agent of mastitis in dairy cows. The pathogenicity of S. aureus may vary; it is able to cause severe clinical mastitis, but most often it is associated with chronic subclinical mastitis. Here, we present the genome assemblies of four S. aureus strains from bovine mastitis.

  1. Genome Sequences of Four Staphylococcus aureus Strains Isolated from Bovine Mastitis.

    Science.gov (United States)

    Kant, Ravi; Taponen, Suvi; Koort, Joanna; Paulin, Lars; Åvall-Jääskeläinen, Silja; Palva, Airi

    2015-01-01

    Staphylococcus aureus is a major causative agent of mastitis in dairy cows. The pathogenicity of S. aureus may vary; it is able to cause severe clinical mastitis, but most often it is associated with chronic subclinical mastitis. Here, we present the genome assemblies of four S. aureus strains from bovine mastitis. PMID:25908141

  2. Development of a method for isolating bovine colostrum mononuclear leukocytes for phenotyping and functional studies.

    Science.gov (United States)

    Meganck, Vanessa; Goddeeris, Bruno M; Stuyven, Edith; Piepers, Sofie; Cox, Eric; Opsomer, Geert

    2014-05-01

    The present study reports a method for isolating bovine colostrum mononuclear cells (CMC) for phenotyping and functional studies. As well as being an important source of immunoglobulins, colostrum also contains leukocytes that may be of greater importance for passive immunity than has previously been thought. Different protocols have been reported for isolating leukocytes from bovine colostrum, although none of these have been validated, and phenotypic analysis of cell populations has not always been performed. In this study, bovine CMC were isolated by density gradient centrifugation. Cell populations were identified by flow cytometry using antibodies against selected bovine cell surface markers and the proliferative capacity of these cells was determined using a (3)H-thymidine proliferation assay. The mean cell count of isolated CMC was 3 × 10(4) and 1 × 10(5) per mL colostrum for the samples used in the flow cytometric assay and the proliferation assay, respectively. A mean of 25.4 ± 17.1% CMC were identified as T lymphocytes, 2.9 ± 3.0% as B lymphocytes and 32.7 ± 13.7% as macrophages. In terms of proliferation, the mean counts per minute were 4.3 × 10(3) and 1.8 × 10(4) for cells cultured in medium only or in the presence of concanavalin A, respectively, showing that CMC are viable and capable of responding to mitogen stimulation. Isolation of CMC and the subsequent phenotypic analysis of the different subpopulations were repeatable, with agreement indices varying between 0.5 and 1.0. Agreement indices for the proliferation assay were estimated at 0.8. PMID:24679458

  3. Characterisation of Staphylococcus aureus isolated from cases of bovine subclinical mastitis in two Uruguayan dairy farms

    OpenAIRE

    de los Santos, R.; Fernández, M.; Carro, S.; P. Zunino

    2014-01-01

    The aim of this work was to identify and to characterise Staphylococcus aureus isolates associated with subclinical mastitis obtained from milk of lactating cows showing a California Mastitis Test (CMT) score result of traces, 1, 2, or 3. Coagulase, hemolysis, presence of capsule, slime formation, biofilm production, autoaggregation, hemagglutination and antibiotic susceptibility were assessed to evaluate S. aureus virulence factors expression potentially associated to bovine subclinical mast...

  4. Draft Genome Sequences of Staphylococcus aureus Strains Isolated from Subclinical Bovine Mastitis in Brazil.

    Science.gov (United States)

    Silva, Danielle Mendes; da Silva, Mônica Pacheco; Vidigal, Pedro M Pereira; Barcelos, Rafael Mazioli; Klein, Raphael Contelli; Aguilar, Ananda Pereira; Fabres-Klein, Mary Hellen; Oliveira, Guilherme; Ribon, Andréa Oliveira Barros

    2016-01-01

    Here, we present the draft genome sequences of four Staphylococcus aureus strains isolated from mastitic milk collected from animals with subclinical manifestations. Three of them were typed as sequence type 126 (ST126), a genotype with no genome sequence available. ST126 is found in several herds of southern Brazil and is described as a bovine pathogen strongly associated with milk around the world. PMID:26893417

  5. Molecular biology of rotaviruses. III. Isolation and characterization of temperature-sensitive mutants of bovine rotavirus.

    OpenAIRE

    Faulkner-Valle, G P; Clayton, A V; McCrae, M A

    1982-01-01

    Twenty-six temperature-sensitive (ts) mutants of United Kingdom tissue culture-adapted bovine rotavirus were isolated and characterized. Fourteen of these mutants were determined to be ts both by efficiency of plating and by virus yield at the nonpermissive temperature of 39.5 degrees C as compared with that at the permissive temperature of 32 degrees C. The remaining mutants were only ts by the criterion of efficiency of plating. High-frequency recombination (gene reassortment) was observed ...

  6. A study of phage- and ribotype patterns of Staphylococcus aureus isolated from bovine mastitis in the Nordic countries

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Wegener, Henrik Caspar; Jensen, N.E.;

    1997-01-01

    This study was conducted to investigate the geographical distribution of phage and ribotypes of Staphylococcus aureus causing bovine mastitis in the 5 Nordic countries. A total of 403 isolates of S. aureus was isolated from 403 different dairy herds. One hundred five strains were isolated in...... the isolates of ribotype 1 belonged to phage type 29/52. This combined type accounted for 17% of all the 403 isolates. These findings show that a large number of different types of S. aureus can be isolated from cases of bovine mastitis. However, few types predominate within different countries. These...... predominating types seem to be specific in each country, however, a single type was common for both Denmark, Sweden and Finland. This could suggest differences in the virulence or in modes of transmission of predominating and rare types of S. aureus associated with bovine mastitis....

  7. Short communication. Genotyping and phylogenetic analysis of bovine viral diarrhea virus (BVDV isolates in Kosovo

    Directory of Open Access Journals (Sweden)

    Izedin Goga

    2014-03-01

    Full Text Available Three serum samples positive in Antigen ELISA BVDV have been tested to characterise genetic diversity of bovine viral diarrhea virus (BVDV in Kosovo. Samples were obtained in 2011 from heifers and were amplified by reverse transcription-polymerase chain reaction, sequenced and analysed by computer-assisted phylogenetic analysis. Amplified products and nucleotide sequence showed that all 3 isolates belonged to BVDV 1 genotype and 1b sub genotype. These results enrich the extant knowledge of BVDV and represent the first documented data about Kosovo BVDV isolates.

  8. Characterisation of Staphylococcus aureus isolated from cases of bovine subclinical mastitis in two Uruguayan dairy farms

    Directory of Open Access Journals (Sweden)

    R de los Santos

    2014-01-01

    Full Text Available The aim of this work was to identify and to characterise Staphylococcus aureus isolates associated with subclinical mastitis obtained from milk of lactating cows showing a California Mastitis Test (CMT score result of traces, 1, 2, or 3. Coagulase, hemolysis, presence of capsule, slime formation, biofilm production, autoaggregation, hemagglutination and antibiotic susceptibility were assessed to evaluate S. aureus virulence factors expression potentially associated to bovine subclinical mastitis isolates. Prevalence of subclinical mastitis along the study was low and did not correlate with months or climatic variables. Most of S. aureus (20 were isolated from milk samples showing a CMT score result of 1. Formation of capsule, slime, biofilms and the occurrence of bacteria aggregation in all the tested isolates converged in the bacterial ability of adherence and persistence in the mammary gland and probably contribute to the further chronicity of the infection and even the colonization of dairy installations. Resistance against a set of commonly used antibiotics was low. The evaluation of virulence factors of S. aureus isolates in the context of subclinical mastitis in dairy farms may be useful to develop precise actions and treatments to control mastitis and to improve animal health and milk production in dairy bovine herds.

  9. Evaluation of antibacterial effect of some Sinai medicinal plant extracts on bacteria isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Gamil S. G. Zeedan

    2014-11-01

    Full Text Available Aim: Bovine mastitis is the most economically important disease affecting dairy cattle worldwide from an economic, diagnostic and public-health point of view. The present study aimed to isolate and identify of bacteria causes mastitis in dairy cows and to evaluate the antibacterial activities of some selected medicinal plants extracts comparing antibiotics used in the treatment of mastitis in Egypt. Materials and Methods: A total of 203 milk samples of dairy cows were collected during the period from February to June 2013 at different Governorates in Egypt. The use clinical inspection and California mastitis test examination were provided efficient diagnostic tool for detection of clinical, subclinical mastitis and apparently normal health cattle. The collected milk samples were cultured on Nutrient, Blood agar, Mannitol salt, Edward’s and MacConkey agar plates supporting the growth of various types of bacteria for their biochemical studies and isolation. The antimicrobial activity of plants extracts (Jasonia montana and Artemisia herb albawith different solvent (ethanol, petroleum ether, chloroform and acetonewere studied in vitro against isolated bacteria from mastitis by paper desk diffusion and minimum inhibitory concentration method (MIC. Results: The prevalence of clinical, subclinical mastitis and normal healthy animals were 34.50%, 24.7% and 40.8% respectively. The major pathogens isolated from collected milk samples were Escherichia coli (22.16%, Staphylococcus aureus (20.19%, Streptococcus spp. (13.3%, Streptococcus agalactiae (12.8%, Streptococcus dysgalactia (0.5%, Pasteurella spp. (2.45%, Klebsiella spp. (1.47%and Pseudomonas spp. (0.45%. The highest antibacterial activity of J. montana plant extracted with acetone solvent against S. agalactiae, E. coli, S. aureus, Klebsiella spp and coagulase-negative Staphylococci with zone of inhibition values ± standard deviation (SD, ranging from 4.33±0.57 to 25.6±0.60 mm. The MIC values

  10. Hyaluronidase in Clinical Isolates of Propionibacterium acnes

    OpenAIRE

    Harmony Tyner; Robin Patel

    2015-01-01

    Objectives. We sought to describe the prevalence of a hyaluronidase gene and hyaluronidase production in 197 clinical isolates of P. acnes; we assessed kinetics of hyaluronidase production in a subset of three isolates. Methods. The hyaluronidase gene was detected using polymerase chain reaction. Hyaluronidase production was detected by growing isolates on BHI agar containing 400 μg/mL hyaluronic acid and 1% albumin and flooding plates with 2 N glacial acetic acid to precipitate unbound hyalu...

  11. Evaluation of the Rapid Mastitis Test for identification of Staphylococcus aureus and Streptococcus agalactiae isolated from bovine mammary glands.

    OpenAIRE

    Watts, J L; Owens, W E

    1988-01-01

    A latex agglutination test system (Rapid Mastitis Test [RMT]; Immucell, Portland, Maine) containing reagents for the identification of Staphylococcus aureus and Streptococcus agalactiae from bovine intramammary infections was evaluated with 527 staphylococcal and 267 streptococcal isolates. The RMT Staphylococcus aureus reagent detected 94.2% of 242 Staphylococcus aureus isolates, 80% of 25 Staphylococcus intermedius isolates, and 42.8% of 21 tube coagulase-positive Staphylococcus hyicus isol...

  12. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan.

    Science.gov (United States)

    Abe, Yuri; Tamura, Tomokazu; Torii, Shiho; Wakamori, Shiho; Nagai, Makoto; Mitsuhashi, Kazuya; Mine, Junki; Fujimoto, Yuri; Nagashima, Naofumi; Yoshino, Fumi; Sugita, Yukihiko; Nomura, Takushi; Okamatsu, Masatoshi; Kida, Hiroshi; Sakoda, Yoshihiro

    2016-01-01

    In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5'-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years.

  13. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan.

    Science.gov (United States)

    Abe, Yuri; Tamura, Tomokazu; Torii, Shiho; Wakamori, Shiho; Nagai, Makoto; Mitsuhashi, Kazuya; Mine, Junki; Fujimoto, Yuri; Nagashima, Naofumi; Yoshino, Fumi; Sugita, Yukihiko; Nomura, Takushi; Okamatsu, Masatoshi; Kida, Hiroshi; Sakoda, Yoshihiro

    2016-01-01

    In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5'-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years. PMID:26400674

  14. Carriage of stx2a differentiates clinical and bovine-biased strains of Escherichia coli O157.

    Directory of Open Access Journals (Sweden)

    Smriti Shringi

    Full Text Available BACKGROUND: Shiga toxin (Stx are cardinal virulence factors of enterohemorrhagic E. coli O157:H7 (EHEC O157. The gene content and genomic insertion sites of Stx-associated bacteriophages differentiate clinical genotypes of EHEC O157 (CG, typical of clinical isolates from bovine-biased genotypes (BBG, rarely identified among clinical isolates. This project was designed to identify bacteriophage-mediated differences that may affect the virulence of CG and BBG. METHODS: Stx-associated bacteriophage differences were identified by whole genome optical scans and characterized among >400 EHEC O157 clinical and cattle isolates by PCR. RESULTS: Optical restriction maps of BBG strains consistently differed from those of CG strains only in the chromosomal insertion sites of Stx2-associated bacteriophages. Multiplex PCRs (stx1, stx2a, and stx2c as well as Stx-associated bacteriophage-chromosomal insertion site junctions revealed four CG and three BBG that accounted for >90% of isolates. All BBG contained stx2c and Stx2c-associated bacteriophage-sbcB junctions. All CG contained stx2a and Stx2a-associated bacteriophage junctions in wrbA or argW. CONCLUSIONS: Presence or absence of stx2a (or another product encoded by the Stx2a-associated bacteriophage is a parsimonious explanation for differential virulence of BBG and CG, as reflected in the distributions of these genotypes in humans and in the cattle reservoir.

  15. Characterisation of bovine viral diarrhoea virus (BVDV) isolates from an outbreak with haemorrhagic enteritis and severe pneumonia.

    Science.gov (United States)

    Yeşilbağ, Kadir; Förster, Christine; Ozyiğit, M Ozgür; Alpay, Gizem; Tuncer, Pelin; Thiel, Heinz-Jürgen; König, Matthias

    2014-02-21

    During 2007 a disease outbreak occurred in cattle in the Marmara region of western Turkey characterised by severe pneumonia and haemorrhagic enteritis in calves. Cases from three farms at different locations were examined and bovine viral diarrhoea virus (BVDV) isolated in all cases. Phylogenetic characterisation of the virus isolates allocated them in a new cluster tentatively named as BVDV-1r.

  16. Characterization of Shiga Toxin-Producing Escherichia coli O157 Isolates from Bovine Carcasses.

    Science.gov (United States)

    Fontcuberta, M; Planell, R; Torrents, A; Sabaté, S; Gonzalez, R; Ramoneda, M; de Simón, M

    2016-08-01

    The main purpose of this study was to determine the prevalence of Escherichia coli O157 on bovine carcasses before and after chilling at a large slaughterhouse located in the city of Barcelona, Spain, to assess the effectiveness of dry chilling on reducing E. coli O157 contamination of carcasses. In addition, the study characterized the E. coli O157 strains isolated in terms of virulence factors, antibiotic susceptibility, and their genetic diversity. Individual bovine carcasses were sampled before (n = 300) and after (n = 300) chilling over an 8-month period. Positive samples for E. coli O157 were subjected to virulence screening by PCR (stx1, stx2, and eaeA genes and the fliCH7 gene), antimicrobial susceptibility testing, and molecular typing by pulsed-field gel electrophoresis. A total of 9.7% (29 of 300) of the nonrefrigerated carcasses examined and 2.3% (7 of 300) of the refrigerated carcasses were positive for E. coli O157. All the isolates were serotype O157:H7, 92% (33 of 36) carried the stx1, stx2, and eaeA genes, and 8% (3 of 36) carried the stx2 and eaeA genes. Antimicrobial susceptibility testing showed a high degree of resistance: 29 strains (81%) were resistant to at least 1 antimicrobial of the 12 antimicrobials tested; 69% (25 of 36) were resistant to 4 or more antimicrobials. Molecular typing by pulsed-field gel electrophoresis found a high diversity of genetic types, implying little cross-contamination in the slaughterhouse. This study confirms that E. coli O157:H7 is present on the carcasses slaughtered in Spain, although its prevalence is reduced by the dry chilling process used. The recovered isolates showed potential pathogenesis and a high degree of multidrug resistance, confirming the importance of bovine meat monitoring.

  17. Characterization of Shiga Toxin-Producing Escherichia coli O157 Isolates from Bovine Carcasses.

    Science.gov (United States)

    Fontcuberta, M; Planell, R; Torrents, A; Sabaté, S; Gonzalez, R; Ramoneda, M; de Simón, M

    2016-08-01

    The main purpose of this study was to determine the prevalence of Escherichia coli O157 on bovine carcasses before and after chilling at a large slaughterhouse located in the city of Barcelona, Spain, to assess the effectiveness of dry chilling on reducing E. coli O157 contamination of carcasses. In addition, the study characterized the E. coli O157 strains isolated in terms of virulence factors, antibiotic susceptibility, and their genetic diversity. Individual bovine carcasses were sampled before (n = 300) and after (n = 300) chilling over an 8-month period. Positive samples for E. coli O157 were subjected to virulence screening by PCR (stx1, stx2, and eaeA genes and the fliCH7 gene), antimicrobial susceptibility testing, and molecular typing by pulsed-field gel electrophoresis. A total of 9.7% (29 of 300) of the nonrefrigerated carcasses examined and 2.3% (7 of 300) of the refrigerated carcasses were positive for E. coli O157. All the isolates were serotype O157:H7, 92% (33 of 36) carried the stx1, stx2, and eaeA genes, and 8% (3 of 36) carried the stx2 and eaeA genes. Antimicrobial susceptibility testing showed a high degree of resistance: 29 strains (81%) were resistant to at least 1 antimicrobial of the 12 antimicrobials tested; 69% (25 of 36) were resistant to 4 or more antimicrobials. Molecular typing by pulsed-field gel electrophoresis found a high diversity of genetic types, implying little cross-contamination in the slaughterhouse. This study confirms that E. coli O157:H7 is present on the carcasses slaughtered in Spain, although its prevalence is reduced by the dry chilling process used. The recovered isolates showed potential pathogenesis and a high degree of multidrug resistance, confirming the importance of bovine meat monitoring. PMID:27497130

  18. Isolation and characterization of bovine parainfluenza virus type 3 from water buffaloes (Bubalus bubalis) in Argentina

    OpenAIRE

    Maidana, Silvina S; Lomonaco, Patricia M; Combessies, Gustavo; Craig, María I; Diodati, Julian; Rodriguez, Daniela; Parreño, Viviana; Zabal, Osvaldo; Konrad, José L; Crudelli, Gustavo; Mauroy, Axel; Thiry, Etienne; Romera, Sonia A

    2012-01-01

    Background Parainfluenza virus type 3 (PIV3) was isolated from dairy buffaloes (Bubalus bubalis) naturally affected with respiratory and reproductive clinical conditions. Results Examination of nasal and vaginal swabs collected from 12 diseased buffaloes led to the isolation of three paramyxovirus isolates from two animals. Antigenic, morphological and biological characteristics of these three isolates were essentially similar to those of members of the Paramyxoviridae family. Antigenic analy...

  19. Antigenic characterization of Brazilian bovine viral diarrhea virus isolates by monoclonal antibodies and cross-neutralization

    Directory of Open Access Journals (Sweden)

    Botton S.A.

    1998-01-01

    Full Text Available Nineteen Brazilian isolates of bovine viral diarrhea virus (BVDV were characterized antigenically with a panel of 19 monoclonal antibodies (mAbs (Corapi WV, Donis RO and Dubovi EJ (1990 American Journal of Veterinary Research, 55: 1388-1394. Eight isolates were further characterized by cross-neutralization using sheep monospecific antisera. Analysis of mAb binding to viral antigens by indirect immunofluorescence revealed distinct patterns of reactivity among the native viruses. Local isolates differed from the prototype Singer strain in recognition by up to 14 mAbs. Only two mAbs - one to the non-structural protein NS23/p125 and another to the envelope glycoprotein E0/gp48 - recognized 100% of the isolates. No isolate was recognized by more than 14 mAbs and twelve viruses reacted with 10 or less mAbs. mAbs to the major envelope glycoprotein E2/gp53 revealed a particularly high degree of antigenic variability in this glycoprotein. Nine isolates (47.3% reacted with three or less of 10 E2/gp53 mAbs, and one isolate was not recognized by any of these mAbs. Virus-specific antisera to eight isolates plus three standard BVDV strains raised in lambs had virus-neutralizing titers ranging from 400 to 3200 against the homologous virus. Nonetheless, many antisera showed significantly reduced neutralizing activity when tested against heterologous viruses. Up to 128-fold differences in cross-neutralization titers were observed for some pairs of viruses. When the coefficient of antigenic similarity (R was calculated, 49 of 66 comparisons (74.24% between viruses resulted in R values that antigenically distinguish strains. Moreover, one isolate had R values suggesting that it belongs to a distinct serologic group. The marked antigenic diversity observed among Brazilian BVDV isolates should be considered when planning diagnostic and immunization strategies.

  20. Properties and antimicrobial susceptibility of Trueperella pyogenes isolated from bovine mastitis in China.

    Science.gov (United States)

    Alkasir, Rashad; Wang, Jianfang; Gao, Jian; Ali, Tariq; Zhang, Limei; Szenci, Ottó; Bajcsy, Árpád Csaba; Han, Bo

    2016-03-01

    Trueperella (T.) pyogenes is an opportunistic pathogen that causes suppurative diseases in domestic animals. In this work, the properties, pathogenesis and phenotypic diversity of T. pyogenes isolates from bovine mastitis were studied. Both pyolysin (plo) and collagen-binding protein (cbp) virulence factor genes were detected by PCR in all T. pyogenes isolates (n = 50). Using the tissue culture plate method, 90% of T. pyogenes isolates were able to form biofilms. The minimum inhibitory concentrations (MICs) of 13 antimicrobials against T. pyogenes isolates were determined. High susceptibility was observed to rifampin (96%), ampicillin (94%), ciprofloxacin (94%), and penicillin (92%), while low susceptibility was found to trimethoprim-sulphamethoxazole (10%) and bacitracin (2%). The intracellular assay revealed that T. pyogenes isolates had different cytopathogenic effects on cells. The high percentage (28.6%) of T. pyogenes isolates suggests that this bacterium is an important contributor to mastitis. Moreover, the high occurrence of multidrug resistance, biofilm production, intracellular survival, and the temporal dynamics of T. pyogenes interactions are key factors for a better understanding of how immunity acts on infections with these bacteria and how they evade immune surveillance, thus highlighting the need for the prudent use of antimicrobial agents in veterinary medicine. PMID:26919137

  1. A bovine G8P[1] group A rotavirus isolated from an asymptomatically infected dog.

    Science.gov (United States)

    Sieg, Michael; Rückner, Antje; Köhler, Christian; Burgener, Iwan; Vahlenkamp, Thomas W

    2015-01-01

    Group A rotaviruses (RVAs) are enteric pathogens with well-documented zoonotic transmissions to humans. The segmented genome of the virus enables reassortment events which might alter host susceptibility and/or disease course. Genetic analysis of rotavirus in dogs has so far only revealed RVAs with the VP7 and VP4 genome constellation G3P[3]. RVA G3P[3] have also been found in cats, humans, monkeys and bats. In the present study, we described an unusual RVA of genotype G8P[1] which was isolated from an asymptomatically infected young dog. The dog did not show signs of diarrhoea. Analysis of full-length segments of VP2, VP6 and VP7 as well as NSP1-NSP5 revealed a typical bovine-like genotype constellation G8-P[1]-I2-Rx-C2-Mx-A3-N2-T6-E2-H3. Phylogenetic analysis supported the hypothesis of an interspecies transmission from a bovine/artiodactyl species or from humans to the young dog. The isolate was likely to represent a multiple reassortant virus. PMID:25304653

  2. Isolation of bovine milk-derived microvesicles carrying mRNAs and microRNAs.

    Science.gov (United States)

    Hata, Taketoshi; Murakami, Kosuke; Nakatani, Hajime; Yamamoto, Yasunari; Matsuda, Tsukasa; Aoki, Naohito

    2010-05-28

    By a series of centrifugation and ultracentrifugation, we could isolate microvesicles with approximately 100 nm in diameter from bovine milk. We also found that approximately 1700 and 1000 ng of total RNA, in which small RNAs were major components, was contained inside the microvesicles isolated from 6 ml of colostrum and mature milk, respectively, despite high RNase activity in the milk. Polyadenylated gene transcripts for major milk proteins and translation elongation factor-1alpha (EF-1alpha) were present in the microvesicles, and integrity of some transcripts was confirmed by real-time PCR targeting 5'- and 3'-ends of mRNA and by in vitro translation analysis. Moreover, a considerable amount of mammary gland and immune-related microRNAs were present in the milk-derived microvesicles. Acidification of milk to mimic gastrointestinal tract did not mostly affected RNA yield and quality. The milk related gene transcripts were detected in cultured cells when incubated with milk-derived microvesicles, suggesting cellular uptake of the microvesicle contents including RNA. Our findings suggest that bovine breast milk contains RNAs capable for being transferred to living cells and involved in the development of calf's gastrointestinal and immune systems. PMID:20434431

  3. Prevalence and Antimicrobial Susceptibility of Coagulase-Negative Staphylococci isolated from Bovine Mastitis

    Directory of Open Access Journals (Sweden)

    Kaliwal B.B.

    2011-08-01

    Full Text Available The study was carried out to investigate the prevalence and antimicrobial susceptibility of Coagulase-Negative Staphylococci isolated from Bovine Mastitis in and around Dharwad region. A total of 310 samples were screened and 180 confirmed Coagulase-Negative Staphylococci were obtained. The antimicrobial susceptibility of Coagulase-Negative Staphylococci against 10 antimicrobial agents was tested using the disc diffusion method. The highest numbers of Coagulase-Negative Staphylococci were susceptible to ceftriaxone 83.88% followed by cefotaxime 79.41%, methicillin 76.47%, ciprofloxacin 73.52%, erythromycin 70.05%, amikacin 66.11%, gentamycin 42.94%, amoxicillin 36.76%, ampicillin 29.41%, and the lowest susceptibility was shown in penicillin 23.23% . The results indicated that the increase in prevalence and antibiotic resistance pattern of the Coagulase-Negative Staphylococci isolated from bovine mastitis exhibited the highest degree of susceptible to ceftriaxone of all the tested antimicrobial agents. [Vet. World 2011; 4(4.000: 158-161

  4. CNS histopathology on bovines with clinical suspicion of BSE in Denmark 2001 to 2011

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre

    During2001 to 2011 a total of 195 bovines were submitted to the instutute with clinical suspicion of having BSE. In two cases BSE was confirmed. The most common differential diagnosis was listeriosis, found in 54% of the cases. Listeriosis was characterized by multifocal, necrotizing, non-suppura...

  5. Hyaluronidase in Clinical Isolates of Propionibacterium acnes

    Directory of Open Access Journals (Sweden)

    Harmony Tyner

    2015-01-01

    Full Text Available Objectives. We sought to describe the prevalence of a hyaluronidase gene and hyaluronidase production in 197 clinical isolates of P. acnes; we assessed kinetics of hyaluronidase production in a subset of three isolates. Methods. The hyaluronidase gene was detected using polymerase chain reaction. Hyaluronidase production was detected by growing isolates on BHI agar containing 400 μg/mL hyaluronic acid and 1% albumin and flooding plates with 2 N glacial acetic acid to precipitate unbound hyaluronic acid, with a zone of clearing representing a positive phenotype. Hyaluronidase production kinetics were measured as a function of hyaluronic acid digestion over time in a liquid medium. Results. A hyaluronidase gene and hyaluronidase production were detected in 100 and 97% of P. acnes isolates, respectively. Hyaluronidase production in liquid medium was detectable after 96 hours of growth. Conclusions. Hyaluronidase production is nearly universal among P. acnes isolates. Three days appear to be required for significant hyaluronidase production in a liquid medium. Detection of hyaluronidase in tissue specimens may be a strategy to differentiate P. acnes infection from colonization when P. acnes is isolated from a clinical specimen.

  6. Staphylococcus aureus isolates from goat and sheep milk seem to be closely related and differ from isolates detected among bovine milk

    Directory of Open Access Journals (Sweden)

    Axel eMerz

    2016-03-01

    Full Text Available Dairy goat and sheep farms suffer severe economic losses due to intramammary infections, with S. aureus representing the main cause of clinical mastitis in small ruminants. In addition, S. aureus contamination of goat and sheep milk may cause staphylococcal food poisoning, as many traditional caprine and ovine milk products are not subjected to pasteurization. Data on virulence and antimicrobial resistance genes, as well as on the clonality of S. aureus detected in goat and sheep milk is scarce. Therefore, it was the aim of this study to determine i spa types and clonal complexes and ii virulence and resistance gene profiles of S. aureus isolated from goat and sheep milk. A total of 162 milk samples from sheep and goats presenting signs of an intramammary infection and 104 bulk milk samples were collected. While low prevalence rates of S. aureus were detected on single animal level, 46% of the bulk tank milk samples from small ruminants were positive for S. aureus. All isolates were spa typed and clonal complexes and virulence and resistance gene patterns were determined using a DNA microarray. Data from 49 S. aureus isolates was included in the statistical analysis and the construction of a SplitsTree. The analyzed isolates could be assigned to eleven clonal complexes, with the large majority of goat and sheep isolates being assigned to CC130 and CC133. The findings of this study suggest that S. aureus shows pronounced adaptation to small ruminants in general, but not to sheep or goats in particular. Although some common characteristics among S. aureus from caprine, ovine, and bovine milk samples were observed, S. aureus from small ruminants seem to form a distinct population. As 67% of the detected S. aureus strains exhibited at least one enterotoxin gene, many caprine or ovine raw milk products may be contaminated with low levels of enterotoxigenic S. aureus, stressing the importance of strict maintenance of the cold chain.

  7. Staphylococcus aureus Isolates from Goat and Sheep Milk Seem to Be Closely Related and Differ from Isolates Detected from Bovine Milk.

    Science.gov (United States)

    Merz, Axel; Stephan, Roger; Johler, Sophia

    2016-01-01

    Dairy goat and sheep farms suffer severe economic losses due to intramammary infections, with Staphylococcus aureus representing the main cause of clinical mastitis in small ruminants. In addition, S. aureus contamination of goat and sheep milk may cause staphylococcal food poisoning, as many traditional caprine and ovine milk products are not subjected to pasteurization. Data on virulence and antimicrobial resistance genes, as well as on the clonality of S. aureus detected in goat and sheep milk is scarce. Therefore, it was the aim of this study to determine (i) spa types and clonal complexes (CC) and (ii) virulence and resistance gene profiles of S. aureus isolated from goat and sheep milk. A total of 162 milk samples from sheep and goats presenting signs of an intramammary infection and 104 bulk milk samples were collected. While low prevalence rates of S. aureus was detected on single animal level, 46% of the bulk tank milk samples from small ruminants were positive for S. aureus. All isolates were spa typed and CC and virulence and resistance gene patterns were determined using a DNA microarray. Data from 49 S. aureus isolates was included in the statistical analysis and the construction of a SplitsTree. The analyzed isolates could be assigned to eleven CC, with the large majority of goat and sheep isolates being assigned to CC130 and CC133. The findings of this study suggest that S. aureus shows pronounced adaptation to small ruminants in general, but not to sheep or goats in particular. Although some common characteristics among S. aureus from caprine, ovine, and bovine milk samples were observed, S. aureus from small ruminants seem to form a distinct population. As 67% of the detected S. aureus strains exhibited at least one enterotoxin gene, many caprine, or ovine raw milk products may be contaminated with low levels of enterotoxigenic S. aureus, stressing the importance of strict maintenance of the cold chain. PMID:27014240

  8. Molecular and antimicrobial susceptibility analyses distinguish clinical from bovine Escherichia coli O157 strains.

    Science.gov (United States)

    Vidovic, Sinisa; Tsoi, Sarah; Medihala, Prabhakara; Liu, Juxin; Wylie, John L; Levett, Paul N; Korber, Darren R

    2013-07-01

    A population-based study combining (i) antimicrobial, (ii) genetic, and (iii) virulence analyses with molecular evolutionary analyses revealed segregative characteristics distinguishing human clinical and bovine Escherichia coli O157 strains from western Canada. Human (n = 50) and bovine (n = 50) strains of E. coli O157 were collected from Saskatchewan and Manitoba in 2006 and were analyzed by using the six-marker lineage-specific polymorphism assay (LSPA6), antimicrobial susceptibility analysis, the colicin assay, plasmid and virulence profiling including the eae, ehxA, espA, iha, stx1, stx2, stx2c, stx2d, stx2d-activatable, stx2e, and stx2f virulence-associated genes, and structure analyses. Multivariate logistic regression and Fisher's exact test strongly suggested that antimicrobial susceptibility was the most distinctive characteristic (P = 0.00487) associated with human strains. Among all genetic, virulence, and antimicrobial determinants, resistance to tetracycline (P coli O157 strains. Among 11 virulence-associated genes, stx2c showed the strongest association with E. coli O157 strains of bovine origin. LSPA6 genotyping showed the dominance of the lineage I genotype among clinical (90%) and bovine (70%) strains, indicating the importance of lineage I in O157 epidemiology and ecology. Population structure analysis revealed that the more-diverse bovine strains came from a unique group of strains characterized by a high degree of antimicrobial resistance and high frequencies of lineage II genotypes and stx2c variants. These findings imply that antimicrobial resistance generated among bovine strains of E. coli O157 has a large impact on the population of this human pathogen.

  9. O-serogroups, eae gene and EAF plasmid in Escherichia coli isolates from cases of bovine mastitis in Brazil.

    Science.gov (United States)

    Correa, M G P; Marin, J M

    2002-03-01

    Mastitis has been recognized for some time as the most costly disease in dairy herds. From March 1997 to August 1998, 2144 samples of bovine mastitic milk were collected, from which 182 Escherichia coli isolates were made, and from which 141 isolates had the somatic antigen (serogroup) determined. Twelve different serogroups were isolated from mastitic milk, and among them were O26, O55, O111 and O119, all of them classic enteropathogenic E. coli (EPEC) serogroups. These represented 40.0% of the isolates. The 20 of 57 isolates tested had plasmids and in dot blot hybridization, nine isolates were positive for an EaeA probe and an EPEC adherence factor (EAF) probe while two isolates were negative for EaeA probe but positive for the EAF probe. The nine isolates were characterized as attaching and effacing (A/E) E. coli (AEEC) isolates.

  10. Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

    KAUST Repository

    Janjanam, Jagadeesh

    2013-10-01

    Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Complete and Closed Genome Sequences of 10 Salmonella enterica subsp. enterica Serovar Anatum Isolates from Human and Bovine Sources

    Science.gov (United States)

    Nguyen, Scott V.; Bono, James L.; Smith, Timothy P. L.; Fields, Patricia I.; Dinsmore, Blake A.; Santovenia, Monica; Kelley, Christy M.; Wang, Rong; Bosilevac, Joseph M.; Harhay, Gregory P.

    2016-01-01

    Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella strains from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present the closed genome and plasmid sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolates from bovine and human sources. PMID:27257192

  12. Complete Genome Sequence of Listeria monocytogenes Strain DPC6895, a Serotype 1/2b Isolate from Bovine Raw Milk.

    Science.gov (United States)

    Casey, Aidan; McAuliffe, Olivia; Coffey, Aidan; Hunt, Karen; Fanning, Seamus; Fox, Edward; Jordan, Kieran

    2015-01-01

    Listeria monocytogenes is a foodborne pathogen and is the causative agent of listeriosis among humans and animals. The draft genome sequence of L. monocytogenes DPC6895, a serotype 1/2b strain isolated from the raw milk of a cow with subclinical bovine mastitis, is reported. PMID:26067969

  13. A comparative study of Staphylococcus aureus strains isolated from bovine subclinical mastitis during 1952-1956 and 1992

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Wegener, Henrik Caspar; Rosdahl, V.T.

    1995-01-01

    Fiftytwo strains of S. aureus isolated from cases of bovine subclinical mastitis in 52 different dairy herds in Denmark, in the peri ods 1952 to 1956 and 1992, were compared with regard to their phage- and EcoRI ribotypes. Furthermore, susceptibility to penicillin and production of fibrinolysin...

  14. Isolation and Culture of Bovine Oviductal Epithelial Cells for Use in the Anatomy and Physiology Laboratory and Undergraduate Research

    Science.gov (United States)

    Way, Amy L.

    2006-01-01

    This article presents methods for the isolation and culture of epithelial cells from the bovine oviduct for use in both research and the teaching laboratory and provides examples of ways that an oviductal cell culture can be incorporated into an undergraduate research program. Cow reproductive tracts are readily available from area butchers, and…

  15. Enterotoxin genes in coagulase-negative and coagulase-positive staphylococci isolated from bovine milk.

    Science.gov (United States)

    de Freitas Guimarães, Felipe; Nóbrega, Diego Borin; Richini-Pereira, Virginia Bodelão; Marson, Pâmela Merlo; de Figueiredo Pantoja, José Carlos; Langoni, Helio

    2013-05-01

    The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20 mL) was collected from all CMT-positive quarters. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes (sea, seb, sec, and sed). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri, S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species (S. saprophyticus ssp. saprophyticus, S. auricularis, S. capitis, and S. chromogenes) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n=85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea, seb, and sec; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb, sec, or sed, whereas S. hyicus isolates showed a greater percentage of sea than sec. Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For

  16. Isolation, characterization, and in vitro evaluation of bovine rumen submucosa films of collagen or chitosan-treated collagen.

    Science.gov (United States)

    Gopal Shankar, K; Udhaya Kumar, S; Sowndarya, S; Suresh Babu, P; Rose, C

    2016-01-01

    Bovine rumen is hitherto considered as an inedible waste of meat industry. The rumen tissues can be used as an alternative source of collagen to produce biocompatible materials for clinical application. In an effort to develop a functional biomaterial from the inedible mammalian tissues, this study aims to isolate and characterize bovine rumen submucosa. Initially, the rumen tissue was sequentially processed using chemical and enzymatic treatment to decellularize, neutralize, stabilize, and to produce a native collagen matrix which is referred as collagen film (COL-F). Thus, prepared matrix was treated with 1% (w/v) chitosan solution to produce a hybrid film which is referred as collagen-chitosan film (COL/CS-F). The comparative study includes the evaluation of physical, chemical, and biological properties of the biofilms prepared. The surface topology of COL-F exhibited a continuous collagenous network with fibrous nature, while the chitosan treatment provided smooth plain surface to the parent film. Incorporation of chitosan in COL-F increased the tensile properties, as well as the thermal stability and durability of the films. The Fourier Transform Infrared spectroscopy results revealed the presence of respective amide peaks, which corresponds to protein (collagen), and the evidence of collagen-chitosan interlinking. The submucosa layer was electrophoretically found to have type I collagen. The X-ray diffraction data showed the presence of amorphous and crystalline peak which attributes to the triple helical structure of collagen in the films. Cytotoxicity studies on the films were performed in vitro using human keratinocytes. The results of cell viability and proliferation demonstrated that COL-F and COL/CS-F exhibit good biocompatibility and therefore can augment cell infiltration and proliferation. However, enhanced cellular activity was observed on the chitosan treated COL-F. These observations demonstrate that the biofilms prepared in this study can be

  17. Distribution of tropomyosin isoforms in different types of single fibers isolated from bovine skeletal muscles.

    Science.gov (United States)

    Oe, M; Ojima, K; Nakajima, I; Chikuni, K; Shibata, M; Muroya, S

    2016-08-01

    To clarify the relationship between myosin heavy chain (MyHC) isoforms and tropomyosin (TPM) isoforms in single fibers, 64 single fibers were isolated from each of bovine three muscles (masseter, semispinalis and semitendinosus). mRNA expressions of MyHC and TPM isoforms were analyzed by real-time PCR. All single fibers from the masseter expressed MyHC-slow. The fibers from the semispinalis expressed both MyHC-slow and 2a. The fibers from the semitendinosus expressed MyHC-slow, 2a and 2x. TPM-1 and TPM-2 were co-expressed in 2a and 2x type fibers, and TPM-2 and TPM-3 were co-expressed in slow type fibers. The expression pattern of TPM isoforms in each fiber type was similar between fibers isolated from different muscles. These results suggest that TPM-1 and TPM-3 isoforms correspond to the function of 2a or 2x type fibers and slow type fibers, respectively, with TPM-2 in common. Furthermore, the patterns of MyHC and TPM isoform combinations did not vary among single fibers isolated from the individual muscles examined. PMID:27105153

  18. Distribution of tropomyosin isoforms in different types of single fibers isolated from bovine skeletal muscles.

    Science.gov (United States)

    Oe, M; Ojima, K; Nakajima, I; Chikuni, K; Shibata, M; Muroya, S

    2016-08-01

    To clarify the relationship between myosin heavy chain (MyHC) isoforms and tropomyosin (TPM) isoforms in single fibers, 64 single fibers were isolated from each of bovine three muscles (masseter, semispinalis and semitendinosus). mRNA expressions of MyHC and TPM isoforms were analyzed by real-time PCR. All single fibers from the masseter expressed MyHC-slow. The fibers from the semispinalis expressed both MyHC-slow and 2a. The fibers from the semitendinosus expressed MyHC-slow, 2a and 2x. TPM-1 and TPM-2 were co-expressed in 2a and 2x type fibers, and TPM-2 and TPM-3 were co-expressed in slow type fibers. The expression pattern of TPM isoforms in each fiber type was similar between fibers isolated from different muscles. These results suggest that TPM-1 and TPM-3 isoforms correspond to the function of 2a or 2x type fibers and slow type fibers, respectively, with TPM-2 in common. Furthermore, the patterns of MyHC and TPM isoform combinations did not vary among single fibers isolated from the individual muscles examined.

  19. Molecular Characterization of the First Bovine Herpesvirus 4 (BoHV-4 Strains Isolated from In Vitro Bovine Embryos production in Argentina.

    Directory of Open Access Journals (Sweden)

    Erika González Altamiranda

    Full Text Available Bovine herpesvirus 4 (BoHV-4 is increasingly considered as responsible for various problems of the reproductive tract. The virus infects mainly blood mononuclear cells and displays specific tropism for vascular endothelia, reproductive and fetal tissues. Epidemiological studies suggest its impact on reproductive performance, and its presence in various sites in the reproductive tract highlights its potential transmission in transfer-stage embryos. This work describes the biological and genetic characterization of BoHV-4 strains isolated from an in vitro bovine embryo production system. BoHV-4 strains were isolated in 2011 and 2013 from granulosa cells and bovine oocytes from ovary batches collected at a local abattoir, used as "starting material" for in vitro production of bovine embryos. Compatible BoHV-4-CPE was observed in the co-culture of granulosa cells and oocytes with MDBK cells. The identity of the isolates was confirmed by PCR assays targeting three ORFs of the viral genome. The phylogenetic analyses of the strains suggest that they were evolutionary unlinked. Therefore it is possible that BoHV-4 ovary infections occurred regularly along the evolution of the virus, at least in Argentina, which can have implications in the systems of in vitro embryo production. Thus, although BoHV-4 does not appear to be a frequent risk factor for in vitro embryo production, data are still limited. This study reveals the potential of BoHV-4 transmission via embryo transfer. Moreover, the high variability among the BoHV-4 strains isolated from aborted cows in Argentina highlights the importance of further research on the role of this virus as an agent with the potential to cause reproductive disease in cattle. The genetic characterization of the isolated strains provides data to better understand the pathogenesis of BoHV-4 infections. Furthermore, it will lead to fundamental insights into the molecular aspects of the virus and the means by which these

  20. Clinical, haematobiochemical, radiographic and ultrasonographic features of traumatic reticuloperitonitis in bovines

    International Nuclear Information System (INIS)

    Study was conducted to compare the clinical, haematobiochemical, radiographic and ultrasonographic features of traumatic reticuloperitonitis in bovines. Clinical cases (4 cows and 17 buffaloes) presented with a history of anorexia, fever, decreased milk yield and loss of defecation/scant faeces, were used. Haematological picture revealed neutrophilic leucocytosis with left shift and blood biochemical status showed elevated levels of total protein, albumin, and fibrinogen. Decreased plasma concentration of sodium, potassium and chloride was observed in majority of the cases. Radiographic examination revealed presence of multiple metallic foreign densities in the reticulum of the bovines. Ultrasonographically, morphological changes of reticular wall and reticulophrenic adhesions in cases of localised peritonitis were visualized. The presence of anechoic fluid without echogenic margins, not restricted to reticulum and sometimes with floating fibrinous shreds was observed in cases of diffuse peritonitis. Ultrasonography in B mode and B+ mode found helpful for the diagnosis of traumatic reticuloperitonitis and differentiation of localised peritonitis from diffuse peritonitis

  1. CNS SPECIES AND ANTIMICROBIAL RESISTANCE IN CLINICAL AND SUBCLINICAL BOVINE MASTITIS

    OpenAIRE

    Persson Waller, K.; Aspán, A; Nyman, A.; Persson, Y.; Grönlund Andersson, U.

    2011-01-01

    Abstract Coagulase-negative staphylococci (CNS) are often associated with bovine mastitis. Knowledge about the relative importance of specific CNS species in different types of mastitis, and differences in antimicrobial resistance among CNS species is, however, scarce. Therefore, the aims of this study were to compare prevalence and antimicrobial susceptibility of CNS species in clinical and subclinical mastitis using material from two national surveys. Overall, S. chromogenes and ...

  2. Production and characterization of monoclonal antibodies to Brazilian isolates of bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    L.C. Kreutz

    2000-12-01

    Full Text Available Three Brazilian isolates of bovine viral diarrhea virus (BVDV, antigenically distinct from the standard North American isolates, were selected to immunize BALB/c mice in order to obtain hybridoma cells secreting anti-BVDV monoclonal antibodies (mAbs. Two hybridoma clones secreting mAbs, reacting specifically with BVDV-infected cells (mAbs 3.1C4 and 6.F11, were selected after five fusions and screening of 1001 hypoxanthine-aminopterin-thymidine-resistant clones. These mAbs reacted in an indirect fluorescent antibody (IFA assay with all 39 South and North American BVDV field isolates and reference strains available in our laboratory, yet failed to recognize other pestiviruses, namely the hog cholera virus. The mAbs reacted at dilutions up to 1:25,600 (ascitic fluid and 1:100 (hybridoma culture supernatant in IFA and immunoperoxidase (IPX staining of BVDV-infected cells but only mAb 3.1C4 neutralized virus infectivity. Furthermore, both mAbs failed to recognize BVDV proteins by IPX in formalin-fixed paraffin-embedded tissues and following SDS-PAGE and immunoblot analysis of virus-infected cells, suggesting they are probably directed to conformational-type epitopes. The protein specificity of these mAbs was then determined by IFA staining of CV-1 cells transiently expressing each of the BVDV proteins: mAb 3.1C4 reacted with the structural protein E2/gp53 and mAb 6.F11 reacted with the structural protein E1/gp25. Both mAbs were shown to be of the IgG2a isotype. To our knowledge, these are the first mAbs produced against South American BVDV isolates and will certainly be useful for research and diagnostic purposes.

  3. Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals and prevalence of lukM-lukF-PV genes by bacteriophages in bovine isolates.

    Science.gov (United States)

    Yamada, Tomoko; Tochimaru, Naoko; Nakasuji, Sachiko; Hata, Eiji; Kobayashi, Hideki; Eguchi, Masashi; Kaneko, Jun; Kamio, Yoshiyuki; Kaidoh, Toshio; Takeuchi, Shotaro

    2005-09-30

    Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals were examined by polymerase chain reaction. LukS and lukF genes were detected in all 48 avian and 72 porcine isolates of S. aureus. LukE and lukD genes, located in a putative staphylococcal pathogenicity island (Sapln3/Saplm3), were recognized in 44 (91.7%) of 48 avian isolates, but these genes were not detected in porcine isolates. In 297 bovine isolates collected from mastitic cow's milk and bulk milk from dairy farms in two regions, lukM and lukF-PV(P83) genes in addition to lukS-lukF and lukE-lukD genes were detected in 100 (62.5%) of the 160 isolates from Ishikawa and in118 (86.1%) of the 137 isolates from Hokkaido. When the lysogeny of S. aureus bovine isolates was examined by treatment with mitomycin C, clearing of the culture due to cell lysis was observed in 34 (91.9%) of 37 lukM-lukF-PV(P83) genes--positive isolates. In addition, we isolated a novel lukM-lukF-PV(P83)-carrying (designated phiLukM), and revealed that the lukM-lukF-PV(P83) genes were located very close to an amidase gene on the temperate phage genomes. These results suggest horizontal transmission of lukM-lukF-PV(P83) genes by temperate bacteriophages in S. aureus of bovine origin.

  4. Isolation and characterization of a strain of Lichtheimia corymbifera (ex Absidia corymbifera from a case of bovine abortion

    Directory of Open Access Journals (Sweden)

    Taddei Simone

    2009-11-01

    Full Text Available Abstract Background Lichtheimia corymbifera (previously Absidia corymbifera is a filamentous zygomycetes belonging to the order Mucorales and to the family Lichtheimiaceae. Members of genus Lichtheimia spp. are cosmopolitan and ubiquitous in nature. Lichtheimia corymbifera is a recognized agent of diseases in man and animals. In cattle it causes abortion and mastitis. Three cases of bovine abortion occurred in a herd located in the Po Valley. Serological examinations were performed on fetal and mother's blood. One of the aborted fetus was referred to our laboratory. The paper describes the isolation and characterization of Lichtheimia corymbifera from a bovine aborted fetus. Methods Serological examinations were performed on fetal and mother's blood. Lesions on fetal tissues and placenta leaded the diagnostic suspect towards a mycotic aetiology. Tissues were then put in culture, and at the same time an histological examination was performed, together with bacteriological and virological tests. The isolate from placenta and fetal tissues was identified and characterized by PCR and RFLP, using the ITS region as a target sequence and AclI restriction site within the amplicon to distinguish Lichtheimia corymbifera among the other fungi. Results Serological, bacteriological and virological tests gave aspecific results. Histological examination evidenced numerous PAS positive hyphae within the necrotic cotiledons and numerous fungal nonseptate hyphae to the GMS stain. Colonies with typical morphological features of fungi grew up on Sabouraud agar from fetal skin and placenta. On the developed colonies the microscopic examination has shown a large number of nonseptate hyphae and sporangia consistent with Mucorales. PCR and RFLP allowed the identification of the isolate as Lichtheimia corymbifera. Conclusion The present report describes the isolation and the molecular characterisation of a fungal isolate from bovine aborted fetus and placenta. The

  5. Isolation and identification of bacterial causes of clinical mastitis in cattle in Sulaimania region

    Directory of Open Access Journals (Sweden)

    S. A. Hussein

    2008-01-01

    Full Text Available A total of 51 cases of bovine clinical mastitis in Sulaimani district were investigated for their bacteriological causative agents; 76 milk samples were cultured on primary and selective media and the isolated bacteria were tested for their susceptibility to antimicrobial agents used in commercial intramammary infusion products. Eighty two bacterial isolates were obtained and further identified using biochemical tests. Escherichia coli was the most common bacteria followed by Staphylococcus aureus, Streptococcus agalactia and coagulase–negative staphylococci. Two other bacterial species (Pseudomonas aeruginosa and Streptococcucs uberis were also isolated but in a lower proportion. Antibacterial susceptibility testing showed that the use of florfenicol, cephalexin and gentamicin may be useful for the treatment of clinical mastitis cases in cows.

  6. Optimization of gradual hemolysis for isolation of hemoglobin from bovine erythrocytes

    Directory of Open Access Journals (Sweden)

    Pravilović Radoslava

    2012-01-01

    Full Text Available In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system provided high yields of hemoglobin (mainly oxyhemoglobin derivate and its separation from the empty erythrocyte membranes (ghosts. Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 83% was performed using 35 mM Na-phosphate/NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The results show that isolation process yielded predominantly to oxyhemoglobin. Kinetic studies showed that maximal concentration of hemoglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min.

  7. Biofilm formation and genotyping of Staphylococcus aureus bovine mastitis isolates: Evidence for lack of penicillin-resistance in Agr-type II strains

    NARCIS (Netherlands)

    Melchior, M.B.; Osch, M.H.J.; Graat, R.; Duijkeren, van E.; Mevius, D.J.; Nielen, M.; Gaastra, W.; Fink, J.

    2009-01-01

    The increasing evidence for a role of biofilm formation in bovine mastitis caused by Staphylococcus aureus led to further investigations on biofilm formation by S. aureus isolates from mastitis in two growth media (TSBg and bovine milk serum). The ability of 99 S. aureus strains that were recently i

  8. Biofilm formation and genotyping of Staphylococcus aureus bovine mastitis isolates: evidence for lack of penicillin-resistance in Agr-type II strains.

    NARCIS (Netherlands)

    Melchior, M.B.; van Osch, M.H.J.; Graat, R.M.; van Duijkeren, E.; Mevius, D.J.; Nielen, M.; Gaastra, W.; Fink-Gremmels, J.

    2009-01-01

    The increasing evidence for a role of biofilm formation in bovine mastitis caused by Staphylococcus aureus led to further investigations on biofilm formation by S. aureus isolates from mastitis in two growth media (TSBg and bovine milk serum). The ability of 99 S. aureus strains that were recently i

  9. Clinical, epidemiological and therapeutic studies on Bovine Papillomatosis in Northern Oases, Egypt in 2008

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    Fayez Awadalla Salib 1 and Haithm Ali Farghali 2

    2011-04-01

    Full Text Available Bovine papillomatosis is a viral disease of cattle characterized clinically by development of multiple benign tumours termed warts. The diagnosis of bovine papillomatosis was confirmed by clinical and pathological examinations of the warts. The prevalence of bovine papillomatosis in Northern Oases was recorded as 4.86%. The prevalence was higher in the females (2.99% than males (1.87%.The prevalence was the highest in cattle less than one year old (2.99%. The infected cattle were examined visually for detection of external parasites and faecal samples were examined for detection of internal parasites. Infestation with ticks was observed in 10 of the 13 confirmed wart infected animals, while fasciola and parasitic gastroenteritis (PGE nematode eggs were demonstrated and quantified in 4 and 5 infected bovids respectively. The statistical correlation between the number of warts and Fasciola eggs, and number of parasitic gastroenteritis (PGE nematode eggs, was 0.6 and 0.89 accordingly. Two therapeutic regimes were evaluated, regimen-I and regimen-II. All cattle treated were completely recovered in days post treatment 15 to 115. We concluded that treatment regimen-I was better than regimen-II when taking into consideration the mean days lapsed for healing and regression of warts of 42 days compared to 83 days for regimen-II. [Veterinary World 2011; 4(2.000: 53-59

  10. Genome Sequences of Klebsiella variicola Isolates from Dairy Animals with Bovine Mastitis from Newfoundland, Canada

    OpenAIRE

    Davidson, Fraser W.; Whitney, Hugh G.; Tahlan, Kapil

    2015-01-01

    Klebsiella variicola was recently reported as an emerging and/or previously misidentified species associated with opportunistic infections in humans. Here, we report the draft genome sequences of K. variicola isolates from two animals with clinical mastitis from a dairy farm in Newfoundland, Canada.

  11. Yeasts isolated from clinical samples of AIDS patients

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    Neves Rejane Pereira

    2002-01-01

    Full Text Available In order to investigate yeasts in oropharyngeal secretion, urine, sputum and inguinal scales from AIDS patients, clinical samples were collected from one hundred patients interned in the Infectious and Parasitic Diseases Sector of the Hospital das Clínicas of the Universidade Federal de Pernambuco and in Hospital Universitário Osvaldo Cruz of the Universidade de Pernambuco. Yeasts were isolated from seventy-two out of one hundred and eight clinical samples. The isolated yeasts were: Candida albicans (sixty-two isolates, Candida tropicalis (four isolates, Candida glabrata (two isolates, Candida parapsilosis (two isolates, Candida krusei (one isolate and Trichosporon pullulans (one isolate.

  12. Conversion from clinically isolated syndrome to multiple sclerosis

    DEFF Research Database (Denmark)

    Kuhle, J; Disanto, G; Dobson, R;

    2015-01-01

    BACKGROUND AND OBJECTIVE: We explored which clinical and biochemical variables predict conversion from clinically isolated syndrome (CIS) to clinically definite multiple sclerosis (CDMS) in a large international cohort. METHODS: Thirty-three centres provided serum samples from 1047 CIS cases...

  13. Biochemical and molecular characterization of Treponema phagedenis-like spirochetes isolated from a bovine digital dermatitis lesion

    OpenAIRE

    Wilson-Welder, Jennifer H.; Elliott, Margaret K.; Zuerner, Richard L.; Bayles, Darrell O.; Alt, David P.; Stanton, Thad B.

    2013-01-01

    Background Bovine papillomatous digital dermatitis (DD) is the leading cause of lameness in dairy cattle and represents a serious welfare and economic burden. Found primarily in high production dairy cattle worldwide, DD is characterized by the development of an often painful red, raw ulcerative or papillomatous lesion frequently located near the interdigital cleft and above the bulbs of the heel. While the exact etiology is unknown, several spirochete species have been isolated from lesion m...

  14. Simultaneous Isolation of Lactoferrin and Lactoperoxidase from Bovine Colostrum by SPEC 70 SLS Cation Exchange Resin

    Science.gov (United States)

    Liang, Yafei; Wang, Xuewan; Wu, Mianbin; Zhu, Wanping

    2011-01-01

    In this work, simultaneous isolation of lactoferrin (Lf) and lactoperoxidase (Lp) from defatted bovine colostrum by one-step cation exchange chromatography with SPEC 70 SLS ion-exchange resin was investigated. A RP-HPLC method for Lf and Lp determination was developed and optimized as the following conditions: detection wavelength of 220 nm, flow rate of 1 mL/min and acetonitrile concentration from 25% to 75% within 20 min. The adsorption process of Lf on SPEC 70 SLS resin was optimized using Lf standard as substrate. The maximum static binding capacity of SPEC 70 SLS resin was of 22.0 mg/g resin at 15 °C, pH 7.0 and adsorption time 3 h. The Lf adsorption process could be well described by the Langmuir adsorption isotherm model, with a maximum adsorption capacity of 21.73 mg/g resin at 15 °C. In batch fractionation of defatted colostrum, the binding capacities of SPEC 70 SLS resin for adsorbing Lf and Lp simultaneously under the abovementioned conditions were 7.60 and 6.89 mg/g resin, respectively, both of which were superior to those of CM Sepharose F.F. or SP Sepharose F.F. resins under the same conditions. As a result, SPEC 70 SLS resin was considered as a successful candidate for direct and economic purification of Lf and Lp from defatted colostrum. PMID:22016715

  15. Simultaneous isolation of lactoferrin and lactoperoxidase from bovine colostrum by SPEC 70 SLS cation exchange resin.

    Science.gov (United States)

    Liang, Yafei; Wang, Xuewan; Wu, Mianbin; Zhu, Wanping

    2011-09-01

    In this work, simultaneous isolation of lactoferrin (Lf) and lactoperoxidase (Lp) from defatted bovine colostrum by one-step cation exchange chromatography with SPEC 70 SLS ion-exchange resin was investigated. A RP-HPLC method for Lf and Lp determination was developed and optimized as the following conditions: detection wavelength of 220 nm, flow rate of 1 mL/min and acetonitrile concentration from 25% to 75% within 20 min. The adsorption process of Lf on SPEC 70 SLS resin was optimized using Lf standard as substrate. The maximum static binding capacity of SPEC 70 SLS resin was of 22.0 mg/g resin at 15 °C, pH 7.0 and adsorption time 3 h. The Lf adsorption process could be well described by the Langmuir adsorption isotherm model, with a maximum adsorption capacity of 21.73 mg/g resin at 15 °C. In batch fractionation of defatted colostrum, the binding capacities of SPEC 70 SLS resin for adsorbing Lf and Lp simultaneously under the abovementioned conditions were 7.60 and 6.89 mg/g resin, respectively, both of which were superior to those of CM Sepharose F.F. or SP Sepharose F.F. resins under the same conditions. As a result, SPEC 70 SLS resin was considered as a successful candidate for direct and economic purification of Lf and Lp from defatted colostrum.

  16. Simultaneous Isolation of Lactoferrin and Lactoperoxidase from Bovine Colostrum by SPEC 70 SLS Cation Exchange Resin

    Directory of Open Access Journals (Sweden)

    Mianbin Wu

    2011-09-01

    Full Text Available In this work, simultaneous isolation of lactoferrin (Lf and lactoperoxidase (Lp from defatted bovine colostrum by one-step cation exchange chromatography with SPEC 70 SLS ion-exchange resin was investigated. A RP-HPLC method for Lf and Lp determination was developed and optimized as the following conditions: detection wavelength of 220 nm, flow rate of 1 mL/min and acetonitrile concentration from 25% to 75% within 20 min. The adsorption process of Lf on SPEC 70 SLS resin was optimized using Lf standard as substrate. The maximum static binding capacity of SPEC 70 SLS resin was of 22.0 mg/g resin at 15 °С, pH 7.0 and adsorption time 3 h. The Lf adsorption process could be well described by the Langmuir adsorption isotherm model, with a maximum adsorption capacity of 21.73 mg/g resin at 15 °С. In batch fractionation of defatted colostrum, the binding capacities of SPEC 70 SLS resin for adsorbing Lf and Lp simultaneously under the abovementioned conditions were 7.60 and 6.89 mg/g resin, respectively, both of which were superior to those of CM Sepharose F.F. or SP Sepharose F.F. resins under the same conditions. As a result, SPEC 70 SLS resin was considered as a successful candidate for direct and economic purification of Lf and Lp from defatted colostrum.

  17. Differentiation of ruminant transmissible spongiform encephalopathy isolate types, including bovine spongiform encephalopathy and CH1641 scrapie.

    Science.gov (United States)

    Jacobs, J G; Sauer, M; van Keulen, L J M; Tang, Y; Bossers, A; Langeveld, J P M

    2011-01-01

    With increased awareness of the diversity of transmissible spongiform encephalopathy (TSE) strains in the ruminant population, comes an appreciation of the need for improved methods of differential diagnosis. Exposure to bovine spongiform encephalopathy (BSE) has been associated with the human TSE, variant Creutzfeldt-Jakob disease, emphasizing the necessity in distinguishing low-risk TSE types from BSE. TSE type discrimination in ruminants such as cattle, sheep, goats and deer, requires the application of several prion protein (PrP)-specific antibodies in parallel immunochemical tests on brain homogenates or tissue sections from infected animals. This study uses in a single incubation step, three PrP-specific antibodies and fluorescent Alexa dye-labelled anti-mouse Fabs on a Western blot. The usual amount of brain tissue needed is 0.5 mg. This multiplex application of antibodies directed towards three different PrP epitopes enabled differential diagnosis of all established main features of classical scrapie, BSE and Nor98-like scrapie in sheep and goats, as well as the currently known BSE types C, H and L in cattle. Moreover, due to an antibody-dependent dual PrP-banding pattern, for the first time CH1641 scrapie of sheep can be reliably discriminated from the other TSE isolate types in sheep. PMID:20943889

  18. Streptococcus uberis strains isolated from the bovine mammary gland evade immune recognition by mammary epithelial cells, but not of macrophages.

    Science.gov (United States)

    Günther, Juliane; Czabanska, Anna; Bauer, Isabel; Leigh, James A; Holst, Otto; Seyfert, Hans-Martin

    2016-01-01

    Streptococcus uberis is frequently isolated from the mammary gland of dairy cattle. Infection with some strains can induce mild subclinical inflammation whilst others induce severe inflammation and clinical mastitis. We compared here the inflammatory response of primary cultures of bovine mammary epithelial cells (pbMEC) towards S. uberis strains collected from clinical or subclinical cases (seven strains each) of mastitis with the strong response elicited by Escherichia coli. Neither heat inactivated nor live S. uberis induced the expression of 10 key immune genes (including TNF, IL1B, IL6). The widely used virulent strain 0140J and the avirulent strain, EF20 elicited similar responses; as did mutants defective in capsule (hasA) or biofilm formation (sub0538 and sub0539). Streptococcus uberis failed to activate NF-κB in pbMEC or TLR2 in HEK293 cells, indicating that S. uberis particles did not induce any TLR-signaling in MEC. However, preparations of lipoteichoic acid (LTA) from two strains strongly induced immune gene expression and activated NF-κB in pbMEC, without the involvement of TLR2. The immune-stimulatory LTA must be arranged in the intact S. uberis such that it is unrecognizable by the relevant pathogen receptors of the MEC. The absence of immune recognition is specific for MEC, since the same S. uberis preparations strongly induced immune gene expression and NF-κB activity in the murine macrophage model cell RAW264.7. Hence, the sluggish immune response of MEC and not of professional immune cells to this pathogen may aid establishment of the often encountered belated and subclinical phenotype of S. uberis mastitis. PMID:26738804

  19. Isolation and Identification of Bovine Viral Diarrhea Virus Shandong Strain%牛病毒性腹泻病毒山东株的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    侯佩莉; 杨宏军; 宋玲玲; 王洪梅; 刘来兴; 何洪彬

    2013-01-01

    牛病毒性腹泻病毒是牛病毒性腹泻-黏膜病的重要病原体,该病毒病是极为复杂、呈多临床类型表现的传染病,给世界养牛业造成了巨大的经济损失.本研究从疑似牛病毒性腹泻-黏膜病的粪便中分离得到一株病毒能使MDBK细胞产生细胞病变,经RT-PCR检测及扩增产物测序进一步证实,该病毒为牛病毒性腹泻-黏膜病病毒,TCID50测定该病毒的滴度为107.15TCID50/ml.该病毒株的分离鉴定为牛病毒性腹泻病毒疫苗的研制奠定了基础.%Bovine viral diarrhea virus ( BVDV) is the causative agent of bovine viral diarrhoea, and is u-biquitous in cattle. It is very complex and presents a wide range of clinical manifestations, which causes large economic loss to cattle industry around the world. In this study, a virus strain causing cytopathic effect of ma-din -darby bovinekidney (MDBK) was isolated from the bovine excrement and identified as BVDV by RT -PCR and sequencing analysis. The virus titer was 10 7.15 TCID50/ml. The isolation and identification of this virus strain laid foundation for exploring its vaccine.

  20. Morphologic characterization of isolated bovine early preantral follicles during short-term individual in vitro culture.

    Science.gov (United States)

    Jorssen, E P A; Langbeen, A; Marei, W F A; Fransen, E; De porte, H F M; Leroy, J L M R; Bols, P E J

    2015-07-15

    To provide new insights in the molecular mechanism controlling preantral follicular development and to unravel the needs to support in vitro follicular development of early-stage preantral follicles (PAFs), there is a need for alternative in vitro bovine follicle culture methods. In this study, we aimed to characterize follicular dynamics using an IVC system of isolated and individually cultured bovine early PAFs during 10 days to generate individual follicle follow-up data. Preantral follicles (ovaries and cultured individually for 10 days. Individual follicle morphology, growth, survival, quality, and cell proliferation were evaluated in time by combining noninvasive and invasive assessment methods. The PAFs were light microscopically evaluated during culture to assess follicular dynamics, stained with neutral red to determine follicle viability, stained with 4',6-diamidino-2-phenylindole and terminal deoxynucleotidyl transferase dUTP nick end labeling to evaluate cell proliferation and follicle quality, and processed for histologic evaluation to assess follicle morphology. On the basis of their morphology, follicles were subdivided in three categories, with category 1 follicles showing the best morphologic features. On Day 0, only category 1 follicles were selected, but follicle categories were reassigned on evaluation Days 1, 2, 4, 7, or 10. Although 67% of the follicles survived 10 days of IVC, the number of follicles exhibiting a normal morphology decreased significantly from Day 7 onward and the apoptotic index increased significantly from Day 10. Both category 1 and 2 follicles showed a significant increase in follicular diameter (Day 10: 21.80 ± 0.86 and 11.82 ± 0.80, respectively). This increase in follicular diameter showed to be correlated with an increase in the total cell number. In conclusion, this culture system showed to support follicular development until Day 10, although the proportion of follicles showing normal morphologic features and the

  1. Polymicrobial biofilms by diabetic foot clinical isolates.

    Science.gov (United States)

    Mottola, Carla; Mendes, João J; Cristino, José Melo; Cavaco-Silva, Patrícia; Tavares, Luís; Oliveira, Manuela

    2016-01-01

    Diabetes mellitus is a major chronic disease that continues to increase significantly. One of the most important and costly complications of diabetes is foot ulceration that may be colonized by pathogenic and antimicrobial resistant bacteria, which may express several virulence factors that could impair treatment success. These bacterial communities can be organized in polymicrobial biofilms, which may be responsible for diabetic foot ulcer (DFU) chronicity. We evaluated the influence of polymicrobial communities in the ability of DFU isolates to produce biofilm, using a microtiter plate assay and a multiplex fluorescent in situ hybridization, at three time points (24, 48, 72 h), after evaluating biofilm formation by 95 DFU isolates belonging to several bacterial genera (Staphylococcus, Corynebacterium, Enterococcus, Pseudomonas and Acinetobacter). All isolates were biofilm-positive at 24 h, and the amount of biofilm produced increased with incubation time. Pseudomonas presented the higher biofilm production, followed by Corynebacterium, Acinetobacter, Staphylococcus and Enterococcus. Significant differences were found in biofilm formation between the three time points. Polymicrobial communities produced higher biofilm values than individual species. Pseudomonas + Enterococcus, Acinetobacter + Staphylococcus and Corynebacterium + Staphylococcus produced higher biofilm than the ones formed by E. faecalis + Staphylococcus and E. faecalis + Corynebacterium. Synergy between bacteria present in dual or multispecies biofilms has been described, and this work represents the first report on time course of biofilm formation by polymicrobial communities from DFUs including several species. The biological behavior of different bacterial species in polymicrobial biofilms has important clinical implications for the successful treatment of these infections.

  2. Investigation of Virulence Genes by PCR in Stapylococcus aureus Isolates Originated from Subclinical Bovine Mastitis in Turkey

    Directory of Open Access Journals (Sweden)

    Murat Karahan, Mehmet Nuri Acik1* and Burhan Cetinkaya

    2011-06-01

    Full Text Available The aim of the present study was to characterize coagulase (coa positive Staphylococcus aureus strains (n=92 isolated from bovine subclinical mastitis in Turkey by PCR amplification of clumping factor A (clfA and protein A (spa genes. All the coa-positive S. aureus isolates were determined to harbor the genes encoding the IgG binding region (spa-IgG and the X region (spa-X of spa. On the other hand, 84 (91.3% isolates were positive for clfA gene. These three genes displayed size polymorphisms. It was concluded that spa gene polymorphisms for S. aureus, when used together with coa-PCR, can be proposed as good alternatives to conventional methods in typing S. aureus isolates of bovine origin which may provide valuable data for the development of effective control strategies against staphylococcal mastitis. The results of the present study showed that S. aureus isolates responsible for the mastitis cases in Turkey were genetically diverse.

  3. Genetic relatedness of Trichomonas vaginalis reference and clinical isolates.

    Science.gov (United States)

    Cornelius, Denise C; Mena, Leandro; Lushbaugh, William B; Meade, John C

    2010-12-01

    We have determined the metronidazole susceptibility status of 20 Trichomonas vaginalis isolates from American Type Culture Collection (ATCC) and assessed the level of genetic relatedness in these isolates using 32 additional T. vaginalis clinical isolates for comparison. These ATCC isolates are commonly used as reference strains in T. vaginalis research and this information provides a rational basis for selection of reference strains for use in comparative studies of T. vaginalis phenotypic and clinical characteristics. PMID:21118935

  4. Generation of bovine (Bos indicus) and buffalo (Bubalus bubalis) adipose tissue derived stem cells: isolation, characterization, and multipotentiality.

    Science.gov (United States)

    Sampaio, R V; Chiaratti, M R; Santos, D C N; Bressan, F F; Sangalli, J R; Sá, A L A; Silva, T V G; Costa, N N; Cordeiro, M S; Santos, S S D; Ambrosio, C E; Adona, P R; Meirelles, F V; Miranda, M S; Ohashi, O M

    2015-01-15

    Adult stem cells are known for their plasticity and their potential to differentiate into several different cell types; these characteristics have implications for cell therapy and reproductive biotechnologies. In this study, we report on the isolation and characterization of mesenchymal stem cells (MSC) derived from bovine and buffalo adipose tissue. Cells isolated using enzymatic digestion of bovine and buffalo adipose-tissue biopsy samples were grown in vitro for at least 15 passages, verifying their capacity to proliferate. These cells were also subjected to immunophenotypic characterization for the presence of CD90, CD105, and CD79, and the absence of CD45, CD34, and CD73, which are positive and negative markers of MSC, respectively. To prove their multipotency, the cells were induced to differentiate into three different cell types, chondrocytes, osteoblasts, and adipocytes, which were stained with tissue-specific dyes (Chondrogenic-Alcian Blue, Osteogenic-Alizarin Red, and Adipogenic-Oil-Red O, respectively) to confirm differentiation. Gene expression analysis of pluripotency-related genes was also conducted. Our results suggest that adipose tissue from bovines and buffalos can be used as a source of MSC, making adipose tissue-derived cells an interesting option for cell therapy and regenerative medicine. Additionally, these findings have implications for reproductive biotechnology because the use of MSC as nuclear donors has been linked to an increase in the efficiency of nuclear transfer.

  5. Reflections on the 24 years durability of an isolate tricuspid bovine pericardium IMC/Braile bioprosthesis.

    Science.gov (United States)

    Bassetto, Solange; Menardi, Antonio Carlos; Alves Junior, Lafaiete; Rodrigues, Alfredo José; Évora, Paulo Roberto Barbosa

    2011-01-01

    We were challenged by the experience of one patient reoperation for a bioprosthetic bovine pericardium degenerative stenosis, 24 years after implantation. This bioprosthesis was implanted due to tricuspid valve bacterial staphylococcal endocarditis after septic abortion.

  6. Dietary requirement for serum-derived bovine immunoglobulins in the clinical management of patients with enteropathy.

    Science.gov (United States)

    Petschow, Bryon W; Burnett, Bruce P; Shaw, Audrey L; Weaver, Eric M; Klein, Gerald L

    2015-01-01

    A variety of human disease conditions are associated with chronic intestinal disorders or enteropathies that are characterized by intestinal inflammation, increased gut permeability, and reduced capacity to absorb nutrients. Such disruptions in the homeostasis of the gastrointestinal (GI) tract can lead to symptoms of abdominal pain and discomfort, bloating, abnormal bowel function, and malabsorption of nutrients. While significant advances have been made in understanding the factors that influence the complex and fragile balance between the gut microbiota, intestinal epithelial cell integrity, and the underlying immune system, effective therapies for restoring intestinal balance during enteropathy are still not available. Numerous studies have demonstrated the ability of oral immunoglobulins to improve weight gain, support gut barrier function, and reduce the severity of enteropathy in animals. More recently, studies in humans provide evidence that serum-derived bovine immunoglobulin/protein isolate is safe and improves nutritional status and GI symptoms in patients with enteropathy associated with irritable bowel syndrome or infection with the human immunodeficiency virus. This review summarizes studies showing the impact of enteropathy on nutritional status and how specially formulated bovine immunoglobulins may help restore intestinal homeostasis and nutritional status in patients with specific enteropathies. Such protein preparations may provide distinct nutritional support required for the dietary management of patients who, because of therapeutic or chronic medical needs, have limited or impaired capacity to digest, absorb, or metabolize ordinary foodstuffs or certain nutrients, or other special medically determined nutrient requirements that cannot be satisfied by changes to the normal diet alone. PMID:25142170

  7. Isolation and characterisation of a ruminant alphaherpesvirus closely related to bovine herpesvirus 1 in a free-ranging red deer

    Directory of Open Access Journals (Sweden)

    Belák Sándor

    2007-09-01

    Full Text Available Abstract Background The genus Varicellovirus of the Herpesviridae subfamily Alphaherpesvirinae includes a cluster of viruses antigenically and genetically related to bovine herpesvirus 1 (BoHV-1: namely bovine herpesvirus 5 (BoHV-5, bubaline herpesvirus 1 (BuHV-1, caprine herpesvirus 1 (CpHV-1, cervid herpesviruses 1 (CvHV-1 and 2 (CvHV-2 and elk herpesvirus 1 (ElkHV-1. Considering the serological relationship between these ruminant alphaherpesviruses, several surveys have studied the occurrence of BoHV-1 related virus infection in wild and domestic ruminant species. In this way, a recent investigation has indicated, in Belgium, a high increase in the serological prevalence of BoHV-1 related virus infection in free-ranging red deer population. In this context, it has been decided to investigate the presence of an alphaherpesvirus spreading in the Belgian free-ranging red deer population. Results The current study reports the first isolation in a free-ranging red deer of a BoHV-1 closely related virus. The isolate was antigenically, genomically and genetically characterised by comparison with several ruminant alphaherpesvirus. Immunofluorescence assays revealed the isolate was antigenically distinct from bovine and caprine alphaherpesviruses. Similarly, BamHI and BstEII restriction analyses demonstrated the genomic difference between the isolate and the other ruminant alphaherpesviruses. Next, the sequencing of selected parts of UL27 and US8 genes showed a high degree of homologies between each BoHV-1 related ruminant alphaherpesvirus and the isolate. Besides the close relationship between all ruminant alphaherpesviruses, the phylogenetic analysis revealed that the isolate clustered with CvHV-1. Conclusion The first isolation of a virus closely related to BoHV-1 in a free-ranging red deer is reported. Data demonstrate that a CvHV-1 strain, named Anlier, circulates in wild red deer in continental Europe. Anlier strain show consistent differences

  8. Identification of amino acid changes in the envelope glycoproteins of bovine viral diarrhea viruses isolated from alpaca that may be involved in host adaptation

    Science.gov (United States)

    Bovine viral diarrhea viruses (BVDV) are most commonly associated with infections of cattle. However, BVDV is often isolated from closely related ruminants with a number of BVDV-1b viruses being isolated from alpacas that were both acutely and persistently infected (PI). The complete nucleotide se...

  9. Comparison of flow cytometry and virus isolation in cell culture for identification of cattle persistently infected with bovine viral diarrhea virus.

    OpenAIRE

    Qvist, P.; Houe, H.; Aasted, B.; Meyling, A

    1991-01-01

    Detection of bovine viral diarrhea virus in 143 blood samples by virus isolation in cell culture and flow cytometry was performed. The material included 37 samples later shown to originate from persistently infected cattle. Thirty-three samples were positive by virus isolation, and all 37 samples were positive by the flow cytometric assay.

  10. Prevalence, genetic diversity, and antimicrobial susceptibility profiles of Staphylococcus aureus isolated from bovine mastitis in Zhejiang Province, China

    Institute of Scientific and Technical Information of China (English)

    Jian-ping LI; Hai-jian ZHOU; Lin YUAN; Ting HE; Song-hua HU

    2009-01-01

    This study was conducted to determine genetic diversity and antimicrobial susceptibility profiles of Staphylococcusaureus recovered from bovine mastitis in Zhejiang Province, China. Out of 3178 quarter milk samples from 846 lactating cows, among which 459 cows (54.3%) were found HMT positive, 890 quarters (28%) were found having subclinical mastitis. From 75 representative S. aureus isolates, 16 distinct types were identified by pulsed-field gel electrophoresis (PFGE). Four major PFGE types (A, B, C, and D) accounted for 82.7% of all isolates, and type A (41.3%) was observed in multiple herds across the studied areas. Each region was found to have a predominant type: Hangzhou type A (64.1%), Ningbo type C (34.5%) and type B (23.1%), Jinhua type D (53.3%), and Taizhou type C (62.5%). Results of antimicrobial susceptibility tests showed that 90.7% of the isolates were resistant to at least one antimicrobial. Resistance to penicillin and ampicillin (77.3%), tetracycline (60.0%), or erythromycin (48.0%) was observed. The bacteria resistant to multiple antibiotics such as penicillin, ampicillin, tetracycline, and erythromycin were commonly found. The information obtained from this study is useful for designing specific control programs for bovine S. aureus mastiffs in this region.

  11. Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

    Science.gov (United States)

    Samrath, Devprabha; Shakya, Sanjay; Rawat, Nidhi; Gilhare, Varsha Rani; Singh, Fateh

    2016-01-01

    Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test. PMID:27051213

  12. Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

    Directory of Open Access Journals (Sweden)

    Devprabha Samrath

    2016-02-01

    Full Text Available Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1 from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK cell line. Further, the virus was identified by agar gel immunodiffusion (AGID test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test.

  13. Complete Genome Sequences of Two Bovine Viral Diarrhea Viruses Isolated from Brain Tissues of Nonambulatory (Downer) Cattle

    OpenAIRE

    Oem, Jae-Ku; Joo, Soo-Kyung; An, Dong-Jun

    2013-01-01

    Here, we report the complete genome sequences of two bovine viral diarrhea viruses (BVDVs) (strains 11F011 and 12F004) isolated from brain tissues from nonambulatory (downer) cattle. The complete genomes of strains 11F011 and 12F004 contain 12,287 nucleotides (nt) with a single large open reading frame and 12,301 nt with a single large open reading frame, respectively. Phylogenetic analysis indicated that these strains belong to the BVDV-2a and -1b genotypes, respectively.

  14. The release and vascular action of bradykinin in the isolated perfused bovine udder.

    Science.gov (United States)

    Zeitlin, I J; Eshraghi, H R

    2002-08-15

    . Thus, in addition to its secretion into milk, BK, together with its precursor and tissue kallikrein, is continuously released into the vasculature of the isolated, perfused, lactating bovine udder. PMID:12181294

  15. Genetic Relatedness of Trichomonas vaginalis Reference and Clinical Isolates

    OpenAIRE

    Cornelius, Denise C.; Mena, Leandro; Lushbaugh, William B.; Meade, John C.

    2010-01-01

    We have determined the metronidazole susceptibility status of 20 Trichomonas vaginalis isolates from American Type Culture Collection (ATCC) and assessed the level of genetic relatedness in these isolates using 32 additional T. vaginalis clinical isolates for comparison. These ATCC isolates are commonly used as reference strains in T. vaginalis research and this information provides a rational basis for selection of reference strains for use in comparative studies of T. vaginalis phenotypic a...

  16. Genotyping of Pasteurella multocida ovine and bovine isolates from Iran based on PCR-RFLP of ompH gene

    Directory of Open Access Journals (Sweden)

    A. Ghanizadeh

    2015-10-01

    Full Text Available Pasteurella multocida (P. multocida, A Gram-negative facultative anaerobic bacterium, is a causative animal pathogen in porcine atrophic rhinitis and avian fowl cholera. The outer membrane of Gram-negative bacteria contains of many different protein in very high copy numbers. One of the major outer membrane, the H proteins have functional as high immunogenicity and antigenicity. In this study to increase information about epidemiology of ovine and bovine P. multocida, the 24 isolates from sheep and nine isolates from cattle were investigated by PCR-RFLP analysis of the ompH gene. In all 33 isolates, digestion of the amplified fragment of ompH gene by using EcoRI, cfoI and HindIII produced 3, 5 and 3 different restriction patterns respectively. Sixteen RFLP patterns were found among 33 investigated P.multocida isolates. This study showed that, the PCR RFLP based on ompH gene is potentially a useful method for typing of P. multocida isolates from sheep and cattle. The RFLP patterns of this gene exhibited extensive restriction site heterogeneity, which may be particularly suitable for fingerprinting of P. multocida isolates.Considering ompH protein as a protective immunogenic moiety of P.ultocida, the results of this study showed a heterogenic bacteria and this means the possibility to produce a multivalent vaccine to be protective against diseases caused by this organism in sheep and cattle in Iran.

  17. Complete Genome Sequence of a Bovine Viral Diarrhea Virus Subgenotype 1e Strain Isolated in Switzerland

    OpenAIRE

    Stalder, Hanspeter; Schweizer, Matthias; Bachofen, Claudia

    2015-01-01

    We sequenced the complete genome of the bovine viral diarrhea virus (BVDV) strain Carlito. It belongs to the subgenotype 1e that is described in Europe only and represents the second most prevalent subgenotype in Switzerland. This is the first report of a full-length sequence of BVDV-1e.

  18. Candida species isolated from various clinical specimens

    OpenAIRE

    Uraz, Güven; Arslan, Seza; Ekener, Serpil

    2000-01-01

    Objectives: The various culture specimens of patients were investigated in our research. Material and methods: 123 Candida strains were isolated. In identification of Candida species that were isolated, germ tube test, growth in Cornmeal-Tween 80 agar and formation of clamydospore, presence of pseudohyphae, carbonhytrate fermentation and assimilation tests, formation of membranes in sabouraud medium, and the tests of ascospore in the mediums with cycloheximide and the test of nitrate were...

  19. Risk factors associated with the antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Daniele C. Beuron

    2014-10-01

    Full Text Available The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7% were culture-positive, and S. aureus comprised 27.77% (n=210 of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18, dry cow treatment for enrofloxacin (OR=2.11 and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57 and penicillin (OR=4.69. In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.

  20. Antibiotic Susceptibility of Coagulase-Negative Staphylococci Isolated from Bovine Subclinical Mastitis in Turkey

    Directory of Open Access Journals (Sweden)

    Beytullah Kenar*, Yahya Kuyucuoğlu and Esra Şeker

    2012-06-01

    Full Text Available A total of 572 California Mastitis Test (CMT positive milk samples were collected from 423 lactating cows on 18 private farms in the Middle Western Anatolia. Coagulase–negative staphylococci colonies and CNS species identification was performed based on conventional biochemical techniques and using the API Staph test. Slime production was detected by Congo Red Agar (CRA method. The antibiotic susceptibility was determined according to the National Committee for Clinical Laboratory Standards guidelines (NCCLS. A total of 67 (11.7% coagulase-negative staphylococci (CNS were isolated from CMT positive milk samples. In total, 11 CNS species: S. epidermidis (n=18, S. simulans (n=14, S. warneri (n=10, S. hominis (n=5, S. chromogenes (n=4, S. caprae (n=4, S. xylosus (n=3, S. haemolyticus (n=3, S. hyicus (n=3, S. cohnii (n=2, and S. capitis (n=1 were identified. The most commonly identified CNS species were Staphylococcus epidermidis (26.8% and Staphylococcus simulans (20.8% followed by Staphylococcus warneri (14.9%. Out of 67 CNS isolates, slime production was found in 37 (55.2% CNS strains. CNS isolates were the most resistance to trimethoprim+sulphamethoxazole (76.2%, erythromycin (73.2%, oxacillin and ampicillin (70.2% followed by penicillin (58.3%, gentamicin (53.8%, tetracycline (52.3%, vancomycin (51.8%, ciprofloxacin (26.9%, cefoxitim (23.9%, and cephalothin (13.5%. These results indicate that CNS species are resistant at high rates to the beta-lactam antibiotics which are intensively used in the prevention and treatment of mastitis without any antibiotic susceptibility test in the Middle Western of Turkey.

  1. Detection of High Pathogenicity Islands in Escherichia coli Isolated from Clinical and Subclinical Bovine Mastitis in Some Areas of China%奶牛乳腺炎源大肠杆菌中耶尔森菌强毒力岛相关基因的检测及序列分析

    Institute of Scientific and Technical Information of China (English)

    徐继英; 杨志强; 陈化琦; 刘俊林; 邢娟; 李建喜; 李宏胜

    2012-01-01

    [目的]研究奶牛乳腺炎源大肠杆菌中耶尔森菌HPI携带情况及其与O血清型的关系,并对部分菌株的相关基因序列进行分析.[方法]从中国北京、内蒙古、甘肃、四川、重庆、云南、贵州等7个省市部分地区1260份临床型和隐性奶牛乳腺炎奶样中分离得到190株大肠杆菌,对分离菌株进行耶尔森菌强毒力岛核心区irp2基因、fyuA基因及HPI毒力岛在大肠杆菌染色体中插入位置的鉴定,分析HPI毒力岛的携带情况及其与分离菌株O血清型之间的关系.[结果]190株大肠杆菌分离株中,irp2基因阳性率为26.31% (50/190),fyuA基因阳性率为18.94% (36/190).50株HPI+分离株中检出asn_tRNA_intB基因32株,阳性率为64% (32/50).本试验克隆的irp2基因(273 bp)、fyuA基因(1071 bp)、asn_tRNA_intB基因(1512 bp)均与已发表序列高度同源,同源性分别在97.1%、98.2%、97.2%以上,且其HPI毒力岛大多位于大肠杆菌染色体的asn-tRNA位点上.[结论]耶尔森菌HPI在奶牛乳腺炎源大肠杆菌中广泛流行分布,但也存在差异,而不同血清型菌株携带HPI的倾向性可能只与特定的血清型有一定的关系.%[Objective] The objective of the experiment is to investigate the present distribution of high pathogenicity island (HPI) in E. coli isolated from cows with clinical and subclinical mastitis in China. [Method] Milk samples from 1 260 cows with clinical and subclinical mastitis in different farms and areas were used to screen Gram-negative bacteria by MacConkey agar. One hundred and ninety E. coli isolates was further identified from the Gram-negative bacteria by microbiological tests. Total DNA extraction from E. coli was used to detect HPI genes by PCR. [ Result] The PCR results showed that 50 out of 190 isolates carried the irp2 gene, 36 isolates were positive for fyuA gene and 32 for intB genes. Sequence analysis of randomly selected PCR products showed that homology of genes irp2, fyu

  2. Biofilm-producing ability and efficiency of sanitizing agents against Prototheca zopfii isolates from bovine subclinical mastitis.

    Science.gov (United States)

    Gonçalves, Juliano Leonel; Lee, Sarah Hwa In; de Paula Arruda, Eurico; Pedroso Galles, Débora; Camargo Caetano, Vinícius; Fernandes de Oliveira, Carlos Augusto; Fernandes, Andrezza Maria; Veiga dos Santos, Marcos

    2015-06-01

    The objectives of the present study were to evaluate (1) the capacity of the microalga Prototheca zopfii isolated from subclinical bovine mastitis cases to form biofilms; and (2) the resistance of these isolates to sanitizing agents. Ten isolates of P. zopfii from cows with subclinical mastitis (somatic cell count>200×10(3) cells/mL), distributed in 5 dairy farms, were evaluated for their capacity to form biofilms in polystyrene microplate assays and stainless steel coupons, at 25°C and 37°C±1°C. Prototheca zopfii were isolated from milk samples via microbiological culture and analyzed by 18S rRNA gene sequencing. Biofilm formation on the coupons was observed by scanning electron microscopy. The resistance to sanitizing agents was assessed using the biofilm-forming P. zopfii isolates in stainless steel coupon assays, which were subjected to 3 sanitizers: peracetic acid, sodium hypochlorite, and iodine solution. To evaluate resistance to the sanitizers, the minimum inhibitory concentration (MIC) technique was performed using decreasing concentrations of the sanitizing agents (20, 10, 5, 2.5, 1.25, 0.625, 0.312, 0.156, 0.078, 0.039, and 0.019 g/L). After inoculating the isolates, all concentrations were evaluated at 3 distinct incubation periods (24, 48, and 72 h) to assess the effect of incubation time on the MIC. Using the polystyrene microplate assays, 1 isolate showed weak biofilm production, 5 moderate, and 4 strong, when incubated at 25°C±1. For isolates incubated at 37°C±1, 6 showed weak biofilm production and 4 moderate. All P. zopfii isolates (n=10) had the capacity to form biofilms on stainless steel coupons. The longer the incubation period of the P. zopfii isolates at different dilutions, the greater the concentrations of sanitizer needed to prevent growth of the microalgae under the tested conditions. We detected a significant effect of sanitizer and time of incubation (24, 48, and 72 h) on MIC values against P. zopfii isolates. The isolates

  3. Genetic characteristics of Japanese clinical Listeria monocytogenes isolates.

    Directory of Open Access Journals (Sweden)

    Satoko Miya

    Full Text Available Listeria monocytogenes causes foodborne illnesses through consumption of ready-to-eat foods. Although 135-201annual listeriosis cases have been estimated in Japan, the details regarding the clinical isolates such as infection source, virulence level, and other genetic characteristics, are not known. In order to uncover the trends of listeriosis in Japan and use the knowledge for prevention measures to be taken, the genetic characteristics of the past human clinical isolates needs to be elucidated. For this purpose, multilocus tandem-repeat sequence analysis (MLTSA and multi-virulence-locus sequence typing (MVLST were used in this study. The clinical isolates showed a variety of genetically distant genotypes, indicating they were from sporadic cases. However, the MVLST profiles of 7 clinical isolates were identical to those of epidemic clone (EC I isolates, which have caused several serious outbreaks in other countries, suggesting the possibility that they have strong virulence potential and originated from a single outbreak. Moreover, 6 Japanese food isolates shared their genotypes with ECI isolates, indicating that there may be risks for listeriosis outbreak in Japan. This is the first investigational study on genetic characteristics of Japanese listeriosis isolates. The listeriosis cases happened in the past are presumably sporadic, but it is still possible that some isolates with strong virulence potential have caused listeriosis outbreaks, and future listeriosis risks also exist.

  4. Investigating the genetic background of bovine digital dermatitis using improved definitions of clinical status.

    Science.gov (United States)

    Schöpke, K; Gomez, A; Dunbar, K A; Swalve, H H; Döpfer, D

    2015-11-01

    Bovine digital dermatitis (DD) is an increasing claw health problem in all cattle production systems worldwide. The objective of this study was to evaluate the use of an improved scoring of the clinical status for DD via M-scores accounting for the dynamics of the disease; that is, the transitions from one stage to another. The newly defined traits were then subjected to a genetic analysis to determine the genetic background for susceptibility to DD. Data consisted of 6,444 clinical observations from 729 Holstein heifers in a commercial dairy herd, collected applying the M-score system. The M-score system is a classification scheme for stages of DD that allows a macroscopic scoring based on clinical inspections of the bovine foot, thus it describes the stages of lesion development. The M-scores were used to define new DD trait definitions with different complexities. Linear mixed models and logistic models were used to identify fixed environmental effects and to estimate variance components. In total, 68% of all observations showed no DD status, whereas 11% were scored as infectious for and affected by DD, and 21% of all observations exhibited an affected but noninfectious status. For all traits, the probability of occurrence and clinical status were associated with age at observation and period of observation. Risk of becoming infected increased with age, and month of observation significantly affected all traits. Identification of the optimal month concerning DD herd status was consistent for all trait definitions; the last month of the trial was identified. In contrast, months exhibiting the highest least squares means of transformed scores differed depending on trait definition. In this respect, traits that can distinguish between healthy, infectious, and noninfectious stages of DD can account for the infectious potential of the herd and can serve as an alert tool. Estimates of heritabilities of traits studied ranged between 0.19 (±0.11) and 0.52 (±0

  5. Isolation of bovine corneal keratan sulfate and its growth factor and morphogen binding.

    Science.gov (United States)

    Weyers, Amanda; Yang, Bo; Solakyildirim, Kemal; Yee, Vienna; Li, Lingyun; Zhang, Fuming; Linhardt, Robert J

    2013-05-01

    Keratan sulfate (KS) is an important glycosaminoglycan that is found in cartilage, reproductive tissues, and neural tissues. Corneal KS glycosaminoglycan is found N-linked to lumican, keratocan and mimecan proteoglycans, and has been widely studied by investigators interested in corneal development and diseases. Recently, the availability of corneal KS has become severely limited, owing to restrictions on the shipment of bovine central nervous system byproducts across international borders in an effort to prevent additional cases of mad cow disease. We report a simple method for the purification of multi-milligram quantities of bovine corneal KS, and characterize its structural properties. We also examined its protein-binding properties, and discovered that corneal KS bound with high affinity to fibroblast growth factor-2 and sonic hedgehog, a growth factor and a morphogen involved in corneal development and healing. PMID:23402351

  6. A microRNA isolation method from clinical samples

    Directory of Open Access Journals (Sweden)

    Sepideh Zununi Vahed

    2016-03-01

    Conclusion: The current isolation method can be applied for most clinical samples including cells, formalin-fixed and paraffin-embedded (FFPE tissues and even body fluids with a wide applicability in molecular biology investigations.

  7. A new bovine conjunctiva model shows that Listeria monocytogenes invasion is associated with lysozyme resistance.

    Science.gov (United States)

    Warren, Jessica; Owen, A Rhys; Glanvill, Amy; Francis, Asher; Maboni, Grazieli; Nova, Rodrigo J; Wapenaar, Wendela; Rees, Catherine; Tötemeyer, Sabine

    2015-08-31

    Listerial keratoconjunctivitis ('silage eye') is a wide spread problem in ruminants causing economic losses to farmers and impacts negatively on animal welfare. It results from direct entry of Listeria monocytogenes into the eye, often following consumption of contaminated silage. An isolation protocol for bovine conjunctival swabbing was developed and used to sample both infected and healthy eyes bovine eyes (n=46). L. monocytogenes was only isolated from one healthy eye sample, and suggests that this organism can be present without causing disease. To initiate a study of this disease, an infection model was developed using isolated conjunctiva explants obtained from cattle eyes post slaughter. Conjunctiva were cultured and infected for 20 h with a range of L. monocytogenes isolates (n=11), including the healthy bovine eye isolate and also strains isolated from other bovine sources, such as milk or clinical infections. Two L. monocytogenes isolates (one from a healthy eye and one from a cattle abortion) were markedly less able to invade conjunctiva explants, but one of those was able to efficiently infect Caco2 cells indicating that it was fully virulent. These two isolates were also significantly more sensitive to lysozyme compared to most other isolates tested, suggesting that lysozyme resistance is an important factor when infecting bovine conjunctiva. In conclusion, we present the first bovine conjunctiva explant model for infection studies and demonstrate that clinical L. monocytogenes isolates from cases of bovine keratoconjunctivitis are able to infect these tissues. PMID:25778543

  8. Sequencing and phylogenetic analysis of the gp51 gene from Korean bovine leukemia virus isolates

    OpenAIRE

    Lee, Eunjung; Kim, Eun-Ju; Joung, Ha-Kyung; Kim, Bo-hye; SONG, Jae-Young; Cho, In-Soo; Lee, Kyoung-Ki; Shin, Yeun-Kyung

    2015-01-01

    Background Bovine Leukemia virus (BLV) infection of cattle has been reported in Korea for more than three decades. However, to date, there have been few studies regarding Korean BLV since 1980s. Thus, the purpose of this study is to perform a diagnosis and molecular characterization of BLV strains circulating in Korea and to estimate genetic diversity of different genotypes of BLV. Method To investigate the distribution of BLV variants in the world and assess the evolutionary history of Korea...

  9. Isolation of bovine corneal keratan sulfate and its growth factor and morphogen binding

    OpenAIRE

    Weyers, Amanda; Yang, Bo; Solakyildirim, Kemal; Yee, Vienna; Li, Lingyun; Zhang, Fuming; Linhardt, Robert

    2013-01-01

    Keratan sulfate (KS) is an important glycosaminoglycan that is found in cartilage, reproductive, and neural tissues. Corneal KS glycosaminoglycan is found N-linked to lumican, keratocan, and mimecan proteoglycans and has been widely studied by investigators interested in corneal development and diseases. Recently, the availability of corneal KS has become severely limited due to restricted the shipment of bovine central nervous system by-products across international borders in efforts to pre...

  10. Isolation and Genetic Analysis of Bovine Viral Diarrhea Virus from Infected Cattle in Indiana

    OpenAIRE

    H. Leon Thacker; Maria Negron; Duane A. Murphy; Raizman, Eran A.; Schnur, Megan E.; Roman M. Pogranichniy

    2011-01-01

    Species and biotype distribution was determined in 44 bovine viral diarrhea virus- (BVDV-) positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL) in Indiana during 2006–2008. BVDV RNA was detected in the 5′-untranslated region and Npro region using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory dis...

  11. Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates

    OpenAIRE

    Meng Dong-Ya; Sun Chang-Jian; Yu Jing-Bo; Ma Jun; Xue Wen-Cheng

    2014-01-01

    To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 ...

  12. Cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf.

    Science.gov (United States)

    Chaplin, P J; Entrican, G; Gelder, K I; Collins, R A

    1996-01-01

    A cDNA encoding a distinct bovine (Bo) interferon (IFN) alpha, designated BoIFN-alpha E, was generated from gut epithelial cells isolated from a rotavirus-infected calf. The BoIFN-alpha E cDNA sequence shared a greater than 90% identity with the other BoIFN-alpha subtypes. The cDNA encoding BoIFN-alpha E has been expressed in insect cells using the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) as a vector. Insect cells infected with recombinant virus secreted a protein with a relative molecular mass of 19,500 into the culture medium not observed in cells infected with wild-type AcMNPV. Supernatants harvested from cultures of insect cells infected with the recombinant AcMNPV encoding IFN-alpha E inhibited the replication of Semliki Forest virus in a bovine cell line and typically showed 10(6) dilution units/ml of antiviral activity. However, differences were observed between the activities of recombinant BoIFN-alpha E and BoIFN-alpha 1 1 on the proliferation of WC1+ gamma/delta T cells. Purified ( > 99%) WC1+ gamma/delta T cells failed to proliferate to IFN-alpha 1 1 or concanavalin A and IFN-alpha E acted as a weak proliferative signal to these cells, demonstrating a functional difference between two closely related BoIFN-alpha subtypes. PMID:8640447

  13. Identification of amino acid changes in the envelope glycoproteins of bovine viral diarrhea viruses isolated from alpaca that may be involved in host adaptation.

    Science.gov (United States)

    Neill, John D; Dubovi, Edward J; Ridpath, Julia F

    2015-09-30

    Bovine viral diarrhea viruses (BVDV) are most commonly associated with infections of cattle. However, BVDV are often isolated from closely related ruminants with a number of BVDV-1b viruses being isolated from alpacas that were both acutely and persistently infected. The complete nucleotide sequence of the open reading frame of eleven alpaca-adapted BVDV isolates and the region encoding the envelope glycoproteins of an additional three isolates were determined. With the exception of one, all alpaca isolates were >99.2% similar at the nucleotide level. The Hercules isolate was more divergent, with 95.7% sequence identity to the other viruses. Sequence similarity of the 14 viruses indicated they were isolates of a single BVDV strain that had adapted to and were circulating through alpaca herds. Hercules was a more distantly related strain that has been isolated only once in Canada and represented a separate adaptation event that possessed the same adaptive changes. Comparison of amino acid sequences of alpaca and bovine-derived BVDV strains revealed three regions with amino acid sequences unique to all alpaca isolates. The first contained two small in-frame deletions near the N-terminus of the E2 glycoprotein. The second was found near the C-terminus of the E2 protein where four altered amino acids were located within a 30 amino acid domain that participates in E2 homodimerization. The third region contained three variable amino acids in the C-terminus of the E(rns) within the amphipathic helix membrane anchor. These changes were found in the polar side of the amphipathic helix and resulted in an increased charge within the polar face. Titration of bovine and alpaca viruses in both bovine and alpaca cells indicated that with increased charge in the amphipathic helix, the ability to infect alpaca cells also increased.

  14. Genomic characterization of Pasteurella multocida HB01, a serotype A bovine isolate from China.

    Science.gov (United States)

    Peng, Zhong; Liang, Wan; Liu, Wenjing; Wu, Bin; Tang, Biao; Tan, Chen; Zhou, Rui; Chen, Huanchun

    2016-04-25

    Pasteurella multocida infects various domestic and feral animals, generally causing clinical disease. To investigate P. multocida disease in cattle, we sequenced the complete genome of P. multocida HB01 (GenBank accession CP006976), a serotype A organism isolated from a cow in China. The genome is composed of a single circular chromosome of 2,416,068 base pairs containing 2212 protein-coding sequences, 6 ribosomal rRNA operons, and 56 tRNA genes. The present study confirms that P. multocida HB01 possesses a more complete metabolic pathway with an intact trichloroacetic acid cycle for anabolism compared with A. pleuropneumoniae and Haemophilus parasuis. This is the first time that this metabolic mechanism of P. multocida has been described. We also identified a full spectrum of genes related to known virulence factors of P. multocida. The differences in virulence factors between strains of different serotypes and origins were also compared. This comprehensive comparative genome analysis will help in further studies of the metabolic pathways, genetic basis of serotype, and virulence of P. multocida. PMID:26827796

  15. BIOFILM DETECTION AND CLINICAL SIGNIFICANCE OF STAPHYLOCOCCUS EPIDERMIDIS ISOLATES

    Directory of Open Access Journals (Sweden)

    Sudha Rani

    2015-04-01

    Full Text Available CONTEXT: Isolates of Staphylococcus epidermidis . AIMS : To detect biofilm among clinical isolates of Staphylococcus epidermidis . MATERIAL AND METHODS : 50 Staphylococcus epidermidis isolates were collected from clinical samples like blood, IV catheter tips, catheterized urine, wound swabs and exudates received from various clinical departments. Biofilm formation was studied in these isolates. The study was carried out over a period o f one year i. e from March 2011 - February 2012. The specimens received were processed by conventional methods. Tissue Culture plate method was used for detection of biofilm. RESULTS: IV catheter tip samples revealed 25%, implant device associated infection s revealed 20%, the Catheterized urine samples showed 16%, blood culture 8%, ventilator associated infections 10%, post - operative wound infections 11% & exudates 5% of Staphylococcus epidermidis isolates. Isolates with O. D. values more than 0. 2 were consi dered as high biofilm producers. 40% of S. epidermidis isolates were weak biofilm producers, 24% were moderate biofilm producers and 36% were high biofilm producers. Isolates from IV catheter tips showed high biofilm formation. CONCLUSIONS: In the modern h ealth care set up, various devices such as IV catheters, urine catheters, shunts, implanted prosthetic devices etc are being increasingly used thereby causing the device associated infections particularly of Staphylococcus epidermidis. Device associated in fections caused by Staphylococcus epidermidis are mainly due to biofilm formation which ultimately makes the treatment difficult.

  16. Full-Length Coding Sequences for 12 Bovine Viral Diarrhea Virus Isolates from Persistently Infected Cattle in a Feedyard in Kansas

    OpenAIRE

    Workman, Aspen M.; Harhay, Gregory P.; Heaton, Michael P; Grotelueschen, Dale M.; Sjeklocha, David; Smith, Timothy P. L.

    2015-01-01

    We report here the full-length coding sequences of 12 bovine viral diarrhea virus (BVDV) isolates from persistently infected cattle in a feedyard in southwest KS. These 12 genomes represent the three major subtypes of BVDV (BVDV-1a, 1b, and 2a) currently circulating in the United States.

  17. Study on some characteristics of Staphylococci isolated from sheep sub clinical mastitis milk in Shahrekord, Iran

    Directory of Open Access Journals (Sweden)

    Azizollah Ebrahimi

    2014-01-01

    Full Text Available Introduction: Staphylococci release a large number of enzymes. Some of these, such as coagulase, beta lactamase, hemolysins and biofilms are considered indices of pathogenicity. The aim of the current study was based on the isolation and identification of Staphylococcus aureus and coagulase negative Staphylococci (CNS strains from sheep sub clinical mastitis and examining their biofilm, beta lactamase, hemolysins production and antibiotic resistance pattern. Materials and methods: 55 Staphylococci strains were isolated from seventy cases of sheep subclinical mastitis. Thirty three were determined as Staphylococcus aureus (60% and 22 (40% as CNS. The hemolytic activity was evaluated by plating Staphylococci strains on 5% bovine blood agar. The biofilm assay was performed by using micro titer plates. Beta Lactamase production was detected by test tube iodometric technique and susceptibility to antimicrobial agents was determined for isolated strains by the disk diffusion method. Results: Twenty six (78.8% S. aureus strains were biofilm producers. For CNS (59.9% strains were positive in biofilm production. Two isolates (6.06%, of S. aureus were α, the same number β and 6 (18.2% isolates were ∂ hemolysin producers. Six isolates of CNS (27.27% were α and ten (45.45% ∂ hemolysin producers. Sixteen S. aureus (48.5% and five CNS (22.72% isolates were positive in beta lactamase production. The isolated Staphylococci show a low sensitivity pattern to methicillin and streptomycin. Discussion and conclusion: A high percentage of strains make α toxin that play a role in S. aureus biofilm formation. Twenty one out of 33 (63.63% isolated Staphylococci were biofilm producers that can have deleterious effects because biofilm formation is thought to play an important role in the survival of virulent strains of Staphylococci. Sixteen out of 33 (48.5% isolated S. aureus were positive in beta lactamase test, Excluding resistant to methicillin, all of these

  18. A comparative study of Staphylococcus aureus strains isolated from bovine subclinical mastitis during 1952-1956 and 1992

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Wegener, Henrik Caspar; Rosdahl, V.T.

    1995-01-01

    Fiftytwo strains of S. aureus isolated from cases of bovine subclinical mastitis in 52 different dairy herds in Denmark, in the peri ods 1952 to 1956 and 1992, were compared with regard to their phage- and EcoRI ribotypes. Furthermore, susceptibility to penicillin and production of fibrinolysin...... were used as additional phenotypic markers. Fortynine strains (94%) could be separated into 12 phage types. Ribotyping assigned the 52 strains to 21 different types. Both methods showed that 57% of the 1950's strains and between 38-45% of the 1992 strains belonged to 3 dominating types. The remaining...... differences. Penicillin resistance only occurred in a single genotype from the 1950's compared to 6 different genotypes among the 1992 strains....

  19. Molecular characterisation of clinical and environmental isolates of Mycobacterium kansasii isolates from South African gold mines.

    Science.gov (United States)

    Kwenda, Geoffrey; Churchyard, Gavin J; Thorrold, Catherine; Heron, Ian; Stevenson, Karen; Duse, Adriano G; Marais, Elsé

    2015-03-01

    Mycobacterium kansasii (M. kansasii) is a major cause of non-tuberculous mycobacterial pulmonary disease in the South African gold-mining workforce, but the source of infection and molecular epidemiology are unknown. This study investigated the presence of M. kansasii in gold and coal mine and associated hostel water supplies and compared the genetic diversity of clinical and environmental isolates of M. kansasii. Five M. kansasii and ten other potentially pathogenic mycobacteria were cultured mainly from showerhead biofilms. Polymerase chain reaction-restriction analysis of the hsp65 gene on 196 clinical and environmental M. kansasii isolates revealed 160 subtype I, eight subtype II and six subtype IV strains. Twenty-two isolates did not show the typical M. kansasii restriction patterns, suggesting that these isolates may represent new subtypes of M. kansasii. In contrast to the clonal population structure found amongst the subtype I isolates from studies in other countries, DNA fingerprinting of 114 clinical and three environmental subtype I isolates demonstrated genetic diversity amongst the isolates. This study demonstrated that showerheads are possible sources of M. kansasii and other pathogenic non-tuberculous mycobacterial infection in a gold-mining region, that subtype I is the major clinical isolate of M. kansasii strain and that this subtype exhibits genetic diversity. PMID:25719478

  20. Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle Infected with Atypical Bovine Spongiform Encephalopathy.

    Science.gov (United States)

    Xerxa, Elena; Barbisin, Maura; Chieppa, Maria Novella; Krmac, Helena; Vallino Costassa, Elena; Vatta, Paolo; Simmons, Marion; Caramelli, Maria; Casalone, Cristina; Corona, Cristiano; Legname, Giuseppe

    2016-01-01

    Prion diseases, such as bovine spongiform encephalopathies (BSE), are transmissible neurodegenerative disorders affecting humans and a wide variety of mammals. Variant Creutzfeldt-Jakob disease (vCJD), a prion disease in humans, has been linked to exposure to BSE prions. This classical BSE (cBSE) is now rapidly disappearing as a result of appropriate measures to control animal feeding. Besides cBSE, two atypical forms (named H- and L-type BSE) have recently been described in Europe, Japan, and North America. Here we describe the first wide-spectrum microarray analysis in whole blood of atypical BSE-infected cattle. Transcriptome changes in infected animals were analyzed prior to and after the onset of clinical signs. The microarray analysis revealed gene expression changes in blood prior to the appearance of the clinical signs and during the progression of the disease. A set of 32 differentially expressed genes was found to be in common between clinical and preclinical stages and showed a very similar expression pattern in the two phases. A 22-gene signature showed an oscillating pattern of expression, being differentially expressed in the preclinical stage and then going back to control levels in the symptomatic phase. One gene, SEL1L3, was downregulated during the progression of the disease. Most of the studies performed up to date utilized various tissues, which are not suitable for a rapid analysis of infected animals and patients. Our findings suggest the intriguing possibility to take advantage of whole blood RNA transcriptional profiling for the preclinical identification of prion infection. Further, this study highlighted several pathways, such as immune response and metabolism that may play an important role in peripheral prion pathogenesis. Finally, the gene expression changes identified in the present study may be further investigated as a fingerprint for monitoring the progression of disease and for developing targeted therapeutic interventions. PMID

  1. Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle Infected with Atypical Bovine Spongiform Encephalopathy.

    Science.gov (United States)

    Xerxa, Elena; Barbisin, Maura; Chieppa, Maria Novella; Krmac, Helena; Vallino Costassa, Elena; Vatta, Paolo; Simmons, Marion; Caramelli, Maria; Casalone, Cristina; Corona, Cristiano; Legname, Giuseppe

    2016-01-01

    Prion diseases, such as bovine spongiform encephalopathies (BSE), are transmissible neurodegenerative disorders affecting humans and a wide variety of mammals. Variant Creutzfeldt-Jakob disease (vCJD), a prion disease in humans, has been linked to exposure to BSE prions. This classical BSE (cBSE) is now rapidly disappearing as a result of appropriate measures to control animal feeding. Besides cBSE, two atypical forms (named H- and L-type BSE) have recently been described in Europe, Japan, and North America. Here we describe the first wide-spectrum microarray analysis in whole blood of atypical BSE-infected cattle. Transcriptome changes in infected animals were analyzed prior to and after the onset of clinical signs. The microarray analysis revealed gene expression changes in blood prior to the appearance of the clinical signs and during the progression of the disease. A set of 32 differentially expressed genes was found to be in common between clinical and preclinical stages and showed a very similar expression pattern in the two phases. A 22-gene signature showed an oscillating pattern of expression, being differentially expressed in the preclinical stage and then going back to control levels in the symptomatic phase. One gene, SEL1L3, was downregulated during the progression of the disease. Most of the studies performed up to date utilized various tissues, which are not suitable for a rapid analysis of infected animals and patients. Our findings suggest the intriguing possibility to take advantage of whole blood RNA transcriptional profiling for the preclinical identification of prion infection. Further, this study highlighted several pathways, such as immune response and metabolism that may play an important role in peripheral prion pathogenesis. Finally, the gene expression changes identified in the present study may be further investigated as a fingerprint for monitoring the progression of disease and for developing targeted therapeutic interventions.

  2. Diversity and Mobility of Integrative and Conjugative Elements in Bovine Isolates of Streptococcus agalactiae, S. dysgalactiae subsp. dysgalactiae, and S. uberis▿ †

    OpenAIRE

    Haenni, Marisa; Saras, Estelle; Bertin, Stéphane; Leblond, Pierre; Madec, Jean-Yves; Payot, Sophie

    2010-01-01

    Bovine isolates of Streptococcus agalactiae (n = 76), Streptococcus dysgalactiae subsp. dysgalactiae (n = 32), and Streptococcus uberis (n = 101) were analyzed for the presence of different integrative and conjugative elements (ICEs) and their association with macrolide, lincosamide, and tetracycline resistance. The diversity of the isolates included in this study was demonstrated by multilocus sequence typing for S. agalactiae and pulsed-field gel electrophoresis for S. dysgalactiae and S. u...

  3. Whole genomic analysis of human and bovine G8P[1] rotavirus strains isolated in Nigeria provides evidence for direct bovine-to-human interspecies transmission.

    Science.gov (United States)

    Komoto, Satoshi; Adah, Mohammed Ignatius; Ide, Tomihiko; Yoshikawa, Tetsushi; Taniguchi, Koki

    2016-09-01

    Bovine group A rotavirus (RVA) G8P[1] strains have been rarely detected in humans. Two Nigerian G8P[1] strains, HMG035 (RVA/Human-tc/NGA/HMG035/1999/G8P[1]) and NGRBg8 (RVA/Cow-tc/NGA/NGRBg8/1998/G8P[1]), were previously suggested to have the VP7, VP4, and NSP1 genes of bovine origin. In order to obtain precise information on the origin and evolution of these G8P[1] strains, the complete nucleotide sequences of the whole genomes of strains HMG035 and NGRBg8 were determined and analyzed in the present study. On whole genomic analysis, strains HMG035 and NGRBg8 were found to be very closely related to each other in all the 11 segments, and were found to have a bovine RVA-like genotype constellation (G8-P[1]-I2-R2-C2-M2-A11-N2-T6-E2-H3). Furthermore, on phylogenetic analysis, each of the 11 genes of strains HMG035 and NGRBg8 appeared to be of bovine origin. Thus, strains HMG035 and NGRBg8 were suggested to be derived from a common origin, and strain NGRBg8 was assumed to represent an example of bovine RVA strains that were transmitted to humans. Our findings provide clear evidence for direct bovine-to-human interspecies transmission of RVA strains. PMID:27302094

  4. Bovine Tuberculosis, A Zoonotic Disease

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    Tarmudji

    2008-12-01

    Full Text Available Bovine tuberculosis is caused by the infection of Mycobacterium tuberculosis var. bovis (M. bovis. This species is one of Mycobacterium tuberculosis complex, can infect wide range of hosts: cattle and other domesticated animals, wild mammals and humans (zoonotic. M. bovis bacterium from infected hosts can be transmitted to other susceptible animals and humans through respiratory excretes and secretion materials. Humans can be infected with M. bovis by ingested M. bovis contaminated animal products, unpasteurised milk from tuberculosis cows or through respiratory route of contaminated aerosol. Bovine tuberculosis at the first stage does not show any clinical sign but as the disease progress in the next stage which may take several months or years, clinical signs may arise, suh as: fluctuative body temperature, anorexia, lost body weight, coughing, oedema of lymph nodes, increased respiratory frequencies. Pathological lesion of bovine tuberculosis is characterised by the formation of granulomas (tubercles, in which bacterial cells have been localised, most in lymph nodes and pulmonum, but can occur in other organs. The granulomas usually arise in small nodules or tubercles appear yellowish either caseus, caseo-calcareus or calcified. In Indonesia, bovine tuberculosis occurred in dairy cattle since 1905 through the imported dairy cows from Holland and Australian. It was unfortunate that until recently, there were not many research and surveilances of bovine tuberculosis conducted in this country, so the distribution of bovine tuberculosis is unknown. Early serological diagnosis can be done on live cattle by means of tuberculin tests under field conditions. Confirmation can be done by isolation and identification of excreted and secreted samples from the slaughter house. Antibiotic treatment and vaccination were uneffective, therefore the effective control of bovine tuberculosis is suggested by tuberculin tests and by slaughtering the selected

  5. Geographical variation in the presence of genes encoding superantigenic exotoxins and beta-hemolysin among Staphylococcus aureus isolated from bovine mastitis in Europe and USA

    DEFF Research Database (Denmark)

    Larsen, H. D.; Aarestrup, Frank Møller; Jensen, N. E.

    2002-01-01

    The object was to examine the geographical variation in the presence of superantigenic exotoxins and beta-hemolysin among epidemiologically independent Staphyirrcoccus aureus isolates from bovine mastitis. A total of 462 S. aureus isolates from nine European countries and USA were examined...... for the presence of genes encoding staphylococcal enterotoxins A-E, and H, toxic shock toxin-1 (TSST-1), and beta-hemolysin, and 128 of these were examined for exfoliative toxins A and B. The detection was done by PCR. Phenotypic methods were used to confirm the PCR-results. None of the 128 isolates carried...... the genes for exfoliative toxin A or B. The total proportion of isolates in which superantigenic exotoxins were detected varied from 2% (one isolate) of the Danish isolates to 65% (32 isolates) of the Norwegian isolates. This marked and highly significant geographical variation was also present...

  6. Coagulase gene polymorphisms detected by PCR in Staphylococcus aureus isolated from subclinical bovine mastitis in Turkey.

    Science.gov (United States)

    Karahan, Murat; Cetinkaya, Burhan

    2007-09-01

    The genetic relatedness of coagulase (coa) positive Staphylococcus aureus isolated from cows with subclinical mastitis in Turkey was investigated by polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) analysis. Among 700 milk samples positive in the California Mastitis Test (CMT), species specific PCR identified 200 (28.6%) isolates as S. aureus and 161 (80.5%) of these isolates were positive for the 3' end of the coa gene by PCR. Most isolates (n=135, 83.9%) produced a single band on coa PCR, with molecular sizes ranging from 500 to 1400bp, whereas a small number of isolates (n=26, 16.1%) yielded two amplification products. Coa RFLP analysis using AluI and Hin6I revealed 23 and 22 band patterns, respectively. The detection of double bands by coa PCR, previously reported in human isolates, suggests that milking personnel can play a role in the transmission of S. aureus. PMID:16901735

  7. Antimicrobial susceptibility of Staphylococcus aureus and characterization of methicillin-resistant Staphylococcus aureus isolated from bovine mastitis in Korea.

    Science.gov (United States)

    Nam, Hyang-Mi; Lee, Ae-Li; Jung, Suk-Chan; Kim, Mal-Nam; Jang, Geum-Chan; Wee, Sung-Hwan; Lim, Suk-Kyung

    2011-02-01

    A total of 402 Staphylococcus aureus isolates from bovine mastitis milk collected during 2003-2009 in Korea were tested for susceptibility to 20 antimicrobial agents. All S. aureus isolates were susceptible to 11 of 20 antimicrobials tested; no resistance was observed against pirlimycin, telithromycin, novobiocin, penicillin/novobiocin, quinupristin/dalfopristin, clindamycin, rifampin, ciprofloxacin, trimethprim/sulfamethoxazol, vancomycin, and linezolid. Over 66% of the S. aureus isolates were resistant to penicillin. Resistance was also seen for gentamicin (11.9%), erythromycin (7.7%), methicillin (oxacillin and cefoxitin, 6.2%), and tetracycline (4.2%). No noticeable change was observed in penicillin, gentamicin, and erythromycin resistance over the 7-year period. Tetracycline resistance appeared to decrease consistently, whereas methicillin resistance was observed from 2005. About 2.7% (11/402) were resistant to three or more antimicrobials. Genotyping of the 17 methicillin-resistant S. aureus (MRSA) isolated from each cow revealed two staphylococcal cassette chromosome mec (SCCmec) types (IV and IVa), three spa types (t286, t324, and untypable), and two sequence types (ST1 and ST72). Eleven of 17 (64.7%) MRSA strains belonged to SCCmec IVa-t324-ST72. The rest of strains belonged to SCCmec IVa-t286-ST1 (n = 3) and SCCmec IV-untypable-ST72 (n = 3). None of the MRSA carried the Panton-Valentine leukocidin gene. These characteristics are the same as those found in community-acquired (CA) MRSA strains prevalent in humans in Korea. Three pulsed-field gel electrophoresis types (A-C) were observed among the 17 MRSA strains examined, and 14 strains belonged to the same pulsed-field gel electrophoresis pattern regardless of their geographical origin and year of isolation. The results of this study provide evidence of CA-MRSA infection in dairy cattle for the first time in Korea. PMID:21034263

  8. Electrophoretic and immunological properties of folate-binding protein isolated from bovine milk

    Energy Technology Data Exchange (ETDEWEB)

    Iwai, Kazuo; Tani, Masako; Fushiki, Tohru (Kyoto Univ. (Japan). Faculty of Agriculture)

    1983-07-01

    Changes of the folate-binding protein (FBP) concentration in bovine milk after parturition were investigated. The FBP was highly purified from mature milk by affinity chromatography. The purified FBP showed a single protein band in polyacrylamide gel electrophoresis and was immunologically homogenous in double immunodiffusion. However, in two-dimensional gel electrophoresis, the FBP was separated into several spots in isoelectric focusing in the first dimension, and each spot also showed two molecular weights in SDS-gel electrophoresis in the second dimension. But these FBP molecules were immunologically identical with each other. The neuraminidase treatment obviously diminished the number of isoelectric points of the FBP. Thus, the variety of FBP molecules was at least partially due to the variability of the sialic acid content in the carbohydrate moieties. Moreover, the milk FBP showed species-specificity among mammals immunologically as well as physicochemically.

  9. Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates

    Directory of Open Access Journals (Sweden)

    Meng Dong-Ya

    2014-01-01

    Full Text Available To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs of DNA gyrase (gyrA and gyrB and topoisomerase IV (parC and parE in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX, intermediate resistant to Levofloxacin (LVX and Sparfloxacin (SFX, and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.

  10. Clinical significance of the isolation of Candidaspecies from hospitalized patients

    Directory of Open Access Journals (Sweden)

    Yankee C. Magalhães

    2015-03-01

    Full Text Available In this study, we isolated and phenotypically identified 108 yeast strains from various clinical specimens collected from 100 hospitalized patients at three tertiary hospitals in São Luís-Maranhão, Brazil, from July to December 2010. The isolates were analyzed for their susceptibility to four of the most widely used antifungal agents in the surveyed hospitals, amphotericin B, fluconazole, 5-flucytosine and voriconazole. The species identified were Candida albicans (41.4%, Candida tropicalis (30.1%, C. glabrata (7.4%, Candida parapsilosis(5.5%, Candida krusei (4.6%, Cryptococcus neoformans (4.6%, Trichosporonspp. (3.7%, Candida norvegensis (0.9%, Rhodotorula glutinis (0.9% and Pichia farinosa (0.9%. A higher isolation rate was observed in the following clinical specimens: urine (54 isolates; 50%, respiratory tract samples (21 isolates; 19.4% and blood (20 isolates; 18.6%. Candida albicans isolates were 100% sensitive to all antifungal agents tested, whereas Candida krusei and Crytococcus neoformans displayed intermediate resistance to 5-flucytosine, with Minimal Inhibitory Concentration (MIC values of 8 mg/mL and 16 mg/mL, respectively. Both strains were also S-DD to fluconazole with an MIC of 16 mg/mL. C. tropicalis was resistant to 5-flucytosine with an MIC of 32 μg/mL. This study demonstrates the importance of identifying the yeast species involved in community and nosocomial infections.

  11. Molecular Characterization of Streptococcus agalactiae Isolated from Bovine Mastitis in Eastern China

    OpenAIRE

    Yongchun Yang; Yinglong Liu; Yunlei Ding; Li Yi; Zhe Ma; Hongjie Fan; Chengping Lu

    2013-01-01

    One hundred and two Streptococcus agalactiae (group B streptococcus [GBS]) isolates were collected from dairy cattle with subclinical mastitis in Eastern China during 2011. Clonal groups were established by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE), respectively. Capsular polysaccharides (CPS), pilus and alpha-like-protein (Alp) family genes were also characterized by molecular techniques. MLST analysis revealed that these isolates were limited to three clo...

  12. Molecular detection and sensitivity to antibiotics and bacteriocins of pathogens isolated from bovine mastitis in family dairy herds of central Mexico.

    Science.gov (United States)

    León-Galván, Ma Fabiola; Barboza-Corona, José E; Lechuga-Arana, A Arianna; Valencia-Posadas, Mauricio; Aguayo, Daniel D; Cedillo-Pelaez, Carlos; Martínez-Ortega, Erika A; Gutierrez-Chavez, Abner J

    2015-01-01

    Thirty-two farms (n = 535 cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY), lactation number (LN), herd size (HS), and number of days in milk (DM) were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH), LN, HS, and DM (P Brevibacterium stationis, B. conglomeratum, and Staphylococcus agnetis. Bacterial isolates were resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized by Bacillus thuringiensis inhibited the growth of multiantibiotic resistance bacteria such as S. agnetis, S. equorum, Streptococcus uberis, Brevibacterium stationis, and Brachybacterium conglomeratum, but they were not active against S. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study. PMID:25815326

  13. Molecular Detection and Sensitivity to Antibiotics and Bacteriocins of Pathogens Isolated from Bovine Mastitis in Family Dairy Herds of Central Mexico

    Directory of Open Access Journals (Sweden)

    Ma. Fabiola León-Galván

    2015-01-01

    Full Text Available Thirty-two farms (n=535 cows located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM and clinical mastitis (CLM were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT (≥3 and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY, lactation number (LN, herd size (HS, and number of days in milk (DM were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH, LN, HS, and DM P<0.01, and correlations between udder quarters from the CMT were around 0.49 P<0.01. Coagulase-negative staphylococci were mainly identified, as well as Staphylococcus aureus, Streptococcus uberis, Brevibacterium stationis, B. conglomeratum, and Staphylococcus agnetis. Bacterial isolates were resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized by Bacillus thuringiensis inhibited the growth of multiantibiotic resistance bacteria such as S. agnetis, S. equorum, Streptococcus uberis, Brevibacterium stationis, and Brachybacterium conglomeratum, but they were not active against S. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study.

  14. Diversity of clinical isolates of Entamoeba histolytica in Japan.

    Science.gov (United States)

    Nozaki, Tomoyoshi; Kobayashi, Seiki; Takeuchi, Tsutomu; Haghighi, Ali

    2006-02-01

    In Japan, amebiasis is domestically transmitted by two major populations: male homosexuals and mentally handicapped persons, which is remarkably different from most other developed countries where Entamoeba dispar infection is predominantly observed. Here we briefly summarize epidemiology of amebiasis in Japan. We also review our current understanding of the diversity of Entamoeba histolytica clinical isolates in Japan, based on polymorphic genetic markers, clinical representations, and in vivo virulence, using an animal model.

  15. Genotypes, Virulence Factors and Antimicrobial Resistance Genes of Staphylococcus aureus Isolated in Bovine Subclinical Mastitis from Eastern China

    Directory of Open Access Journals (Sweden)

    Javed Memon§, Yongchun Yang§, Jam Kashifa, Muhammad Yaqoob, Rehana Buriroa, Jamila Soomroa, Wang Liping and Fan Hongjie*

    2013-11-01

    Full Text Available This study was carried out to determine the genotypes, virulence factors and antimicrobial resistance traits of 34 Staphylococcus aureus isolated from subclinical mastitis in Eastern China. Minimal inhibitory concentration (MIC results showed resistance to erythromycin in all isolates. A high frequency of Methicillin resistant S. aureus (MRSA; 29% was observed and these isolates were also highly resistant to penicillin, oxacillin, oxytetracycline and chloramphenicol than methicillin sensitive S. aureus (MSSA isolates. Thirteen pathogenic factors and seven resistance genes including mecA and blaZ gene were checked through PCR. The spaX gene was found in all isolates, whereas cna, spaIg, nuc, clfA, fnbpB, hlA, hlB and seA were present in 35, 79, 85, 59, 35, 85, 71 and 38% isolates, respectively. Nine isolates carried a group of 8 different virulence genes. Moreover, macrolide resistance genes ermB and ermC were present in all isolates. High resistance rate against methicillin was found but no isolate was positive for mecA gene, whereas blaZ and tetK were detected in 82 and 56% isolates, respectively. Genes; fnbpA, seB, seC, seD, dfrK and tetM were not found in any isolate. The statistical association between phenotypic resistance and virulence genes showed, clfA, fnbpB, hlB and seA, were potentially associated with penicillin G, ciprofloxacin, methicillin, chloramphenicol, trimethoprim and oxytetracycline resistance (P≤0.05. REP-PCR based genotyping showed seven distinct genotypes (A-G prevalent in this region. This study reports the presence of multidrug resistant S. aureus in sub-clinical mastitis which were also highly virulent that could be a major obstacle in the treatment of mastitis in this region of China.

  16. Antibiotic Susceptibilities and Serotyping of Clinical Streptococcus Agalactiae Isolates

    Directory of Open Access Journals (Sweden)

    Altay Atalay

    2011-11-01

    Full Text Available Objective: Streptococcus agalactiae (Group B streptococci, GBS are frequently responsible for sepsis and meningitis seen in the early weeks of life. GBS may cause perinatal infection and premature birth in pregnant women. The aim of this study was to serotype GBS strains isolated from clinical samples and evaluate their serotype distribution according to their susceptibilities to antibiotics and isolation sites. Material and Methods: One hundred thirty one S. agalactiae strains isolated from the clinical samples were included in the study. Of the strains, 99 were isolated from urine, 20 from soft tissue, 10 from blood and 2 from vaginal swab. Penicillin G and ceftriaxone susceptibilities of GBS were determined by the agar dilution method. Susceptibilities to erythromycin, clindamycin, vancomycin and tetracycline were determined by the Kirby-Bauer method according to CLSI criteria. Serotyping was performed using the latex aglutination method using specific antisera (Ia, Ib, II-VIII. Results: While in 131 GBS strains, serotypes VII and VIII were not detected, the most frequently isolated serotypes were types Ia (36%, III (30.5% and II (13% respectively. Serotype Ia was the most frequently seen serotype in all samples. All GBS isolates were susceptible to penicilin G, ceftriaxone and vancomycin. Among the strains, tetracycline, erythromycin and clindamycin resistance rates were determined as 90%, 14.5%, and 13% respectively. Conclusion: Penicillin is still the first choice of treatment for the infections with all serotypes of S. agalactiae in Turkey.

  17. Asperger syndrome, violent thoughts and clinically isolated syndrome.

    Science.gov (United States)

    Vanderbruggen, N; Van Geit, N; Bissay, V; Zeeuws, D; Santermans, L; Baeken, C

    2010-12-01

    A young man, 23 years old, with a clinically isolated syndrome (CIS), presented violent thoughts during a neurological consultation. He was diagnosed with Asperger Syndrome based on a psychiatric and (neuro)psychological examination. Possible risk factors for acting-out and the implications for treatment, if CIS would evolve to MS, are discussed based on a review of the literature.

  18. Imunogenicidade de isolados de herpesvírus bovino 5 como candidatos à vacina Immunogenicity of bovine herpesvirus 5 isolates as vaccine candidates

    Directory of Open Access Journals (Sweden)

    Luiz Felipe Lourenço de Souza

    2009-02-01

    Full Text Available O herpesvírus bovino 5 (BoHV-5 é o agente da menigoencefalite herpética bovina. A doença neurológica, associada à infecção pelo BoHV-5, apresenta altas taxas de letalidade em bovinos jovens e está disseminada no Brasil. A prevenção das perdas causadas pela infecção pelo herpesvírus está baseada, principalmente, na imunização dos animais. Nesse sentido, foi delineada uma comparação entre isolados de BoHV-5, buscando selecionar o isolado mais antigênico para a formulação de vacinas. As formulações inativadas foram produzidas com os isolados ISO9898292, SV507, SV163, 1807 e EVI145 e administradas a cinco grupos de 10 ovelhas cada, que receberam duas doses vacinais por via intramuscular com intervalo de 21 dias. Foram realizadas coletas de sangue para análise de presença de anticorpos por soroneutralização e acompanhamento dos animais até o 63° dia após a primo-vacinação. Foram observados dois picos na curva de anticorpos, o primeiro no dia 14, após a vacinação, quando os títulos médios de anticorpos variaram entre 23,1 e 138,6. O segundo pico foi observado 14 dias após a revacinação, quando os títulos médios variaram entre 301,3 e 1017,5. No 42° dia após a revacinação, foi observada variação de título entre 82,4 e 305,9. A diferença entre as médias de títulos de anticorpos de cada grupo de animais sugere uma menor antigenicidade do isolado ISO9898292 em relação aos demais, demonstrando uma possível variação antigênica entre os isolados. Todos os isolados, com exceção do ISO9898292, mostraram-se imunogênicos para a indução de anticorpos.Herpesvirus bovine 5 (BoHV-5 is the agent of bovine herpetic menigoencephalitis. The neurological disease associated with the infection is highly lethal in young cattle and it is widespread in Brazil. Control of the clinical signs caused by herpesviruses is based mainly on the immunization of cattle. A comparative study was performed among Brazilian Bo

  19. Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

    Directory of Open Access Journals (Sweden)

    Griffiths T Daniel

    2009-05-01

    Full Text Available Abstract Background The Retinal Pigmented Epithelium (RPE is juxtaposed with the photoreceptor outer segments of the eye. The proximity of the photoreceptor cells is a prerequisite for their survival, as they depend on the RPE to remove the outer segments and are also influenced by RPE cell paracrine factors. RPE cell death can cause a progressive loss of photoreceptor function, which can diminish vision and, over time, blindness ensues. Degeneration of the retina has been shown to induce a variety of retinopathies, such as Stargardt's disease, Cone-Rod Dystrophy (CRD, Retinitis Pigmentosa (RP, Fundus Flavimaculatus (FFM, Best's disease and Age-related Macular Degeneration (AMD. We have cultured primary bovine RPE cells to gain a further understanding of the mechanisms of RPE cell death. One of the cultures, named tRPE, surpassed senescence and was further characterized to determine its viability as a model for retinal diseases. Results The tRPE cell line has been passaged up to 150 population doublings and was shown to be morphologically similar to primary cells. They have been characterized to be of RPE origin by reverse transcriptase PCR and immunocytochemistry using the RPE-specific genes RPE65 and CRALBP and RPE-specific proteins RPE65 and Bestrophin. The tRPE cells are also immunoreactive to vimentin, cytokeratin and zonula occludens-1 antibodies. Chromosome analysis indicates a normal diploid number. The tRPE cells do not grow in suspension or in soft agar. After 3H thymidine incorporation, the cells do not appear to divide appreciably after confluency. Conclusion The tRPE cells are immortal, but still exhibit contact inhibition, serum dependence, monolayer growth and secrete an extra-cellular matrix. They retain the in-vivo morphology, gene expression and cell polarity. Additionally, the cells endocytose exogenous melanin, A2E and purified lipofuscin granules. This cell line may be a useful in-vitro research model for retinal

  20. Investigation of risk factors of bovine mastitis in Ethiopia; Isolation of mastitis causing agents and determination of the content of somatic cells in milk

    OpenAIRE

    Frese, Mathias Lutz

    2010-01-01

    In this thesis, the risk factors of bovine mastitis in different milk production systems in Ethiopia were investigated. Furthermore, mastitis causing agents were isolated after California Mastitis Test (CMT) was used as the field test. Somatic cells were counted and compared with the CMT. Low milk production and low quality of milk are apparently related to a lack of proper hygienic measures throughout the farm clusters.

  1. Antiviral Activity of Bacillus sp. Isolated from the Marine Sponge Petromica citrina against Bovine Viral Diarrhea Virus, a Surrogate Model of the Hepatitis C Virus

    OpenAIRE

    Clarice Weis Arns; Cláudia Beatriz Afonso de Menezes; Bárbara Pereira da Silva; Eduardo Furtado Flores; Fabiana Fantinatti-Garboggini; Marina Aiello Padilla; Juliana Cristina Santiago Bastos; Luciana Konecny Kohn

    2013-01-01

    The Hepatitis C virus causes chronic infections in humans, which can develop to liver cirrhosis and hepatocellular carcinoma. The Bovine viral diarrhea virus is used as a surrogate model for antiviral assays for the HCV. From marine invertebrates and microorganisms isolated from them, extracts were prepared for assessment of their possible antiviral activity. Of the 128 tested, 2 were considered active and 1 was considered promising. The best result was obtained from the extracts produced fro...

  2. Molecular Screening of Staphylococcal Enterotoxin B Gene in Clinical Isolates

    Directory of Open Access Journals (Sweden)

    Ali Karami

    2011-01-01

    Full Text Available Objective: The role of staphylococcal enterotoxin B (SEB in food poisoning is well known,however its role in other diseases remains to be explored. The aim of this study is the molecularscreening and characterization of the SEB gene in clinically isolated strains.Materials and Methods: In this experimentally study, 300 Staphylococcus aureus (S.aureus strains isolated from clinical samples were assayed. The isolated strains wereconfirmed by conventional bacteriological methods. Polymerase chain reaction (PCRwas used to determine the enterotoxin B (ent B gene. Assessment of toxin productionin all strains that contained the ent B gene was then performed. Finally, using specificantibody against SEB, a Western-blot was applied to confirm detection of enterotoxin Bproduction.Results: Results indicated that only 5% of the 300 clinically isolated S. aureus containedthe ent B gene. All strains which contained the ent B gene produced a proteinous enterotoxinB. The results of sequence determination of the PCR product were compared withthe gene bank database and 98% similarity was achieved. The results of the Western-blotconfirmed that enterotoxin B was produced in strains that contained the ent B gene.Conclusion: The results of this study indicate that 5% of clinically isolated S. aureusstrains produce enterotoxin B. Considering that the enterotoxin B is an important superantigen,it is possible that a delay in diagnosis and lack of early proper treatment can causean incidence of late complications, particularly in staphylococcal chronic infections. Forthis reason, it is suggested that in addition to detecting bacteria, an enterotoxin B detectiontest should be performed to control its toxigenicity.

  3. Evaluation of Candida strains isolated from clinical specimens

    Directory of Open Access Journals (Sweden)

    Hafize Sav

    2014-01-01

    [¤]METHODS[|]January 2011 to June 2012, Candida strains were isolated from 3905 clinical specimen. In identification of Candida species that were isolated, germ tube test, growth in Cornmeal-Tween 80 agar and formation of clamydospore, presence of pseudohyphae, carbonhytrate fermentation and assimilation tests, and the test of nitrate were studied.[¤]RESULTS[|]Finally 1122 Candida strains were isolated from 3905 various clinical specimens. The distribution of clinical specimens were as fallows: 556 from bronchoalveolar lavage (49.6%, 271 from sputum (24.2%, 114 from blood (10.2%, 51 vaginal swabs (4.6%, 50 from urine (4.4%, 30 from tissue (2.6%, 22 from endotracheal tracheal aspirate (ETA (1.9%, nine from pleural mai (0.80%, six from peritoneal fluid (0.53%, four from gastric fluid(0.35%,three from stool(0.28%,two from abscess (0.18%,three from nail (0.26%, one from cerebrospinal fluid (0.10%. From these clinical samples 848 C. albicans (75.6%, 143 C. glabrata (12.8%, 40 C. parapsilosis, (3.57%, 33 C. krusei (2.94%, 33 C. kefyr (2.94%, 19 C. tropicalis (1.7% were isolated. Other strains were identified as C. lusitania, C. lipolytica, C. norvegensis, C. pelliculosa ve C. zeylanoides.[¤]CONCLUSION[|] It was concluded that C.albicans has still been the most frequent species among Candida isolates of in our hospital; however, the incidence of non-albicans species have increased.[¤

  4. In vitro Synergistic Antimicrobial Activity of Romanian Propolis and Antibiotics against Escherichia coli Isolated from Bovine Mastitis

    Directory of Open Access Journals (Sweden)

    Mihaela NICULAE

    2015-12-01

    Full Text Available The study was aimed to characterize the chemical composition and the antimicrobial activity of Romanian propolis ethanolic extracts (EEP against antibiotic-sensitive and antibiotic-resistant E. coli strains isolated from bovine mastitis. The preliminary antimicrobial screening was performed by a disk diffusion method, followed by determination of minimum inhibitory concentrations (MIC and minimum bactericidal concentrations (MBC based on broth microdilution assay; further, the synergistic action of propolis with antimicrobial drugs was assessed by a disk diffusion method on agar containing subinhibitory concentrations of propolis. For the chemical characterisation of EEP, the flavonoids (flavones/flavonols, flavanones/dihydroflavonols and total phenolics were evaluated by spectrophotometric methods. The phenolic compounds of these extracts were also determined using HPLC. The results indicated for Romanian propolis ethanolic extracts the typical poplar composition profile with flavonoids and phenolic acids as main biological active compounds, with chromatographic analysis data confirmed also spectrophotometrically. In addition, positively correlated with the chemical composition, a strong antimicrobial efficacy was exhibited towards E. coli strains, along with interesting synergistic interaction with antibiotics that can be further investigated to obtain propolis-based formulation with antibacterial properties. Subsequent in vitro and in vivo studies evaluating the safety and efficacy are intended to consider propolis in veterinary therapeutic protocols.

  5. Inhibition of S-fimbria-mediated adhesion to human ileostomy glycoproteins by a protein isolated from bovine colostrum.

    Science.gov (United States)

    Ouwehand, A C; Conway, P L; Salminen, S J

    1995-01-01

    The aim of this study was to isolate and purify the component in bovine colostrum which is responsible for the inhibition of S-fimbria-mediated adhesion of Escherichia coli. Whey from defatted colostrum was fractionated by ultrafiltration, and the < 100K, < 30K, and < 10K fractions and the colostral whey were tested for inhibition of in vitro adhesion of radiolabelled S-fimbria-bearing E. coli to human ileostomy glycoproteins, which provide a model for human intestinal mucus. The inhibiting compound was purified from a dialyzed < 30K fraction with an anion exchange column which was eluted with a NaCl gradient (0 to 1.0 M). The compound was found to be a heat-resistant but pepsin-sensitive protein with an Mr of approximately 18,000 and an isoelectric point of approximately 5.75. The protein appears to block receptor sites for S-fimbriae on ileostomy glycoproteins, with steric hindrance being the most likely mechanism. Analysis of the amino acid sequence of the amino terminus of the 18K protein showed similarity with the sequence of beta-lactoglobulin. PMID:7591156

  6. Homogeneity of Danish environmental and clinical isolates of Shewanella algae

    DEFF Research Database (Denmark)

    Vogel, Birte Fonnesbech; Holt, H.M.; Gerner-Smidt, P.;

    2000-01-01

    Danish isolates of Shewanella algae constituted by whole-cell protein profiling a very homogeneous group, and no clear distinction was seen between strains from the marine environment and strains of clinical origin. Although variation between all strains was observed by ribotyping and random...... amplified polymorphic DNA analysis, no clonal relationship between infective strains was found. From several patients, clonally identical strains of S. algae were reisolated up to 8 months after the primary isolation, indicating that the same strain may be able to maintain the infection....

  7. Prevalence of 16S rRNA Methylase Gene rmtB Among Escherichia coli Isolated from Bovine Mastitis in Ningxia, China.

    Science.gov (United States)

    Yu, Ting; He, Tao; Yao, Hong; Zhang, Jin-Bao; Li, Xiao-Na; Zhang, Rong-Ming; Wang, Gui-Qin

    2015-09-01

    The aim of this study is to understand the prevalence and molecular characterization of 16S rRNA methylase gene, rmtB, among Escherichia coli strains isolated from bovine mastitis in China. A total of 245 E. coli isolates were collected from bovine mastitis in China between 2013 and 2014 and were screened for 16S rRNA methylase genes (armA, rmtA, rmtB, rmtC, rmtD, rmtE, and npmA) by polymerase chain reaction. About 5.3% (13/245) of the isolates carried the rmtB gene; the isolates were highly resistant to amikacin. Thirteen rmtB-positive strains were analyzed for the presence of extended-spectrum β-lactamase genes (bla(TEM), bla(CTX-M), bla(OXA), and bla(SHV)). All the isolates harbored both bla(TEM-1) and bla(CTX-M-15) genes and two of the isolates were also positive for bla(OXA-1). Pulsed-field gel electrophoresis (PFGE) analysis indicated that the nine rmtB-positive strains belonging to ST10 from one farm showed the similar PFGE pattern, indicating a clonal expansion in this farm. S1-PFGE and Southern blotting showed that 12 isolates harbored the rmtB gene in plasmids of two different sizes (≈45 kb [n=10] and ≈48 kb [n=2]), while only 1 strain harbored the rmtB gene in the chromosome. These plasmids were transferable by conjugation studies, and two isolates from two respective farms carried the same size of plasmid, suggesting that the horizontal transmission of plasmids also contributed to the spread of rmtB gene. This is the first report of prevalence of the 16S rRNA methylase gene rmtB among E. coli isolated from bovine mastitis in China, and rmtB-carrying E. coli may pose a threat to the treatment of bovine mastitis.

  8. Detection and Isolation of Digital Dermatitis Treponemes from Bovine Pressure Sores.

    Science.gov (United States)

    Clegg, S R; Crosby-Durrani, H E; Bell, J; Blundell, R; Blowey, R W; Carter, S D; Evans, N J

    2016-05-01

    Pressure sores cause severe pain and discomfort in hospitalized people and in farmed cattle and are often infected with unknown bacteria. Pressure sores occur on the upper legs of 6-10% of recumbent cattle and are generally considered to be caused by constant pressure, commonly on bony areas of the limbs. This study analyzed pressure sores taken from the upper limbs of 14 cattle using isolation in culture and nested polymerase chain reaction (PCR) to detect treponemes associated with digital dermatitis (DD). A 100% association of DD treponemes with the pressure sores was demonstrated, but treponemes were shown not to be part of the normal skin microbiota. Immunohistochemistry showed an association of DD treponemes with lesions and particularly with the hair follicles in lesions, identifying the bacteria deep within wounds, thereby suggesting that they could contribute to lesion pathogenesis. The bacteria isolated from the pressure sore lesions were similar or identical on analysis of the 16S rRNA gene to those found in DD foot lesions in cattle, suggesting the same bacteria can infect multiple lesions. Indeed, the results of this study suggest that these spirochaetal bacteria may be expanding in host range and in their ability to colonize different tissues and contribute to a range of disease manifestations in farm animals. PMID:27040650

  9. Molecular identification of Sporothrix clinical isolates in China.

    Science.gov (United States)

    Liu, Ting-ting; Zhang, Ke; Zhou, Xun

    2014-01-01

    In this study, we investigated the molecular phylogeny of 64 clinical isolates which were identified as Sporothrix schenckii sensu lato by morphological identification. All of the strains were isolates from patients from several provinces in China. The phylogeny was inferred by DNA sequence analyses based on datasets of the ribosomal internal transcribed spacer (ITS) and a combined ITS and partial β-tubulin region. Reference sequences were retrieved from GenBank. Results showed that all of the isolates were clustered in a distinct clade with a type of Sporothrix globosa. Our analysis showed that S. globosa is the causal agent of the tested sporotrichosis in China, rather than S. schenckii that was generally believed to be the case. The existence of S. schenckii in China remains to be confirmed. This study improved our understanding of the distribution of the species in S. schenckii complex.

  10. Clinical evaluation of isolated abutment teeth in removable partial dentures

    Directory of Open Access Journals (Sweden)

    Zarrati S

    2011-02-01

    Full Text Available "nBackground and Aims: Nowadays, removable partial dentures are applied to patients who are not able to use dental implants or fixed prosthesis. Although based on the studies the users of removable partial dentures are in the risk of plaque accumulation and unacceptable changes such as gingivitis, periodontitis and mobility in abutment tooth. It is not clear whether the negative effects of removable partial dentures are more on the isolated teeth which are a kind of abutment adjacent to endentulous area in both sides. The purpose of this study was to investigate the clinical condition of isolated abutment teeth without splinting in comparison to control abutment from the aspects of B.O.P (bleeding on probing, mobility, pocket depth and gingivitis."nMaterials and Methods: In this cross-sectional study, the prepared questionnaires were filled out by 50 patients who received removable partial dentures in department of removable prosthodontics of dental school of Tehran University of Medical Sciences. The patients had isolated abutment tooth and did not have any systemic disease. The obtained data were analyzed. Using Wilcoxon, exact Fisher and Kruskal-Wallis test."nResults: B.O.P (P=0.004, pocket depth (P=0.035, and mobility (P<0.001 in isolated abutments were more than those in control abutments, but there were not significant differences in the degree of caries (P=0.083 and gingivitis (P=0.07."nConclusion: This study showed that clinical condition of isolated abutments is worse than that of control abutments. More attention should be paid to healthiness of isolated teeth without splinting and periodic follow ups should be done in these cases.

  11. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Science.gov (United States)

    Celandroni, Francesco; Salvetti, Sara; Gueye, Sokhna Aissatou; Mazzantini, Diletta; Lupetti, Antonella; Senesi, Sonia; Ghelardi, Emilia

    2016-01-01

    The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  12. Antibiotic Susceptibilities of Acinetobacter Baumanii Strains Isolated from Clinical Samples

    OpenAIRE

    Harun Aðca

    2013-01-01

         Aim :  In this study it was aimed to investigate the antibiotic susceptibilities of Acinetobacter baumanii strains isolated from various clinical samples sent to Tavsanli State Hospital Microbiology Laboratory retrospectively. Material and Method: All of the cultures were examined for the agent and antibiotic susceptibilities. For the identification of bacteria, various chemical tests and BBL Crystal E/NF (Beckton Dickinson, ABD) system was used. Antibiotic susce...

  13. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Directory of Open Access Journals (Sweden)

    Francesco Celandroni

    Full Text Available The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  14. Homogeneity of Danish Environmental and Clinical Isolates of Shewanella algae

    OpenAIRE

    Vogel, Birte Fonnesbech; Holt, Hanne Marie; Gerner-Smidt, Peter; Bundvad, Anemone; Søgaard, Per; Gram, Lone

    2000-01-01

    Danish isolates of Shewanella algae constituted by whole-cell protein profiling a very homogeneous group, and no clear distinction was seen between strains from the marine environment and strains of clinical origin. Although variation between all strains was observed by ribotyping and random amplified polymorphic DNA analysis, no clonal relationship between infective strains was found. From several patients, clonally identical strains of S. algae were reisolated up to 8 months after the prima...

  15. Primary isolation of Mycobacterium bovis from bovine tissues: conditions for maximising the number of positive cultures.

    Science.gov (United States)

    Corner, L A L; Gormley, E; Pfeiffer, D U

    2012-04-23

    In studies of Mycobacterium bovis infection in animals a definitive diagnosis requires the isolation of the organism. However, the optimum conditions for the primary isolation of M. bovis have not been determined. The aim of this study was to determine for primary isolation of M. bovis, (a) the incubation time required to achieve maximum sensitivity (i.e., the number of positive samples identified), (b) the effect of decontaminants on bacterial growth rates, and (c) the influence of media and the number of slopes of media on the number of positive samples detected. Two agar-based media, modified Middlebrook 7H11 (7H11) and tuberculosis blood agar (B83), and an egg-based medium, Stonebrink's (SB) were compared. Three decontaminants, 2% (w/v) sodium hydroxide (NaOH), 0.75% (w/v) and 0.075% (w/v) cetylpyridinium chloride (CPC, also called hexadecylpyridinium chloride, HPC) and 0.5% (w/v) benzalkonium chloride (BC) were evaluated against treatment with sterile distilled water. The inoculated media slopes were incubated for up to 15 weeks. Colonies first appeared after 2 weeks on all media types and 75% of positive slopes were identified by 8 weeks. An incubation time of 15 weeks was required to identify all positive samples. The slowest growth was associated with inocula that contained the fewest viable bacilli. The time to the appearance of colonies was influenced by medium type: the median time to detection of colonies was 28 days on 7H11 and B83, and 36 days on SB. However, SB returned the greatest number of positive samples. Decontamination procedures increased the minimum incubation time required to detect positive cultures, probably due to the toxic effect of the decontaminants. Increasing the number of inoculated slopes resulted in an increased number of positive samples and a decreased time to the detection of colonies. Overall, the detection of M. bovis was significantly influenced by the choice of media, the decontaminant and the duration of incubation of

  16. Antibiotic Susceptibility of Coagulase-Negative Staphylococci Isolated from Bovine Subclinical Mastitis in Turkey

    OpenAIRE

    Beytullah Kenar*, Yahya Kuyucuoğlu and Esra Şeker

    2012-01-01

    A total of 572 California Mastitis Test (CMT) positive milk samples were collected from 423 lactating cows on 18 private farms in the Middle Western Anatolia. Coagulase–negative staphylococci colonies and CNS species identification was performed based on conventional biochemical techniques and using the API Staph test. Slime production was detected by Congo Red Agar (CRA) method. The antibiotic susceptibility was determined according to the National Committee for Clinical Laboratory Standards...

  17. Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Bruno F. Robles

    2014-04-01

    Full Text Available The objectives of the study were to evaluate the presence/production of beta-lactamases by both phenotypic and genotypic methods, verify whether results are dependent of bacteria type (Staphylococcus aureus versus coagulase-negative Staphylococcus - CNS and verify the agreement between tests. A total of 200 bacteria samples from 21 different herds were enrolled, being 100 CNS and 100 S. aureus. Beta-lactamase presence/detection was performed by different tests (PCR, clover leaf test - CLT, Nitrocefin disk, and in vitro resistance to penicillin. Results of all tests were not dependent of bacteria type (CNS or S. aureus. Several S. aureus beta-lactamase producing isolates were from the same herd. Phenotypic tests excluding in vitro resistance to penicillin showed a strong association measured by the kappa coefficient for both bacteria species. Nitrocefin and CLT are more reliable tests for detecting beta-lactamase production in staphylococci.

  18. Genetic diversity of multidrug resistant Staphylococcus aureus isolated from clinical and non clinical samples in Egypt.

    Science.gov (United States)

    Bendary, M M; Solyman, S M; Azab, M M; Mahmoud, N F; Hanora, A M

    2016-01-01

    In recent years, the increasing incidence of diseases caused by Staphylococcus aureus (S. aureus) has been noted in the university hospitals of El-Sharkia and Assuit governorates - Egypt. Therefore, we studied the genetic relatedness of multidrug resistant S. aureus isolates from different sources in the above mentioned governorates. One hundred and fifty six S. aureus isolates were divided into 5 different groups, 1 non clinical isolates from different food products and 4 different clinical isolates of human and animal sources in the 2 different governorates. Epidemiological characteristics of 156 S. aureus isolates were determined by phenotypic methods including quantitative antibiogram typing and biofilm production. Genetic typing of 35 multidrug resistant (MDR) isolates (7 from each group) based on 16S rRNA gene sequence, virulence and antimicrobial resistance gene profiles was done. The genetic relatedness of the highest virulent strain from each group was detected based on different single locus sequence typing and multi-locus sequence typing (MLST). S. aureus strains isolated from different sources and geographical areas showed high diversity. The genetic typing revealed different sequence types and different sequences of coa and spa genes. S. aureus isolates were found highly diverse in Egypt. PMID:27609475

  19. Extended biofilm susceptibility assay for Staphylococcus aureus bovine mastitis isolates: evidence for association between genetic makeup and biofilm susceptibility.

    Science.gov (United States)

    Melchior, M B; van Osch, M H J; Lam, T J G M; Vernooij, J C M; Gaastra, W; Fink-Gremmels, J

    2011-12-01

    Staphylococcus aureus is one of the most prevalent causes of bovine mastitis. The antimicrobial treatment of this disease is currently based on antimicrobial susceptibility tests according to Clinical and Laboratory Standards Institute standards. However, various authors have shown a discrepancy between the results of this standard susceptibility test and the actual cure rate of the applied antimicrobial treatment. Increasing evidence suggests that in vivo biofilm formation by Staph. aureus, which is not assessed in the antimicrobial susceptibility tests, is associated with this problem, resulting in disappointing cure rates, especially for infections of longer duration. Previous data obtained with a limited number of strains showed that the extended biofilm antimicrobial susceptibility (EBS) assay reveals differences between strains, which cannot be derived from a standard susceptibility test or from a 24-h biofilm susceptibility test. The objective of this study was to test a collection of Staph. aureus bovine mastitis strains in the EBS assay and to model the effect of antimicrobial exposure, duration of antimicrobial exposure, and genotype profile of the strains on antimicrobial susceptibility. With the results from a previous study with the same collection of strains, the effect of genotype represented by accessory gene regulator gene (agr-type), the presence of insertional sequence 257 (IS257), intercellular adhesion (ica), and the β-lactamase (blaZ) gene were entered as explanatory factors in a logistic regression model. The agr locus of Staph. aureus controls the expression of most of the virulence factors, represses the transcription of several cell wall-associated proteins, and activates several exoproteins during the post-exponential phase. The IS257 gene has been related to biofilm formation in vitro and was found earlier in 50% of the agr-type 2 strains. The ica gene cluster encodes for the production of an extracellular polysaccharide adhesin, termed

  20. Isolation Frequency Characteristics of Candida Species from Clinical Specimens

    Science.gov (United States)

    Kim, Ga-Yeon; Jeon, Jae-Sik

    2016-01-01

    Candida spp. is an invasive infectious fungus, a major risk factor that can increase morbidity and mortality in hospitalized patients. In this study, 2,508 Candida spp. were isolated from various clinical specimens collected from university hospitals from July 2011 to October 2014. They were identified in order to determine isolation frequencies and characteristics by specimen, gender, age group, year, season, and month. The strain-specific isolation rate of Candida spp. is in the order of Candida albicans (1,218 strains, 48.56%), Candida glabrata (416 strains, 16.59%), Candida utilis (305 strains, 12.16%), Candida tropicalis (304 strains, 12.12%), and Candida parapsilosis (116 strains, 4.63%) and these five species accounted for more than 94% of the total strains. Of the specimens, Candida spp. were most frequently isolated from urine-catheter, followed by urine-voided, blood, sputum, other, open pus, vaginal discharge, Tip, ear discharge, bronchial aspiration and bile, in that order. Looking at the age distribution, the detection rate of patients in their 60s and older was significantly higher at 75.8% (1,900/2,508). The detection rate of patients in their 20s and younger was shown to be very low at 2.55% (64/2,508). By year, the detection rate of non-albicans Candida spp. showed a tendency to gradually increase each year compared with C. albicans. As isolation of Candida spp. from clinical samples at the specie level can vary depending on characteristics of the patient, sample, season, etc., continual studies are required. PMID:27433120

  1. Isolation Frequency Characteristics of Candida Species from Clinical Specimens.

    Science.gov (United States)

    Kim, Ga-Yeon; Jeon, Jae-Sik; Kim, Jae Kyung

    2016-06-01

    Candida spp. is an invasive infectious fungus, a major risk factor that can increase morbidity and mortality in hospitalized patients. In this study, 2,508 Candida spp. were isolated from various clinical specimens collected from university hospitals from July 2011 to October 2014. They were identified in order to determine isolation frequencies and characteristics by specimen, gender, age group, year, season, and month. The strain-specific isolation rate of Candida spp. is in the order of Candida albicans (1,218 strains, 48.56%), Candida glabrata (416 strains, 16.59%), Candida utilis (305 strains, 12.16%), Candida tropicalis (304 strains, 12.12%), and Candida parapsilosis (116 strains, 4.63%) and these five species accounted for more than 94% of the total strains. Of the specimens, Candida spp. were most frequently isolated from urine-catheter, followed by urine-voided, blood, sputum, other, open pus, vaginal discharge, Tip, ear discharge, bronchial aspiration and bile, in that order. Looking at the age distribution, the detection rate of patients in their 60s and older was significantly higher at 75.8% (1,900/2,508). The detection rate of patients in their 20s and younger was shown to be very low at 2.55% (64/2,508). By year, the detection rate of non-albicans Candida spp. showed a tendency to gradually increase each year compared with C. albicans. As isolation of Candida spp. from clinical samples at the specie level can vary depending on characteristics of the patient, sample, season, etc., continual studies are required.

  2. Platelet aggregation responses and virus isolation from platelets in calves experimentally infected with type I or type II bovine viral diarrhea virus.

    Science.gov (United States)

    Walz, P H; Bell, T G; Grooms, D L; Kaiser, L; Maes, R K; Baker, J C

    2001-10-01

    Altered platelet function has been reported in calves experimentally infected with type II bovine viral diarrhea virus (BVDV). The purpose of the present study was to further evaluate the ability of BVDV isolates to alter platelet function and to examine for the presence of a virus-platelet interaction during BVDV infection. Colostrum-deprived Holstein calves were obtained immediately after birth, housed in isolation, and assigned to 1 of 4 groups (1 control and 3 treatment groups). Control calves (n = 4) were sham inoculated, while calves in the infected groups (n = 4 for each group) were inoculated by intranasal instillation with 10(7) TCID50 of either BVDV 890 (type II), BVDV 7937 (type II), or BVDV TGAN (type I). Whole blood was collected prior to inoculation (day 0) and on days 4, 6, 8, 10, and 12 after inoculation for platelet function testing by optical aggregometry by using adenosine diphosphate and platelet activating factor. The maximum percentage aggregation and the slope of the aggregation curve decreased over time in BVDV-infected calves; however, statistically significant differences (Freidman repeated measures ANOVA on ranks, P infected with the type II BVDV isolates. Bovine viral diarrhea virus was not isolated from control calves, but was isolated from all calves infected with both type II BVDV isolates from days 4 through 12 after inoculation. In calves infected with type I BVDV, virus was isolated from 1 of 4 calves on days 4 and 12 after inoculation and from all calves on days 6 and 8 after inoculation. Altered platelet function was observed in calves infected with both type II BVDV isolates, but was not observed in calves infected with type I BVDV. Altered platelet function may be important as a difference in virulence between type I and type II BVDV infection.

  3. Isolation of an inactive bovine heart cAMP-dependent protein kinase holoenzyme containing bound cAMP

    International Nuclear Information System (INIS)

    Bovine heart Type II cAMP-dependent protein kinase (PK) was purified to homogeneity as determined by SDS-PAGE. The purification steps were DEAE-cellulose, ammonium sulfate precipitation, phenyl-Sepharose, alumina C-γ, and HPLC-DEAE. The last step resolved two distinct peaks of cAMP dependent kinase activity (activity ratios = 0.05 - 0.10) eluting at approximately 250 (Peak 1) and 275 (Peak 2) mM NaCl. When subjected to HPLC-gel permeation they had the same Stoke's radii. Linear sucrose gradients gave S/sub 20,w/ values of 7.73 S and 7.25 S for Peaks 1 and 2, respectively. It was determined by integrating the areas of scanned SDS-PAGE bands that regulatory and catalytic subunits were present in equimolar amounts in both Peaks 1 and 2. The ratios of equilibrium [3H] cAMP binding to kinase activity for the two peaks were the same. cAMP was present in trace amounts in Peak 1 but in stoichiometric amounts in Peak 2 (between 25 and 75 % saturation of cAMP binding sites). Although SDS-PAGE analysis ruled out conversions by proteolysis or autophosphorylation-dephosphorylation, Peak 1 could be partially converted to Peak 2 by the addition of subsaturating amounts of cAMP, 3H] cAMP binding and in kinase activation (as indicated by a larger Hill coefficient) of Peak 2 relative to Peak 1 also supported the presence of cAMP in Peak 2. In conclusion, two forms of inactive PK have been isolated, one of which is a ternary complex of PK holoenzyme and cAMP. This complex could represent a cellular form of the enzyme which is primed for activation

  4. Est10: A Novel Alkaline Esterase Isolated from Bovine Rumen Belonging to the New Family XV of Lipolytic Enzymes.

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    María Cecilia Rodríguez

    Full Text Available A metagenomic fosmid library from bovine rumen was used to identify clones with lipolytic activity. One positive clone was isolated. The gene responsible for the observed phenotype was identified by in vitro transposon mutagenesis and sequencing and was named est10. The 367 amino acids sequence harbors a signal peptide, the conserved secondary structure arrangement of alpha/beta hydrolases, and a GHSQG pentapeptide which is characteristic of esterases and lipases. Homology based 3D-modelling confirmed the conserved spatial orientation of the serine in a nucleophilic elbow. By sequence comparison, Est10 is related to hydrolases that are grouped into the non-specific Pfam family DUF3089 and to other characterized esterases that were recently classified into the new family XV of lipolytic enzymes. Est10 was heterologously expressed in Escherichia coli as a His-tagged fusion protein, purified and biochemically characterized. Est10 showed maximum activity towards C4 aliphatic chains and undetectable activity towards C10 and longer chains which prompted its classification as an esterase. However, it was able to efficiently catalyze the hydrolysis of aryl esters such as methyl phenylacetate and phenyl acetate. The optimum pH of this enzyme is 9.0, which is uncommon for esterases, and it exhibits an optimal temperature at 40 °C. The activity of Est10 was inhibited by metal ions, detergents, chelating agents and additives. We have characterized an alkaline esterase produced by a still unidentified bacterium belonging to a recently proposed new family of esterases.

  5. Genome sequences of serotype A6 Mannheimia haemolytica isolates D174 and D38 recovered from bovine pneumonia

    Science.gov (United States)

    Here we report two genomes, one complete and one draft, from virulent bovine strains of Mannheimia haemolytica(strains D174 and D38)serotype A2 recovered prior to the field usage of modern antimicrobial drugs....

  6. Production and assay of cellulolytic enzyme activity of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya abbatoir, Indonesia

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    W. P. Lokapirnasari

    2015-03-01

    Full Text Available Aim: This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4-β-D-glucanase, exo-(1,4-β-Dglucanase, and β-glucosidase, at optimum temperature and optimum pH of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia. Materials and Methods: To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase, and β-glucosidase. Results: Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5. Conclusion: E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase and β-glucosidase.

  7. Characterization of integrons in Burkholderia cepacia clinical isolates

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    Linda Furlanis

    2010-03-01

    Full Text Available Burkholderia cepacia is an opportunistic pathogen able to colonize the airways of Cystic Fibrosis (CF patients, frequently developing chronic infections. In 20% of cases these infections cause severe and poorly controlled pathological situations because of the intrinsic antibiotic resistance expressed by the microorganism. CF patients are often subjected to antibiotic therapy: this facilitates the acquisition of antibiotic resistance determinants by the infecting bacteria. Integrons are mobile genetic elements that are widespread in bacterial populations and favor the acquisition of gene cassettes coding for these determinants.The presence of class 1 integrons was investigated by PCR with primers specific for the 5’ and 3’ ends in Burkholderia isolates recovered from patients in treatment at the CF center of Friuli Venezia Giulia. The same integron, carrying an uncommon allelic form (Ib of the aacA4 gene in its cassette array and conferring resistance to some aminoglycosides, was found in two independent isolates (different RAPD profiles infecting two different patients. In both isolates the integron was carried by plasmids and was still present 3 and 6 years later the first finding. Despite the exchange of integrons between bacterial pathogens is fully described, these items were not frequently found in Burkholderia isolates. Although the clinical relevance of the integron we identified is low (a single gene cassette encoding a widespread resistance,we feel concerned that these genetic elements begin to circulate in this bacterial species, as this could make more and more troublesome the treatment of infections notoriously difficult to eradicate.

  8. Antibiotic Susceptibilities of Acinetobacter Baumanii Strains Isolated from Clinical Samples

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    Harun Aðca

    2013-01-01

    Full Text Available      Aim :  In this study it was aimed to investigate the antibiotic susceptibilities of Acinetobacter baumanii strains isolated from various clinical samples sent to Tavsanli State Hospital Microbiology Laboratory retrospectively. Material and Method: All of the cultures were examined for the agent and antibiotic susceptibilities. For the identification of bacteria, various chemical tests and BBL Crystal E/NF (Beckton Dickinson, ABD system was used. Antibiotic susceptibilities were investigated according to CLSI criteria on Mueller Hinton agar by disc diffusion method. Results: There were 74 strains isolated and identified as Acinetobacter baumanii. Most of the strains were isolated from  tracheal aspirate specimens (46 % Most of the strains were isolated from nosocomial infections. Antibiotic resistance was high among strains. The most susceptible antibiotic was gentamicin (30%. Discussion: To prevent the development of resistance, antibiotics should be used carefully in appropriate doses and time, empirical  antibiotherapy should be determined for each centre according to resistance rates of the centre and should be regulated according to the antibiogram results. Increasing resistance rates in Acinetobacter strains leads to the usage of new alternative antibiotics.  

  9. Detection of Enterotoxigenic Potential and Determination of Clonal Profile in Staphylococcus aureus and Coagulase-Negative Staphylococci Isolated from Bovine Subclinical Mastitis in Different Brazilian States.

    Science.gov (United States)

    Mello, Priscila Luiza; Moraes Riboli, Danilo Flávio; Pinheiro, Luiza; de Almeida Martins, Lisiane; Vasconcelos Paiva Brito, Maria Aparecida; Ribeiro de Souza da Cunha, Maria de Lourdes

    2016-01-01

    Epidemiological studies have identified Staphylococcus aureus as the most common agent involved in food poisoning. However, current research highlights the importance of toxigenic coagulase-negative staphylococci (CoNS) isolated from food. The aim of this study was to characterize Staphylococcus spp. isolated from cows with bovine subclinical mastitis regarding the presence of genes responsible for the production of staphylococcal enterotoxins and of the tst-1 gene encoding toxic shock syndrome toxin 1, and to determine the clonal profile of the isolates carrying any of the genes studied. A total of 181 strains isolated in different Brazilian states, including the South, Southeast, and Northeast regions, were analyzed. The sea gene was the most frequent, which was detected in 18.2% of the isolates, followed by seb in 7.7%, sec in 14.9%, sed in 0.5%, see in 8.2%, seg in 1.6%, seh in 25.4%, sei in 6.6%, and ser in 1.6%. The sej, ses, set, and tst-1 genes were not detected in any of the isolates. The typing of the isolates by pulsed-field gel electrophoresis revealed important S. aureus and S. epidermidis clusters in different areas and the presence of enterotoxin genes in lineages isolated from animals that belong to herds located geographically close to each other. PMID:27092525

  10. [Taxonomic study of clinic isolates of Trichophyton in Rosario, Argentina].

    Science.gov (United States)

    Tartabini, Mirta L; Bonino, Guillermo S; Racca, Liliana; Luque, Alicia G

    2013-01-01

    Due to the pleomorphism and cultural variability displayed by species of the genus Trichophyton, the identification methods based solely on morphological features are usually insufficient for their classification. The goal of the present work was to test a set of phenotypic methods in order to identify fungal isolates that belong to the aforementioned genus. These methods were based on a molecular taxonomic technique used as standard. Clinical isolates (56) were used as samples along with 6 reference strains. Macro and micromorphological studies were performed as well as biochemical and physiological tests such as in vitro hair perforation, nutritional requirements in Trichophyton agar media, urease production and growth on bromocresol purple-milk. solids-glucose (BCP-MS-G) agar. Additionally, PCR fingerprinting using the (GACA)4 primer was employed. As a result of the PCR method, specific profiles were observed for Microsporum canis, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton interdigitale. Identical profiles were obtained for Arthroderma benhamiae y Trichophyton erinacei. Of the total number of clinical isolates, 39 matched the T. rubrum profile while 13 corresponded to A. benhamiae and 4 to T. interdigitale. The most useful phenotypic test to differentiate between T. rubrum and T. mentagrophytes complex strains was alkalinization of the BCP-MS-G medium. Phenotypic tests did not allow differentiation among the T. mentagrophytes complex species. On the other hand, the molecular technique allowed characterization of T. rubrum isolates as well as of those observed in our study and included in the T. mentagrophytes complex: T. interdigitale and Trichophyton sp., the anamorph of A. benhamiae.

  11. Comparison of genomic and antimicrobial resistance features of latex agglutination test-positive and latex agglutination test-negative Staphylococcus aureus isolates causing bovine mastitis.

    Science.gov (United States)

    Moser, A; Stephan, R; Corti, S; Johler, S

    2013-01-01

    The dairy industry suffers massive economic losses due to staphylococcal mastitis in cattle. The Staphaureux latex agglutination test (Oxoid, Basel, Switzerland) was reported to lead to negative results in 54% of bovine Staphylococcus aureus strains, and latex-negative strains are thought to be less virulent than Staphaurex latex-positive strains. However, comparative information on virulence and resistance profiles of these 2 groups of Staph. aureus is scarce. Our objective was to associate the latex agglutination phenotype of Staph. aureus strains isolated from bovine mastitis milk with data on clonal complexes, virulence genes, and antibiotic resistance to (1) determine the virulence profiles of the Staphaureux test positive and Staphaurex test negative groups, and (2) provide data needed to improve treatment of bovine mastitis and to identify potential vaccine targets. Seventy-eight Staph. aureus strains isolated from 78 cows on 57 Swiss farms were characterized. Latex agglutination was tested by Staphaureux kit, and resistance profiles were generated by disk diffusion. A DNA microarray was used to assign clonal complexes (CC) and to determine virulence and resistance gene profiles. By the Staphaureux test, 49% of the isolates were latex-positive and 51% were latex-negative. All latex-negative strains were assigned to CC151, whereas latex-positive strains were assigned to various clonal complexes, including CC97 (n=16), CC8 (n=10), CC479 (n=5), CC20 (n=4), CC7 (n=1), CC9 (n=1), and CC45 (n=1). Although the latex-negative isolates were susceptible to all antimicrobial agents tested, 24% of latex-positive isolates were classified as intermediate with regard to cefalexin-kanamycin and 13% were resistant to both ampicillin and penicillin. Microarray profiles of latex-negative isolates were highly similar, but differed largely from those of latex-positive isolates. Although the latex-negative group lacked several enterotoxin genes and sak, it exhibited significantly

  12. Correlation of hypothetical virulence traits of two Streptococcus uberis strains with the clinical manifestation of bovine mastitis.

    Science.gov (United States)

    Tassi, Riccardo; McNeilly, Tom N; Sipka, Anja; Zadoks, Ruth N

    2015-01-01

    Streptococcus uberis is a common cause of clinical and subclinical mastitis in dairy cattle. Several virulence mechanisms have been proposed to contribute to the species' ability to cause disease. Here, virulence characteristics were compared between S. uberis strains FSL Z1-048, which consistently caused clinical mastitis in a challenge model, and FSL Z1-124, which consistently failed to cause disease in the same model, to ascertain whether in vitro virulence characteristics were related to clinical outcome. Macrophages derived from bovine blood monocytes failed to kill FSL Z1-048 whilst reducing survival of FSL Z1-124 by 42.5%. Conversely, blood derived polymorphonuclear cells caused more reduction (67.1 vs. 44.2%, respectively) in the survival of FSL Z1-048 than in survival of FSL Z1-124. After 3 h of coincubation with bovine mammary epithelial cell line BME-UV1, 1000-fold higher adherence was observed for FSL Z1-048 compared to FSL Z1-124, despite presence of a frame shift mutation in the sua gene of FSL Z1-048 that resulted in predicted truncation of the S. uberis Adhesion Molecule (SUAM) protein. In contrast, FSL Z1-124 showed higher ability than FSL Z1-048 to invade BME-UV1 cells. Finally, observed biofilm formation by FSL Z1-124 was significantly greater than for FSL Z1-048. In summary, for several hypothetical virulence characteristics, virulence phenotype in vitro did not match disease phenotype in vivo. Evasion of macrophage killing and adhesion to mammary epithelial cells were the only in vitro traits associated with virulence in vivo, making them attractive targets for further research into pathogenesis and control of S. uberis mastitis. PMID:26497306

  13. Antimicrobial sensitivity profile of Staphylococcus spp. Isolated from clinical mastitis

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    Thamires Martins

    2012-12-01

    Full Text Available Inflammation of the mammary gland, which is also known as mastitis, occupies a prominent place among the diseases that affect dairy cattle, having a great economic importance in the dairy sector. Mastitis may have different origins, however, infectious mastitis is the most frequent and represents a risk to public health due to the propagation of microorganisms through milk. Staphylococcus spp. are considered the microorganisms that cause the greatest losses in milk production, being that Staphylococcus aureus is the pathogen of major importance because they present high resistence to antimicrobials. Empirical treatment, without prior identification of the pathogens and their resistance profile, may contribute to the emergence of multidrug-resistant strains and risk the efficiency of the antimicrobial. In that scenery, the study aimed to evaluate the resistance profile of Staphylococcus spp. against some antimicrobials used in the treatment of cows with clinical mastitis. The study was conducted on a property in the state of São Paulo from January 2011 to June 2012. We evaluated 29 lactating cows that present clinical mastitis in, at least, one mammary quarter. The diagnosis of clinical mastitis was performed by evaluating the clinical signs and also by Tamis test. Samples of milk from mammary quarters were collected aseptically in sterile tubes for microbiological evaluation. Microorganisms were isolated on sheep blood agar 5% and Sabouraud agar with chloramphenicol. The sensitivity profile of Staphylococcus spp. to the antibiotics ampicillin, cephalexin, ceftiofur, cefaclor, gentamicin, kanamycin, neomycin, penicillin G and oxacillin, was tested by disk diffusion test on Mueller-Hinton agar. From a total of 106 samples of milk analyzed, 64 (60.38% presented microbiological growth, being observed isolation of Streptococcus spp. 29 (34.52%, Staphylococcus spp. 28 (33.33%, Corynebacterium spp. 17 (20.24%, filamentous fungi 4 (4.76%, yeast 4 (4

  14. Fatores de virulência em linhagens de Escherichia coli isoladas de mastite bovina Virulence factors in Escherichia coli strains isolated from bovine mastitis

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    M.G. Ribeiro

    2006-10-01

    Full Text Available Avaliou-se a ocorrência de fatores de virulência e do sorotipo O157:H7 em 120 linhagens de Escherichia coli, isoladas de 80 casos de mastite clínica bovina e 40 de mastite subclínica. Verificou-se alfa-hemolisina em oito (6,7% linhagens, isoladas de cinco casos de mastite clínica e três de mastite subclínica e em nenhuma das estirpes detectou-se enteroemolisina. A presença de sideróforos foi encontrada em 11 (9,2% linhagens, sete de mastite clínica e quatro de subclínica. Em duas (1,7% estirpes isoladas de mastite subclínica, identificou-se enterotoxina STa. Observou-se efeito citopático em células vero compatível com a produção de verotoxina-VT em cinco (4,2% linhagens, duas de mastite clínica e três subclínicas. Em uma (0,8% linhagem isolada de mastite clínica, detectou-se efeito citopático compatível com o fator necrosante citotóxico. Nenhuma estirpe apresentou-se sorbitol-negativa no MacConkey-sorbitol, tampouco aglutinou com o sorotipo O157:H7. Os antimicrobianos mais efetivos foram polimixina B (97,5% e norfloxacina (95,8%. Observou-se multi-resistência a dois ou mais antimicrobianos em 24 (20% estirpes, principalmente com o uso de ampicilina e ceftiofur.The occurrence of different virulence factors and O157:H7 serotype investigation in 120 Escherichia coli strains isolated from clinical (80 cases and subclinical (40 cases bovine mastitis was evaluated. Alpha-haemolysin was detected in 8 (6.7% strains (5 clinical and 3 subclinical cases. None strain showed enterohaemolysin production. E. coli growth under iron restriction conditions (siderophores production was observed in 11 (9.2% strains (7 clinical and 4 subclinical cases. STa enterotoxin was detected in 2 (1.7% strains from subclinical cases. Cytotoxic effect in vero cells compatible with verotoxin-VT production was observed in 5 (4.2% strains (2 clinical and 3 subclinical cases. One strain (0.8% isolated from clinical mastitis showed cytophatic effect in vero

  15. Isolation and proliferation of umbilical cord tissue derived mesenchymal stem cells for clinical applications.

    Science.gov (United States)

    Van Pham, Phuc; Truong, Nhat Chau; Le, Phuong Thi-Bich; Tran, Tung Dang-Xuan; Vu, Ngoc Bich; Bui, Khanh Hong-Thien; Phan, Ngoc Kim

    2016-06-01

    Umbilical cord (UC) is a rich source of rapidly proliferating mesenchymal stem cells (MSCs) that are easily cultured on a large-scale. Clinical applications of UC-MSCs include graft-versus-host disease, and diabetes mellitus types 1 and 2. UC-MSCs should be isolated and proliferated according to good manufacturing practice (GMP) with animal component-free medium, quality assurance, and quality control for their use in clinical applications. This study developed a GMP standard protocol for UC-MSC isolation and culture. UC blood and UC were collected from the same donors. Blood vasculature was removed from UC. UC blood was used as a source of activated platelet rich plasma (aPRP). Small fragments (1-2 mm(2)) of UC membrane and Wharton's jelly were cut and cultured in DMEM/F12 medium containing 1 % antibiotic-antimycotic, aPRP (2.5, 5, 7.5 and 10 %) at 37 °C in 5 % CO2. The MSC properties of UC-MSCs at passage 5 such as osteoblast, chondroblast and adipocyte differentiation, and markers including CD13, CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105, and HLA-DR were confirmed. UC-MSCs also were analyzed for karyotype, expression of tumorigenesis related genes, cell cycle, doubling time as well as in vivo tumor formation in NOD/SCID mice. Control cells consisted of UC-MSCs cultured in DMEM/F12 plus 1 % antibiotic-antimycotic, and 10 % fetal bovine serum (FBS). All UC-MSC (n = 30) samples were successfully cultured in medium containing 7.5 and 10 % aPRP, 92 % of samples grew in 5.0 % aPRP, 86 % of samples in 2.5 % aPRP, and 72 % grew in 10 % FBS. UC-MSCs in these four groups exhibited similar marker profiles. Moreover, the proliferation rates in medium with PRP, especially 7.5 and 10 %, were significantly quicker compared with 2.5 and 5 % aPRP or 10 % FBS. These cells maintained a normal karyotype for 15 sub-cultures, and differentiated into osteoblasts, chondroblasts, and adipocytes. The analysis of pluripotent cell markers showed UC-MSCs maintained

  16. Leukocidin genes lukF-P83 and lukM are associated with taphylococcus aureus clonal complexes 151, 479 and 133 isolated from bovine udder infections in Thuringia, Germany

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    Schlotter Katharina

    2012-05-01

    Full Text Available Abstract Staphylococcus aureus is one of the most important causal agents of bovine mastitis. Population studies on bovine Staphylococcus aureus isolates have identified considerable genetic heterogeneity among strains causing mastitis. The aim of this study was to investigate the contribution of different clonal complexes and the occurrence of virulence factors and resistance determinants within bovine Staphylococcus aureus isolates. A total of 189 Staphylococcus aureus isolates obtained from milk samples of 34 dairy herds in the German Federal State of Thuringia were characterised by microarray technology. The isolates were assigned to eleven different clonal complexes with CC151, CC479 and CC133 being dominant (together 80.5%. The methicillin resistance gene mecA was found in four isolates (2.1%, which belonged to CC398. Enterotoxin genes could be detected in 79.3% of analysed Staphylococcus aureus and 19 isolates (10.1% harboured a distinct allele of the toxic shock syndrome toxin gene, tst-RF122. The most striking finding of the present study was that almost all except one isolate (151/152 belonging to CC151, CC479 and CC133 harboured the leukocidin genes lukF-P83 and lukM, whereas no further isolates from other lineages possessed these genes. The consistent occurrence of lukF-P83/lukM in the dominating clonal complexes suggests an essential role of this leukocidin in the etiology of bovine mastitis.

  17. Leukocidin genes lukF-P83 and lukM are associated with Taphylococcus aureus clonal complexes 151, 479 and 133 isolated from bovine udder infections in Thuringia, Germany.

    Science.gov (United States)

    Schlotter, Katharina; Ehricht, Ralf; Hotzel, Helmut; Monecke, Stefan; Pfeffer, Martin; Donat, Karsten

    2012-01-01

    Staphylococcus aureus is one of the most important causal agents of bovine mastitis. Population studies on bovine Staphylococcus aureus isolates have identified considerable genetic heterogeneity among strains causing mastitis. The aim of this study was to investigate the contribution of different clonal complexes and the occurrence of virulence factors and resistance determinants within bovine Staphylococcus aureus isolates.A total of 189 Staphylococcus aureus isolates obtained from milk samples of 34 dairy herds in the German Federal State of Thuringia were characterised by microarray technology.The isolates were assigned to eleven different clonal complexes with CC151, CC479 and CC133 being dominant (together 80.5%). The methicillin resistance gene mecA was found in four isolates (2.1%), which belonged to CC398. Enterotoxin genes could be detected in 79.3% of analysed Staphylococcus aureus and 19 isolates (10.1%) harboured a distinct allele of the toxic shock syndrome toxin gene, tst-RF122. The most striking finding of the present study was that almost all except one isolate (151/152) belonging to CC151, CC479 and CC133 harboured the leukocidin genes lukF-P83 and lukM, whereas no further isolates from other lineages possessed these genes.The consistent occurrence of lukF-P83/lukM in the dominating clonal complexes suggests an essential role of this leukocidin in the etiology of bovine mastitis.

  18. Frequency and Antimicrobial Susceptibility of Coagulasenegative Staphylococci Isolated from Bovine Mastitis in Dairy Farms from Tolima, Colombia

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    María del Pilar Sánchez Bonilla

    2015-09-01

    Full Text Available Objective: To determine the antimicrobial presence and susceptibility of the coagulase-negative staphylococci group (ECN, for its initials in Spanish in some cattle farms from Tolima, Colombia. Materials and methods: Using the California test for the diagnosis of mastitis (CMT, for its initials in Spanish, 484 quarters belonging to 121 cows from five small production ranches from a region of central Tolima were evaluated. CMT positive samples were cultivated for bacterial isolation. The ECN found were tested for susceptibility to the antibiotics. The results were analyzed with descriptive statistics. Results: 252 (52% quarters did not show any type of reaction to the CMT, nor did they show any clinical signs of mastitis, therefore they were considered free of the disease. From the quarters, 73 (15% turned positive for CMT and bacteriological culture. From these, 36 strains of ECN were isolated (7.4% of the total of quarters; S. aureus-ECP, 28 (5.8%; Streptococcus spp., 6 (1.2%.; Escherichia coli, 2 (0.4%, and Klebsiella pneumoniae, 2 (0.2%. The clinical and subclinical mastitis in the quarter occurred in 1.4% and 13.6%, respectively. In 5 (1.0% of the quarters, clinical mastitis caused by ECN was found and subclinical mastitis in 31 (6.4%. 61% of the ECN strains were resistant to penicillin, and 58%, to tetracycline; 97% were sensitive to cefoperazone. Conclusion: The ECN group, considered a global emergent of mastitis, is evidenced with high frequency in ranches from Tolima, Colombia, causing clinical and subclinical mastitis with varied response to antimicrobials.

  19. Isolation, Identification, and Antibiotic Susceptibility Testing of Salmonella from Slaughtered Bovines and Ovines in Addis Ababa Abattoir Enterprise, Ethiopia: A Cross-Sectional Study.

    Science.gov (United States)

    Kebede, Abe; Kemal, Jelalu; Alemayehu, Haile; Habte Mariam, Solomon

    2016-01-01

    Salmonellae are ubiquitous, found in animals, humans, and the environment, a condition which facilitates transmission and cross contamination. Salmonella enterica serotypes exert huge health and economic impacts due to their virulence or carriage of antibiotic resistance traits. To address this significant issues with regard to public health, availability of adequate information on the prevalence and antibiotic resistance patterns of Salmonella, and establishment of adequate measures to control contamination and infection are needed. A cross-sectional study was conducted to assess the level of Salmonella infection in slaughtered bovines and ovines at Addis Ababa abattoir. Samples were collected randomly and processed for identification and antimicrobial susceptibility testing of Salmonella spp. From 280 animals examined, 13 (4.64%) (8 bovines and 5 ovines) were positive, with most samples (12/13, 92%) comprising Salmonella Dublin. Very high level of resistance to some antibiotics used in human medicine was detected. Most isolates were susceptible to gentamycin and amikacin. Nine (69%) of all isolates were resistant to multiple antibiotics. Serotyping revealed 12 of 13 isolates to be of the Dublin serotype with 9,12:g,p:- antigenic formula. This study emphasizes the importance of improving the evisceration practice during slaughtering and restricting the use of antibiotics in farm animals. PMID:27660816

  20. Isolation, Identification, and Antibiotic Susceptibility Testing of Salmonella from Slaughtered Bovines and Ovines in Addis Ababa Abattoir Enterprise, Ethiopia: A Cross-Sectional Study

    Directory of Open Access Journals (Sweden)

    Abe Kebede

    2016-01-01

    Full Text Available Salmonellae are ubiquitous, found in animals, humans, and the environment, a condition which facilitates transmission and cross contamination. Salmonella enterica serotypes exert huge health and economic impacts due to their virulence or carriage of antibiotic resistance traits. To address this significant issues with regard to public health, availability of adequate information on the prevalence and antibiotic resistance patterns of Salmonella, and establishment of adequate measures to control contamination and infection are needed. A cross-sectional study was conducted to assess the level of Salmonella infection in slaughtered bovines and ovines at Addis Ababa abattoir. Samples were collected randomly and processed for identification and antimicrobial susceptibility testing of Salmonella spp. From 280 animals examined, 13 (4.64% (8 bovines and 5 ovines were positive, with most samples (12/13, 92% comprising Salmonella Dublin. Very high level of resistance to some antibiotics used in human medicine was detected. Most isolates were susceptible to gentamycin and amikacin. Nine (69% of all isolates were resistant to multiple antibiotics. Serotyping revealed 12 of 13 isolates to be of the Dublin serotype with 9,12:g,p:- antigenic formula. This study emphasizes the importance of improving the evisceration practice during slaughtering and restricting the use of antibiotics in farm animals.

  1. Isolation, Identification, and Antibiotic Susceptibility Testing of Salmonella from Slaughtered Bovines and Ovines in Addis Ababa Abattoir Enterprise, Ethiopia: A Cross-Sectional Study

    Science.gov (United States)

    Kebede, Abe; Alemayehu, Haile

    2016-01-01

    Salmonellae are ubiquitous, found in animals, humans, and the environment, a condition which facilitates transmission and cross contamination. Salmonella enterica serotypes exert huge health and economic impacts due to their virulence or carriage of antibiotic resistance traits. To address this significant issues with regard to public health, availability of adequate information on the prevalence and antibiotic resistance patterns of Salmonella, and establishment of adequate measures to control contamination and infection are needed. A cross-sectional study was conducted to assess the level of Salmonella infection in slaughtered bovines and ovines at Addis Ababa abattoir. Samples were collected randomly and processed for identification and antimicrobial susceptibility testing of Salmonella spp. From 280 animals examined, 13 (4.64%) (8 bovines and 5 ovines) were positive, with most samples (12/13, 92%) comprising Salmonella Dublin. Very high level of resistance to some antibiotics used in human medicine was detected. Most isolates were susceptible to gentamycin and amikacin. Nine (69%) of all isolates were resistant to multiple antibiotics. Serotyping revealed 12 of 13 isolates to be of the Dublin serotype with 9,12:g,p:- antigenic formula. This study emphasizes the importance of improving the evisceration practice during slaughtering and restricting the use of antibiotics in farm animals.

  2. Molecular Characterization and Toxicity Confirmation of LukM/F'-PV Producing Staphylococcus aureus Isolated from Bovine Mastitis Samples in Mysore, India.

    Science.gov (United States)

    Padmaja, R J; Halami, P M

    2013-09-01

    The widespread status of subclinical condition of bovine mastitis is often associated with the production of leukotoxin M/F'-PV producing Staphylococcus aureus. The present study aims for the profiling of such leukotoxin producers through conventional and molecular methods in parallel to their leukotoxicity. The incidence of this particular pathogen was assessed in mastitis infected Holstein-Friesian cattle, where eight isolates of staphylococci were found to be present in 20 % of collected samples. Being intermediately resistant to vancomycin, they showed characteristic double zone hemolysis on 7 % sheep blood agar and typical type II reaction for coagulase test indicating the pathogenic attributes. Further with RAPD-PCR and 16S rDNA-RFLP, epidemiological specificity and genotypic relatedness of isolates to S. aureus was confirmed. Subsequently, the presence of leukotoxin (lukM) gene in native isolates was detected by leukotoxin gene specific PCR. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay evaluated for secreted leukotoxin in cell free supernatant was estimated to be 223 toxic units which had an LD50 cytotoxic activity on bovine neutrophil. Thus, the data acquired during study can be of prime diagnostic method for timely and accurate analysis of subclinical mastitis samples which goes undetected at consumer level. PMID:24426122

  3. Isolation and identification of bovine viral diarrhea virus from swine%猪源牛病毒性腹泻病毒的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    杨小燕; 魏春华; 刘建奎; 戴爱玲; 李晓华

    2011-01-01

    从福建省某猪场疑似猪瘟的发病仔猪采集病料,处理后接种于牛肾细胞(MDBK),分离到1株病毒,并对其进行了系统鉴定.结果显示,病料接种MDBK细胞72 h后产生了明显的细胞病变;该分离毒株可以被牛病毒性腹泻病毒(BVDV)阳性血清中和,中和效价为1:106;该分离毒株对氯仿、乙醚、胰酶、酸敏感,不耐热,56℃30 min可被灭活,PCR检测该分离毒株的细胞培养液,结果可扩增出325 bp的片段,该片段的核苷酸序列与BVDV NADL株标准种毒的同源性为89.2%.结果表明,该分离毒株为猪源牛病毒性腹泻病毒.%One strain of virus was isolated from the infected swine in Fujian Province by cultivating it in Madin-Darby bovine kidney(MDBK) cells, then it was systematically identified by a series of methods. In result,the isolated virus caused typical cytopathogenic effect in the MDBK cells at 72 hours after infection.The isolated virus could be neutralized by the positive sera with bovine viral diarrhea virus at titer 1: 106.The isolate was sensitive to chloroform, ether, trypsin, acid and temperature, and could be inactivated at 56 ℃ for 30 minutes. A DNA fragment of 325 bp was amplified by PCR from the MDBK cells infected with the isolated virus. The identity of this gene sequence to the isolated virus and NADL strain from GenBank was 89.2 %. It was concluded from the results that the isolated virus from the infected swine is a bovine viral diarrhea virus.

  4. Isolation of a cDNA clone for the type I regulatory subunit of bovine cAMP-dependent protein kinase.

    OpenAIRE

    Lee, D C; Carmichael, D F; Krebs, E G; McKnight, G S

    1983-01-01

    A cDNA clone for the type I regulatory subunit (RI) of cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) was isolated from bovine testis by a differential screening method. mRNA coding for RI was enriched 50- to 100-fold by polysome immunoadsorption chromatography with affinity-purified rabbit anti-RI and protein A-Sepharose. Poly(A)+ RNA from these polysomes was utilized to construct a cDNA library in pBR322, and this library was screened for hybridization to 32P-la...

  5. Investigation of biofilm formation in clinical isolates of Staphylococcus aureus.

    Science.gov (United States)

    Cassat, James E; Smeltzer, Mark S; Lee, Chia Y

    2014-01-01

    Invasive methicillin-resistant Staphylococcus aureus (MRSA) infections are often characterized by recalcitrance to antimicrobial therapy, which is a function not only of widespread antimicrobial resistance among clinical isolates, but also the capacity to form biofilms. Biofilms consist of ordered populations of bacterial colonies encased in a polysaccharide and/or proteinaceous matrix. This unique physiologic adaptation limits penetration of antimicrobial molecules and innate immune effectors to the infectious focus, increasing the likelihood of treatment failure and progression to chronic infection. Investigation of mechanisms of biofilm formation and dispersal, as well as the physiologic adaptations to the biofilm lifestyle, is therefore critical to developing new therapies to combat MRSA infections. In this chapter, we describe two in vitro methods for the investigation of staphylococcal biofilm formation, a microtiter plate-based assay of biofilm formation under static conditions and a flow cell-based assay of biofilm formation under fluid shear. We also detail an in vivo murine model of catheter-associated biofilm formation that is amenable to imaging and microbiologic analyses. Special consideration is given to the conditions necessary to support biofilm formation by clinical isolates of S. aureus. PMID:24085698

  6. Comparative genomics and the role of lateral gene transfer in the evolution of bovine adapted Streptococcus agalactiae

    OpenAIRE

    Richards, Vincent P; Lang, Ping; Bitar, Paulina D. Pavinski; Lefebure, Tristan; Schukken, Ynte H.; Zadoks, Ruth N.; Michael J Stanhope

    2011-01-01

    In addition to causing severe invasive infections in humans, Streptococcus agalactiae, or group B Streptococcus (GBS), is also a major cause of bovine mastitis. Here we provide the first genome sequence for S. agalactiae isolated from a cow diagnosed with clinical mastitis (strain FSL S3-026). Comparison to eight S. agalactiae genomes obtained from human disease isolates revealed 183 genes specific to the bovine strain. Subsequent polymerase chain reaction (PCR) screening for the presence/abs...

  7. Comparison of type I and type II bovine viral diarrhea virus infection in swine.

    OpenAIRE

    Walz, P H; Baker, J. C.; Mullaney, T P; Kaneene, J B; Maes, R K

    1999-01-01

    Some isolates of type II bovine viral diarrhea virus (BVDV) are capable of causing severe clinical disease in cattle. Bovine viral diarrhea virus infection has been reported in pigs, but the ability of these more virulent isolates of type II BVDV to induce severe clinical disease in pigs is unknown. It was our objective to compare clinical, virologic, and pathologic findings between type I and type II BVDV infection in pigs. Noninfected control and BVDV-infected 2-month-old pigs were used. A ...

  8. Development of a set of multiplex standard polymerase chain reaction assays for the identification of infectious agents from aborted bovine clinical samples.

    Science.gov (United States)

    Tramuta, Clara; Lacerenza, Daniela; Zoppi, Simona; Goria, Mariella; Dondo, Alessandro; Ferroglio, Ezio; Nebbia, Patrizia; Rosati, Sergio

    2011-07-01

    The current study describes the development of a set of 5 multiplex polymerase chain reaction (mPCR) assays for the simultaneous detection of abortive infection agents in bovine fetal tissues, including Brucella spp., Leptospira spp., and Campylobacter fetus (mPCR1); Hammondia heydorni, Neospora caninum, and Toxoplasma gondii (mPCR2); Coxiella burnetii and Chlamydophila psittaci (mPCR3); Mycoplasma bovis, Mycoplasma bovigenitalium, and Ureaplasma diversum (mPCR4); and Bovine viral diarrhea virus (BVDV) and Bovine herpesvirus-1 (BoHV-1; mPCR5). The protocol was tested on different tissue samples collected from 50 aborted bovine fetuses, and it showed that out of the 50 fetuses, 7 (14%, mPCR2) were PCR-positive for N. caninum, 4 (8%, mPCR5) were PCR-positive for BVDV, and 2 (4%, mPCR4) were PCR-positive for U. diversum. The results obtained by using each multiplex PCR were 100% concordant with those obtained by using the respective PCR assays targeting single genes on the same specimens. Moreover, all multiplex PCR assays on clinical samples were compared with reference methods, obtaining a perfect accordance in all samples and confirming the validity of the set of multiplex PCR assays. The proposed set of multiplex PCR assays is, therefore, suitable for the simultaneous detection of the main infectious agents responsible for bovine abortion.

  9. Udder quarter risk factors associated with prevalence of bovine clinical mastitis

    OpenAIRE

    Nakov Dimitar; Trajcev Metodija

    2012-01-01

    A cross sectional study was carried out to estimate prevalence of clinical mastitis on udder quarters level and to determinate the quarter risk factors associated with the development of clinical mastitis during lactation.The individual risk factors included assessments of parity, season of year when case of clinical mastitis was occurred, conformation characteristics of udder quarters and teats and distance from front and rear teat end to the floor. Cows with clinical mastitis were detected ...

  10. Biochemical and molecular characterization of Treponema phagedenis-like spirochetes isolated from a bovine digital dermatitis lesion

    Science.gov (United States)

    Bovine papillomatous digital dermatitis (DD) is the leading cause of lameness in dairy cattle and represents a serious welfare and economic burden. Found primarily in high production dairy cattle worldwide, DD is characterized by the development of an often painful red, raw ulcerative or papillomato...

  11. Comparison of the Immune Response Between a Pair of NCP and CP Bovine Viral Diarrhea Virus (BVDV) Type 1 Isolates

    Science.gov (United States)

    Aim: Bovine viral diarrhea virus (BVDV) is a major pathogen of cattle causing severe respiratory and reproductive disease. BVDV vaccines remain an important part of the control strategy. Previous work has described higher antibody responses in animals infected with a noncytopathic (NCP) BVDV when ...

  12. The Neuroprotective Potential of Rho-Kinase Inhibition in Promoting Cell Survival and Reducing Reactive Gliosis in Response to Hypoxia in Isolated Bovine Retina

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    Aizhan Alt

    2013-07-01

    Full Text Available Aims: To investigate the outcomes of Rho-kinase inhibition in the electrophysiological ex vivo model of the isolated perfused vertebrate retina under hypoxia. Methods: Bovine retinas were perfused with an oxygen saturated nutrient solution with or without the Rho-kinase inhibitor H-1152P. The retinas were stimulated repeatedly until stable amplitudes were reached and the electroretinogram was recorded at five minute intervals. Hypoxia was induced for 15, 30, and 45 minutes, after which the oxygen saturation was restored. The extent of the cell damage and glial reactivity was determined by Ethidium homodimer-1 staining, immunohistochemistry, and Western blot. Results: Hypoxia caused a time-dependent reduction of the b-wave amplitudes, which could not be prevented by the H-1152P. Although the Rho-kinase inhibitor maintained higher b-wave amplitudes, these effects did not reach statistical significance. Hypoxia also resulted in an increase in cell damage and the activation of the glial cells in the untreated retinas whereas the administration of H-1152P significantly reduced the extent of these events. Conclusion: H-1152P exerted a neuroprotective effect against necrosis on the isolated bovine retina under hypoxia together with a reduction in glial cell reactivity. However, the inhibitor could not prevent the hypoxia induced retinal dysfunction possibly due to the interference with synaptic modulation.

  13. Occurrence of enterotoxin genes and macrorestriction analysis of Staphylococcus aureus isolated from bovine mastitis and bulk-tank milk samples in Italy. An epidemiological study

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    Roberto Rosmini

    2010-01-01

    Full Text Available The goal of the study was to genotypically compare S. aureus isolates from mastitis milk and raw milk to identify therelation between strains and to assess the enterotoxigenicity of the isolates. Eighty-three Staphylococcus aureus isolatesrecovered from cows and bulk tank milk of five farms in northern Italy were compared genotypically. The genes for theenterotoxins A, D, G and I, but not for B, C, E and H and the toxic shock syndrome toxin 1 (TSST-1, were detected byPCR amplification. Macrorestriction analysis with the restrictions enzyme SmaI revealed 14 pulsed-field gel electrophoresispatterns. These were in part different from each other only in a few fragments and thus displayed a closeclonal relation. The results of the present investigation showed that identical or closely related clones seemed to beresponsible for the cases of bovine mastitis in the farms investigated and partly responsible for contamination of bulktank milk.

  14. Antiviral Activity of Bacillus sp. Isolated from the Marine Sponge Petromica citrina against Bovine Viral Diarrhea Virus, a Surrogate Model of the Hepatitis C Virus

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    Clarice Weis Arns

    2013-04-01

    Full Text Available The Hepatitis C virus causes chronic infections in humans, which can develop to liver cirrhosis and hepatocellular carcinoma. The Bovine viral diarrhea virus is used as a surrogate model for antiviral assays for the HCV. From marine invertebrates and microorganisms isolated from them, extracts were prepared for assessment of their possible antiviral activity. Of the 128 tested, 2 were considered active and 1 was considered promising. The best result was obtained from the extracts produced from the Bacillus sp. isolated from the sponge Petromica citrina. The extracts 555 (500 µg/mL, SI>18 and 584 (150 µg/mL, SI 27 showed a percentage of protection of 98% against BVDV, and the extract 616, 90% of protection. All of them showed activity during the viral adsorption. Thus, various substances are active on these studied organisms and may lead to the development of drugs which ensure an alternative therapy for the treatment of hepatitis C.

  15. Visual pathways involvement in clinically isolated syndrome in children

    Institute of Scientific and Technical Information of China (English)

    Vladislav; Voitenkov; Natalia; Skripchenko; Andrey; Klimkin

    2015-01-01

    AIM: To investigate extent and nature of visual pathways involvement in children with clinically isolated syndrome(CIS).METHODS: Forty-seven patients(age 11-17y) with CIS, which later proved to be multiple sclerosis(MS)onset, and 30 controls underwent visual evoked potentials(VEP) investigation within 12 d from the appearance of the first signs of disease. Latency and amplitude of P100 peak were compared with normative data and between groups.RESULTS: In 58% patients, including those without signs of retrobulbar neuritis, significant slowing of conduction along the central visual pathways(P100latency lengthening) is seen. P100 amplitudes drop(signs of axonal damage) are registered less frequently(29% cases).CONCLUSION: The results indicate that visual pathways are often affected in the MS onset; mostly demyelination signs are seen. Despite MRI significance for MS diagnostic, VEPs proved to be still effective in early diagnosis of MS in children.

  16. Bovine rhinitis viruses are common in U.S. cattle with bovine respiratory disease.

    Science.gov (United States)

    Hause, Ben M; Collin, Emily A; Anderson, Joe; Hesse, Richard A; Anderson, Gary

    2015-01-01

    Bovine rhinitis viruses (BRV) are established etiological agents of bovine respiratory disease complex however little research into their epidemiology and ecology has been published for several decades. In the U.S., only bovine rhinitis A virus 1 (BRAV1) has been identified while bovine rhinitis A virus 2 (BRAV2) and bovine rhinitis B virus (BRBV) were previously only identified in England and Japan, respectively. Metagenomic sequencing of a nasal swab from a bovine respiratory disease (BRD) diagnostic submission from Kansas identified contigs with approximately 90% nucleotide similarity to BRAV2 and BRBV. A combination of de novo and templated assemblies using reference genomes yielded near complete BRAV2 and BRBV genomes. The near complete genome of bovine rhinitis A virus 1 (BRAV1) was also determined from a historical isolate to enable further molecular epidemiological studies. A 5'-nuclease reverse transcription PCR assay targeting the 3D polymerase gene was designed and used to screen 204 archived BRD clinical specimens. Thirteen (6.4%) were positive. Metagenomic sequencing of six positive samples identified mixed BRAV1/BRAV2, BRAV1/BRBV and BRAV2/BRBV infections for five samples. One sample showed infection only with BRAV1. Seroprevalence studies using a cell culture adapted BRBV found immunofluorescence assay-reactive antibodies were common in the herds analyzed. Altogether, these results demonstrate that BRV infections are common in cattle with respiratory disease and that BRAV1, BRAV2 and BRBV co-circulate in U.S. cattle and have high similarity to viruses isolated more than 30 years ago from diverse locations.

  17. Resistance to β-lactam antibiotic may influence nanH gene expression in Trueperella pyogenes isolated from bovine endometritis.

    Science.gov (United States)

    Zhang, De-Xian; Tian, Kai; Han, Li-Mei; Wang, Qiu-Xia; Liu, Yao-Chuan; Tian, Chun-Lian; Liu, Ming-Chun

    2014-01-01

    Virulence could be modulated by many instinctive and environmental factors including oxygen, osmolarity and antimicrobial agents. This study aimed to investigate the correlation between drug resistance and the nanH expression in Trueperella pyogenes (T. pyogenes). Minimum inhibitory concentrations (MICs) of 6 β-lactam antimicrobial agents (penicillin G, amoxicillin, oxacillin, cefazolin, ceftiofur, and ampicillin) against T. pyogenes were tested by standard broth dilution method according to the protocols of the Clinical and Laboratory Standards Institute (CLSI), and real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) was selected to investigate the mRNA expression levels of the nanH in T. pyogenes. All the isolates were resistant to atleast 2 of antimicrobial agents, and multidrug resistance (resistance to atleast 3 antimicrobials) was observed in 84.38% (27/32) of isolates. The mRNA expression levels of the nanH were significantly higher in comparison with that in ATCC19411, as the resistance profile enlarged, the nanH mRNA expression levels decreased in T. pyogenes. These results indicated that β-lactam antibiotic resistance in T. pyogenes may alter the expression of the nanH. PMID:24803199

  18. Genomes of Two Clinical Isolates of Mycobacterium tuberculosis from Odisha, India

    Science.gov (United States)

    Majid, Mohammad; Qureshi, Asifa; Yerra, Priyadarshini; Kumar, Ashutosh; Kumar, Mandala Kiran; Tiruvayipati, Suma; Baddam, Ramani; Shaik, Sabiha; Srikantam, Aparna

    2014-01-01

    We report whole-genome sequences of two clinical isolates of Mycobacterium tuberculosis isolated from patients in Odisha, India. The sequence analysis revealed that these isolates are of an ancestral type and might represent some of the “pristine” isolates in India that have not admixed with other lineages. PMID:24652981

  19. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

    OpenAIRE

    A.H. Maghsood; Sissons, J.; Rezaian, M.; Nolder, D; Warhurst, D.; Khan, Naveed Ahmed

    2005-01-01

    The majority of the keratitis-causing Acanthamoeba isolates are genotype T4. In an attempt to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence of this genotype, Acanthamoeba genotypes were determined for 13 keratitis isolates and 12 environmental isolates from Iran. Among 13 clinical isolates, eight (61.5 %) belonged to T4, two (15.3 %) belonged to T3 and three (23 %) belonged to the T2 genotype. In contrast, the majorit...

  20. Intramammary administration of platelet concentrate as an unconventional therapy in bovine mastitis: first clinical application.

    Science.gov (United States)

    Lange-Consiglio, A; Spelta, C; Garlappi, R; Luini, M; Cremonesi, F

    2014-10-01

    Bovine udder infections induce a variety of changes in gene expression of different growth factors that may suggest their possible role in glandular tissue protection or repair processes. Growth factors and also chemokines and cytokines may act synergistically to increase the infiltration of neutrophils and macrophages to promote angiogenesis, fibroplasia, matrix deposition, and, ultimately, re-epithelialization. Considering the vast applications, typically in human medicine, of platelet concentrate (PC) and its ease of preparation, the aim of our study was to evaluate an alternative therapy to stimulate the regeneration of glandular tissue, administering a concentration in excess of the growth factors contained in the PC. In each one of the 3 farms examined in the trial, PC was prepared from donor cows in good health, free from infections, and with no records of medications administered during the previous 2 mo. The platelet produced in one farm was used only for treating the cows of the same farm in a heterologous way. A total of 229 mastitic quarters were divided in 3 groups: antibiotic group (treated with intramammary antibiotic), antibiotic and PC group (treated intramammarily with antibiotics in association with PC), and PC group (treated with intramammary PC alone). The diagnosis of mastitis was based on somatic cell count and bacteriological evaluation of the milk from the affected quarter. Platelet concentrate, alone or in association with antibiotic, was used for 3 consecutive days as an unconventional therapy in bovine acute and chronic mastitis. Our data show that the associated action of antibiotic and PC performed significantly better than the antibiotic alone, either for the recovery of the affected mammary quarters or for somatic cell count reduction. In the same way, the association antibiotic plus PC showed significantly fewer relapses compared with the antibiotic alone, either for acute or chronic mastitis. The treatment with only PC did not show

  1. Testing the biocompatibility of a glutathione-containing intra-ocular irrigation solution by using an isolated perfused bovine retina organ culture model - an alternative to animal testing.

    Science.gov (United States)

    Januschowski, Kai; Zhour, Ahmad; Lee, Albert; Maddani, Ramin; Mueller, Sebastien; Spitzer, Martin S; Schnichels, Sven; Schultheiss, Maximilian; Doycheva, Deshka; Bartz-Schmidt, Karl-Ulrich; Szurman, Peter

    2012-03-01

    The effects of a glutathione-containing intra-ocular irrigation solution, BSS Plus©, on retinal function and on the survival of ganglion cells in whole-mount retinal explants were studied. Evidence is provided that the perfused ex vivo bovine retina can serve as an alternative to in vivo animal testing. Isolated bovine retinas were prepared and perfused with an oxygen-saturated standard irrigation solution, and an electroretinogram was recorded to assess retinal function. After stable b-waves were detected, the isolated retinas were perfused with BSS Plus for 45 minutes. To investigate the effects of BSS Plus on photoreceptor function, 1mM aspartate was added to the irrigation solution in order to obtain a-waves, and the ERG trace was monitored for 75 minutes. For histological analysis, isolated whole retinal mounts were stored for 24 hours at 4°C, in the dark. The percentages of cell death in the retinal ganglion cell layer and in the outer and inner nuclear layers were estimated by using an ethidium homodimer-1 stain and the TUNEL assay. General swelling of the retina was examined with high-resolution optical coherence tomography. During perfusion with BSS Plus, no significant changes in a-wave and b-wave amplitudes were recorded. Retinas stored for 24 hours in BSS Plus showed a statistically significant smaller percentage (52.6%, standard deviation [SD] = 16.1%) of cell death in the retinal ganglion cell layer compared to the control group (69.6%, SD = 3.9, p = 0.0031). BSS Plus did not seem to affect short-term retinal function, and had a beneficial effect on the survival of retinal ganglion cells. This method for analysing the isolated perfused retina represents a valuable alternative for testing substances for their retinal biocompatibility and toxicity. PMID:22558975

  2. Failed detection of Bovine viral diarrhea virus 2 subgenotype a (BVDV-2a) by direct fluorescent antibody test on tissue samples due to reduced reactivity of field isolates to raw anti-BVDV antibody.

    Science.gov (United States)

    Yan, Lifang; Pace, Lanny W; Baughman, Brittany; Wilson, Floyd D; Zhang, Shuping; Zhang, Michael Z

    2016-03-01

    Bovine viral diarrhea virus 1 (BVDV-1) is associated with mild or subclinical infections, whereas BVDV-2 is frequently implicated in outbreaks of severe thrombocytopenia and acute fatal disease. In the present study, the carcass of a beef breed cow and tissue samples of a beef calf were received for laboratory diagnosis. Both animals exhibited severe clinical signs compatible with thrombocytopenia or hemorrhagic syndrome. Direct fluorescent antibody test (DFAT) failed to detect BVDV antigen in the tissue specimens of both cases. However, immunohistochemistry (IHC) revealed the presence of BVDV antigen in oral and esophageal mucosa and Peyer patches of the beef breed cow. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) detected BVDV-2 in selected tissues of both animals. Subsequently, BVDV was isolated from both cases and subjected to genetic and serologic characterizations. Mutations in the 5'-untranslated genomic region (5'-UTR) primer and probe binding sites and the E2 gene were associated with reduced efficiency of an established real-time RT-PCR assay and amino acid alterations in the E2 glycoprotein, respectively. Both viral isolates were classified by real-time RT-PCR and phylogenetic analysis as BVDV-2 subgenotype a. Unlike BVDV reference strains Singer and 125c, the isolates cross-reacted with anti-BVDV-1 and anti-BVDV-2 reference sera, indicating antigenic variations in field isolates. The isolates also showed reduced reactivity to porcine anti-BVDV antiserum (the raw serum used to produce BVDV DFA conjugate). In summary, data from the present investigation indicated that genetic and antigenic variations affected the performance of detection assays, especially DFAT, highlighting the need for regular evaluation and modification of BVDV tests. PMID:26965235

  3. Molecular Detection and Sensitivity to Antibiotics and Bacteriocins of Pathogens Isolated from Bovine Mastitis in Family Dairy Herds of Central Mexico

    OpenAIRE

    Ma. Fabiola León-Galván; Barboza-Corona, José E.; A. Arianna Lechuga-Arana; Mauricio Valencia-Posadas; Aguayo, Daniel D.; Carlos Cedillo-Pelaez; Erika A. Martínez-Ortega; Gutierrez-Chavez, Abner J.

    2015-01-01

    Thirty-two farms (n = 535 cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages f...

  4. Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis - doi: 10.4025/actascibiolsci.v32i4.6276 Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis - doi: 10.4025/actascibiolsci.v32i4.6276

    Directory of Open Access Journals (Sweden)

    Ulisses de Pádua Pereira

    2010-11-01

    Full Text Available Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS, isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus by phenotypic tests were tested with the APISTAPH kit (Biomériux-France, for precise identification of species. Phenotypic tests revealed 338 strains (98.25% as S. aureus, three strains (0.86% as Staphylococcus hyicus, and three microorganisms (0.86% as Staphylococcus intermedius. PCR demonstrated that 338 (98.25% strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83% was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS, isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose

  5. 牛传染性鼻气管炎病毒的分离鉴定及gD基因序列分析%Isolation, Identification and Sequence Analysis of gD Gene of Infectious Bovine Rhinotracheitis Virus

    Institute of Scientific and Technical Information of China (English)

    杨有武

    2013-01-01

    [目的]从疑似牛传染性鼻气管炎病毒感染的牛鼻腔棉拭子中分离出1株病毒,对其进行鉴定和序列分析.[方法]从疑似牛传染性鼻气管炎病毒感染的牛鼻腔棉拭子中分离到1株病毒,将其命名为NM株.通过PCR、MDBK细胞病变观察、中和试验、动物回归试验、gD基因序列分析对其进行鉴定.[结果]确定该分离株为牛传染性鼻气管炎病毒强毒株.NM株病毒能使MDBK细胞产生牛传染性鼻气管炎病毒典型性细胞病变.Reed-Muench法测定NM株病毒的TCID50为10-6.0/ml.NM株病毒与IBRV阳性血清发生中和反应,而与IBRV阴性血清不发生中和反应.NM株病毒能使8月龄IBRV抗体阴性牛致病,表现出牛传染性鼻气管炎的典型临床症状.gD基因序列鉴定结果表明NM株病毒与IBRV K22株的同源性最高.[结论]该研究可为IBR诊断试剂与疫苗的研制奠定基础.%[Objective] The research aimed to isolate a strain of virus from nasal swab samples of cattle which might be infected by infectious bovine rhinotracheitis(IBR) and make the identification and sequencing analysis. [Method] A strain of virus was isolated from nasal swab samples of cattle which might be infected by infectious bovine rhinotracheitis( IBR) and it was named as NM strain. The isolated strain was i-dentified by PCR, MDBK cell lesion observation, neutralization test, animal regression test and gD gene sequence analysis. [ Result] The isolated strain was confirmed as virulent strain of infectious bovine rhinotracheitis virus. NM strain made MDBK cell to produce typical cell lesion of infectious bovine rhinotracheitis virus. TCID50 of NM strain determined by Reed-Muench method was 10-6.0/ml. NM strain could make neutralization test with IBRV positive serum, but it didn't react with IBRV negative serum. IBRV antibody-free calves with the age of 8 months showed typical clinical symptoms of IBR after inoculation of NM strain. The sequencing analysis results showed

  6. Intestinal carriage of Campylobacter jejuni and Campylobacter coli among cattle from South-western Norway and comparative genotyping of bovine and human isolates by amplified-fragment length polymorphism

    Directory of Open Access Journals (Sweden)

    Vardund T

    2006-06-01

    Full Text Available Abstract In a survey conducted in 1999–2001, the carriage of thermotolerant Campylobacters in cattle was investigated, and the genetic diversity of C. jejuni within one herd was examined and compared with human isolates. C. jejuni, C. coli and other thermotolerant Campylobacter spp. were isolated from intestinal contents from 26%, 3% and 2% of 804 cattle, respectively. The carriage rate was higher in calves (46% than in adults (29%. Twenty-nine C. jejuni isolates from one herd and 31 human isolates from the study area were genotyped with amplified-fragment length polymorphism (AFLP. Eighty-three % of the bovine isolates fell into three distinct clusters with 95–100% similarity, persistent in the herd for 5–10 months. Among human isolates, 58% showed >90% similarity with bovine isolates. The results show that cattle are a significant and stable reservoir for C. jejuni in the study area. Transmission between individuals within the herd may be sufficient to maintain a steady C. jejuni population independent of environmental influx. The results of this study have provided new information on C. jejuni and C. coli transmission, and also on the carriage in cattle, genotypes stability and similarity between bovine and human isolates.

  7. Characterization of bovine viral diarrhea virus isolates resistant to a novel antiviral compound obtained from persistently infected calves

    Science.gov (United States)

    The objective of this research was to characterize isolates resistant to a novel antiviral compound (DB772) isolated from persistently infected (PI) calves treated with the compound. Viral isolates were obtained from four Angus-cross beef calves (A,B,C,D) persistently infected with BVDV type 1 or 2 ...

  8. Genotypic and phenotypic detection of capsular polysaccharides in Staphylococcus aureus isolated from bovine intramammary infections in Argentina

    Directory of Open Access Journals (Sweden)

    C. Camussone

    2012-09-01

    Full Text Available Staphylococcus aureus (n=157 isolated from intramammary infections in Argentine dairy areas were evaluated for presence of cap5 and cap8 loci. Isolates carrying cap5 and cap8 were serotyped using specific antisera. Sixty four percent of the isolates were genotyped as cap5 or cap8 and 50% of them expressed CP5 or 8.

  9. Phylogenetic relationships among Streptococcus agalactiae isolated from piscine, dolphin, bovine and human sources: a dolphin and piscine lineage associated with a fish epidemic in Kuwait is also associated with human...

    Science.gov (United States)

    Group B Streptococcus agalactiae (GBS) causes of infections in multiple animals. This study examined the genetic relatedness of piscine, dolphin, and human GBS isolates and bovine GBS reference strains from different geographical regions using serological and multilocus sequence typing (MLST) techni...

  10. Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

    DEFF Research Database (Denmark)

    Christensen, Henrik; Angen, Øystein; Olsen, John E.;

    2004-01-01

    Strains deviating in key phenotypic characters, mainly isolated from cases of bovine pneumonia in five European countries, were genotyped in order to examine their genotypic relationship with Pasteurella multocida. Twenty-two strains of Pasteurella avium biovar 2, including variants in indole, xy...

  11. Aislamiento y caracterización del virus de la diarrea viral bovina en un ternero con síndrome purpúrico Characterization of bovine viral diarrhea virus isolated from cattle with hemorrhagic syndrome

    Directory of Open Access Journals (Sweden)

    A Gollán

    2006-01-01

    Full Text Available El Virus de la Diarrea Viral Bovina (VDVB produce en el ganado bovino numerosas patologías que van desde pérdidas reproductivas hasta afecciones de poca significación clínica en el aparato digestivo. Se ha reportado variabilidad entre las cepas de VDVB, que se manifiesta por la existencia de los biotipos citopatogénicos (CP y no citopatogénicos (NCP, y los tipos virales I y II. El presente trabajo describe los hallazgos patológicos y virológicos en un ternero que clínicamente exhibió trombocitopenia y diarrea. La cepa viral aislada (334/3 fue caracterizada molecularmente por secuenciación de la región 5' no-codificante (5' RNC. Los análisis realizados revelaron un 90-98% de homología con las cepas de referencia tipo I, no encontrándose cambios asociados a las cepas VDVB tipo II.The Bovine Viral Diarrhea (BVDV virus causes numerous pathologies that range from reproductive losses to infections of little clinical significance in the bovine digestive tract. Variation have been reported among the strains of the BVDV, which are classified into two biotypes; cytopathogenic (CP and non-cytopathogenic (NCP, and the viral types I and II. This work describes the pathological findings in a calf with diarrhea and severe thrombocytopenia. The strain isolated (334/3 was molecularly characterized by sequencing of the 5’non-coding region (5’ NCR. These analyses revealed 90-98% homologies with reference strains type I strains and the changes associated with BVDV type II, were not found.

  12. Bovine Herpesvirus 4 infections and bovine mastitis

    NARCIS (Netherlands)

    Wellenberg, Gerardus Johannus

    2002-01-01

    Mastitis is an often occurring disease in dairy cattle with an enormous economic impact for milk producers worldwide. Despite intensive research, which is historically based on the detection of bacterial udder pathogens, still around 20-35% of clinical cases of bovine mastitis have an unknown aetiol

  13. New antimicrobial drug resistance and epidemiological typing patterns of Staphylococci from clinical isolates and raw meats.

    Science.gov (United States)

    Lee, Do Kyung; Hwang, Jae Ung; Baek, Eun Hye; Lee, Kang Oh; Kim, Kyung Jae; Ha, Nam Joo

    2008-08-01

    The antimicrobial susceptibilities of Staphylococcus isolated from clinical isolates and raw meats were tested for six different antimicrobial agents that are in widespread clinical use in Korea and four new antimicrobials, linezolid, quinupristin/dalfopristin, daptomycin, and tigecycline. And this study analyzed the mecA genes and genetic patterns of MRSA by performing epidemiological studies using the PCR method. 46%, 51%, and 79% of clinical isolates were identified as MRSA in 1998, 1999, and 2005, respectively, and the mecA gene was detected in 82% of these isolates. Of the 133 staphylococci isolated from raw meats, 18% of the isolates were found to be resistant to methicillin, but none of these isolates showed the presence of the mecA gene. New antimicrobials, which have rarely or not yet been used in Korean hospitals, showed high activity against all staphylococcal isolates including methicillin-resistant isolates. The randomly amplified polymorphic DNA (RAPD) patterns of MRSA isolates differed significantly between clinical isolates and raw meat isolates. PMID:18787791

  14. Factors affecting cure when treating bovine clinical mastitis with cephalosporin-based intramammary preparations

    OpenAIRE

    Bradley, A. J.; Green, M. J.

    2009-01-01

    Data were collated for an independent scientific analysis from 2 international, multicenter studies that had compared the efficacy of 3 different cephalosporin-containing intramammary preparations in the treatment of clinical mastitis in dairy cattle [cefalexin (first generation) in combination with kanamycin; cefquinome (fourth generation); and cefoperazone (third generation)]. Quarters were assessed using standard bacteriological techniques before treatment and at 16 and 25 d posttreatment....

  15. Experimental infection of colostrum-deprived calves with bovine viral diarrhea virus type 1a isolated from free-ranging white-tailed deer (Odocoileus virginianus)

    Science.gov (United States)

    Raizman, Eran A.; Pogranichniy, Roman M.; Levy, Michel; Negron, Maria; Van Alstine, William

    2011-01-01

    The objective of this study was to experimentally infect calves with bovine viral diarrhea virus (BVDV) isolated from free-ranging white-tailed deer. Twelve colostrum-deprived male Holstein calves were used. Eight were inoculated intranasally with a BVDV type 1a isolated from free-ranging white-tailed deer, and the other four were inoculated with the cell culture medium only and served as a control group. Whole blood, saliva, and nasal and rectal secretions were collected on days 0, 3, 7, 10, 14, 17, and 21 after inoculation for virus isolation and real-time reverse-transcriptase polymerase chain reaction (RT-PCR). On days 14 and 21, 4 calves in the infected group and 2 in the control group were euthanized; multiple tissue samples were collected for histopathologic study. Histopathologic changes included thymic atrophy and lymphoid depletion of the Peyer’s patches in all 8 infected calves. The RT-PCR gave positive results with the buffy coat of all 8 infected calves, the nasal samples of 7, and the saliva samples of 2. Virus neutralization testing of the serum gave positive results for 4 of the 8 infected calves, and enzyme-linked immunosorbent assay of the serum gave positive results for 3. All of the samples from the control calves yielded negative results. PMID:21461198

  16. Isolation and characterization of a specific endogenous Na/sup +/, K/sup +/-ATPase inhibitor from bovine adrenal

    Energy Technology Data Exchange (ETDEWEB)

    Tamura, M.; Lam, T.T.; Inagami, T.

    1988-06-14

    In order to identify a specific endogenous Na/sup +/,K/sup +/-ATPase inhibitor which could possibly be related to salt-dependent hypertension, the authors looked for substances in the methanol extract of bovine whole adrenal which show all of the following properties: (i) inhibitory activity for Na/sup +/,K/sup +/-ATPase; (ii) competitive displacing activity against (/sup 3/H)ouabain binding to the enzyme; (iii) inhibitory activity for /sup 86/Rb uptake into intact human erythrocytes; and (iv) cross-reactivity with sheep anti-digoxin-specific antibody. After stepwise fractionation of the methanol extract of bovine adrenal glands by chromatography on a C/sub 18/ open column, a 0-15% acetonitrile fraction was fractionated by high-performance liquid chromatography on a Zorbax octadecylsilane column. One of the most active fractions in 0-15% acetonitrile was found to exhibit all of the four types of the activities. It was soluble in water and was distinct from various substances which have been known to inhibit Na/sup +/,K/sup +/-ATPase. These results strongly suggest that this water-soluble nonpeptidic Na/sup +/,K/sup +/-ATPase inhibitor may be a specific endogenous regulator for the ATPase.

  17. Isolation and proliferation of umbilical cord tissue derived mesenchymal stem cells for clinical applications.

    Science.gov (United States)

    Van Pham, Phuc; Truong, Nhat Chau; Le, Phuong Thi-Bich; Tran, Tung Dang-Xuan; Vu, Ngoc Bich; Bui, Khanh Hong-Thien; Phan, Ngoc Kim

    2016-06-01

    Umbilical cord (UC) is a rich source of rapidly proliferating mesenchymal stem cells (MSCs) that are easily cultured on a large-scale. Clinical applications of UC-MSCs include graft-versus-host disease, and diabetes mellitus types 1 and 2. UC-MSCs should be isolated and proliferated according to good manufacturing practice (GMP) with animal component-free medium, quality assurance, and quality control for their use in clinical applications. This study developed a GMP standard protocol for UC-MSC isolation and culture. UC blood and UC were collected from the same donors. Blood vasculature was removed from UC. UC blood was used as a source of activated platelet rich plasma (aPRP). Small fragments (1-2 mm(2)) of UC membrane and Wharton's jelly were cut and cultured in DMEM/F12 medium containing 1 % antibiotic-antimycotic, aPRP (2.5, 5, 7.5 and 10 %) at 37 °C in 5 % CO2. The MSC properties of UC-MSCs at passage 5 such as osteoblast, chondroblast and adipocyte differentiation, and markers including CD13, CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105, and HLA-DR were confirmed. UC-MSCs also were analyzed for karyotype, expression of tumorigenesis related genes, cell cycle, doubling time as well as in vivo tumor formation in NOD/SCID mice. Control cells consisted of UC-MSCs cultured in DMEM/F12 plus 1 % antibiotic-antimycotic, and 10 % fetal bovine serum (FBS). All UC-MSC (n = 30) samples were successfully cultured in medium containing 7.5 and 10 % aPRP, 92 % of samples grew in 5.0 % aPRP, 86 % of samples in 2.5 % aPRP, and 72 % grew in 10 % FBS. UC-MSCs in these four groups exhibited similar marker profiles. Moreover, the proliferation rates in medium with PRP, especially 7.5 and 10 %, were significantly quicker compared with 2.5 and 5 % aPRP or 10 % FBS. These cells maintained a normal karyotype for 15 sub-cultures, and differentiated into osteoblasts, chondroblasts, and adipocytes. The analysis of pluripotent cell markers showed UC-MSCs maintained

  18. The genetic diversity of clinical isolates of Malassezia pachydermatis from dogs and cats.

    Science.gov (United States)

    Aizawa, T; Kano, R; Nakamura, Y; Watanabe, S; Hasegawa, A

    2001-08-01

    Molecular investigation of 110 clinical isolates of non-lipid-dependent Malassezia pachydermatis from dogs and cats was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD analysis indicated that the clinical isolates of M. pachydermatis constituted four distinct genetic types (A, B, C and D). Moreover, the results from CHS2 gene analysis completely agreed with those from the RAPD analyses. The clinical isolates of M. pachydermatis were obtained from normal external ears, lesions of atopic dermatitis, flea allergic dermatitis, otitis externa, pyoderma and seborrheic dermatitidis in dogs and cats. Type A consisted of 93 clinical isolates as well as the ex-neotype strain of M. pachydermatis. The isolates of type A M. pachydermatis originated from lesions of all kinds of diseases. They were predominant on dog and cat skin. The other types, B, C, and D were isolated mainly from otitis externa. PMID:11556762

  19. A search for RNA insertions and NS3 gene duplication in the genome of cytopathic isolates of bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    V.L. Quadros

    2006-07-01

    Full Text Available Calves born persistently infected with non-cytopathic bovine viral diarrhea virus (ncpBVDV frequently develop a fatal gastroenteric illness called mucosal disease. Both the original virus (ncpBVDV and an antigenically identical but cytopathic virus (cpBVDV can be isolated from animals affected by mucosal disease. Cytopathic BVDVs originate from their ncp counterparts by diverse genetic mechanisms, all leading to the expression of the non-structural polypeptide NS3 as a discrete protein. In contrast, ncpBVDVs express only the large precursor polypeptide, NS2-3, which contains the NS3 sequence within its carboxy-terminal half. We report here the investigation of the mechanism leading to NS3 expression in 41 cpBVDV isolates. An RT-PCR strategy was employed to detect RNA insertions within the NS2-3 gene and/or duplication of the NS3 gene, two common mechanisms of NS3 expression. RT-PCR amplification revealed insertions in the NS2-3 gene of three cp isolates, with the inserts being similar in size to that present in the cpBVDV NADL strain. Sequencing of one such insert revealed a 296-nucleotide sequence with a central core of 270 nucleotides coding for an amino acid sequence highly homologous (98% to the NADL insert, a sequence corresponding to part of the cellular J-Domain gene. One cpBVDV isolate contained a duplication of the NS3 gene downstream from the original locus. In contrast, no detectable NS2-3 insertions or NS3 gene duplications were observed in the genome of 37 cp isolates. These results demonstrate that processing of NS2-3 without bulk mRNA insertions or NS3 gene duplications seems to be a frequent mechanism leading to NS3 expression and BVDV cytopathology.

  20. Influence of saliva and mucin on the adhesion of Candida oral clinical isolates

    OpenAIRE

    Seabra, C.; Botelho, C.M.; Oliveira, Ana C. N.; Henriques, Mariana

    2015-01-01

    Objectives: This research work intends to clarify the role of artificial saliva, in particularly the role of mucin, a salivary protein, on the surface properties and adhesion ability of Candida spp. oral clinical isolates to abiotic surfaces. Methods: Four oral clinical isolates of Candida spp. were used: two Candida albicans strains (AC; AM) and two Candida parapsilosis strains (AD; AM2). The strains were isolated from patients using oral prosthesis. The microorganisms were cultured in the a...

  1. Mutant prevention concentrations of daptomycin for Enterococcus faecium clinical isolates.

    Science.gov (United States)

    Sinel, Clara; Jaussaud, Clara; Auzou, Michel; Giard, Jean-Christophe; Cattoir, Vincent

    2016-10-01

    Owing to the emergence of vancomycin-resistant Enterococcus faecium, treatment of enterococcal infections has become challenging. Although spontaneous in vitro resistance frequencies are low, the emergence of resistance is increasingly reported during daptomycin therapy. The mutant selection window (MSW), comprised between the minimum inhibitory concentration (MIC) and the mutant prevention concentration (MPC), corresponds to the concentration range within which resistant mutants may be selected. Since no data are available for enterococci, the aim of this study was to determine MPCs and MSWs for 12 representative E. faecium clinical isolates. MICs and MPCs were determined by broth microdilution and agar dilution methods, respectively. A basic MSW-derived pharmacodynamic analysis was also performed using mean maximum plasma concentration (Cmax) values obtained with dosages from 4 to 12 mg/kg. MICs and MPCs of daptomycin ranged from 0.5 to 4 mg/L and from 2 to 32 mg/L, respectively, with no correlation between them. The wideness of MSWs ranged from 2× to 32× MIC. Mean plasma Cmax values of daptomycin were calculated from 55 to 174.5 mg/L when using a dosage from 4 to 12 mg/kg. All Cmax values were above the MPCs whatever the dosage. Taking into account the protein binding of daptomycin (ca. 90%), the unbound fraction Cmax was just within the MSW in 67-92% of strains at recommended dosages (4-6 mg/kg) and was above the MPC for the majority of strains only with the highest dosage (12 mg/kg). This study shows that free daptomycin Cmax values usually fell into MSWs when using lower dosages (<10 mg/kg).

  2. Oral Contraceptives and Multiple Sclerosis/Clinically Isolated Syndrome Susceptibility.

    Directory of Open Access Journals (Sweden)

    Kerstin Hellwig

    Full Text Available The incidence of multiple sclerosis (MS is rising in women.To determine whether the use of combined oral contraceptives (COCs are associated with MS risk and whether this varies by progestin content.We conducted a nested case-control study of females ages 14-48 years with incident MS or clinically isolated syndrome (CIS 2008-2011 from the membership of Kaiser Permanente Southern California. Controls were matched on age, race/ethnicity and membership characteristics. COC use up to ten years prior to symptom onset was obtained from the complete electronic health record.We identified 400 women with incident MS/CIS and 3904 matched controls. Forty- percent of cases and 32% of controls had used COCs prior to symptom onset. The use of COCs was associated with a slightly increased risk of MS/CIS (adjusted OR = 1.52, 95%CI = 1.21-1.91; p<0.001. This risk did not vary by duration of COC use. The association varied by progestin content being more pronounced for levenorgestrol (adjusted OR = 1.75, 95%CI = 1.29-2.37; p<0.001 than norethindrone (adjusted OR = 1.57, 95%CI = 1.16-2.12; p = 0.003 and absent for the newest progestin, drospirenone (p = 0.95.Our findings should be interpreted cautiously. While the use of some combination oral contraceptives may contribute to the rising incidence of MS in women, an unmeasured confounder associated with the modern woman's lifestyle is a more likely explanation for this weak association.

  3. Excretion of (3H)prednisolone in clinically normal and experimentally infected bovine udders

    Energy Technology Data Exchange (ETDEWEB)

    Geleta, J.N.; Shimoda, W.; Mercer, H.D.

    1984-08-01

    The excretion rate of (3H)prednisolone from clinically normal and experimentally infected udders of 10 lactating cows was studied. Each quarter of 6 cows was injected with a single dose of (3H)prednisolone mixed with non-radioactive prednisolone equivalent to 10 mg in 10 ml of peanut oil base. Each of the remaining 4 cows was given 40 mg of nonradioactive prednisolone and (3H)prednisolone in 60% ethanol IV. Control and postadministration samples of blood, milk, and urine were examined for radioactivity. The effects of (3H)prednisolone were evaluated in the same cows, first in clinically normal udders, then 2 weeks later in udders experimentally infected with Streptococcus agalactiae. Absorption and elimination of prednisolone were the same before and after induced infection. Within 3 hours after intramammary injection, 95% of the labeled prednisolone was absorbed systemically, less than 5% of this dose was recovered in milk, and 29% was excreted in urine. After IV injection of (3H)prednisolone, less than 0.2% of the total radioactivity was recovered in milk and less than 46% was excreted in urine. Clinical mastitis induced by S agalactiae was moderate. Circulating blood leukocytes and somatic cells in the milk of normal cows remained essentially unchanged. The leukocyte response to induced infection was rapid in blood and milk. Large numbers of leukocytes were noticed in the milk and a severe leukopenia occurred. Prednisolone treatment did not alter the number of somatic cells in milk or reduce the inflammatory response of experimentally infected cows.

  4. Excretion of [3H]prednisolone in clinically normal and experimentally infected bovine udders

    International Nuclear Information System (INIS)

    The excretion rate of [3H]prednisolone from clinically normal and experimentally infected udders of 10 lactating cows was studied. Each quarter of 6 cows was injected with a single dose of [3H]prednisolone mixed with non-radioactive prednisolone equivalent to 10 mg in 10 ml of peanut oil base. Each of the remaining 4 cows was given 40 mg of nonradioactive prednisolone and [3H]prednisolone in 60% ethanol IV. Control and postadministration samples of blood, milk, and urine were examined for radioactivity. The effects of [3H]prednisolone were evaluated in the same cows, first in clinically normal udders, then 2 weeks later in udders experimentally infected with Streptococcus agalactiae. Absorption and elimination of prednisolone were the same before and after induced infection. Within 3 hours after intramammary injection, 95% of the labeled prednisolone was absorbed systemically, less than 5% of this dose was recovered in milk, and 29% was excreted in urine. After IV injection of [3H]prednisolone, less than 0.2% of the total radioactivity was recovered in milk and less than 46% was excreted in urine. Clinical mastitis induced by S agalactiae was moderate. Circulating blood leukocytes and somatic cells in the milk of normal cows remained essentially unchanged. The leukocyte response to induced infection was rapid in blood and milk. Large numbers of leukocytes were noticed in the milk and a severe leukopenia occurred. Prednisolone treatment did not alter the number of somatic cells in milk or reduce the inflammatory response of experimentally infected cows

  5. Genotypic and phenotypic comparison of Escherichia coli from uterine infections with different outcomes: clinical metritis in the cow and pyometra in the bitch.

    Science.gov (United States)

    Henriques, Sofia; Silva, Elisabete; Lemsaddek, Abdelhak; Lopes-da-Costa, Luís; Mateus, Luisa

    2014-05-14

    Escherichia coli uterine infection originates different clinical outcomes in the canine and bovine species. Here, E. coli strains isolated from bovine clinical metritis and canine pyometra cases were analyzed by PFGE, screened for 33 virulence factor (VF) genes and for phylogenetic grouping. Bovine and canine E. coli isolates presented a low degree of genetic similarity. Canine E. coli strains belonged to phylogenetic group B2 and presented a high number of VF genes, whereas bovine E. coli strains belonged to phylogenetic groups B1 and A and had a low number of VF genes. In conclusion, E. coli strains isolated from cow clinical metritis had a low potential of virulence. In contrast, bitch pyometra E. coli isolates had a high virulence potential, which might be relevant in the pathogenesis of pyometra. These differences between canine and bovine E. coli isolates may partially explain the different outcomes of the uterine infection in the two species. PMID:24598134

  6. Bovine viral diarrhea virus in postweaned calves in a feedlot after vaccination and from fatal respiratory cases: isolation and differentiation of MLV BVDV and field strains

    Science.gov (United States)

    Viral infections are important etiologies in BRD cases. Calves at stocker/feedlot entry usually receive modified live viral (MLV) vaccines containing bovine herpesvirus-1 (BoHV-1), parainfluenza-3 virus (PI3V), bovine viral diarrhea viruses (BVDV), and bovine respiratory syncytial virus (BRSV). In...

  7. ETIOLOGY OF THE BOVINE CLINICAL MASTITIS IN GOIÂNIA ETIOLOGIA DA MASTITE CLÍNICA BOVINA NA BACIA LEITEIRA DE GOIÂNIA

    Directory of Open Access Journals (Sweden)

    Paulo César Moreira

    2007-09-01

    Full Text Available

    There is an estimate that the mastitis in dairy herds causes production losses between 5 and 35%, equivalent from 85 to 500 million dollars per year. 231 milk samples from 231 cows on different stages of lactation, with clinic mastitis, from 35 farms of Goiânia, were analyzed in order to map the pathogens implicated in these process and to discover the microorganisms with major prevalence. All the samples had positive growth. The principal agents were Staphylococcus coagulase positive (32.90%, Streptococcus sp. (22.07%, Pseudomonas sp. (12.12%, Enterobacter sp. (10.38%, Corynebacterium sp. (8.65%, Escherichia coli (8.22%, Bacillus sp. (8.22%, Proteus sp. (6.49%, Klebsiella sp. (4.32% and Staphylococcus coagulase negative (3.46%. The Nocardia genus was isolated in 0.86% of the cases.

    KEY-WORDS: Bovine mastitis; etiology; isolated microorganisms.

    Estima-se que a presença de mastite bovina em rebanhos produtores provoque perdas de produção entre 5 e 35%, o que equivale de 85 a 500 milhões de dólares ao ano. Com o objetivo de mapear os patógenos envolvidos nesse processo e evidenciar os microrganismos com maior freqüência foram examinadas amostras de leite de 231 vacas, em diferentes estágios de lactação, que apresentaram sinais de mastite clínica e eram pertencentes a 35 propriedades rurais da bacia leiteira de Goiânia. Todas as amostras tiveram crescimento bacteriano positivo. Os principais agentes isolados foram o Staphylococcus coagulase positiva (32,90%, Streptococcus sp. (22,07%, Pseudomonas sp. (12,12%, Enterobacter sp. (10,38%, Corynebacterium sp. (8,65%, Escherichia coli (8,22%, Bacillus sp. (8,22%,

  8. Distribution of Phylogenetic Groups among Extraintestinal Pathogenic Escherichia coli isolates from Bovine in Heilongjiang%黑龙江奶牛肠外致病大肠埃希菌种系分类群研究

    Institute of Scientific and Technical Information of China (English)

    佟春玉; 曹宏伟; 朱战波; 刘振华; 崔玉东

    2013-01-01

    研究黑龙江省奶牛主要肠外致病(奶牛乳房炎、子宫内膜炎)大肠埃希菌的种系分类群。用PCR扩增具有种系分类群标记的chuA、yjaA基因和TSPE4.C2 DNA片段的方法,对黑龙江省84株来源于奶牛乳房炎和41株来源于奶牛子宫内膜炎的大肠埃希菌进行种系进化群分类。奶牛乳房炎所检测菌株中A进化种系群占11.90%。B1种系进化群占88.10%。奶牛子宫内膜炎所检测菌株中A种系进化群占14.63%,B1种系进化亚群占85.37%。未检测到属于B2和D群的肠外强毒力株。大肠杆菌分离株接种小鼠剖检可见明显病变。通过遗传进化分析,引起黑龙江奶牛肠外致病大肠埃希菌主要属于B1种系进化群,为奶牛乳房炎和子宫内膜炎疫苗的研制提供理论依据。%Researched distribution of phylogenetic groups among extraintestinal pathogenic Escherichia coli isolates from bovine mastitis,Bovine endometritis in Heilongjiang. 84 E.coli isdates from bovine mastitis and 41 E.coli isdates from Bovine endometritis in Heilongjiang were PCR amplified for genetic markers of chuA,yjaA and TSPE4.C2. The result showed that 11.90% E.coli isolates belonged to group A and 88.10%to group B1 from bovine mastitis,14.63%E.coli isolates belonged to group A and 85.37%to group B1 from Bovine endometritis,no-one belonged to B2 and D groups. Obviously pathological changes were observed in internal organs of dissected experiment mouse. Extraintestinal pathogenic E.coli isolates from bovine for the predominant Phylogenetic group was group B1,this research provided a theoretical basis to prepare bovine mastitis and endometritis vaccine.

  9. PER, CTX-M, TEM and SHV Beta-lactamases in Clinical Isolates of Klebsiella pneumoniae Isolated from Tehran, Iran

    Directory of Open Access Journals (Sweden)

    Leila Nasehi

    2010-06-01

    Full Text Available Objective(sDifferent types of extended spectrum beta-lactamases (ESBLs are encountered in the clinical settings worldwide. There are a few studies regarding the prevalence of ESBL genes among Klebsiella pneumoniae isolates at Tehran especially those of blaPER and blaCTX. The aim of this study was to determine the prevalence of blaSHV, blaTEM ,blaPER and blaCTX genes among clinical K. pneumoniae of different hospitals in Tehran.Materials and MethodsTwo hundred isolates of K. pneumoniae were received from different clinical specimens. The susceptibility of the isolates to 10 different antibiotics was examined by disk diffusion test. The MICs for ceftazidime were also determined using micro-broth dilution assay. Isolates showing MIC 4 μg/ml for ceftazidime were screened for ESBL production by phenotypic confirmatory test (PCT and subjected to PCR for studied genes. Variation among four amplified genes was evaluated using PCR-RFLP.ResultsBy disk diffusion test, resistance to ceftazidime and cefotaxime were 34.7% and 33.5% respectively. However, all strains were susceptible to imipenem. Eighty isolates showed MICs≥ 4 μg/ml for ceftazidime of which 77 (96% were positive for ESBL in PCT. The prevalence of blaSHV, blaCTX-M, blaTEM and blaPER among these isolates were 26%, 24.5%, 18% and 7.5%, respectively. No variation was detected in the genes by PCR-RFLP.ConclusionAs far as we know this is the first report of the blaPER-1 in K. pneumoniae in Iran. The blaCTX-M was the second most common gene detected among the ESBL positive isolates of K. pneumoniae. For rapid identification of ESBL producing isolates it was recommended that clinical laboratories adopt simple test based on CLSI recommendation for confirming ESBL production in enterobacterial species.

  10. Evaluation of phenotypic and genotypic methods for epidemiological typing of Staphylococcus aureus isolates from bovine mastitis in Denmark

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Wegener, H. C.; Rosdahl, V. T.

    1995-01-01

    The value of five different typing methods (antibiogram typing, biotyping, phage typing, plasmid profiling and restriction fragment length polymorphism of the gene encoding 16S and 23S ribosomal RNA (ribotyping)), in discriminating 105 Staphylococcus aureus strains from bovine milk samples obtained...... (75%) of the 20 antibiotic resistant strains. Biotyping assigned the strains to 14 different types, with the most common type accounting for 25.7% of the strains. Ninety eight (93.3%) strains could be typed by phages, assigning them to 19 different phage types. The predominant phage type accounted...... for 31.4% of the strains. Eight different plasmid profiles was observed among 24 (23%) strains harbouring plasmids. Ribotyping yielded 30 different types, with the most common accounting far 29.5% of the strains. The single most discriminatory typing method was ribotyping (0.863) followed by biotyping (0...

  11. PER, CTX-M, TEM and SHV Beta-lactamases in Clinical Isolates of Klebsiella pneumoniae Isolated from Tehran, Iran

    OpenAIRE

    Leila Nasehi; Fereshteh Shahcheraghi; Vajihe Sadat Nikbin; Shoeib Nematzadeh

    2010-01-01

    Objective(s)Different types of extended spectrum beta-lactamases (ESBLs) are encountered in the clinical settings worldwide. There are a few studies regarding the prevalence of ESBL genes among Klebsiella pneumoniae isolates at Tehran especially those of blaPER and blaCTX. The aim of this study was to determine the prevalence of blaSHV, blaTEM ,blaPER and blaCTX genes among clinical K. pneumoniae of different hospitals in Tehran.Materials and MethodsTwo hundred isolates of K. pneumoniae were ...

  12. Genetic diversity of clinical isolates of Bacillus cereus using multilocus sequence typing

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    Pruckler James M

    2008-11-01

    Full Text Available Abstract Background Bacillus cereus is most commonly associated with foodborne illness (diarrheal and emetic but is also an opportunistic pathogen that can cause severe and fatal infections. Several multilocus sequence typing (MLST schemes have recently been developed to genotype B. cereus and analysis has suggested a clonal or weakly clonal population structure for B. cereus and its close relatives B. anthracis and B. thuringiensis. In this study we used MLST to determine if B. cereus isolates associated with illnesses of varying severity (e.g., severe, systemic vs. gastrointestinal (GI illness were clonal or formed clonal complexes. Results A retrospective analysis of 55 clinical B. cereus isolates submitted to the Centers for Disease Control and Prevention between 1954 and 2004 was conducted. Clinical isolates from severe infections (n = 27, gastrointestinal (GI illness (n = 18, and associated isolates from food (n = 10 were selected for analysis using MLST. The 55 isolates were diverse and comprised 38 sequence types (ST in two distinct clades. Of the 27 isolates associated with serious illness, 13 clustered in clade 1 while 14 were in clade 2. Isolates associated with GI illness were also found throughout clades 1 and 2, while no isolates in this study belonged to clade 3. All the isolates from this study belonging to the clade 1/cereus III lineage were associated with severe disease while isolates belonging to clade1/cereus II contained isolates primarily associated with severe disease and emetic illness. Only three STs were observed more than once for epidemiologically distinct isolates. Conclusion STs of clinical B. cereus isolates were phylogenetically diverse and distributed among two of three previously described clades. Greater numbers of strains will need to be analyzed to confirm if specific lineages or clonal complexes are more likely to contain clinical isolates or be associated with specific illness, similar to B. anthracis and

  13. Study on Clinical Bovine Dermatophilosis and its Potential Risk Factors in North Western Ethiopia

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    Meseret Admassu and Sefinew Alemu

    2011-02-01

    Full Text Available A cross-sectional study of dermatophilosis was undertaken from October 2008 to March 2009 on 3456 cattle (3181 indigenous zebu and 275 Holestien-zebu cross with the aim of determining prevalence and associated risk factors in urban and periurban areas of Bahir Dar, north western Ethiopia. Culturing of Dermatiphilus congolensis and Giemsa staining were the techniques used. Thirty six of 3456 examined animals (1.04% had clinical dermatophilosis. Prevalence was higher in cross bred (5.5% than in indigenous zebu (0.7% cattle, in male cattle (1.7% than in female (0.8% , in adults (1.2% than in young (0.8% age groups, in wet (1.6% than in dry season (0.5%, and in cattle infested with tick (2.7% than cattle with no tick infestation (0.4%. Statistically significant difference (p# 0.05 was observed in the prevalence between breeds of cattle, between age groups, between wet and dry seasons, and between cattle with and without tick infestation. Amblyoma variegatum was identified. The study indicated dermatophilosis is a potential determinant factor for the dairy development strategy started through cross breeding in the study area. Tick control especially on crossbred cattle is suggested to reduce the risk of dermatophilosis.

  14. Diversity and antifungal susceptibility of Norwegian Candida glabrata clinical isolates

    Science.gov (United States)

    Andersen, Kari-Mette; Kristoffersen, Anne Karin; Ingebretsen, André; Vikholt, Katharina Johnsen; Örtengren, Ulf Thore; Olsen, Ingar; Enersen, Morten; Gaustad, Peter

    2016-01-01

    Background Increasing numbers of immunocompromised patients have resulted in greater incidence of invasive fungal infections with high mortality. Candida albicans infections dominate, but during the last decade, Candida glabrata has become the second highest cause of candidemia in the United States and Northern Europe. Reliable and early diagnosis, together with appropriate choice of antifungal treatment, is needed to combat these challenging infections. Objectives To confirm the identity of 183 Candida glabrata isolates from different human body sites using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and VITEK®2, and to analyze isolate protein profiles and antifungal susceptibility. The minimum inhibitory concentration (MIC) of seven antifungal drugs was determined for the isolates to elucidate susceptibility. Design A total of 183 C. glabrata isolates obtained between 2002 and 2012 from Norwegian health-care units were analyzed. For species verification and differentiation, biochemical characterization (VITEK®2) and mass spectrometry (MALDI–TOF) were used. MIC determination for seven antifungal drugs was undertaken using E-tests®. Results Using VITEK®2, 92.9% of isolates were identified as C. glabrata, while all isolates (100%) were identified as C. glabrata using MALDI-TOF. Variation in protein spectra occurred for all identified C. glabrata isolates. The majority of isolates had low MICs to amphotericin B (≤1 mg/L for 99.5%) and anidulafungin (≤0.06 mg/L for 98.9%). For fluconazole, 18% of isolates had MICs >32 mg/L and 82% had MICs in the range ≥0.016 mg/L to ≤32 mg/L. Conclusions Protein profiles and antifungal susceptibility characteristics of the C. glabrata isolates were diverse. Clustering of protein profiles indicated that many azole resistant isolates were closely related. In most cases, isolates had highest susceptibility to amphotericin B and anidulafungin. The results confirmed previous observations of high

  15. Acute exposure to ergot alkaloids from endophyte-infected tall fescue does not alter absorptive or barrier function of the isolated bovine ruminal epithelium.

    Science.gov (United States)

    Foote, A P; Penner, G B; Walpole, M E; Klotz, J L; Brown, K R; Bush, L P; Harmon, D L

    2014-07-01

    Ergot alkaloids in endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum) have been shown to cause a reduction in blood flow to the rumen epithelium as well as a decrease in volatile fatty acids (VFA) absorption from the washed rumen of steers. Previous data also indicates that incubating an extract of endophyte-infected tall fescue seed causes an increase in the amount of VFA absorbed per unit of blood flow, which could result from an alteration in the absorptive or barrier function of the rumen epithelium. An experiment was conducted to determine the acute effects of an endophyte-infected tall fescue seed extract (EXT) on total, passive or facilitated acetate and butyrate flux across the isolated bovine rumen as well as the barrier function measured by inulin flux and tissue conductance (G t ). Flux of ergovaline across the rumen epithelium was also evaluated. Rumen tissue from the caudal dorsal sac of Holstein steers (n=6), fed a common diet, was collected and isolated shortly after slaughter and mounted between two halves of Ussing chambers. In vitro treatments included vehicle control (80% methanol, 0.5% of total volume), Low EXT (50 ng ergovaline/ml) and High EXT (250 ng ergovaline/ml). Results indicate that there is no effect of acute exposure to ergot alkaloids on total, passive or facilitated flux of acetate or butyrate across the isolate bovine rumen epithelium (P>0.51). Inulin flux (P=0.16) and G t (P>0.17) were not affected by EXT treatment, indicating no alteration in barrier function due to acute ergot alkaloid exposure. Ergovaline was detected in the serosal buffer of the High EXT treatment indicating that the flux rate is ~0.25 to 0.44 ng/cm2 per hour. Data indicate that specific pathways for VFA absorption and barrier function of the rumen epithelium are not affected by acute exposure to ergot alkaloids from tall fescue at the concentrations tested. Ergovaline has the potential to be absorbed from the rumen of cattle that

  16. Identification of Prototheca Zopfii from Bovine Mastitis

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    F Zaini

    2012-08-01

    Full Text Available Background: The aim of this study was identification of the epidemiology of Prototheca zopfii species from the milk samples of dairy cattle in Isfahan, central Iran.Methods: Milk samples were obtained from 230 dairy cattle, 130 with and 100 without mastitis, in Isfahan. The samples were cultured in Prototheca Isolation Medium (PIM and Sabouraud's dextrose agar. All P. zopfii isolates were identified by morphological and biochemical methods. Then, as a confirmatory test they were examined by genotype-specific PCR.Results: Four P. zopfii strains (3.07% were isolated from the 130 samples of dairy cattle with clinical mastitis and there was no isolation from totally 100 samples of healthy bovines without mastitis. Specific PCR product (about 946 bp was detected in four isolates.Conclusion: It seems that P. zopfii genotype II plays a key role in affecting bovine mastitis that confirmed other previous studies. Our study was the first, which identified the Prototheca species by traditional and molecular methods in Iran and Middle East as well.

  17. Evaluation of PCR assays for the detection of Campylobacter fetus in bovine preputial scrapings and the identification of subspecies in South African field isolates

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    T. Schmidt

    2010-05-01

    Full Text Available As a result of the high lability and slow growth of Campylobacter fetus subspecies, the laboratory diagnosis of bovine genital campylobacteriosis has always been difficult. This is especially true under South African conditions, where farms are far apart, laboratories are only present in major centres and there are high ambient temperatures. In order to overcome the shortcomings associated with traditional diagnostic methods, the implementation of a molecular assay was sought. This work describes how a previously published PCR assay (MG3F / MG4R primers was adapted, optimised and applied in the diagnostic laboratory to test preputial samples directly for the presence of Campylobacter fetus. Field evaluation of the assay revealed an analytical sensitivity and specificity of 85.7 % and 99 %, respectively. Subsequent genotyping and phenotyping of a diverse collection of South African field isolates revealed that South Africa has an unexpected and previously unreported high incidence of Campylobacter fetus subsp. venerealis biovar intermedius strains. These strains were not identified correctly by the subspecies-specific primer set evaluated. Until such time that cost-effective genotyping methods are available to diagnostic laboratories in South Africa, and other countries with these atypical Campylobacter fetus subsp. venerealis strains, the need for bacterial culture will persist. Identification to subspecies level of isolates at present remains dependent upon a single phenotypic criterion, namely tolerance to 1 % glycine.

  18. Occurrence of genes coding for MSCRAMM and biofilm-associated protein Bap in Staphylococcus spp. isolated from bovine subclinical mastitis and relationship with somatic cell counts.

    Science.gov (United States)

    Zuniga, Eveline; Melville, Priscilla A; Saidenberg, André B S; Laes, Marco A; Gonsales, Fernanda F; Salaberry, Sandra R S; Gregori, Fabio; Brandão, Paulo E; dos Santos, Franklin G B; Lincopan, Nilton E; Benites, Nilson R

    2015-12-01

    This study aimed to elucidate aspects of the epidemiology of bovine subclinical mastitis through the assessment of genes encoding MSCRAMM (microbial surface components recognizing adhesive matrix molecules - a group of adhesins) and protein Bap (implicated in biofilm formation), in coagulase-positive (CPS) and coagulase-negative (CNS) Staphylococcus isolated from subclinical mastitis. Milk samples were collected for microbiological exams, somatic cell count (SCC) and a survey of the genes coding for MSCRAMM (cna, eno, ebpS, fnbA, fnbB and fib) and biofilm-associated protein Bap (bap) in 106 Staphylococcus spp. isolates using PCR. The frequencies of occurrence of eno (82.1%), fnbA (72.6%), fib (71.7%) and bap (56.6%) were higher (P bap gene in CNS compared with CPS suggests that in these species biofilm formation is an important mechanism for the persistence of the infection. The medians of the SCCs in the samples where eno, fnbA, fib and bap genes were detected were higher compared with Staphylococcus without the assessed genes (P < 0.05) and negative samples (P < 0.01), which indicated that the presence of these MSCRAMM may be related to a higher intensity of the inflammatory process.

  19. Molecular Characterization and Computational Modelling of New Delhi Metallo-β-Lactamase-5 from an Escherichia coli Isolate (KOEC3) of Bovine Origin.

    Science.gov (United States)

    Purkait, D; Ahuja, A; Bhattacharjee, U; Singha, A; Rhetso, K; Dey, T K; Das, S; Sanjukta, R K; Puro, K; Shakuntala, I; Sen, A; Banerjee, A; Sharma, I; Bhatta, R S; Mawlong, M; Guha, C; Pradhan, N R; Ghatak, S

    2016-06-01

    Emergence of antimicrobial resistance mediated through New Delhi metallo-β-lactamases (NDMs) is a serious therapeutic challenge. Till date, 16 different NDMs have been described. In this study, we report the molecular and structural characteristics of NDM-5 isolated from an Escherichia coli isolate (KOEC3) of bovine origin. Using PCR amplification, cloning and sequencing of full blaNDM gene, we identified the NDM type as NDM-5. Cloning of full gene in E. coli DH5α and subsequent assessment of antibiotic susceptibility of the transformed cells indicated possible role of native promoter in expression blaNDM-5. Translated amino acid sequence had two substitutions (Val88Leu and Met154Leu) compared to NDM-1. Theoretically deduced isoelectric pH of NDM-5 was 5.88 and instability index was 36.99, indicating a stable protein. From the amino acids sequence, a 3D model of the protein was computed. Analysis of the protein structure elucidated zinc coordination and also revealed a large binding cleft and flexible nature of the protein, which might be the reason for broad substrate range. Docking experiments revealed plausible binding poses for five carbapenem drugs in the vicinity of metal ions. In conclusion, results provided possible explanation for wide range of antibiotics catalyzed by NDM-5 and likely interaction modes with five carbapenem drugs. PMID:27570310

  20. Occurrence of genes coding for MSCRAMM and biofilm-associated protein Bap in Staphylococcus spp. isolated from bovine subclinical mastitis and relationship with somatic cell counts.

    Science.gov (United States)

    Zuniga, Eveline; Melville, Priscilla A; Saidenberg, André B S; Laes, Marco A; Gonsales, Fernanda F; Salaberry, Sandra R S; Gregori, Fabio; Brandão, Paulo E; dos Santos, Franklin G B; Lincopan, Nilton E; Benites, Nilson R

    2015-12-01

    This study aimed to elucidate aspects of the epidemiology of bovine subclinical mastitis through the assessment of genes encoding MSCRAMM (microbial surface components recognizing adhesive matrix molecules - a group of adhesins) and protein Bap (implicated in biofilm formation), in coagulase-positive (CPS) and coagulase-negative (CNS) Staphylococcus isolated from subclinical mastitis. Milk samples were collected for microbiological exams, somatic cell count (SCC) and a survey of the genes coding for MSCRAMM (cna, eno, ebpS, fnbA, fnbB and fib) and biofilm-associated protein Bap (bap) in 106 Staphylococcus spp. isolates using PCR. The frequencies of occurrence of eno (82.1%), fnbA (72.6%), fib (71.7%) and bap (56.6%) were higher (P < 0.0001) compared with the other assessed genes (cna, ebpS and fnbB). The higher frequency of occurrence (P < 0.005) of the bap gene in CNS compared with CPS suggests that in these species biofilm formation is an important mechanism for the persistence of the infection. The medians of the SCCs in the samples where eno, fnbA, fib and bap genes were detected were higher compared with Staphylococcus without the assessed genes (P < 0.05) and negative samples (P < 0.01), which indicated that the presence of these MSCRAMM may be related to a higher intensity of the inflammatory process. PMID:26318876

  1. Isolation of chlorhexidine-resistant Pseudomonas aeruginosa from clinical lesions.

    OpenAIRE

    Nakahara, H; Kozukue, H

    1982-01-01

    The chlorhexidine resistance of 317 strains of Pseudomonas aeruginosa isolated from hospital patients was determined. The distribution pattern of their susceptibility to chlorhexidine clearly revealed two peaks, and the frequency of resistance to chlorhexidine was 84.2%.

  2. Isolation and Characterization of Tetracycline-Resistant Clinical Isolates of Helicobacter pylori

    OpenAIRE

    Kwon, Dong H.; Kim, J.J.; Lee, M.; Yamaoka, Y.; Kato, M; Osato, M. S.; El-Zaatari, F. A. K.; David Y Graham

    2000-01-01

    Tetracycline is an important component of combination therapies for Helicobacter pylori eradication. Twenty-nine tetracycline-resistant isolates requiring MICs ranging from 4 to 16 μg/ml were isolated from Korean (22 of 460) and Japanese (7 of 105) patients. Interestingly, all of the 29 tetracycline-resistant isolates exhibited cross-resistance to metronidazole, and the cross-resistance was transferred to tetracycline-sensitive H. pylori strains.

  3. Zinc Salts Inactivate Clinical Isolates of Herpes Simplex Virus In Vitro

    OpenAIRE

    Arens, Max; Travis, Sharon

    2000-01-01

    Using a standard plaque assay and clinical isolates of herpes simplex virus (HSV), we have tested the ability of zinc salts to inactivate HSV. Virus was treated by incubation at 37°C with zinc salts in morpholinepropanesulfonic acid-buffered culture medium and was then diluted and plated onto CV-1 cells for detection and quantitation of remaining infectious virus. Of 10 randomly chosen clinical isolates (five HSV type 1 [HSV-1] isolates and five HSV-2 isolates), seven were inactivated >98% by...

  4. Screening of medicinal plants for antibacterial activities on Staphylococcus aureus strains isolated from bovine mastitis Screening de plantas medicinais com atividade antimicrobiana contra cepas de Staphylococcus aureus isoladas de mastite bovina

    Directory of Open Access Journals (Sweden)

    Marisa A. N. Diaz

    2010-11-01

    Full Text Available Staphylococcus aureus is the main causative agent of bovine mastitis. The activity of several extracts from ten medicinal plants traditionally used in Brazil as antiseptic was investigated against fifteen strains of Staphylococcus aureus isolated from animals with mastitis manifestation by the disc diffusion method and broth microdilution assay. The interference of the extracts on cell in the form of adherent colonies was also evaluated. MIC values ranged from 0.5 mg/mL to 1.0 mg/mL and biofilm inhibitory concentration (BIC were between 0.25 mg/mL and 0.8 mg/mL. Results revealed the potential of extracts of Senna macranthera, Artemisia absinthium, Cymbopogon nardus and Baccharis dracunculifolia as antibacterial agents against S. aureus strains isolated from bovine mastitis and support the possible use of these phytotherapic agents in the clinical management of the disease.Staphylococcus aureus é o principal agente causador de mastite bovina. A atividade de diversos extratos de dez plantas medicinais tradicionalmente usadas no Brasil como anti-sépticas foi investigada contra quinze cepas de Staphylococcus aureus isoladas de animais com manifestação de mastite pelo método de difusão em ágar e ensaio de microdiluição. A interferência dos extratos na célula bacteriana em forma de colônias aderidas também foi avaliada. Os valores de MIC variaram de 0.5 mg/mL a 1.0 mg/mL e a concentração inibitória de biofilme (BIC variou de 0.25 mg/mL a 0.8 mg/mL. Os resultados revelaram o potencial dos extratos de Senna macranthera, Artemisia absinthium, Cymbopogon nardus e Baccharis dracunculifolia como agentes antibacterianos contra cepas de S. aureus isolados de mastite bovina e suportam o possível uso destas plantas no manejo clínico da doença.

  5. Antimicrobial Susceptibility and Clonality of Clinical Ureaplasma Isolates in the United States.

    Science.gov (United States)

    Fernández, Javier; Karau, Melissa J; Cunningham, Scott A; Greenwood-Quaintance, Kerryl E; Patel, Robin

    2016-08-01

    Ureaplasma urealyticum and Ureaplasma parvum are pathogens involved in urogenital tract and intrauterine infections and also in systemic diseases in newborns and immunosuppressed patients. There is limited information on the antimicrobial susceptibility and clonality of these species. In this study, we report the susceptibility of 250 contemporary isolates of Ureaplasma (202 U. parvum and 48 U. urealyticum isolates) recovered at Mayo Clinic, Rochester, MN. MICs of doxycycline, azithromycin, ciprofloxacin, tetracycline, erythromycin, and levofloxacin were determined by broth microdilution, with MICS of the last three interpreted according to CLSI guidelines. Levofloxacin resistance was found in 6.4% and 5.2% of U. parvum and U. urealyticum isolates, respectively, while 27.2% and 68.8% of isolates, respectively, showed ciprofloxacin MICs of ≥4 μg/ml. The resistance mechanism of levofloxacin-resistant isolates was due to mutations in parC, with the Ser83Leu substitution being most frequent, followed by Glu87Lys. No macrolide resistance was found among the 250 isolates studied; a single U. parvum isolate was tetracycline resistant. tet(M) was found in 10 U. parvum isolates, including the single tetracycline-resistant isolate, as well as in 9 isolates which had low tetracycline and doxycycline MICs. Multilocus sequence typing (MLST) performed on a selection of 46 isolates showed high diversity within the clinical Ureaplasma isolates studied, regardless of antimicrobial susceptibility. The present work extends previous knowledge regarding susceptibility to antimicrobial agents, resistance mechanisms, and clonality of Ureaplasma species in the United States. PMID:27246773

  6. Mixed Infections and Rifampin Heteroresistance among Mycobacterium tuberculosis Clinical Isolates.

    Science.gov (United States)

    Zheng, Chao; Li, Song; Luo, Zhongyue; Pi, Rui; Sun, Honghu; He, Qingxia; Tang, Ke; Luo, Mei; Li, Yuqing; Couvin, David; Rastogi, Nalin; Sun, Qun

    2015-07-01

    Mixed infections and heteroresistance of Mycobacterium tuberculosis contribute to the difficulty of diagnosis, treatment, and control of tuberculosis. However, there is still no proper solution for these issues. This study aimed to investigate the potential relationship between mixed infections and heteroresistance and to determine the high-risk groups related to these factors. A total of 499 resistant and susceptible isolates were subjected to spoligotyping and 24-locus variable-number tandem repeat methods to analyze their genotypic lineages and the occurrence of mixed infections. Two hundred ninety-two randomly selected isolates were sequenced on their rpoB gene to examine mutations and heteroresistance. The results showed that 12 patients had mixed infections, and the corresponding isolates belonged to Manu2 (n = 8), Beijing (n = 2), T (n = 1), and unknown (n = 1) lineages. Manu2 was found to be significantly associated with mixed infections (odds ratio, 47.72; confidence interval, 9.68 to 235.23; P < 0.01). Four isolates (1.37%) were confirmed to be heteroresistant, which was caused by mixed infections in three (75%) isolates; these belonged to Manu2. Additionally, 3.8% of the rifampin-resistant isolates showing no mutation in the rpoB gene were significantly associated with mixed infections (χ(2), 56.78; P < 0.01). This study revealed for the first time that Manu2 was the predominant group in the cases of mixed infections, and this might be the main reason for heteroresistance and a possible mechanism for isolates without any mutation in the rpoB gene to become rifampin resistant. Further studies should focus on this lineage to clarify its relevance to mixed infections.

  7. Phenotypic and molecular characterization of clinically isolated Escherichia coli

    Directory of Open Access Journals (Sweden)

    Vaishnavi Chetana

    2010-07-01

    Full Text Available All diarrheagenic Escherichia coli carry at least one virulence-related property. Stool samples from 244 patients having acute or persistent diarrhea received after the exclusion of routine enteropathogens were investigated. Purely or predominantly isolated E. coli (n = 100 were subjected to serotyping, of which only 25 were typable. They belonged to 14 different O-serogroups comprising 5 O153, 4 O102, 3 O25, 2 each of O130 and O169, and 1 each of O1, O8, O15, O37, O86, O101, O127, O143, and O160. The typable E. coli isolates along with 5 other untypable isolates were investigated for molecular markers, such as intimin (eae, enterohemolysin (EhlyA, a-hemolysin, heat-labile enterotoxins (LT, heat-stable enterotoxins (STa, verotoxins (VT1 and VT2, invasivity (ial, enteroaggregative E. coli (EAEC gene (EAGG, and enterotoxin (EAST. Two of the isolates (O153 and O86 were positive for enterohemolysin phenotypically and 5 for β-hemolysin both phenotypically and genotypically. Interestingly, 16.6% of the randomly isolated E. coli were O153, a serogroup common in cattle, and 10% belonged to EAEC pathotype of which two-thirds had the EAST gene, which is quite frequent in these strains. Additionally, there was one strain (O153 that was positive for EAST only. Between the two 0130:H6 strains isolated, one belonged to EAEC serogroup. None of the E. coli isolated were positive for verotoxins, eae, LT1, STa, and ial. Data obtained emphasize the need for additional research into the role of eae gene and other putative factors affecting the virulence of diarrheagenic E. coli in India.

  8. [Isolation and characterization of Escherichia coli O157 in bovine meat products and cattle in the province of Tucuman].

    Science.gov (United States)

    Jure, María A; Condorí, Marina S; Pérez Terrazzino, Gabriela; Catalán, Mariana G; López Campo, Alejandro; Zolezzi, Gisella; Chinen, Isabel; Rivas, Marta; Castillo, Marta

    2015-01-01

    Escherichia coli O157 is an emergent pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. Meat products constitute an important transmission source of this microorganism. The aims of this study were to characterize E. coli O157 isolated from cattle and meat products collected from abattoirs and retail stores, to establish the clonal relatedness among regional isolates and to compare them with those in the national database. Between 2004 and 2013, 169 minced meat, 35 sausage and 216 carcass samples were analyzed. Thirteen E. coli O157 isolates were identified; 6 of which were O157:H7 and characterized as stx2c(vh-a)/eae/ehxA (n = 5) and stx2/eae/ehxA (n = 1). The 7 remaining isolates were non-toxigenic E. coli strains, and serotyped as O157:NT (n = 4), O157:NM (n = 1), O157:ND (n = 1) and O157:H16 (n = 1). The strains yielded different XbaI-PFGE patterns. Compared to the E. coli O157 isolates in the National Database, none of these patterns have been previously detected in strains of different origin in Argentina.

  9. Isolation of bovine coronavirus (BCoV in monolayers of HmLu-1 cells Isolamento de coronavírus bovino (BCoV em monocamada de cultura de células da linhagem HmLu-1

    Directory of Open Access Journals (Sweden)

    José Antonio Jerez

    2005-09-01

    Full Text Available Isolation of BCoV was performed in monolayers of HmLu-1 cells, using 20 fecal samples from clinical cases of diarrhea in calves. Samples were positive for BCoV by means of hemagglutination / hemagglutination inhibition (HA/HI. Up to the fifth serial passage, 13 of these isolates presented syncytial cytopathic effect, similar to Kakegawa standard strain. When isolates were submitted to seroneutralization with gammaglobulin anti-bovine coronavirus, 8 of them were considered to be positive, once cytopathic effect was neutralized. After titration and seroneutralization in microplates, only three of them were confirmed as positive. The lineage HmLu-1 showed higher permissivity to BCoV isolation, but the low intensity of viral replication demonstrated that new methodologies should be developed for this purpose and then submitted to confirmation of isolation by means of seroneutralization.A partir de 20 amostras fecais de bezerros, com quadro clínico de diarréia, positivas para BCoV por hemaglutinação/inibição da hemaglutinação (HA/HI, procedeu-se o isolamento viral em monocamadas de células da linhagem HmLu-1. Até a quinta passagem seriada, 13 apresentaram efeito citopático do tipo sincicial, semelhante à amostra padrão Kakegawa de BCoV. Ao serem submetidas a uma reação de soroneutralização com gamaglobulina anti-coronavírus bovino, 8 delas foram consideradas positivas, uma vez que o efeito citopático foi neutralizado. Ao serem tituladas e submetidas à reação de soroneutralização em microplacas, apenas 3 delas puderam ser confirmadas como positivas. As células da linhagem HmLu-1 mostraram-se permissivas para o isolamento de BCoV, todavia a baixa intensidade na replicação viral demonstrou ser necessário o desenvolvimento de novas metodologias para se poder alcançar esse intuito e a confirmação do isolamento por soroneutralização.

  10. Beyond semantics: a clinical and theoretical study of isolation.

    Science.gov (United States)

    Killingmo, B

    1990-01-01

    Theories of Freud, Piaget, Fonagy and Kristeva have been drawn on to argue that the concept of isolation should not be restricted to a delimited defence mechanism associated with obsessional/compulsive neurosis, as is the usual psychoanalytic conception. From a semiological point of view isolation may be seen as a disturbance of the relationship between signifier and the signified embracing affective communication ranging from the most primitive bodily expression via sound and intonation to a semantic level where affects are expressed through the content of words and ideas. As affects isolated on a pre-verbal level are deprived of expression on a semantic-symbolic level, they are not likely to be modified by content of interpretations alone. They can only be fully activated and brought into therapeutic dialogue through the quality of the speech sound of the analyst.

  11. Genome analysis of environmental and clinical P. aeruginosa isolates from sequence type-1146.

    Directory of Open Access Journals (Sweden)

    David Sánchez

    Full Text Available The genomes of Pseudomonas aeruginosa isolates of the new sequence type ST-1146, three environmental (P37, P47 and P49 and one clinical (SD9 isolates, with differences in their antibiotic susceptibility profiles have been sequenced and analysed. The genomes were mapped against P. aeruginosa PAO1-UW and UCBPP-PA14. The allelic profiles showed that the highest number of differences were in "Related to phage, transposon or plasmid" and "Secreted factors" categories. The clinical isolate showed a number of exclusive alleles greater than that for the environmental isolates. The phage Pf1 region in isolate SD9 accumulated the highest number of nucleotide substitutions. The ORF analysis of the four genomes assembled de novo indicated that the number of isolate-specific genes was higher in isolate SD9 (132 genes than in isolates P37 (24 genes, P47 (16 genes and P49 (21 genes. CRISPR elements were found in all isolates and SD9 showed differences in the spacer region. Genes related to bacteriophages F116 and H66 were found only in isolate SD9. Genome comparisons indicated that the isolates of ST-1146 are close related, and most genes implicated in pathogenicity are highly conserved, suggesting a genetic potential for infectivity in the environmental isolates similar to the clinical one. Phage-related genes are responsible of the main differences among the genomes of ST-1146 isolates. The role of bacteriophages has to be considered in the adaptation processes of isolates to the host and in microevolution studies.

  12. Caracterização preliminar de amostras do vírus da Diarréia Viral Bovina (BVDV isoladas no Brasil Preliminary characterization of brazilian isolates of bovine viral diarrhea virus (BVDV

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    Sônia A. Botton

    1998-04-01

    ção contra o vírus.This article reports the preliminary characterization of 19 Brazilian bovine viral diarrhea virus (BVDV isolates, regarding the biological, antigenic and molecular properties. Eleven viruses were isolated from bovine fetuses, six were obtained from blood of animals from herds with reproductive problems, and two were isolated from clinical cases of gastroenteric disease. The clinical cases affected young animals and were characterized by diarrhea, oronasal and digestive erosions and ulceration, and occasional digestive bleeding and vulvar petechial hemorrhage. Sixteen isolates (84.2%, including those obtained from fetuses and clinical cases, were of the non-cytopathic (ncp biotype. Replication of three isolates (15.8% in tissue culture was characterized by appearance of cellular vacuolation and progressive destruction of the monolayers. Analysis of these isolates after cloning revealed a mixed population of cytopathic (cp and non-cytopathic viruses. Analysis of viral polypeptides by SDS-PAGE followed by "Western immunoblot" revealed the production of the non-structural protein NS3/p80 in cells infected with the cp viruses. In contrast, generation of NS3/p80 was not observed in cells infected with the ncp isolates, which only expressed the precursor polypeptide NS23/p125. Analysis of reactivity with a panel of 15 monoclonal antibodies (MAbs revealed a marked antigenic variability among the isolates, mainly in the envelope glycoprotein E2/gp53. Although one MAb to this glycoprotein recognized 18 isolates (94.7%, the other nine E2/gp53 MAbs recognized zero to 57.9% of the isolates. The marked antigenic diversity observed among the brazilian BVDV isolates may have important implications on diagnosis and immunization strategies.

  13. Proteomic Analysis of Bovine Nucleolus

    Institute of Scientific and Technical Information of China (English)

    Amrutlal K.Patel; Doug Olson; Suresh K. Tikoo

    2010-01-01

    Nucleolus is the most prominent subnuclear structure, which performs a wide variety of functions in the eu-karyotic cellular processes. In order to understand the structural and functional role of the nucleoli in bovine cells,we analyzed the proteomie composition of the bovine nueleoli. The nucleoli were isolated from Madin Darby bo-vine kidney cells and subjected to proteomie analysis by LC-MS/MS after fractionation by SDS-PAGE and strongcation exchange chromatography. Analysis of the data using the Mascot database search and the GPM databasesearch identified 311 proteins in the bovine nucleoli, which contained 22 proteins previously not identified in theproteomic analysis of human nucleoli. Analysis of the identified proteins using the GoMiner software suggestedthat the bovine nueleoli contained proteins involved in ribosomal biogenesis, cell cycle control, transcriptional,translational and post-translational regulation, transport, and structural organization.

  14. Genome Sequence of Vibrio cholerae G4222, a South African Clinical Isolate

    OpenAIRE

    le Roux, Wouter J.; Chan, Wai Yin; De Maayer, Pieter; Venter, Stephanus N.

    2013-01-01

    Vibrio cholerae, a Gram-negative pathogen autochthonous to the aquatic environment, is the causative agent of cholera. Here, we report the complete genome sequence of V. cholerae G4222, a clinical isolate from South Africa.

  15. 我国部分地区奶牛乳房炎源大肠杆菌生物学特性及耐药性分析%Characterization and Antibiotic Resistance of Ninety-five Strains of Escherichia coli Isolated from Bovine with Mastitis in Some Regions of China

    Institute of Scientific and Technical Information of China (English)

    徐继英; 刘俊林; 李先波; 霍生东; 杨志强

    2012-01-01

    调查奶牛乳房炎源大肠杆菌的某些生物学特性及其耐药状况,以提高药物疗效,减少牛乳中药物的残留.本研究从国内7个省、市、自治区部分地区患乳房炎奶牛的乳样中分离纯化与鉴定出95株大肠杆菌(Escherichia coli),并对大肠杆菌分离株进行O血清型鉴定、小鼠(Mus musculus)致病性试验以及抗菌药物敏感性分析.研究结果显示,95株大肠杆菌共鉴定出37种血清型,覆盖了54株分离株,另有2株自凝,39株未鉴定出型,较常见血清型为093和09;大肠杆菌分离株接种小白鼠剖检可见明显病变;95株大肠杆菌对16种抗菌药物中的8种药物耐药率超过50%,青霉素的耐药率甚至达到100%,同一菌株最多耐药14种,最少耐药2种,耐药6种以上菌株占到51.58%.表明,奶牛乳房炎源大肠杆菌分离株血清型比较复杂,且对多种药物产生不同程度的耐药性,存在着严重的多重耐药情况.本研究为奶牛乳房炎疫苗的研制和乳房炎的临床治疗提供理论依据.%The aim of the study was to investigate that characterization and antibiotic resistance of Escherichia coli isolated from bovine mastitis in some areas of China for raising therapeutic effect and decreasing drug residues. Milk samples of clinical and subclinical bovine mastitis were collected from seven provinces in China and a total of 95 E. Coli isolates were further identified by microbiological tests. Following, serotype, pathogenicity and antibiotic resistance of 95 strains were also studied. The results showed that for the 95 E. Coli strains, 54 isolates were identified as 37 serogroups, 2 isolates were self-clumpinged and 39 isolates could not be serotyped, 093 and 09 serotypes were the most common serogroups. Obviously pathological changes were observed in internal organs of dissected experiment mouse (Mus musculus). Among the 95 strains, the rate of drug fast to 8 out of 16 antibacterial drugs exceeded 50%, and a same

  16. Antimicrobial Susceptibility of Staphylococci Isolated from Clinical Mastitis in Dairy Cows in Two Regions of Kosovo

    OpenAIRE

    DRITON SYLEJMANI; AVNI ROBAJ; AFRIM HAMIDI

    2015-01-01

    This study was undertaken to evaluate antibiotic susceptibility of 26 strains of Coagulase Negative Staphylococci (CNS) and 18 strains of Staphylococcus aureus isolated from samples of dairy cows with clinical mastitis. The isolates species of staphylococci were identified by using coagulase test (using rabbit plasma) and API Staph system (bioMėrieux SA) which differentiates S. aurues from other staphylococci. Isolates were tested for antimicrobial susceptibility on Mueller Hinton...

  17. Clinical and diagnostic developments of a gamma interferon release assay for use in bovine tuberculosis control programs

    Science.gov (United States)

    Currently the Bovigam assay is used as an official supplemental test within the bovine tuberculosis eradication program. This assay measures interferon-gamma (IFN-gamma) produced by lymphocytes in response to specific antigens. The objectives of the present study were to evaluate two Mycobacterium ...

  18. Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis

    Directory of Open Access Journals (Sweden)

    Poliana de Castro Melo

    2013-01-01

    Full Text Available Biofilm formation is considered to be a selective advantage for Staphylococcus aureus mastitis isolates by facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers. The objective of this study was to determine the sensitivity and specificity of three techniques for the detection of S. aureus biofilm-positive strains. Two phenotypic tests, including growth on microtitre plates and Congo red agar, were compared with a PCR technique using 94 S. aureus strains obtained from cows with subclinical mastitis from two farms in the state of São Paulo. These strains were characterised by in vitro slime production on Congo red agar, biofilm formation on microtitre plates and the presence of the icaA and icaD genes. The results revealed that 85% of the isolates tested produced slime on the Congo red agar, 98.9% of the isolates produced biofilms in vitro by adhering to sterile 96-well "U" bottom polystyrene tissue culture plates, and 95.7% of the isolates carried the icaA and icaD genes. The results of the phenotypic tests for biofilm formation were compared with those of the molecular analysis, and the sensitivity and specificity of the Congo red agar test were 88.9% and 100%, respectively, while those of the microtitre plate test were 100% and 25%, respectively. When the phenotypic methods for the detection of biofilm producers, namely growth on microtitre plates and Congo red agar, were compared, the sensitivity and specificity were 86% and 100%, respectively. Therefore, growth on Congo red agar and the microtitre plate test are methods that could be used to determine whether an isolate has the potential for biofilm production.

  19. DETECTION OF STX2 COLIPHAGES IN STRAIN OF ESCHERICHIA COLI ISOLATED FROM BOVINE FLEECE AND MILK FILTERS

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    G. Delle Donne

    2011-01-01

    Full Text Available Lambda(4-phage vectors of gene codifying for synthesis of Shiga-toxins are suspected to be involved in the virulence evolution of Vero-Toxin producing Escherichia coli(VTEC. Herds of domestic or wild ruminants are reservoirs of these bacteria, but excretion with faeces is more frequent in groups of heifers and feeder calves. Studies have shown that slurries produced by infected herds are often positive for VTEC and that Stx2 carrying lambda coliphages can be isolated. These viruses can induce lysogenic cycles only in some strain of Escherichia coli and the Stx gene is then integrated in the bacterial chromosome. When these bacteria also posses other virulence traits, like those responsible for the intimate attachment to the enteric mucosal cells (eae or saa the recombinant strains might became pathogen for humans. Our research was aimed at detecting the coliphages form ten Stx2 positive strains isolated in our previous studies. We have included strains, possessing or not the ‘eae’ genes. In addition we have used other isolates originating from slaughterhouses, with the aim of evaluating their susceptibility to the isolated 4-phages. Following induction of a lytic cycle with mitomycin C, the strains were screened by hybridization of plaque blots with Stx2 probes. The purified extracts of eight of the ten strains produced plaque/halos of lysis in cultures of susceptible strains, thus showing these strains were infected by inducible phages, but only one proved to be Stx2 carrier. Attempt to obtain new lysogens using the purified Stx2 phage with other strains ‘eae’ positive and STx negative isolated from slaughterhouses were unsuccessful. Stx2 lysogens were obtained only using the reference strains DM1187.

  20. Kinetics and saturation of light-induced near-infrared scattering changes in isolated bovine rod outer segments.

    Science.gov (United States)

    Reichert, J; Hofmann, K P

    1981-01-01

    The axial and radial shrinkage of bovine rod outer segments, monitored by near-infrared scattering changes (P-signal), is investigated in dependence on the intensity of the activating flash. Suspensions of axially oriented and randomly oriented rod outer segments were measured. In the latter case, axial and radial effects are superimposed to another. The following results are obtained: 1. The axial signal (Pa, tau approximately 10 ms) and the radial signal (Pr, tau = 40-100 ms), simultaneously measured on axially oriented rod outer segments, are similarly saturated with a half-saturation at a rhodopsin turnover of 3.5%. 2. For the saturation of the signal amplitude, measured on randomly oriented rod outer segments, a good fit is obtained by: P(rho) approximately 1 - e-beta rho, rho: relative rhodopsin turnover by the flash; beta is found in the range 23 less than or equal to beta less than or equal to 27 in all measurements. 3. The kinetics of the signal, also measured on the isotropic sample, depends on the rhodopsin turnover, the apparent time constant becoming faster with increasing turnover, The distortion of the signal cannot be fitted by a sum of exponentials with a fixed set of time constants. The signals from the isotropic sample are fitted by a phenomenological model. It introduces three first order processes concatenated in series; the first step is assumed as a rhodopsin transition inducing the two further processes. The distortion of the signals with increasing rho is then described assuming a rho-dependent quenching of this induction, according to the measured amplitude saturation. The time constants remain thereby unchanged. The fit yields the values in 2/k = 4, 11, and 45 ms with mean square deviations of 20%. PMID:7326357

  1. COMPARISON OF TWO METHODS OR DETECTION OF SECRETED ASPARTYL PROTEINASE IN URINARY ISOLATES OF CANDIDA SPECIES

    Directory of Open Access Journals (Sweden)

    Santosh Patil

    2014-04-01

    Results: Out of 150 candida species isolated from clinical urine samples 86.6% were C. tropicalis, followed by C. albicans (11.3%, C. glabrata (1.33% and C. dubliniensis (0.6%. SAP detection using Bovine serum agar was 29.3% where as using bovine haemoglobin agar was 18.6%. Conclusions: So we conclude that the bovine serum agar is far superior to bovine haemoglobin agar for detection of SAP. [Natl J Med Res 2014; 4(2.000: 119-121

  2. Clinical course of isolated stable angina due to coronary heart disease

    NARCIS (Netherlands)

    Poole-Wilson, Philip A.; Voko, Zoltan; Kirwan, Bridget-Anne; de Brouwer, Sophie; Dunselman, Peter H. J. M.; Lubsen, Jacobus

    2007-01-01

    Aims To describe the clinical course of patients with stable angina due to coronary heart disease without a history of cardiovascular (CV) events or revascutarization (isolated angina). Methods and results Of 7665 patients in a trial comparing long-acting nifedipine with placebo, 2170 (28%) had isol

  3. Aspergillus pragensis sp nov discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi

    DEFF Research Database (Denmark)

    Lyskova, Pavlina; Hubka, Vit; Kolarik, Miroslav;

    2014-01-01

    The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of beta-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspe...

  4. Complete Genome Sequence of the Campylobacter ureolyticus Clinical Isolate RIGS 9880

    DEFF Research Database (Denmark)

    Miller, William G; Yee, Emma; On, Stephen L W;

    2015-01-01

    The emerging pathogen Campylobacter ureolyticus has been isolated from human and animal genital infections, human periodontal disease, domestic and food animals, and from cases of human gastroenteritis. We report the whole-genome sequence of the human clinical isolate RIGS 9880, which is the first...

  5. Detection of Intracellular Adhesion (ica Gene and Biofilm Formation Staphylococcus aureus Isolates from Clinical Blood Cultures

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    Mohsen Mirzaee

    2015-10-01

    Full Text Available Background: In fact the biofilms are composed of bacterial cells living inmulticellular structures such as tissues and organs embedded within a self-produced matrix of extracellular polymeric substance (EPS. Ability to attach and biofilm formation are the most important virulence factors Staphylococcus aureus isolates. The aims of this study were to detect intracellular adhesion (ica locus and its relation to the biofilm formation phenotype in clinical isolates of S. aureus isolated from bloodcultures.Methods: A total of 31 clinical S. aureus isolates were collected from Loghman Hospital of Tehran, Iran. In vitro biofilm formation ability was determined by microliter tissue culture plates. All clinical isolates were examined for determination the ica locus by using PCR method.Results: Twelve (38.7% of the isolates were strong biofilm producers. The results showed that 18(80.6% of the isolates carried icaD gene, whereas the prevalence of icaA, icaB and icaC were 51.6%, 45.1% and 77.4% respectively.Conclusions: S. aureus clinical isolates have different ability to form biofilm. This may be caused by the differences in the expression of biofilm related genes, genetic make-up and physiological conditions.

  6. Antimicrobial sensitivity pattern among clinical isolates of Escherichia coli in tertiary care centre of Northern India

    OpenAIRE

    Ranjana Malhotra; Rama Sikka; Uma Chaudhary

    2016-01-01

    Background: Rampant rise in resistance to various antimicrobials among clinical isolates of Escherichia coli is an issue of serious concern, especially in developing countries. It not only causes failure in treatment but also pose a health burden over the society at large. Methods: Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion method on Muller Hinton agar as per Clinical and Laboratory Standard Institute recommendations over 100 isolates of E. coli. Isola...

  7. Prevalence of dermatophytes and other fungal agents isolated from clinical samples

    OpenAIRE

    Kannan P.; Janaki C; Selvi G

    2006-01-01

    The common cause of skin infections are dermatophytes and opportunistic fungi. Aim of this study was to isolate and identify the fungal agents from clinical samples from patients with different mycoses. Clinical samples from 165 patients were subjected to potassium hydroxide (KOH) examination and culture isolation; causative agents were identified macroscopically and microscopically. All the 165 specimens were KOH positive and 110/165 (66.7%) samples were culture positive. Of these, hi...

  8. Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

    OpenAIRE

    Seung-Bae Lee

    2016-01-01

    Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV) and sweet bee venom (SBV) against Candida albicans (C. albicans) clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC) strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC) assays were performed by ...

  9. Prevalence, conservation and functional analysis of Yersinia and Escherichia CRISPR regions in clinical Pseudomonas aeruginosa isolates

    OpenAIRE

    Cady, K. C.; White, A. S.; Hammond, J. H.; Abendroth, M. D.; Karthikeyan, R. S. G.; Lalitha, P.; Zegans, M E; O'Toole, G. A.

    2011-01-01

    Here, we report the characterization of 122 Pseudomonas aeruginosa clinical isolates from three distinct geographical locations: Dartmouth Hitchcock Medical Center in New Hampshire, USA, the Charles T. Campbell Eye Microbiology Lab at the University of Pittsburgh Medical Center, USA, and the Aravind Eye Hospital in Madurai, India. We identified and located clustered regularly interspaced short palindromic repeats (CRISPR) in 45/122 clinical isolates and sequenced these CRISPR, finding that Ye...

  10. MALDI-TOF mass spectrometry proteomic based identification of clinical bacterial isolates

    Directory of Open Access Journals (Sweden)

    Ashutosh Panda

    2014-01-01

    Full Text Available Background & objectives: Pathogenic bacteria often cause life threatening infections especially in immunocompromised individuals. Therefore, rapid and reliable species identification is essential for a successful treatment and disease management. We evaluated a rapid, proteomic based technique for identification of clinical bacterial isolates by protein profiling using matrix-assisted laser desorption-ionization time - of - flight mass spectrometry (MALDI-TOF MS. Methods: Freshly grown bacterial isolates were selected from culture plates. Ethanol/formic acid extraction procedure was carried out, followed by charging of MALDI target plate with the extract and overlaying with α-cyano-4 hydroxy-cinnamic acid matrix solution. Identification was performed using the MALDI BioTyper 1.1, software for microbial identification (Bruker Daltonik GmbH, Bremen, Germany. Results: A comparative analysis of 82 clinical bacterial isolates using MALDI -TOF MS and conventional techniques was carried out. Amongst the clinical isolates, the accuracy at the species level for clinical isolates was 98.78%. One out of 82 isolates was not in accordance with the conventional assays because MALDI-TOF MS established it as Streptococcus pneumoniae and conventional methods as Streptococcus viridans. Interpretation & conclusions: MALDI - TOF MS was found to be an accurate, rapid, cost-effective and robust system for identification of clinical bacterial isolates. This innovative approach holds promise for earlier therapeutic intervention leading to better patient care.

  11. Antimicrobial susceptibility of Acinetobacter clinical isolates and emerging antibiogram trends for nosocomial infection management

    Directory of Open Access Journals (Sweden)

    Muhammad Sohail

    2016-06-01

    Full Text Available Abstract: Introduction: The drug resistant Acinetobacter strains are important causes of nosocomial infections that are difficult to control and treat. This study aimed to determine the antimicrobial susceptibility patterns of Acinetobacter strains isolated from different clinical specimens obtained from patients belonging to different age groups. METHODS: In total, 716 non-duplicate Acinetobacter isolates were collected from the infected patients admitted to tertiary-care hospitals at Lahore, Pakistan, over a period of 28 months. The Acinetobacter isolates were identified using API 20E, and antimicrobial susceptibility testing was performed and interpreted according to Clinical and Laboratory Standards Institute (CLSI guidelines. RESULTS: The isolation rate of Acinetobacter was high from the respiratory specimens, followed by wound samples. Antibiotic susceptibility analyses of the isolates revealed that the resistance to cefotaxime and ceftazidime was the most common, in 710 (99.2% specimens each, followed by the resistance to gentamicin in 670 (93.6% isolates, and to imipenem in 651 (90.9% isolates. However, almost all isolates were susceptible to tigecycline, colistin, and polymyxin B. CONCLUSIONS: The present study showed the alarming trends of resistance of Acinetobacter strains isolated from clinical specimens to the various classes of antimicrobials. The improvement of microbiological techniques for earlier and more accurate identification of bacteria is necessary for the selection of appropriate treatments.

  12. Complete Genome Sequences of Three African Foot-and-Mouth Disease Viruses from Clinical Samples Isolated in 2009 and 2010.

    Science.gov (United States)

    Van Borm, Steven; Rosseel, Toon; Haegeman, Andy; Fana, Mpolokang Elliot; Seoke, Latoa; Hyera, Joseph; Matlho, George; Vandenbussche, Frank; De Clercq, Kris

    2016-01-01

    The complete genome sequences of three foot-and-mouth disease viruses (one virus of each serotype SAT1, SAT2 and O) were directly sequenced from RNA extracted from clinical bovine samples, demonstrating the feasibility of full-genome sequencing from strong positive samples taken from symptomatic animals. PMID:27151795

  13. Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

    Directory of Open Access Journals (Sweden)

    Seung-Bae Lee

    2016-03-01

    Full Text Available Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV and sweet bee venom (SBV against Candida albicans (C. albicans clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti- fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from 62.5 μg/ mL to 125 μg/mL for BV and from 15.63 μg/mL to 62.5 μg/mL for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

  14. Frequency and Antimicrobial Susceptibility of Coagulasenegative Staphylococci Isolated from Bovine Mastitis in Dairy Farms from Tolima, Colombia

    OpenAIRE

    María del Pilar Sánchez Bonilla; Norma Patricia Gutiérrez Murillo

    2015-01-01

    Objective: To determine the antimicrobial presence and susceptibility of the coagulase-negative staphylococci group (ECN, for its initials in Spanish) in some cattle farms from Tolima, Colombia. Materials and methods: Using the California test for the diagnosis of mastitis (CMT, for its initials in Spanish), 484 quarters belonging to 121 cows from five small production ranches from a region of central Tolima were evaluated. CMT positive samples were cultivated for bacterial isolation. The ECN...

  15. Clinical evaluation of isolated abutment teeth in removable partial dentures

    OpenAIRE

    Zarrati S; Baheri Mghadam T; Nematollahi F

    2011-01-01

    "nBackground and Aims: Nowadays, removable partial dentures are applied to patients who are not able to use dental implants or fixed prosthesis. Although based on the studies the users of removable partial dentures are in the risk of plaque accumulation and unacceptable changes such as gingivitis, periodontitis and mobility in abutment tooth. It is not clear whether the negative effects of removable partial dentures are more on the isolated teeth which are a kind of abutment adjacent to ...

  16. Molecular epidemiology of Mycobacterium tuberculosis clinical isolates in Southwest Ireland.

    LENUS (Irish Health Repository)

    Ojo, Olabisi O

    2010-10-01

    Tuberculosis has had significant effects on Ireland over the past two centuries, causing persistently higher morbidity and mortality than in neighbouring countries until the last decade. This study describes the results of genotyping and drug susceptibility testing of 171 strains of Mycobacterium tuberculosis complex isolated between January 2004 and December 2006 in a region of Ireland centred on the city of Cork. Spoligotype comparisons were made with the SpolDB4 database and clustered 130 strains in 23 groups, forty-one strains showed unique Spoligotyping patterns. The commonest spoligotypes detected were ST0137 (X2) (16.9%), and ST0351 (15.8%) (\\'U\\' clade). The major spoligotype clades were X (26.2%), U (19.3%), T (15.2%), Beijing (5.9%), Haarlem (4.7%), LAM (4.1%), BOVIS (1.75%), with 12.9% unassigned strains. A 24-locus VNTR genotyping produced 15 clusters containing 49 isolates, with high discrimination index (HGDI>0.99). A combination of Spoligotyping and VNTR reduced the number of clustered isolates to 47 in 15 clusters (27.5%). This study identified ST351 as common among Irish nationals, and found a low rate of drug resistance with little evidence of transmission of drug resistant strains. Strain clustering was significantly associated with age under 55 years and Irish nationality. Only strains of Euro-American lineage formed clusters. Molecular typing did not completely coincide with the results of contact investigations.

  17. Diogenes syndrome or isolated syllogomania? Four heterogeneous clinical cases.

    Science.gov (United States)

    Zuliani, Giovanni; Soavi, Cecilia; Dainese, Anna; Milani, Paola; Gatti, Marino

    2013-08-01

    Diogenes syndrome (DS) is an acquired behavioural disturbance more often affecting elderly patients, but possible in all ages. It is characterised by social withdrawal, extreme self and house neglect, tendency to hoard any kind of objects/rubbish (syllogomania), and rejection against external help for lack of concern about one's condition. It is considered infrequent, but with quite high mortality. DS might be divided into several forms including Active (the patient gathers objects outside and accumulates them inside his house), Passive (patient invaded by his own rubbish), "à deux" (DS sharing between two people), and "under-threshold" (DS "blocked" by precocious intervention). Four cases are here presented. In case 1 (passive DS) alcoholism and cognitive impairment could be trigger factors for DS, predisposed by a "personality alteration". In case 2 (active, "à trois") superimposed psychosis could be the trigger, borderline intelligence being the predisposing factor. In case 3 (active), fronto-parietal internal hyperostosis might support an organic aetiology. Finally, case 4 was an example of isolated syllogomania in patient with evolving Alzheimer's dementia. Despite being heterogeneous, our casuistry suggest that DS can develop in both sexes, is prevalent in geriatric age and often associated with cognitive impairment/psychiatric disturbances, which are not specific, nor sufficient to justify DS. Isolated syllogomania only shares the characteristic hoarding with DS; although cognitive impairment might be present, the other DS typical aspects (social isolation, help refusal, characterial aspects, personal hygiene neglect) are absent. PMID:23846849

  18. Biofilm formation, hemolysin production and antimicrobial susceptibilities of Streptococcus agalactiae isolated from the mastitis milk of dairy cows in Shahrekord district, Iran

    OpenAIRE

    Azizollah Ebrahimi; Azar Moatamedi; Sharareh Lotfalian; Pejhman Mirshokraei

    2014-01-01

    Streptococcus agalactiae is a major contagious pathogen causing bovine sub-clinical mastitis. The present investigation was carried out to determine some phenotypic characteristics of the S. agalactiae strains isolated from bovine mastitis cases in dairy cows of Shahrekord in the west-center of Iran. One hundred eighty California mastitis test (CMT) positive milk samples were bacteriologically studied. A total of 31 (17.2%) S. agalactiae isolated. Twenty eight (90.3%) of the isolates were bio...

  19. [Phospholipase, proteinase and hemolytic activities of Candida albicans isolates obtained from clinical specimens].

    Science.gov (United States)

    Yenişehirli, Gülgün; Bulut, Yunus; Tunçoglu, Ebru

    2010-01-01

    This study was aimed to determine the phospholipase, proteinase and hemolytic activities of Candida albicans strains isolated from clinical specimens. A total of 147 C. albicans strains isolated from blood (n = 29), respiratory specimens (n = 44), urine (n = 52), pus (n = 17) and stool (n = 5) were included in the study. Proteinase and phospholipase activities were determined in 81% and 76% of C. albicans isolates, respectively. All C. albicans isolates revealed beta-hemolytic activity on Sabouraud dextrose agar supplemented with 7% fresh sheep blood and 3% glucose. Phospholipase and proteinase positivity were highest among the respiratory isolates. Proteinase activity of respiratory (93%) and blood (83%) isolates were statistically significantly higher than that of urine (77%; p = 0.032), pus (65%; p = 0.007) and stool isolates (60%; p = 0.026). While phospholipase activity showed statistically significant difference between respiratory (84%) and pus (53%) isolates (p = 0.014), no statistically significant difference was determined for blood (79%), urine (75%) and stool (80%) isolates (p > 0.05). Two blood isolates with 4+ proteinase activity and 3 urine isolates with 3+ proteinase activity were phospholipase negative. One urine isolate with 4+ phospholipase activity and 4 with 3+ phospholipase activity were proteinase negative. Phospholipase and proteinase negative 1 isolate from stool and 1 isolate from pus were found to have 4+ hemolytic activity. In conclusion, besides proteinase and phospholipase enzyme activities, hemolytic activity may play an important role for the C.albicans infections. The pathogenetic role of these virulence factors should be evaluated by further clinical studies.

  20. Isolating and using sections of bovine mesenteric artery and vein as a bioassay to test for vasoactivity in the small intestine.

    Science.gov (United States)

    Klotz, James L; Barnes, Adam J

    2014-10-07

    Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that system. Changes in blood flow to the small intestine can result in effects on the absorptive functions of the organ. Particular interest in toxins liberated from feedstuffs through fermentative and digestive processes has developed in ruminants as an area where productive efficiencies could be improved. The video associated with this article describes an in vitro bioassay developed to screen compounds for vasoactivity in isolated cross-sections of bovine mesenteric artery and vein using a multimyograph. Once the blood vessels are mounted and equilibrated in the myograph, the bioassay itself can be used: as a screening tool to evaluate the contractile response or vasoactivity of compounds of interest; determine the presence of receptor types by pharmacologically targeting receptors with specific agonists; determine the role of a receptor with the presence of one or more antagonists; or determine potential interactions of compounds of interest with antagonists. Through all of this, data are collected real-time, tissue collected from a single animal can be exposed to a large number of different experimental treatments (an in vitro advantage), and represents vasculature on either side of the capillary bed to provide an accurate picture of what could be happening in the afferent and efferent blood supply supporting the small intestine.

  1. Application of restriction fragment length polymorphism analysis to simple and rapid genotyping of bovine viral diarrhea virus strains isolated in Japan.

    Science.gov (United States)

    Seki, Yoshihisa; Seimiya, Yukio M; Motokawa, Masato; Yaegashi, Gakuji; Nagai, Makoto; Hayashi, Michiko

    2008-04-01

    The E2 regions of 177 bovine viral diarrhea virus (BVDV) strains isolated in Japan between 1957 and 2006 were analyzed for genotyping. The strains were classified into 8 genotypes (1a, 1b, 1c, 1d, 1e, 1f, So and 2a) based on the phylogenetic analysis. The restriction fragment length polymorphism (RFLP) analysis of the RT-PCR products using 6 selected enzymes (Apo I, Mly I, BstAP I, Pvu II, Ear I, EcoR V) disclosed the cutting patterns classified into 11 groups (I-XI), each of that consisted of strains belonging to a single genotype. Namely, groups-I and -II were composed by genotype-1a strains, groups-III and -IV by 1b strains, and groups-V and -VI by 1c strains. Other groups-VII, -VIII, -IX, -X and -XI comprised genotypes-1d, -1e, -1f, -So and -2a strains, respectively. The results suggest that the RFLP analysis can simply and rapidly differentiate the 8 genotypes of BVDV strains.

  2. Investigation of the Drug Resistance of Staphylococcus aureus Isolates from Bovine Mastiffs of Xinjiang%新疆牛源金黄色葡萄球菌抗菌素敏感性调查

    Institute of Scientific and Technical Information of China (English)

    高攀; 王登峰; 李建军; 王治才; 简子健

    2012-01-01

    [Objective]In order to understand the current status of the antimicrobial resistance of bovine Staphylococcus aureus isolates and provide a reference for clinical drug treatment. [ Method ] The antibiotic sensitive of 143 Saphylococcus aureus strains, which were isolated in 16 dairy farms in six districts of Changji, Hutubi, Kuitun, Urumqi, Yili and Korla were tested by using the disk diffusion method. [Result]The results showed that more than 70% of the strains resisted to erythromycin, penicillin, cotrimoxazole, doxycycline and tetracycline. About 40% of the strains resisted to chloramphenicol, ciprofloxacin and gentamicin. Furthermore, 67.1% of the strains resisted to more than five kinds of antibiotics and 10. 5% of the strains resisted to 10 kinds of antibiotics. In addition, 19. 6% of the strains were methicillin -resistant Staphylococcus aureus. [Conclusion]The survey indicates that Staphylococcus aureus isolated from bovine mastitis in Xinjiang resisted to multiple types of antibiotics that clinical veterinary medicine was allowed to be used in current situation and the treatment of Staphylococcus aureus mastitis at present is not desirable in our region. Arising and spreading of the muliti - resistant strains threats the development of dairy farming and milk products security, and suggest that antibiotic therapy would base on the resistant data of epidemic strains.%[目的]了解新疆奶牛乳房炎中金黄色葡萄球菌的药物敏感性情况,为该病治疗的临床用药提供参考.[方法]2009年以来对乌鲁木齐市、昌吉、呼图壁、奎屯、伊犁和库尔勒6地区16个奶牛养殖场乳房炎奶样分离金黄色葡萄球菌,用纸片扩散法进行抗菌素敏感性检测.[结果]共分离143株金黄色葡萄球菌,临床药敏试验显示,新疆70%以上分离株对红霉素、青霉素、复方新诺明、多西环素、四环素耐药,40%的菌株对氯霉素、环丙沙星和庆大霉素耐药;并且67.1%菌株同时对5

  3. Phenotypic and Genotypic Characterization of Enterotoxigenic Escherichia coli Clinical Isolates from Northern Colombia, South America

    Directory of Open Access Journals (Sweden)

    Julio A. Guerra

    2014-01-01

    Full Text Available Enterotoxigenic Escherichia coli (ETEC are major causes of childhood diarrhea in low and middle income countries including Colombia, South America. To understand the diversity of ETEC strains in the region, clinical isolates obtained from northern Colombia children were evaluated for multiple locus sequencing typing, serotyping, classical and nonclassical virulence genes, and antibiotic susceptibility. Among 40 ETEC clinical isolates evaluated, 21 (52.5% were positive for LT gene, 13 (32.5% for ST gene, and 6 (15% for both ST and LT. The most prevalent colonization surface antigens (CS were CS21 and CFA/I identified in 21 (50% and 13 (32.5% isolates, respectively. The eatA, irp2, and fyuA were the most common nonclassical virulence genes present in more than 60% of the isolates. Ampicillin resistance (80% of the strains was the most frequent phenotype among ETEC strains followed by trimethoprim-sulfamethoxazole resistance (52.5%. Based on multiple locus sequencing typing (MLST, we recognize that 6 clonal groups of ETEC clinical isolates circulate in Colombia. ETEC clinical isolates from children in northern Colombia are highly diverse, yet some isolates circulating in the community belong to well-defined clonal groups that share a unique set of virulence factors, serotypes, and MLST sequence types.

  4. Phenotypic and genotypic characterization of enterotoxigenic Escherichia coli clinical isolates from northern Colombia, South America.

    Science.gov (United States)

    Guerra, Julio A; Romero-Herazo, Yesenia C; Arzuza, Octavio; Gómez-Duarte, Oscar G

    2014-01-01

    Enterotoxigenic Escherichia coli (ETEC) are major causes of childhood diarrhea in low and middle income countries including Colombia, South America. To understand the diversity of ETEC strains in the region, clinical isolates obtained from northern Colombia children were evaluated for multiple locus sequencing typing, serotyping, classical and nonclassical virulence genes, and antibiotic susceptibility. Among 40 ETEC clinical isolates evaluated, 21 (52.5%) were positive for LT gene, 13 (32.5%) for ST gene, and 6 (15%) for both ST and LT. The most prevalent colonization surface antigens (CS) were CS21 and CFA/I identified in 21 (50%) and 13 (32.5%) isolates, respectively. The eatA, irp2, and fyuA were the most common nonclassical virulence genes present in more than 60% of the isolates. Ampicillin resistance (80% of the strains) was the most frequent phenotype among ETEC strains followed by trimethoprim-sulfamethoxazole resistance (52.5%). Based on multiple locus sequencing typing (MLST), we recognize that 6 clonal groups of ETEC clinical isolates circulate in Colombia. ETEC clinical isolates from children in northern Colombia are highly diverse, yet some isolates circulating in the community belong to well-defined clonal groups that share a unique set of virulence factors, serotypes, and MLST sequence types.

  5. Attaching and effacing Escherichia coli isolates from Danish children: clinical significance and microbiological characteristics

    DEFF Research Database (Denmark)

    Jensen, C; Ethelberg, S; Olesen, B;

    2007-01-01

    This study describes the prevalence, clinical manifestations and microbiological characteristics of attaching and effacing Escherichia coli isolates, i.e., enteropathogenic E. coli (EPEC) belonging to the classical EPEC serotypes, non-EPEC attaching and effacing E. coli (A/EEC) and verocytotoxin......-producing E. coli (VTEC), isolated in a case-control study of Danish children aged ... encoding the bundle-forming pilus (bfpA) and enteroaggregative heat-stable enterotoxin (astA) were rare among all isolates, and seemed to be of limited pathogenic importance in this population. Virulence characterisation of A/EEC isolates did not reveal any significant differences between cases...

  6. No Polymorphism in Plasmodium falciparum K13 Propeller Gene in Clinical Isolates from Kolkata, India

    Directory of Open Access Journals (Sweden)

    Moytrey Chatterjee

    2015-01-01

    Full Text Available Molecular markers associated with artemisinin resistance in Plasmodium falciparum are yet to be well defined. Recent studies showed that polymorphisms in K13 gene are associated with artemisinin resistance. The present study was designed to know the pattern of polymorphisms in propeller region of K13 gene among the clinical isolates collected from urban Kolkata after five years of ACT implementation. We collected 59 clinical isolates from urban Kolkata and sequenced propeller region of K13 gene in 51 isolates successfully. We did not find any mutation in any isolate. All patients responded to the ACT, a combination of artesunate + sulphadoxine-pyrimethamine. The drug regimen is still effective in the study area and there is no sign of emergence of resistance against artemisinin as evidenced by wild genotype of K13 gene in all isolates studied.

  7. Characterization of bacteriophages infecting clinical isolates of Pseudomonas aeruginosa stored in a culture collection

    Directory of Open Access Journals (Sweden)

    C.C.S. Zanetti

    2013-08-01

    Full Text Available Some clinical isolates of Pseudomonas aeruginosa stored in our culture collection did not grow or grew poorly and showed lysis on the culture plates when removed from the collection and inoculated on MacConkey agar. One hypothesis was that bacteriophages had infected and killed those clinical isolates. To check the best storage conditions to maintain viable P. aeruginosa for a longer time, clinical isolates were stored at various temperatures and were grown monthly. We investigated the presence of phage in 10 clinical isolates of P. aeruginosa stored in our culture collection. Four strains of P. aeruginosa were infected by phages that were characterized by electron microscopy and isolated to assess their ability to infect. The best condition to maintain the viability of the strains during storage was in water at room temperature. Three Siphoviridae and two Myoviridae phages were visualized and characterized by morphology. We confirmed the presence of bacteriophages infecting clinical isolates, and their ability to infect and lyse alternative hosts. Strain PAO1, however, did not show lysis to any phage. Mucoid and multidrug resistant strains of P. aeruginosa showed lysis to 50% of the phages tested.

  8. Use of MALDI-TOF MS for Identification of Nontuberculous Mycobacterium Species Isolated from Clinical Specimens

    Directory of Open Access Journals (Sweden)

    María Concepción Mediavilla-Gradolph

    2015-01-01

    Full Text Available The aim of this study was to compare the results obtained for identification by MALDI-TOF of nontuberculous mycobacteria (NTM isolated in clinical samples with those obtained by GenoType Mycobacterium CM/AS (common mycobacteria/additional species. A total of 66 Mycobacterium isolates from various clinical specimens (mainly respiratory were tested in this study. They were identified using MALDI-TOF Bruker from strains isolated in Lowenstein, following the recommended protocol of heat inactivation and extraction, and were simultaneously analyzed through hybridization by GenoType Mycobacterium from liquid culture MGIT. Our results showed that identification by MALDI-TOF was correct in 98.4% (65/66 of NTM isolated in our clinical practice (M. avium, M. intracellulare, M. abscessus, M. chelonae, M. fortuitum, M. mucogenicum, M. kansasii, and M. scrofulaceum. MALDI-TOF was found to be an accurate, rapid, and cost-effective system for identification of mycobacteria species.

  9. Prevalence and molecular characterization of clinical isolates of Escherichia coli expressing an AmpC phenotype

    DEFF Research Database (Denmark)

    Jørgensen, Rikke Lind; Nielsen, Jesper Boye; Friis-Møller, Alice;

    2010-01-01

    OBJECTIVES: To establish the prevalence of the AmpC beta-lactamase phenotype in clinical isolates of Escherichia coli and characterize the genetic resistance mechanisms causing the observed phenotype. METHODS: Clinical E. coli (n = 74) with reduced susceptibility to third-generation cephalosporin...

  10. High resolution genotyping of clinical Aspergillus flavus isolates from India using microsatellites.

    Directory of Open Access Journals (Sweden)

    Shivaprakash M Rudramurthy

    Full Text Available BACKGROUND: Worldwide, Aspergillus flavus is the second leading cause of allergic, invasive and colonizing fungal diseases in humans. However, it is the most common species causing fungal rhinosinusitis and eye infections in tropical countries. Despite the growing challenges due to A. flavus, the molecular epidemiology of this fungus has not been well studied. We evaluated the use of microsatellites for high resolution genotyping of A. flavus from India and a possible connection between clinical presentation and genotype of the involved isolate. METHODOLOGY/PRINCIPAL FINDINGS: A panel of nine microsatellite markers were selected from the genome of A. flavus NRRL 3357. These markers were used to type 162 clinical isolates of A. flavus. All nine markers proved to be polymorphic displaying up to 33 alleles per marker. Thirteen isolates proved to be a mixture of different genotypes. Among the 149 pure isolates, 124 different genotypes could be recognized. The discriminatory power (D for the individual markers ranged from 0.657 to 0.954. The D value of the panel of nine markers combined was 0.997. The multiplex multicolor approach was instrumental in rapid typing of a large number of isolates. There was no correlation between genotype and the clinical presentation of the infection. CONCLUSIONS/SIGNIFICANCE: There is a large genotypic diversity in clinical A. flavus isolates from India. The presence of more than one genotype in clinical samples illustrates the possibility that persons may be colonized by multiple genotypes and that any isolate from a clinical specimen is not necessarily the one actually causing infection. Microsatellites are excellent typing targets for discriminating between A. flavus isolates from various origins.

  11. Occurrence of qnrA-positive clinical isolates in French teaching hospitals during 2002-2005.

    Science.gov (United States)

    Cambau, E; Lascols, C; Sougakoff, W; Bébéar, C; Bonnet, R; Cavallo, J-D; Gutmann, L; Ploy, M-C; Jarlier, V; Soussy, C-J; Robert, J

    2006-10-01

    Bacteria harbouring the novel qnrA plasmid-mediated mechanism of quinolone resistance have been described in different countries, but the frequency of their occurrence has not been investigated. In total, 1,468 clinical isolates of Enterobacteriaceae with quinolone resistance or extended-spectrum beta-lactamase (ESBL) phenotypes were collected from eight teaching hospitals in France during 2002-2005 and screened for qnrA. Overall, 28 isolates (22 Enterobacter cloacae, three Klebsiella pneumoniae, one Citrobacter freundii, one Klebsiella oxytoca and one Proteus mirabilis) were positive for qnrA, representing 1.9% of all isolates, 3.3% of ESBL-producing isolates (22% of the E. cloacae isolates) and 0% of non-ESBL-producing isolates. The prevalence of qnrA among consecutive ESBL-producing isolates in 2004 from the eight hospitals was 2.8% (18/639). Of the qnrA-positive isolates, 100% were intermediately-resistant or resistant to nalidixic acid, and 75% to ciprofloxacin. Twenty-one of the 22 qnrA-positive E. cloacae isolates were obtained from two hospitals in the Paris area, and molecular typing and plasmid content analysis showed clonal relationships for five, three and two isolates, respectively. The qnrA genetic environment was similar to that of the In36 integron. The remaining two isolates had qnrA variants (30 and 29 nucleotide differences, respectively, compared with the original sequence) and an unknown genetic environment. The ESBL gene associated with qnrA was bla(SHV-12) in most of the isolates, but bla(PER-1) and bla(SHV-2a) were found in two isolates. In France, it appears that qnrA-positive isolates are predominantly E. cloacae isolates producing SHV-12, and may be associated with the dissemination of an In36-like integron. PMID:16961639

  12. In vitro antimicrobial activity of Combretum molle (Combretaceae) against Staphylococcus aureus and Streptococcus agalactiae isolated from crossbred dairy cows with clinical mastitis.

    Science.gov (United States)

    Regassa, Fekadu; Araya, Mengistu

    2012-08-01

    Following the rapidly expanding dairy enterprise, mastitis has remained the most economically damaging disease. The objective of this study was mainly to investigate the in vitro antibacterial activities of ethanol extracts of Combretum molle (R.Br.Ex.G.Don) Engl & Diels (Combretaceae) against antibiotic-resistant and susceptible Staphylococcus aureus and Streptococcus agalactiae isolated from clinical cases of bovine mastitis using agar disc diffusion method. The leaf and bark extracts showed antibacterial activity against S. aureus at concentrations of 3 mg/ml while the stem and seed extract did not show any bioactivity. Although both leaf and bark extracts were handled in the same manner, the antibacterial activity of the bark extract against the bacterial strains had declined gradually to a lower level as time advanced after extraction. The leaf extract had sustained bioactivity for longer duration. The susceptibility of the bacteria to the leaf extract is not obviously different between S. aureus and S. agalactiae. Also, there was no difference in susceptibility to the leaf extract between the antibiotic-resistant and antibiotic-sensitive bacteria. Further phytochemical and in vivo efficacy and safety studies are required to evaluate the therapeutic value of the plant against bovine mastitis.

  13. Biofilm Formation and Motility Depend on the Nature of the Acinetobacter baumannii Clinical Isolates.

    Science.gov (United States)

    Vijayakumar, Saranya; Rajenderan, Sangeetha; Laishram, Shakti; Anandan, Shalini; Balaji, Veeraraghavan; Biswas, Indranil

    2016-01-01

    Acinetobacter baumannii is a nosocomial pathogen involved in various infections ranging from minor soft-tissue infections to more severe infections such as ventilator-associated pneumonia and bacteremia. The severity and the type of infections depend on the genetic and phenotypic variations of the strains. In this study, we compared the extent of biofilm formation and motility displayed by 60 multidrug-resistant A. baumannii clinical strains isolated from blood and sputum samples from patients from Southern India. Our results showed that isolates from the sputum samples formed significantly more robust biofilm compared to the blood isolates. On the other hand, we observed that the blood isolates were more motile than the sputum isolates. To the best of our knowledge, this is the first study that systematically evaluated the correlation between these two phenotypic traits and the nature of the isolates. PMID:27252939

  14. Polymorphic Amplified Typing Sequences and Pulsed-Field Gel Electrophoresis Yield Comparable Results in the Strain Typing of a Diverse Set of Bovine Escherichia coli O157:H7 Isolates

    Directory of Open Access Journals (Sweden)

    Indira T. Kudva

    2012-01-01

    Full Text Available Polymorphic amplified typing sequences (PATS, a PCR-based Escherichia coli O157:H7 (O157 strain typing system, targets insertions-deletions and single nucleotide polymorphisms at XbaI and AvrII restriction enzyme sites, respectively, and the virulence genes (stx1, stx2, eae, hlyA in the O157 genome. In this study, the ability of PATS to discriminate O157 isolates associated with cattle was evaluated. An in-depth comparison of 25 bovine O157 isolates, from different geographic locations across Northwest United States, showed that about 85% of these isolates shared the same dendogram clade by PATS and pulsed-field gel electrophoresis (PFGE, irrespective of the restriction enzyme sites targeted. The Pearson’s correlation coefficient, r, calculated at about 0.4, 0.3, and 0.4 for XbaI-based, AvrII-based and combined-enzymes PATS and PFGE similarities, respectively, indicating that these profiles shared a good but not high correlation, an expected inference given that the two techniques discriminate differently. Isolates that grouped differently were better matched to their locations using PATS. Overall, PATS discriminated the bovine O157 isolates without interpretive biases or sophisticated analytical software, and effectively complemented while not duplicating PFGE. With its quick turnaround time, PATS has excellent potential as a convenient tool for early epidemiological or food safety investigations, enabling rapid notification/implementation of quarantine measures.

  15. MRI criteria for MS in patients with clinically isolated syndromes

    DEFF Research Database (Denmark)

    Montalban, X.; Tintore, M.; Swanton, J.;

    2010-01-01

    In recent years, criteria for the diagnosis of multiple sclerosis (MS) have changed, mainly due to the incorporation of new MRI criteria. While the new criteria are a logical step forward, they are complex and-not surprisingly-a good working knowledge of them is not always evident among neurologi......In recent years, criteria for the diagnosis of multiple sclerosis (MS) have changed, mainly due to the incorporation of new MRI criteria. While the new criteria are a logical step forward, they are complex and-not surprisingly-a good working knowledge of them is not always evident among...... isolated syndrome. Within the European multicenter collaborative research network that studies MRI in MS (MAGNIMS), a workshop was held in London in November 2007 to review information that may simplify the existing MS diagnostic criteria, while maintaining a high specificity that is essential to minimize...

  16. Experimental arthritis induced by a clinical Mycoplasma fermentans isolate

    Directory of Open Access Journals (Sweden)

    Giono Silvia

    2002-06-01

    Full Text Available Abstract Background Mycoplasma fermentans has been associated with rheumatoid arthritis. Recently, it was detected in the joints and blood of patients with rheumatoid arthritis, but it is not clear yet how the bacteria enter the body and reach the joints. The purpose of this study was to determine the ability of M. fermentans to induce experimental arthritis in rabbits following inoculation of the bacteria in the trachea and knee joints. Methods P-140 and PG-18 strains were each injected in the knee joints of 14 rabbits in order to evaluate and compare their arthritogenicity. P-140 was also injected in the trachea of 14 rabbits in order to test the ability of the bacteria to reach the joints and induce arthritis. Results M. fermentans produced an acute arthritis in rabbits. Joint swelling appeared first in rabbits injected with P-140, which caused a more severe arthritis than PG-18. Both strains were able to migrate to the uninoculated knee joints and they were detected viable in the joints all along the duration of the experiment. Changes in the synovial tissue were more severe by the end of the experiment and characterized by the infiltration of neutrophils and substitution of adipose tissue by connective tissue. Rabbits intracheally injected with P-140 showed induced arthritis and the bacteria could be isolated from lungs, blood, heart, kidney, spleen, brain and joints. Conclusion M. fermentans induced arthritis regardless of the inoculation route. These findings may help explain why mycoplasmas are commonly isolated from the joints of rheumatic patients.

  17. The role of active efflux in antibiotic - resistance of clinical isolates of Helicobacter pylori

    Directory of Open Access Journals (Sweden)

    Falsafi T

    2009-01-01

    Full Text Available Purpose: In gram-negative bacteria, active efflux pumps that excrete drugs can confer resistance to antibiotics however, in Helicobacter pylori this role is not well established. The purpose of this study is to evaluate the role of active efflux in resistance of H. pylori isolates to antibiotics. Materials and Methods: Twelve multiple antibiotic resistant (MAR isolates resistant to at least four antibiotics, including β-lactams, metronidazole, tetracycline, erythromycin, and ciprofloxacin; three resistant to only β-lactams, and two hyper-susceptible isolates, were obtained from screening of 96 clinical isolates of H. pylori . Their minimal inhibitory concentrations (MICs for antibiotics and ethidium-bromide (EtBr were compared in the presence- and absence of a proton-conductor, carbonyl cyanide-m chlorophenyl-hydrazone (CCCP using agar-dilution and disc diffusion. Drug accumulation studies for EtBr and antibiotics were assessed in the presence and absence of CCCP using spectrofluorometry. Results: MIC of EtBr for eight MAR-isolates was decreased two- to four-folds in the presence of CCCP, of which five showed reduced MICs for β-lactam, metronidazole, tetracycline, and ciprofloxacin with CCCP. Accumulation of EtBr by the MAR-isolates was rapid and not dependant on the pattern of multiple resistance. Antibiotic accumulation assay confirmed the presence of energy-dependant efflux of β-lactam, metronidazole, tetracycline, and ciprofloxacin, but no erythromycin in five MAR isolates. Energy-dependant efflux of EtBr or antibiotics was not observed for four MAR-isolates, and three isolates were resistant only to β-lactams. Conclusion: Energy-dependant efflux plays a role in the resistance of H. pylori clinical isolates to structurally unrelated antibiotics in a broadly specific multidrug efflux manner. Difference in the efflux potential of MAR isolates may be related to the presence or absence of functional efflux-pumps in diverse H. pylori

  18. Detection of a Novel Bovine Lymphotropic Herpesvirus

    OpenAIRE

    Rovnak, Joel; Quackenbush, Sandra L.; Reyes, Richard A.; Baines, Joel D.; Parrish, Colin R.; Casey, James W.

    1998-01-01

    Degenerate PCR primers which amplify a conserved region of the DNA polymerase genes of the herpesvirus family were used to provide sequence evidence for a new bovine herpesvirus in bovine B-lymphoma cells and peripheral blood mononuclear cells (PBMC). The sequence of the resultant amplicon was found to be distinct from those of known herpesvirus isolates. Alignment of amino acid sequences demonstrated 70% identity with ovine herpesvirus 2, 69% with alcelaphine herpesvirus 1, 65% with bovine h...

  19. Storage of bovine isolated follicles: a new alternative way to improve the recovery rate of viable embryos from ovarian follicles of slaughtered cows.

    Science.gov (United States)

    Pavlok, A; Cech, S; Kubelka, M; Lopatárová, M; Holý, L; Jindra, M

    2006-11-01

    The vitality of bovine oocytes stored in isolated follicles was examined. The aim of this work was to prolong the time of in vitro manipulation of oocytes before their maturation and develop a new alternative of oocyte "capacitation" to improve the quality of in vitro produced embryos. Follicles were dissected from the ovaries of slaughtered cows; subsequently, follicles were divided according to their diameter into three categories (2-3, 3-4 and 4-6 mm), and stored at 17-18 degrees C for 24 or 48 h in a modified tissue culture medium-199 (TCM-199) with reduced pH. After that time, the cumulus-oocyte complexes (COCs) were isolated, matured, fertilized, and embryos cultured in vitro for a total of 9 days. The percentage of total blastocysts, and hatched blastocysts developed from oocytes, initially kept ("capacitated") for 24h at 17-18 degrees C, within follicles of 3-6mm size categories, were significantly higher than that oocytes of the control [of control oocytes] (44.9 and 30.3% versus 36.2 and 20.4%, respectively). The oocytes of follicles stored for 48 h at 17-18 degrees C already had decreased developmental capacity. Interesting data were obtained when COCs of the 3-4 and 4-6 categories were additionally divided into two subgroups according to their presumed developmental history (originating from the supposed growing "fit" in contrast to the supposed regressing "unfit" follicles). The higher improvement in the rate of hatched blastocysts from 24h stored oocytes was observed only in the subgroup originated from "fit" COCs (15.3 versus 25.0%, and 20.0 versus 34.4%, in the 3-4 and 4-6mm categories, respectively). The transfer of 26 blastocysts (developed of follicles kept for 24h at 17-18 degrees C) to 26 recipient heifers resulted in 18 pregnancies. Storage of follicles at 17-18 degrees C in vitro resulted not only in recovery of higher numbers of blastocysts of better quality but also facilitated the safe transport of follicles for a long distance. The

  20. Application and validation of PFGE for serovar identification of Leptospira clinical isolates.

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    Renee L Galloway

    Full Text Available Serovar identification of clinical isolates of Leptospira is generally not performed on a routine basis, yet the identity of an infecting serovar is valuable from both epidemiologic and public health standpoints. Only a small number of reference laboratories worldwide have the capability to perform the cross agglutinin absorption test (CAAT, the reference method for serovar identification. Pulsed-field gel electrophoresis (PFGE is an alternative method to CAAT that facilitates rapid identification of leptospires to the serovar level. We employed PFGE to evaluate 175 isolates obtained from humans and animals submitted to the Centers for Disease Control and Prevention (CDC between 1993 and 2007. PFGE patterns for each isolate were generated using the NotI restriction enzyme and compared to a reference database consisting of more than 200 reference strains. Of the 175 clinical isolates evaluated, 136 (78% were identified to the serovar level by the database, and an additional 27 isolates (15% have been identified as probable new serovars. The remaining isolates yet to be identified are either not represented in the database or require further study to determine whether or not they also represent new serovars. PFGE proved to be a useful tool for serovar identification of clinical isolates of known serovars from different geographic regions and a variety of different hosts and for recognizing potential new serovars.

  1. Isolation, speciation, and antibiogram of clinically relevant non-diphtherial Corynebacteria (Diphtheroids

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    B S Reddy

    2012-01-01

    Full Text Available Purpose: Coryneform or the non-diphtherial Corynebacterium species largely remains a neglected group with the traditional consideration of these organisms as contaminants. This concept, however, is slowly changing in the light of recent observations. This study has been done to find out the species distribution and antibiogram of various members of the clinically relevant Coryneform group, isolated from various clinical materials. Materials and Methods: One hundred and fourteen non-duplicate isolates of diphtheroids from various clinical isolates were selected for the study. The isolates were identified to the species level by using a battery of tests; and antimicrobial susceptibility was tested by using a combination of Clinical and Laboratory Standards Institute (CLSI and the British Society for Antimicrobial Chemotherapy (BSAC guidelines, in the absence of definitive CLSI guidelines. Results: Corynebacterium amycolatum was the predominant species (35.9% in our series followed by the CDC Group G organisms (15.7%. Each of the remaining 19 species comprised of less than 10% of the isolates. More than half the total isolates were resistant to the penicillins, erythromycin, and clindamycin; while excellent activity (all the strains being susceptible was shown by vancomycin, linezolid, and tigecycline. Chloramphenicol and tetracycline also had good activity in inhibiting more than 80% of the isolates. Multiply drug resistance was exhibited by all the species. Conclusion: This study was an attempt to establish the clinical significance of coryneform organisms. The high level of resistance shown by this group to some of the common antibacterial agents highlights the importance of processing these isolates in select conditions to guide the clinicians towards an appropriate therapy.

  2. Greater Diversity of Shiga Toxin-Encoding Bacteriophage Insertion Sites among Escherichia coli O157:H7 Isolates from Cattle than in Those from Humans▿

    Science.gov (United States)

    Besser, Thomas E.; Shaikh, Nurmohammad; Holt, Nicholas J.; Tarr, Phillip I.; Konkel, Michael E.; Malik-Kale, Preeti; Walsh, Coilin W.; Whittam, Thomas S.; Bono, James L.

    2007-01-01

    Escherichia coli O157:H7, a zoonotic human pathogen for which domestic cattle are a reservoir host, produces a Shiga toxin(s) (Stx) encoded by bacteriophages. Chromosomal insertion sites of these bacteriophages define three principal genotypes (clusters 1 to 3) among clinical isolates of E. coli O157:H7. Stx-encoding bacteriophage insertion site genotypes of 282 clinical and 80 bovine isolates were evaluated. A total of 268 (95.0%) of the clinical isolates, but only 41 (51.3%) of the bovine isolates, belonged to cluster 1, 2, or 3 (P < 0.001). Thirteen additional genotypes were identified in isolates from both cattle and humans (four genotypes), from only cattle (seven genotypes), or from only humans (two genotypes). Two other markers previously associated with isolates from cattle or with clinical isolates showed similar associations with genotype groups within bovine isolates; the tir allele sp-1 and the Q933W allele were under- and overrepresented, respectively, among cluster 1 to 3 genotypes. Stx-encoding bacteriophage insertion site typing demonstrated that there is broad genetic diversity of E. coli O157:H7 in the bovine reservoir and that numerous genotypes are significantly underrepresented among clinical isolates, consistent with the possibility that there is reduced virulence or transmissibility to humans of some bovine E. coli O157:H7 genotypes. PMID:17142358

  3. Differential expression of putative drug resistance genes in Mycobacterium tuberculosis clinical isolates.

    Science.gov (United States)

    González-Escalante, Laura; Peñuelas-Urquides, Katia; Said-Fernández, Salvador; Silva-Ramírez, Beatriz; Bermúdez de León, Mario

    2015-12-01

    Understanding drug resistance in Mycobacterium tuberculosis requires an integrated analysis of strain lineages, mutations and gene expression. Previously, we reported the differential expression of esxG, esxH, infA, groES, rpmI, rpsA and lipF genes in a sensitive M. tuberculosis strain and in a multidrug-resistant clinical isolate. Here, we have evaluated the expression of these genes in 24 clinical isolates that belong to different lineages and have different drug resistance profiles. In vitro, growth kinetics analysis showed no difference in the growth of the clinical isolates, and thus drug resistance occurred without a fitness cost. However, a quantitative reverse transcription PCR analysis of gene expression revealed high variability among the clinical isolates, including those with similar drug resistance profiles. Due to the complexity of gene regulation pathways and the wide diversity of M. tuberculosis lineages, the use of gene expression as a molecular signature for drug resistance is not straightforward. Therefore, we recommend that the expression of M. tuberculosis genes be performed individually, and baseline expression levels should be verified among several different clinical isolates, before any further applications of these findings.

  4. Aspergillus pragensis sp. nov. discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi.

    Science.gov (United States)

    Hubka, Vit; Lyskova, Pavlina; Frisvad, Jens C; Peterson, Stephen W; Skorepova, Magdalena; Kolarik, Miroslav

    2014-08-01

    The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of β-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspected and proven onychomycosis, one from otitis externa, and two associated with probable invasive aspergillosis. The results showed that one Aspergillus candidus isolate was the cause of otitis externa, and both isolates obtained from sputa of patients with probable invasive aspergillosis were reidentified as A. carneus (sect. Terrei) and A. flavus (sect. Flavi). Three isolates from nail scrapings were identified as A. tritici, a verified agent of nondermatophyte onychomycosis. One isolate from toenail was determined to be A. candidus and the two isolates belonged to a hitherto undescribed species, Aspergillus pragensis sp. nov. This species is well supported by phylogenetic analysis based on β-tubulin and calmodulin gene and is distinguishable from other members of sect. Candidi by red-brown reverse on malt extract agar, slow growth on Czapek-Dox agar and inability to grow at 37°C. A secondary metabolite analysis was also provided with comparison of metabolite spectrum to other species. Section Candidi now encompasses five species for which a dichotomous key based on colony characteristics is provided. All clinical isolates were tested for susceptibilities to selected antifungal agents using the Etest and disc diffusion method. Overall sect. Candidi members are highly susceptible to common antifungals.

  5. A GENETIC STUDY TO DIFFERENTIAL HA/CA MRSA ISOLATED FROM CLINICAL CASES IN IRAQ HOSPITALS.

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    ISRAA MOHAMED SAFI AL- KADMY

    2013-01-01

    Full Text Available Methicillin-resistant Staphylococcus aureus (MRSA has been a major cause of nosocomial infections since the 1960s . Currently, MRSA is divided into two subgroups: the healthcare associated MRSA (HA-MRSA and community associated MRSA(CA-MRSA, CA-MRSA infections have been increasing. The most common of these infections present in soft skin. The aim of this study to different between CA and HA MRSA in clinical isolates of Baghdad hospitals.Methods: clinical isolates were collected from patients with different infections, Simple laboratory testing followed by the complementary API Staph, followed by antibiotic sensitivity and D-test, and finally by using PCR technique, detection of this genes : mecA, PVL, SCCmec IV and V .Results: A total of 105 S.aureus found 104 methicillin-resistant Staphylococcus aureus (MRSA strains, after a D-test , S.aureus divided to two group: CA and HA –MRSA, where the ratio of CA 18(17.1% out of 105 isolates, while HA reached 87(82.8%. MRSA was characterized by PCR amplification mecA gene, 104(99.04% isolates out of 105 gave positive result, all isolates of HA carry mecA gene, while 17 out of 18 isolates of CA carry mecA gene which was CA-MRSA and one isolates was CA-MSSA . All isolates 18(100% of CA gave positive result in risk factors PVL gene, while for detection of SCCmec IV 17 (94.4% out of 18 isolates of CA gave positive result, and finally two isolates of CA-MRSA gave positive result in SCCmec V gene .Conclusions: This is the first report in Iraq for the emergence of CA isolates especially CA-MRSA which is responsible for the majority of infection in soft tissue and skin abscesses , are likely to be sensitive to clindamycin.

  6. Oral Serum-Derived Bovine Immunoglobulin/Protein Isolate Has Immunomodulatory Effects on the Colon of Mice that Spontaneously Develop Colitis

    Science.gov (United States)

    Maijó, Mònica; Polo, Javier; Campbell, Joy M.; Russell, Louis; Crenshaw, Joe D.; Weaver, Eric; Moretó, Miquel

    2016-01-01

    Dietary immunoglobulin concentrates prepared from animal plasma can modulate the immune response of gut-associated lymphoid tissue (GALT). Previous studies have revealed that supplementation with serum-derived bovine immunoglobulin/protein isolate (SBI) ameliorates colonic barrier alterations in the mdr1a-/- genetic mouse model of IBD. Here, we examine the effects of SBI on mucosal inflammation in mdr1a-/- mice that spontaneously develop colitis. Wild type (WT) mice and mice lacking the mdr1a gene (KO) were fed diets supplemented with either SBI (2% w/w) or milk proteins (Control diet), from day 21 (weaning) until day 56. Leucocytes in mesenteric lymph nodes (MLN) and in lamina propria were determined, as was mucosal cytokine production. Neutrophil recruitment and activation in MLN and lamina propria of KO mice were increased, but were significantly reduced in both by SBI supplementation (p animals, but SBI prevented these changes (both p < 0.05). In the colon of KO mice, there was an increased production of mucosal pro-inflammatory cytokines such as IL-2 (2-fold), IL-6 (26-fold) and IL-17 (19-fold), and of chemokines MIP-1β (4.5-fold) and MCP-1 (7.2-fold). These effects were significantly prevented by SBI (p < 0.05). SBI also significantly increased TGF-β secretion in the colon mucosa, suggesting a role of this anti-inflammatory cytokine in the modulation of GALT and the reduction of the severity of the inflammatory response during the onset of colitis. PMID:27139220

  7. Sympathetic nerve activity in normal and cystic follicles from isolated bovine ovary: local effect of beta-adrenergic stimulation on steroid secretion

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    Ortega Hugo H

    2011-05-01

    Full Text Available Abstract Cystic ovarian disease (COD is an important cause of abnormal estrous behavior and infertility in dairy cows. COD is mainly observed in high-yielding dairy cows during the first months post-partum, a period of high stress. We have previously reported that, in lower mammals, stress induces a cystic condition similar to the polycystic ovary syndrome in humans and that stress is a definitive component in the human pathology. To know if COD in cows is also associated with high sympathetic activity, we studied isolated small antral (5mm, preovulatory (10mm and cystic follicles (25mm. Cystic follicles which present an area 600 fold greater compared with preovulatory follicles has only 10 times less concentration of NE as compared with small antral and preovulatory follicles but they had 10 times more NE in follicular fluid, suggesting a high efflux of neurotransmitter from the cyst wall. This suggestion was reinforced by the high basal release of recently taken-up 3H-NE found in cystic follicles. While lower levels of beta-adrenergic receptor were found in cystic follicles, there was a heightened response to the beta-adrenergic agonist isoproterenol and to hCG, as measured by testosterone secretion. There was however an unexpected capacity of the ovary in vitro to produce cortisol and to secrete it in response to hCG but not to isoproterenol. These data suggest that, during COD, the bovine ovary is under high sympathetic nerve activity that in addition to an increased response to hCG in cortisol secretion could participate in COD development.

  8. Oral Serum-Derived Bovine Immunoglobulin/Protein Isolate Has Immunomodulatory Effects on the Colon of Mice that Spontaneously Develop Colitis

    Science.gov (United States)

    Maijó, Mònica; Polo, Javier; Campbell, Joy M.; Russell, Louis; Crenshaw, Joe D.; Weaver, Eric; Moretó, Miquel

    2016-01-01

    Dietary immunoglobulin concentrates prepared from animal plasma can modulate the immune response of gut-associated lymphoid tissue (GALT). Previous studies have revealed that supplementation with serum-derived bovine immunoglobulin/protein isolate (SBI) ameliorates colonic barrier alterations in the mdr1a-/- genetic mouse model of IBD. Here, we examine the effects of SBI on mucosal inflammation in mdr1a-/- mice that spontaneously develop colitis. Wild type (WT) mice and mice lacking the mdr1a gene (KO) were fed diets supplemented with either SBI (2% w/w) or milk proteins (Control diet), from day 21 (weaning) until day 56. Leucocytes in mesenteric lymph nodes (MLN) and in lamina propria were determined, as was mucosal cytokine production. Neutrophil recruitment and activation in MLN and lamina propria of KO mice were increased, but were significantly reduced in both by SBI supplementation (p < 0.05). The increased neutrophil recruitment and activation observed in KO mice correlated with increased colon oxidative stress (p < 0.05) and SBI supplementation reduced this variable (p < 0.05). The Tact/Treg lymphocyte ratios in MLN and lamina propria were also increased in KO animals, but SBI prevented these changes (both p < 0.05). In the colon of KO mice, there was an increased production of mucosal pro-inflammatory cytokines such as IL-2 (2-fold), IL-6 (26-fold) and IL-17 (19-fold), and of chemokines MIP-1β (4.5-fold) and MCP-1 (7.2-fold). These effects were significantly prevented by SBI (p < 0.05). SBI also significantly increased TGF-β secretion in the colon mucosa, suggesting a role of this anti-inflammatory cytokine in the modulation of GALT and the reduction of the severity of the inflammatory response during the onset of colitis. PMID:27139220

  9. In Vitro Antimicrobial Activity of Plant-Derived Diterpenes against Bovine Mastitis Bacteria

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    Rodrigo C. S. Veneziani

    2013-07-01

    Full Text Available We evaluated the antibacterial activity of three diterpenes isolated from natural sources against a panel of microorganisms responsible for bovine mastitis. ent-Copalic acid (CA was the most active metabolite, with promising MIC values (from 1.56 to 6.25 µg mL−1 against Staphylococcus aureus (ATCC and clinical isolate, Staphylococcus epidermidis, Streptococcus agalactiae, and Streptococcus dysgalactiae. We conducted time-kill assays of CA against S. aureus, a commensal organism considered to be a ubiquitous etiological agent of bovine mastitis in dairy farms worldwide. In the first 12 h, CA only inhibited the growth of the inoculums (bacteriostatic effect, but its bactericidal effect was clearly noted thereafter (between 12 and 24 h. In conclusion, CA should be considered for the control of several Gram-positive bacteria related to bovine mastitis.

  10. Zearalenone and Its Derivatives α-Zearalenol and β-Zearalenol Decontamination by Saccharomyces cerevisiae Strains Isolated from Bovine Forage

    Science.gov (United States)

    Keller, Luiz; Abrunhosa, Luís; Keller, Kelly; Rosa, Carlos Alberto; Cavaglieri, Lilia; Venâncio, Armando

    2015-01-01

    Zearalenone (ZEA) and its derivatives are mycotoxins with estrogenic effects on mammals. The biotransformation for ZEA in animals involves the formation of two major metabolites, α- and β-zearalenol (α-ZOL and β-ZOL), which are subsequently conjugated with glucuronic acid. The capability of Saccharomyces cerevisiae strains isolated from silage to eliminate ZEA and its derivatives α-ZOL and β-ZOL was investigated as, also, the mechanisms involved. Strains were grown on Yeast Extract-Peptone-Dextrose medium supplemented with the mycotoxins and their elimination from medium was quantified over time by HPLC-FL. A significant effect on the concentration of ZEA was observed, as all the tested strains were able to eliminate more than 90% of the mycotoxin from the culture medium in two days. The observed elimination was mainly due to ZEA biotransformation into β-ZOL (53%) and α-ZOL (8%) rather than to its adsorption to yeast cells walls. Further, the biotransformation of α-ZOL was not observed but a small amount of β-ZOL (6%) disappeared from culture medium. ZEA biotransformation by yeasts may not be regarded as a full detoxification process because both main end-products are still estrogenic. Nonetheless, it was observed that the biotransformation favors the formation of β-ZOL which is less estrogenic than ZEA and α-ZOL. This metabolic effect is only possible if active strains are used as feed additives and may play a role in the detoxification performance of products with viable S. cerevisiae cells. PMID:26308051

  11. Zearalenone and Its Derivatives α-Zearalenol and β-Zearalenol Decontamination by Saccharomyces cerevisiae Strains Isolated from Bovine Forage

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    Luiz Keller

    2015-08-01

    Full Text Available Zearalenone (ZEA and its derivatives are mycotoxins with estrogenic effects on mammals. The biotransformation for ZEA in animals involves the formation of two major metabolites, α- and β-zearalenol (α-ZOL and β-ZOL, which are subsequently conjugated with glucuronic acid. The capability of Saccharomyces cerevisiae strains isolated from silage to eliminate ZEA and its derivatives α-ZOL and β-ZOL was investigated as, also, the mechanisms involved. Strains were grown on Yeast Extract-Peptone-Dextrose medium supplemented with the mycotoxins and their elimination from medium was quantified over time by HPLC-FL. A significant effect on the concentration of ZEA was observed, as all the tested strains were able to eliminate more than 90% of the mycotoxin from the culture medium in two days. The observed elimination was mainly due to ZEA biotransformation into β-ZOL (53% and α-ZOL (8% rather than to its adsorption to yeast cells walls. Further, the biotransformation of α-ZOL was not observed but a small amount of β-ZOL (6% disappeared from culture medium. ZEA biotransformation by yeasts may not be regarded as a full detoxification process because both main end-products are still estrogenic. Nonetheless, it was observed that the biotransformation favors the formation of β-ZOL which is less estrogenic than ZEA and α-ZOL. This metabolic effect is only possible if active strains are used as feed additives and may play a role in the detoxification performance of products with viable S. cerevisiae cells.

  12. Expression of Multidrug Efflux Pump SmeDEF by Clinical Isolates of Stenotrophomonas maltophilia

    OpenAIRE

    Alonso, Ana; Jose L Martinez

    2001-01-01

    The presence of the multidrug efflux pump SmeDEF was assessed in a collection of clinical isolates of Stenotrophomonas maltophilia. All isolates encoded this pump, as demonstrated by PCR. Forty-seven percent of the strains overproduced a protein of the same size that was immunoreactive against an anti-SmeF antibody, and 33% overexpressed the gene semD when they were tested by reverse transcription-PCR. A correlation between smeDEF overexpression and antibiotic resistance was observed.

  13. PHENOTYPIC DETECTION OF CARBAPENEM RESISTANCE IN CLINICAL ISOLATES OF ACINETOBACTER BAUMANII IN KANCHIPURAM

    OpenAIRE

    Sivasankari S; Senthamarai; Anitha C; Apurba Sastry; Sandhya bhatt; Kumudhavathi M.S; Amshavadhani S.K

    2014-01-01

    Acinctobacter species are common non fermentative gram negative bacilli isolated in clinical laboratory most frequently encountered species. Acinetobacter resistance is develop due to acquired resistance. Because of frequent multidrug resistance isolates carbapenems have become important for treating resistant strains. There is a need for rapid screening & detection of MBL in Acinetobacter to modify the treatment. The present study was aim to determine the resistance of A.baumanii complese to...

  14. Characteristics of CDC group 1 and group 1-like coryneform bacteria isolated from clinical specimens.

    OpenAIRE

    Funke, G; Lucchini, G M; Pfyffer, G E; Marchiani, M; von Graevenitz, A

    1993-01-01

    Fifteen strains of CDC group 1 coryneform and biochemically similar bacteria were isolated from clinical specimens. Of the 15 strains isolated, 11 were derived from abscesses and purulent lesions, mostly from the upper part of the body, and 3 were grown from blood cultures. Nine strains were associated with mixed anaerobic but no other aerobic flora. Seven strains exhibited the classical biochemical profile of CDC coryneform group 1; however, eight strains were unable to reduce nitrate and we...

  15. In Vitro Activities of 15 Antimicrobial Agents against Clinical Isolates of South African Enterococci

    OpenAIRE

    Struwig, M. C.; Botha, P. L.; Chalkley, L. J.

    1998-01-01

    The activities of a panel of currently available antibiotics and the investigational agents LY 333328, linezolid, CL 331,002, CL 329,998, moxifloxacin (BAY 12-8039), trovafloxacin, and quinupristin-dalfopristin against 274 clinical isolates of enterococci were determined. No vancomycin resistance or β-lactamase production was observed. Except for 12 isolates (all non-Enterococcus faecalis) showing reduced susceptibility to quinupristin-dalfopristin (MIC, ≥4 μg/ml), the new agents exhibited pr...

  16. Antibacterial Activity of Twenty Iranian Plant Extracts Against Clinical Isolates of Helicobacter pylori

    OpenAIRE

    Farahnaz Nariman; Fereshteh Eftekhar; Zohreh Habibi; Sadegh Massarrat; Reza Malekzadeh

    2009-01-01

    Objective(s)Due to increasing emergence of drug-resistance in Helicobacter pylori isolates, traditional plants arepotentially valuable sources of novel anti-H. pylori agents. In this research, anti-H. pylori activity of theorganic extracts of twenty native Iranian plants was determined against ten clinical isolates of H. pylori.Materials and MethodsDisc diffusion was used to determine the biological activity of 20 plant extracts as well as 8 antibioticscommonly used to treat H. pylori infecti...

  17. Prevalence of Extended –Spectrum-Beta-Lactamase-Producing Klebsiella Pneumonia Isolates from Clinical Samples

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    Alizade, H. (MSc

    2014-06-01

    Full Text Available Background and Objective: Klebsiella pneumonia (K.pneumonia is one of the common causes of nosocomial infections. The aim of this research was to determine the prevalence of beta-lactamase genes and phenotypic confirmation of extended–spectrum-beta-lactamase (ESBL producing K.pneumonia isolates from clinical samples. Material and Methods: In this study, 122 K.pneumonia were isolated from clinical specimens of Khoramabad city and were confirmed by standard bacteriological tests. The presence of ESBL enzymes was detected by combined disk diffusion method. PCR assay with specific primers was used to determine blaSHV, blaTEM, blaCTX-15 and blaCTX-M genes in the confirmed isolates. Results: of 122 K.pneumonia isolates, 78 (64.18% were positive for ESBL, using disk diffusion method. According to antibiogram results, 10.65% of isolates were resistant to cefotaxime, 3.27% to ceftazidime and 68.03% to both antibiotics. Ninety isolates (64.18% considered as ESBLs isolates, at the same time, with being resistant to cefotaxime and ceftazidime were also sensitive to cefotaxime-clavulanic acid and ceftazidime-clavulanic acid. In PCR assays, blaCTX-15, blaSHV, blaCTX-M and blaTEM genes were detected in 78.68%, 40.16%, 26.22% and 22.13% of isolates, respectively. Ten resistant patterns of genes were detected. Conclusion: The significance percentage of antibiotic resistant genes of K.pneumonia isolates from clinical samples in Khoramabad city had ESBLs genes; CTX-M category was the most prevalent encoding genes of these enzymes. Keywords: Klebsiella Pneumonia, Extended-Spectrum Beta-Lactamase, Antibiotic Resistance

  18. Candida species biofilm and Candida albicans ALS3 polymorphisms in clinical isolates.

    Science.gov (United States)

    Bruder-Nascimento, Ariane; Camargo, Carlos Henrique; Mondelli, Alessandro Lia; Sugizaki, Maria Fátima; Sadatsune, Terue; Bagagli, Eduardo

    2014-01-01

    Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Candida isolates obtained from patients attended at a Brazilian tertiary public hospital (Botucatu, Sao Paulo). C. albicans ALS3 gene polymorphism was also evaluated by determining the number of repeated motifs in the central domain. Of the 198 total biofilm-positive isolates, 72 and 126 were considered as low and high biofilm producers, respectively. Biofilm production by C. albicans was significantly lower than that by non-albicans isolates and was most frequently observed in C. tropicalis. Biofilm production was more frequent among bloodstream isolates than other clinical sources, in urine, the isolates displayed a peculiar distribution by presenting two distinct peaks, one containing biofilm-negative isolates and the other containing isolates with intense biofilm production. The numbers of tandem-repeat copies per allele were not associated with biofilm production, suggesting the evolvement of other genetic determinants.

  19. Candida species biofilm and Candida albicans ALS3 polymorphisms in clinical isolates

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    Ariane Bruder-Nascimento

    2014-12-01

    Full Text Available Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Candida isolates obtained from patients attended at a Brazilian tertiary public hospital (Botucatu, Sao Paulo. C. albicans ALS3 gene polymorphism was also evaluated by determining the number of repeated motifs in the central domain. Of the 198 total biofilm-positive isolates, 72 and 126 were considered as low and high biofilm producers, respectively. Biofilm production by C. albicans was significantly lower than that by non-albicans isolates and was most frequently observed in C. tropicalis. Biofilm production was more frequent among bloodstream isolates than other clinical sources,in urine, the isolates displayed a peculiar distribution by presenting two distinct peaks, one containing biofilm-negative isolates and the other containing isolates with intense biofilm production. The numbers of tandem-repeat copies per allele were not associated with biofilm production, suggesting the evolvement of other genetic determinants.

  20. Biological activity of Cryptococcus neoformans and Cryptococcus gattii from clinical and environmental isolates

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    Antonio Marcio Barbosa Junior

    2013-06-01

    Full Text Available INTRODUCTION: Cryptococcus neoformans and Cryptococcus gattii are encapsulated basidiomycetous yeasts with worldwide distribution. They cause cryptococcosis with features of systemic infection, affecting the central nervous system, lungs and skin in humans and animals. These fungi present numerous virulence factors that allow them to invade the host and multiply, among which extracellular enzyme capacity and microbial adaptation to different temperatures are worth mentioning. OBJECTIVE: To evaluate the production of protease and investigate possible differences in thermotolerance and urease activity in clinical and environmental yeast isolates. MATERIAL AND METHODS: Culture methods and Pz analysis were applied to assess urease and protease, whereas the optical density method was used to analyze biological activity in thermotolerance. RESULTS: There was no significant results as to microbial growth at the tested temperatures (25º, 37º and 42ºC. It was observed that clinical specimens grew better than environmental ones at elevated temperatures. As to C. neoformans, the moderate production of urease enzyme prevailed in both clinical and environmental isolates within 24h or 48h. Moreover, there was significant production on the seventh day of reading. The best reading time for viewing protease production in both isolates and species was the seventh day: 96% clinical samples and 94% environmental isolates. CONCLUSION: Further studies are required in order to investigate the virulence factors of C. neoformans and C. gattii cerebrospinal isolates from patients with meningoencephalitis and environmental samples from Sergipe. Furthermore, a higher technical accuracy and statistical precision are indispensable.

  1. Physicochemical properties of elastase isolated from clinical Pseudomonas Aeruginosa

    International Nuclear Information System (INIS)

    Purified elastase was obtained from clinical Pseudomonas Aeruginosa (P.A.-283). The enzyme showed not only elasto lytic activity, but also a broad proteolytic activity against various proteins. The activity of the enzyme on collagen and gelatin was also observed. The optimum pH for elastase was 7.8 to 8.0 for both the proteolytic and elasto lytic activities. The elastase was stable in a pH range from 6.6 to 9.0. Optimum temperature for proteolytic and elasto lytic activities was 40 and inhibition of elastase occurs at 80 . The D10 value of the P.A-283 was found to be 0.11 kGy. Increasing the dose level value of gamma-irradiation decrease the proteolytic activity in the culture filtrate reaching only 16% at the dose level 0.5 kGy. Chelating agents and some metal ions inhibited both proteolytic and elasto lytic activities. Selective inhibition of elasto lytic activity was observed in high concentrations of sodium and ammonium salts without concurrent decrease in the proteolytic activity of the enzyme.4 fig., 3 tab

  2. Cloning and sequencing of the bovine gastrin gene

    DEFF Research Database (Denmark)

    Lund, T; Rehfeld, J F; Olsen, Jørgen

    1989-01-01

    In order to deduce the primary structure of bovine preprogastrin we therefore sequenced a gastrin DNA clone isolated from a bovine liver cosmid library. Bovine preprogastrin comprises 104 amino acids and consists of a signal peptide, a 37 amino acid spacer-sequence, the gastrin-34 sequence followed...

  3. Biochemical and Molecular Analysis of Staphylococcus aureus Clinical Isolates from Hospitalized Patients

    Directory of Open Access Journals (Sweden)

    Amit Karmakar

    2016-01-01

    Full Text Available Staphylococcus aureus is opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 100 Staphylococcus aureus isolates were obtained from clinical samples derived from hospitalized patients. The presumptive Staphylococcus aureus clinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species specific 16S rRNA primer pairs and finally 100 isolates were found to be positive as Staphylococcus aureus. Screened isolates were further analyzed by several microbiological diagnostics tests including gelatin hydrolysis, protease, and lipase tests. It was found that 78%, 81%, and 51% isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolated Staphylococcus aureus strains with respect to different antimicrobial agents revealed resistance pattern ranging from 57 to 96%. Our study also shows 70% strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection of mecA, nuc, and hlb genes and 70%, 84%, and 40% were found to harbour mecA, nuc, and hlb genes, respectively. The current investigation is highly important and informative for the high level multidrug resistant Staphylococcus aureus infections inclusive also of methicillin and vancomycin.

  4. Clinical significance of the isolation of Candida species from hospitalized patients.

    Science.gov (United States)

    Magalhães, Yankee C; Bomfim, Maria Rosa Q; Melônio, Luciane C; Ribeiro, Patrícia C S; Cosme, Lécia M; Rhoden, Cristianne R; Marques, Sirlei G

    2015-03-01

    In this study, we isolated and phenotypically identified 108 yeast strains from various clinical specimens collected from 100 hospitalized patients at three tertiary hospitals in São Luís-Maranhão, Brazil, from July to December 2010. The isolates were analyzed for their susceptibility to four of the most widely used antifungal agents in the surveyed hospitals, amphotericin B, fluconazole, 5-flucytosine and voriconazole. The species identified were Candida albicans (41.4%), Candida tropicalis (30.1%), C. glabrata (7.4%), Candida parapsilosis (5.5%), Candida krusei (4.6%), Cryptococcus neoformans (4.6%), Trichosporon spp . (3.7%), Candida norvegensis (0.9%), Rhodotorula glutinis (0.9%) and Pichia farinosa (0.9%). A higher isolation rate was observed in the following clinical specimens: urine (54 isolates; 50%), respiratory tract samples (21 isolates; 19.4%) and blood (20 isolates; 18.6%). Candida albicans isolates were 100% sensitive to all antifungal agents tested, whereas Candida krusei and Crytococcus neoformans displayed intermediate resistance to 5-flucytosine, with Minimal Inhibitory Concentration (MIC) values of 8 mg/mL and 16 mg/mL, respectively. Both strains were also S-DD to fluconazole with an MIC of 16 mg/mL. C. tropicalis was resistant to 5-flucytosine with an MIC of 32 μg/mL. This study demonstrates the importance of identifying the yeast species involved in community and nosocomial infections.

  5. Glucose & sodium chloride induced biofilm production & ica operon in clinical isolates of staphylococci

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    Astha Agarwal

    2013-01-01

    Full Text Available Background & objectives: All colonizing and invasive staphylococcal isolates may not produce biofilm but may turn biofilm producers in certain situations due to change in environmental factors. This study was done to test the hypothesis that non biofilm producing clinical staphylococci isolates turn biofilm producers in presence of sodium chloride (isotonic and high concentration of glucose, irrespective of presence or absence of ica operon. Methods: Clinical isolates of 100 invasive, 50 colonizing and 50 commensal staphylococci were tested for biofilm production by microtiter plate method in different culture media (trypticase soy broth alone or supplemented with 0.9% NaCl/ 5 or 10% glucose. All isolates were tested for the presence of ica ADBC genes by PCR. Results: Biofilm production significantly increased in the presence of glucose and saline, most, when both glucose and saline were used together. All the ica positive staphylococcal isolates and some ica negative isolates turned biofilm producer in at least one of the tested culture conditions. Those remained biofilm negative in different culture conditions were all ica negative. Interpretation & conclusions: The present results showed that the use of glucose or NaCl or combination of both enhanced biofilm producing capacity of staphylococcal isolates irrespective of presence or absence of ica operon.

  6. Differentiation of clinical Mycobacterium tuberculosis complex isolates by their GyrB polymorphism

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    Abass N

    2010-01-01

    Full Text Available Purpose: To evaluate the reliability of the gyrB PCR-RFLP technique in differentiating clinical Mycobacterium tuberculosis complex isolates. Materials and Methods: A primer pair MTUB-f and MTUB-r for M. tuberculosis complex (MTBC was used to differentiate 79 mycobacterial isolates by specific amplification of the 1,020-bp fragment of the gyrB gene (gyrB-PCR1. The MTBC isolates were further differentiated using a set of specific primers MTUB-756-Gf and MTUB-1450-Cr that allowed selective amplification of the gyrB fragment specific for M. tuberculosis (gyrB-PCR2. The DNA polymorphisms in the 1,020-bp gyrB fragment for 7 M. tuberculosis strains confirmed by PCR as well as 2 reference strains; M. tuberculosis H37Rv and M. bovis BCG were analyzed with the restriction enzyme Rsa1. Results: Seventy-seven (97.5% isolates were positive for gyrB-PCR1 and thus identified as members of M. tuberculosis complex (MTBC and two (2.6% isolates were negative and identified as Mycobacteria other than tuberculosis (MOTT. All the M. tuberculosis isolates showed the typical M. tuberculosis specific Rsa1 RFLP patterns (100, 360, 560-bp while 360 and 480-bp fragments were generated from M. bovis BCG. Conclusion: The gyrB PCR-RFLP using the endonuclease Rsa1 can be used to differentiate M. tuberculosis from M. bovis in clinical isolates.

  7. Use of mitochondrial and ribosomal DNA polymorphisms to classify clinical and soil isolates of Histoplasma capsulatum.

    OpenAIRE

    Spitzer, E. D.; Lasker, B A; Travis, S J; Kobayashi, G. S.; Medoff, G

    1989-01-01

    We have developed an improved scheme for the classification of environmental and clinical isolates of Histoplasma capsulatum that is based on analysis of mitochondrial DNA (mtDNA) and ribosomal DNA (rDNA). Strains were initially divided into mtDNA groups according to restriction digests of whole-cell DNA and Southern hybridization with cloned mtDNA probes. Strains within a mtDNA class could be further grouped by polymorphisms in rDNA. The majority of soil and clinical isolates from the United...

  8. A comparison of three media for isolation of Nocardia species from clinical specimens.

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    Ayyar S

    1992-04-01

    Full Text Available The study was carried out to compare the efficacy of three media namely Modified Thayer Martin medium, McClung′s carbon free broth with paraffin bait and paraffin agar in isolating Nocardia species from clinical specimens. Two hundred and seventy six clinical specimens from 245 cases were studied which included cases of bronchopulmonary and systemic infections and cases of mycetoma. Paraffin agar was found to be an inexpensive and selective medium for isolation of Nocardia species when compared with Modified Thayer Martin medium and paraffin bait techniques.

  9. First insight into the genotypic diversity of clinical Mycobacterium tuberculosis isolates from Gansu Province, China.

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    Jie Liu

    Full Text Available BACKGROUND: Investigations of Mycobacterium tuberculosis genetic diversity in China have indicated a significant regional distribution. The aim of this study was to characterize the genotypes of clinical M. tuberculosis isolates obtained from Gansu, which has a special geographic location in China. METHODOLOGY/PRINCIPAL FINDINGS: A total of 467 clinical M. tuberculosis strains isolated in Gansu Province were genotyped by 15-locus mycobacterial interspersed repetitive units-variable number tandem repeats (MIRU-VNTR and spoligotyping. The results showed that 445 isolates belonged to six known spoligotype lineages, whereas 22 isolates were unknown. The Beijing genotype was the most prevalent (87.58%, n = 409, while the shared type 1 was the dominant genotype (80.94%, n = 378. The second most common lineage was the T lineage, with 25 isolates (5.35%, followed by the H lineage with 5 isolates (1.07%, the MANU family (0.64%, 3 isolates, the U family (0.43%, 2 isolates and the CAS lineage with 1 isolate (0.21%. By using the VNTR15China method, we observed 15 groups and 228 genotypes among the 467 isolates. We found no association between the five larger groups (including the Beijing genotype and sex, age, or treatment status, and there was no noticeable difference in the group analysis in different areas. In the present study, seven of the 15 MIRU-VNTR loci were highly or moderately discriminative according to their Hunter-Gaston discriminatory index. CONCLUSIONS/SIGNIFICANCE: The Beijing genotype is the predominant genotype in Gansu province. We confirm that VNTR15China is suitable for typing Beijing strains in China and that it has a better discriminatory power than spoligotyping. Therefore, the use of both methods is the most suitable for genotyping analysis of M. tuberculosis.

  10. Staphylococcus aureus Clinical Isolates: Antibiotic Susceptibility, Molecular Characteristics, and Ability to Form Biofilm

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    N. Indrawattana

    2013-01-01

    Full Text Available Periodic monitoring of Staphylococcus aureus characteristics in a locality is imperative as their drug-resistant variants cause treatment problem. In this study, antibiograms, prevalence of toxin genes (sea-see, seg-ser, seu, tsst-1, eta, etb, and etd, PFGE types, accessory gene regulator (agr groups, and ability to form biofilm of 92 S. aureus Thailand clinical isolates were investigated. They were classified into 10 drug groups: groups 1–7 (56 isolates were methicillin resistant (MRSA and 8–10 (36 isolates were methicillin sensitive (MSSA. One isolate did not have any toxin gene, 4 isolates carried one toxin gene (seq, and 87 isolates had two or more toxin genes. No isolate had see, etb, or tsst-1; six isolates had eta or etd. Combined seg-sei-sem-sen-seo of the highly prevalent egc locus was 26.1%. The seb, sec, sel, seu, and eta associated significantly with MSSA; sek was more in MRSA. The sek-seq association was 52.17% while combined sed-sej was not found. Twenty-three PFGE types were revealed, no association of toxin genes with PFGE types. All four agr groups were present; agr group 1 was predominant (58.70% but agr group 2 strains carried more toxin genes and were more frequent toxin producers. Biofilm formation was found in 72.83% of the isolates but there was no association with antibiograms. This study provides insight information on molecular and phenotypic markers of Thailand S. aureus clinical isolates which should be useful for future active surveillance that aimed to control a spread of existing antimicrobial resistant bacteria and early recognition of a newly emerged variant.

  11. Isolation and characterization of Wharton’s jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

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    Cardoso Tereza C

    2012-05-01

    Full Text Available Abstract Background The possibility for isolating bovine mesenchymal multipotent cells (MSCs from fetal adnexa is an interesting prospect because of the potential for these cells to be used for biotechnological applications. Bone marrow and adipose tissue are the most common sources of MSCs derived from adult animals. However, little knowledge exists about the characteristics of these progenitors cells in the bovine species. Traditionally most cell cultures are developed in two dimensional (2D environments. In mammalian tissue, cells connect not only to each other, but also support structures called the extracellular matrix (ECM. The three-dimensional (3D cultures may play a potential role in cell biotechnology, especially in tissue therapy. In this study, bovine-derived umbilical cord Wharton’s jelly (UC-WJ cells were isolated, characterized and maintained under 3D-free serum condition as an alternative of stem cell source for future cell banking. Results Bovine-derived UC-WJ cells, collected individually from 5 different umbilical cords sources, were successfully cultured under serum-free conditions and were capable to support 60 consecutive passages using commercial Stemline® mesenchymal stem cells expansion medium. Moreover, the UC-WJ cells were differentiated into osteocytes, chondrocytes, adipocytes and neural-like cells and cultured separately. Additionally, the genes that are considered important embryonic, POU5F1 and ITSN1, and mesenchymal cell markers, CD105+, CD29+, CD73+ and CD90+ in MSCs were also expressed in five bovine-derived UC-WJ cultures. Morphology of proliferating cells typically appeared fibroblast-like spindle shape presenting the same viability and number. These characteristics were not affected during passages. There were 60 chromosomes at the metaphase, with acrocentric morphology and intense telomerase activity. Moreover, the proliferative capacity of T cells in response to a mitogen stimulus was suppressed when

  12. In Vitro Activities of Amphotericin B, Terbinafine, and Azole Drugs against Clinical and Environmental Isolates of Aspergillus terreus Sensu Stricto

    Science.gov (United States)

    Fernández, Mariana S.; Rojas, Florencia D.; Cattana, María E.; Sosa, María de los Ángeles; Iovannitti, Cristina A.; Giusiano, Gustavo E.

    2015-01-01

    The antifungal susceptibilities of 40 clinical and environmental isolates of A. terreus sensu stricto to amphotericin B, terbinafine, itraconazole, and voriconazole were determined in accordance with CLSI document M38-A2. All isolates had itraconazole and voriconazole MICs lower than epidemiologic cutoff values, and 5% of the isolates had amphotericin B MICs higher than epidemiologic cutoff values. Terbinafine showed the lowest MICs. No significant differences were found when MICs of clinical and environmental isolates were compared. PMID:25824228

  13. Copy Numbers of Telomeric Repeat Sequences of Human Herpesvirus 6B in Clinical Isolates: Possibility of Mixed Infections

    OpenAIRE

    KATO, Yuri; Ihira, Masaru; Umeda, Mami; Higashimoto, Yuki; Kawamura, Yoshiki; Ohashi, Masahiro; Ishi, Junichi; Yoshikawa, Tetsushi

    2014-01-01

    In order to determine whether mixed infections of human herpesvirus 6B (HHV-6B) occur in immunocompetent and immunocompromised individuals, we examined the copy numbers of telomeric repeat sequences (TRS) of clinical isolates. In clinical isolates obtained from patients with exanthem subitum caused by primary HHV-6B infection, PCR products with HHV-6B TRS ranging between 400 and 800 bp were amplified. PCR products of various sizes were amplified in four clinical isolates from drug-induced hyp...

  14. Molecular epidemiology of nontuberculous mycobacteria isolates from clinical and environmental sources of a metropolitan city.

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    Ali Akbar Velayati

    Full Text Available While NTM infection is mainly acquired from environmental exposure, monitoring of environmental niches for NTM is not a routine practice. This study aimed to find the prevalence of environmental NTM in soil and water in four highly populated suburbs of Tehran, Iran.A total of 4014 samples from soil and water resources were collected and studied. Sediments of each treated sample were cultured in Lowenstein-Jensen medium and observed twice per week for growth rate, colony morphology, and pigmentation. Colonies were studied with phenotypic tests. Molecular analysis was performed on single colonies derived from subculture of original isolates. Environmental samples were compared with 34 NTM isolates from patients who were residents of the study locations.Out of 4014 samples, mycobacteria were isolated from 862 (21.4% specimens; 536 (62.1% belonged to slow growing mycobacteria (SGM and 326 (37.8% were rapid growing mycobacteria (RGM. The five most frequent NTM were M. farcinogens (105/862; 12.1%, M. fortuitum (72/862; 8.3%, M. senegalense (58/862; 6.7%, M. kansasii (54/862; 6.2%, and M. simiae (46/862; 5.3%. In total, 62.5% (539/862 of mycobacterial positive samples were isolated from water and only 37.4% (323/862 of them were isolated from soil samples (P<0.05. Out of 5314 positive clinical samples for mycobacteria, 175 (3.2% isolates were NTM. The trend of NTM isolates increased from 1.2% (13 out of 1078 in 2004 to 3.8% (39 out of 1005 in 2014 (P = 0.0001. The major clinical isolates were M. simiae (51; 29.1%, M. kansasii (26; 14.8%, M. chelonae (28; 16%, and M. fortuitum (13; 7.4%.Comparing the distribution pattern of environmental NTM isolates with clinical isolates suggests a possible transmission link, but this does not apply to all environmental NTM species. Our study confirms an increasing trend of NTM isolation from clinical samples that needs further investigation.

  15. Prevalence of aac(3-IIa gene among clinical isolates of uropathogenic Escherichia coli in Delfan, Lorestan

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    Somayeh Momeni Mofrad

    2013-09-01

    Full Text Available Backgrounds: Uropathogenic Escherichia coli strains are the predominant causative organisms of urinary tract infections (UTIs. Aminoglycosides are clinically useful antibiotics with bactericidal activity against this bacterium. The most common mechanism for resistance to these antibiotics are mediated through production of aminoglycoside modifying enzymes (AMEs. The most common of these enzymes are Aminoglycoside Acetyltransferases (AACs. The epidemiology of the dominant type of these enzymes, AAC(3-II, varies from region to region. The aim of this study was to determine the antimicrobial susceptibility pattern with a focus on aminoglycosides and the prevalence of aac(3-IIa gene among clinical isolates of uropathogenic Escherichia coli obtained from Delfan, Lorestan, Iran. Materials and Methods: In this descriptive study, a total of 100 uropathogenic Escherichia coli isolates were collected from BoAli hospital in Delfan city, Lorestan, from July to November 2010. Antibiotic susceptibility patterns of the isolates were determined using disk diffusion method according to Clinical and Laboratory Standards Institute CLSI guidelines. Prevalence of aac(3-IIa gene was determined by PCR and the relationship between resistance phenotypes to aminoglycosides and presence of aac(3-IIa gene was evaluated. Results: Among the 100 tested isolates, maximal resistance was seen to ampicillin (85%; whereas, no resistance to imipenem was found. Sixty percent of the isolates demonstrated resistance to at least one of the tested aminoglycosides. Resistance rate towards these agents were as followed: gentamicin 39%, kanamycin 26%, neomycin 31% and amikacin 1%. Forty–four isolates (44% harbored the aac(3-IIa gene. The maximal rate of gene presence (36 isolates, 92.3% was detected in strains with gentamicin resistant phenotype (39 isolates, 39%. Conclusion: On the basis of our findings, use of antibiotics such as nitrofurantoin, amikacin or imipenem are recommended for

  16. [The Weaver syndrome in cattle. Clinical, biochemical and pathologico-anatomic studies in a Braunvieh/Brown Swiss cow with bovine progressive degenerative myeloencephalopathy].

    Science.gov (United States)

    Braun, U; Ehrensperger, F; Bracher, V

    1987-01-01

    The clinical signs and pathological lesions, consistent with Bovine Progressive Degenerative Myeloencephalopathy (BPDME), are described in a heifer. The animal was an 18-month-old Braunvieh-/Brown Swiss crossbreed with 50% Brown Swiss. Both the sire as well as the dam were related to the bull "B.". The only neurological signs were a posterior ataxia. Mobility and coordination of the front legs were normal. The CK activity of the cerebrospinal fluid was enhanced. There was a moderate to severe degeneration of the white matter consisting of marked axonal degradation and distension and degradation of myelin sheats in all parts of the spinal cord as well as in the medulla oblongata. The lesions were more severe in the descending than in the ascending tracts.

  17. HIGH VARIABILITY OF HUMAN CYTOMEGALOVIRUS UL150 OPEN READING FRAME IN LOW-PASSAGED CLINICAL ISOLATES

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective To investigate the polymorphism of human cytomegalovirus (HCMV) UL150 open reading frame(ORF) in low-passaged clinical isolates, and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection.Methods PCR was performed to amplify the entire HCMV UL150 ORF region of 29 clinical isolates, which hadbeen proven containing detectable HCMV-DNA using fluorescence quantitative PCR. PCR amplifcation products weresequenced directly, and the data were analyzed.Results Totally 25 among 29 isolates were amplified, and 18 isolates were sequenced successfully. HCMVUL150 ORF sequences derived from congenitally infected infants were high variability. The UL150 ORF in all 18 clinical isolates shifted backward by 8 nucleotides leading to frame-shift, and contained a single nucleotide deletion at nucleotide position 226 compared with that of Toledo strain. The nucleotide diversity was 0. 1% to 6. 8% and the amino acid diversity was 0. 2% to 19. 2% related to Toledo strain. However, the nucleotide diversity was 0. 1% to 6.4% and amino acid diversity was 0. 2% to 8.3% by compared with Merlin strain. Compared with Toledo, 4 new cysteine residues and 13 additional posttranslational modification sites were observed in UL150 putative proteins of clinical isolates. Moreover, the UL150 putative protein contained an additional transmembrane helix at position of 4-17 amino acid related to Toledo.Conclusion HCMV UL150 ORF and deduced amino acid sequences of clinical strains are hypervariability. No obvious linkage between the polymorphism and different pathogenesis of congenital HCMV infection is found.

  18. Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

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    Ronak Bakhtiari

    2011-10-01

    Full Text Available Resistance to b-lactam antibiotics by gramnegative bacteria, especially Escherichia coli (E. coli, is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly E. coli is via the production of extended spectrum beta lactamase (ESBLs enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk. Five hundred clinical samples were collected and 200 E. coli isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on E. coli isolates. A confirmation test (Combined Disk method was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64% E. coli isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8% isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.

  19. Meningoencefalite por Herpesvirus bovino 5 em Minas Gerais: relato de caso clínico Meningoencephalitis by Bovine herpesvirus 5 in Minas Gerais state: clinical case report

    Directory of Open Access Journals (Sweden)

    H.M. Aquino Neto

    2009-02-01

    Full Text Available Relata-se um caso de meningoencefalite causada por Herpesvirus bovino 5 (BoHV-5 heritabilityem uma vaca com cinco anos de idade. O animal manifestou quadro clínico inicial de síndrome medular baixa, caracterizada por incoordenação dos membros pélvicos, sinais estes ainda não descritos para a enfermidade. Dentro de pouco tempo a doença evoluiu para síndrome cerebral, e o óbito ocorreu seis dias após o inicio dos sintomas. Na histopatologia, evidenciou-se meningoencefalite difusa, não supurada, e a confirmação do diagnóstico foi feita por reação em cadeia de polimerase e sequenciamento do segmento parcial da glicoproteína G do vírus. O trabalho confirma a presença do BoHV-5 em Minas Gerais, descreve características clínicas novas para a enfermidade e ressalta sua importância no diagnóstico diferencial das neuropatias bovinas.A clinical case of meningoencephalitis by Bovine herpesvirus 5 (BoHV-5 in a five-year-old cow was reported. The disease began with low spinal cord signs, characterized by incoordination, and these symptoms had never been related to this illness before. Signs of a brain syndrome were observed and the cow died in six days. At the histopathology, a spread non-supurative meningoencephalitis was diagnosed, and the virus identification was made by PCR and partial sequence of the glycoprotein G. This study confirm the BoHV-5 presence in the State of Minas Gerais, Brazil, describes new clinic characteristics, and show the importance of the disease in the differentiate diagnosis with others bovine central nervous system affections.

  20. Class 1 integron and Imipenem Resistance in Clinical Isolates of Pseudomonas aeruginosa: Prevalence and Antibiotic Susceptibility

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    M Milani

    2010-12-01

    Full Text Available Background and Objectives: Pseudomonas aeruginosa is one of the most important causative agents of nosocomial infections especially in ICU and burn units. P. aeruginosa infections are normally difficult to eradicate due to acquired resistance to many antibiotics. Recent appearance of carbapenem resistant P. aeruginosa isolates is considered a major healthcare problem. The present study was conducted to detect class 1 integron and antibiotic susceptibility profiles of imipenem-sensitive and resistant clinical isolates of P. aeruginosa."nMaterials and Methods: Antibiotic susceptibility profiles and minimum inhibitory concentration against imipenem was studied in 160 clinical isolates of P. aeruginosa by disk agar diffusion method and Etest, respectively. Detection of class 1 integron was performed by the PCR method. Demographic and microbiological data were compared between imipenem susceptible and non-susceptible isolates by the SPSS software."nResults: PCR results showed that 90 (56.3% of P. aeruginosa isolates carried class 1 integron. Antibiotic susceptibility results revealed that 93 (58.1% were susceptible and 67 (41.9% were non-susceptible to imipenem. Comparison of antibiotic susceptibility patterns showed high level of drug resistance among imipenem non-susceptible isolates. We found that MDR phenotype, presence of class 1 integron and hospitalization in ICU and burn units were significantly associated with imipenem non-susceptible isolates."nConclusion: The high frequency of imipenem resistance was seen among our P. aeruginosa isolates. Since carbapenems are considered as the last drugs used for treatment of P. aeruginosa infections, it is crucial to screen imipenem non-susceptible isolates in infection control and optimal therapy.

  1. Protein-Based Classifier to Predict Conversion from Clinically Isolated Syndrome to Multiple Sclerosis.

    Science.gov (United States)

    Borràs, Eva; Cantó, Ester; Choi, Meena; Maria Villar, Luisa; Álvarez-Cermeño, José Carlos; Chiva, Cristina; Montalban, Xavier; Vitek, Olga; Comabella, Manuel; Sabidó, Eduard

    2016-01-01

    Multiple sclerosis is an inflammatory, demyelinating, and neurodegenerative disease of the central nervous system. In most patients, the disease initiates with an episode of neurological disturbance referred to as clinically isolated syndrome, but not all patients with this syndrome develop multiple sclerosis over time, and currently, there is no clinical test that can conclusively establish whether a patient with a clinically isolated syndrome will eventually develop clinically defined multiple sclerosis. Here, we took advantage of the capabilities of targeted mass spectrometry to establish a diagnostic molecular classifier with high sensitivity and specificity able to differentiate between clinically isolated syndrome patients with a high and a low risk of developing multiple sclerosis. Based on the combination of abundances of proteins chitinase 3-like 1 and ala-β-his-dipeptidase in cerebrospinal fluid, we built a statistical model able to assign to each patient a precise probability of conversion to clinically defined multiple sclerosis. Our results are of special relevance for patients affected by multiple sclerosis as early treatment can prevent brain damage and slow down the disease progression.

  2. Antimicrobial Susceptibility of Staphylococci Isolated from Clinical Mastitis in Dairy Cows in Two Regions of Kosovo

    Directory of Open Access Journals (Sweden)

    DRITON SYLEJMANI

    2015-12-01

    Full Text Available This study was undertaken to evaluate antibiotic susceptibility of 26 strains of Coagulase Negative Staphylococci (CNS and 18 strains of Staphylococcus aureus isolated from samples of dairy cows with clinical mastitis. The isolates species of staphylococci were identified by using coagulase test (using rabbit plasma and API Staph system (bioMėrieux SA which differentiates S. aurues from other staphylococci. Isolates were tested for antimicrobial susceptibility on Mueller Hinton agar by dick diffusion method according to the Clinical Laboratory Standards Institute (CLSI/NCCLS. The findings of this study showed that the most effective in vitro drugs for S. aureus strains isolated from cows with clinical mastitis were amoxycillin/clavulanic acid (83.3%, tetracycline (77.8%, gentamycin (77.8% and trimethoprim (66.7% and for CNS strains trimethoprim (88.5%, amoxicillin-clavulanic acid (84.6%, tetracycline (77% and gentamicin (69.2% while the isolates of S. aureus and CNS were found to be more resistant to penicillin (55.5% and 53.8%, respectively, streptomycin (50% and 46.1%, respectively and ampicillin (38.9% and 46.1%, respectively.

  3. Multilocus sequence typing of a global collection of Pasteurella multocida isolates from cattle and other host species demonstrates niche association

    Directory of Open Access Journals (Sweden)

    Lainson F Alex

    2011-05-01

    Full Text Available Abstract Background Pasteurella multocida causes disease in many host species throughout the world. In bovids, it contributes to bovine respiratory disease (BRD and causes haemorrhagic septicaemia (HS. Previous studies have suggested that BRD-associated P. multocida isolates are of limited diversity. A multilocus sequence typing (MLST scheme for P. multocida was used to determine whether the low levels of diversity reported are due to the limited discriminatory power of the typing method used, restricted sample selection or true niche association. Bovine respiratory isolates of P. multocida (n = 133 from the UK, the USA and France, collected between 1984 and 2008 from both healthy and clinically affected animals, were typed using MLST. Isolates of P. multocida from cases of HS, isolates from other host species and data from the MLST database were used as comparison. Results Bovine respiratory isolates were found to be clonal (ISA 0.45 with 105/128 belonging to clonal complex 13 (CC13. HS isolates were not related to bovine respiratory isolates. Of the host species studied, the majority had their own unique sequence types (STs, with few STs being shared across host species, although there was some cross over between porcine and bovine respiratory isolates. Avian, ovine and porcine isolates showed greater levels of diversity compared to cattle respiratory isolates, despite more limited geographic origins. Conclusions The homogeneity of STs of bovine respiratory P. multocida observed, and the differences between these and P. multocida subpopulations from bovine non-respiratory isolates and non-bovine hosts may indicate niche association.

  4. Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species

    Directory of Open Access Journals (Sweden)

    Danielly Beraldo dos Santos Silva

    2016-03-01

    Full Text Available The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51 which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candidaspecies known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G and two new nonsynonymous mutations in the isolates of Candida krusei - A497C (Y166S and G1570A (G524R. The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. kruseidemands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

  5. Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species.

    Science.gov (United States)

    Silva, Danielly Beraldo dos Santos; Rodrigues, Luana Mireli Carbonera; Almeida, Adriana Araújo de; Oliveira, Kelly Mari Pires de; Grisolia, Alexéia Barufatti

    2016-03-01

    The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candida species known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei--A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. krusei demands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates. PMID:26982177

  6. Slime production and antibiotic susceptibility in staphylococci isolated from clinical samples

    Directory of Open Access Journals (Sweden)

    Seza Arslan

    2007-02-01

    Full Text Available A total of 187 isolates from several clinical specimens were identified to species level as 129 Staphylococcus aureus strains and 58 coagulase-negative staphylococci (CNS strains by the API Staph System (Biomerieux. Slime production was detected both by the conventional Christensen's method as well as by the Congo red agar method. Seventy-two strains of staphylococci isolates (38.5% were found to be slime producers by Christensen's test tube method whereas 58 strains (31% were slime positive with Congo red agar method. There was no statistically significant difference between the two methods for the detection of slime production (P > 0.05. Susceptibility of isolates against antimicrobial agents was tested by the disk diffusion method. Staphylococcal species had resistance to one or more antibiotics. Among the various antimicrobial agents, oxacillin (71.1% and erythromycin (47.1% showed higher resistance than most of the agents used against all isolates. Oxacillin resistant S. aureus (ORSA and oxacillin resistant coagulase-negative staphylococci (ORCNS, 97 (75.2% and 36 (62.1% respectively were frequently observed in strains isolated from clinical materials. Among the ORSA strains, two strains were resistant to vancomycin. Moreover, 96 (74.4% of 129 S. aureus strains were positive for blactamase enzyme. However, 78 (81.25% of 96 b-lactamase positive S. aureus strains were b-lactamase positive ORSA isolates, but none of them had vancomycin resistance.

  7. Comparison of Virulence Gene Identification, Ribosomal Spacer PCR, and Pulsed-Field Gel Electrophoresis for Typing of Staphylococcus aureus Strains Isolated from Cases of Subclinical Bovine Mastitis in the United States.

    Science.gov (United States)

    Adkins, Pamela R F; Middleton, John R; Fox, Lawrence K

    2016-07-01

    Staphylococcus aureus is one of the most important pathogens causing contagious mastitis in dairy cattle worldwide. The objectives of this study were to determine if recently described S. aureus genotype B was present among previously characterized isolates from cases of bovine intramammary infection in the United States and to compare pulsed-field gel electrophoresis (PFGE) to the combination of ribosomal spacer PCR (RS-PCR) and virulence gene identification for typing of S. aureus strains. The hypothesis was that isolates that were previously characterized as contagious would be identified as genotype B and that the results of the two strain-typing methods would be comparable. Isolates were selected from a collection of S. aureus isolates from eight dairy farms. Mammary quarter milk somatic cell count (SCC) and N-acetyl-β-d-gluconaminidase (NAGase) activity data were known and used to evaluate strain pathogenicity. RS-PCR was performed with conventional gel electrophoresis, and PCR was used for toxin gene identification. RS-PCR patterns were associated with a specific virulence gene pattern, as previously reported. Five RS-PCR banding patterns were identified. None of the isolates were characterized as genotype B. No association between RS-PCR types and milk SCC was found; however, NAGase activity was significantly higher in milk from mammary glands infected with RS-PCR banding type 1 (RSP type 1) than in milk from those infected with RSP type 2. The discriminatory power values were 1.0 and 0.46 for PFGE and RS-PCR, respectively. These data suggest that genotype B may have a limited geographic distribution and that PFGE is more discriminatory than RS-PCR performed with conventional gel electrophoresis for typing of S. aureus isolates of bovine origin.

  8. Comparison of Virulence Gene Identification, Ribosomal Spacer PCR, and Pulsed-Field Gel Electrophoresis for Typing of Staphylococcus aureus Strains Isolated from Cases of Subclinical Bovine Mastitis in the United States.

    Science.gov (United States)

    Adkins, Pamela R F; Middleton, John R; Fox, Lawrence K

    2016-07-01

    Staphylococcus aureus is one of the most important pathogens causing contagious mastitis in dairy cattle worldwide. The objectives of this study were to determine if recently described S. aureus genotype B was present among previously characterized isolates from cases of bovine intramammary infection in the United States and to compare pulsed-field gel electrophoresis (PFGE) to the combination of ribosomal spacer PCR (RS-PCR) and virulence gene identification for typing of S. aureus strains. The hypothesis was that isolates that were previously characterized as contagious would be identified as genotype B and that the results of the two strain-typing methods would be comparable. Isolates were selected from a collection of S. aureus isolates from eight dairy farms. Mammary quarter milk somatic cell count (SCC) and N-acetyl-β-d-gluconaminidase (NAGase) activity data were known and used to evaluate strain pathogenicity. RS-PCR was performed with conventional gel electrophoresis, and PCR was used for toxin gene identification. RS-PCR patterns were associated with a specific virulence gene pattern, as previously reported. Five RS-PCR banding patterns were identified. None of the isolates were characterized as genotype B. No association between RS-PCR types and milk SCC was found; however, NAGase activity was significantly higher in milk from mammary glands infected with RS-PCR banding type 1 (RSP type 1) than in milk from those infected with RSP type 2. The discriminatory power values were 1.0 and 0.46 for PFGE and RS-PCR, respectively. These data suggest that genotype B may have a limited geographic distribution and that PFGE is more discriminatory than RS-PCR performed with conventional gel electrophoresis for typing of S. aureus isolates of bovine origin. PMID:27194685

  9. Absence of acetohydroxy acid synthetase in a clinical isolate of Neisseria gonorrhoeae requiring isoleucine and valine.

    OpenAIRE

    Lerner, S A; Friedman, E L; Dudek, E J; Kominski, G; Bohnhoff, M.; Morello, J A

    1980-01-01

    A clinical isolate of Neisseria gonorrhoeae with an unusual growth requirement for isoleucine and valine lacked the activity of acetohydroxy acid synthetase, one of the enzymes required for the biosynthesis of these amino acids. A spontaneous mutant which no longer required isoleucine and valine had acquired this enzymatic activity.

  10. Total Protein Profile and Drug Resistance in Candida albicans Isolated from Clinical Samples

    Directory of Open Access Journals (Sweden)

    Kamal Uddin Zaidi

    2016-01-01

    Full Text Available This study was done to assess the antifungal susceptibility of clinical isolates of Candida albicans and to evaluate its total protein profile based on morphological difference on drug resistance. Hundred and twenty clinical isolates of C. albicans from various clinical specimens were tested for susceptibility against four antifungal agents, namely, fluconazole, itraconazole, amphotericin B, and ketoconazole. A significant increase of drug resistance in clinical isolates of C. albicans was observed. The study showed 50% fluconazole and itraconazole resistance at 32 μg mL−1 with a MIC50 and MIC90 values at 34 and 47 and 36 and 49 μg mL−1, respectively. All isolates were sensitive to amphotericin B and ketoconazole. The SDS-PAGE protein profile showed a prevalent band of ~52.5 kDa, indicating overexpression of gene in 72% strains with fluconazole resistance. Since the opportunistic infections of Candida spp. are increasing along with drug resistance, the total protein profile will help in understanding the evolutionary changes in drug resistance and also to characterize them.

  11. Genome Sequence of the Historical Clinical Isolate Burkholderia pseudomallei PHLS 6.

    Science.gov (United States)

    D'haeseleer, Patrik; Johnson, Shannon L; Davenport, Karen W; Chain, Patrick S; Schoeniger, Joe; Ray, Debjit; Sinha, Anupama; Williams, Kelly P; Peña, José; Branda, Steven S; El-Etr, Sahar

    2016-01-01

    Here, we present the draft genome sequence of Burkholderia pseudomallei PHLS 6, a virulent clinical strain isolated from a melioidosis patient in Bangladesh in 1960. The draft genome consists of 39 contigs and is 7,322,181 bp long. PMID:27365360

  12. Correlation of MIC value and disk inhibition zone diameters in clinical Legionella pneumophila serogroup 1 isolates

    NARCIS (Netherlands)

    Bruin, J.P. de; Diederen, B.M.; Ijzerman, E.P.; Boer, J.W. den; Mouton, J.W.

    2013-01-01

    OBJECTIVES: Routine use of disk diffusion tests for detecting antibiotic resistance in Legionella pneumophila has not been described. The goal of this study was to determine the correlation of MIC values and inhibition zone diameter (MDcorr) in clinical L. pneumophila isolates. METHODS: Inhibition z

  13. Sensitivity patterns of pseudomonas aeruginosa isolates obtained from clinical specimens in peshawar

    International Nuclear Information System (INIS)

    Pseudomonas aeruginosa (P. aeruginosa) is a highly virulent opportunistic pathogen and a leading cause of nosocomial infections.Affected patients are often hospitalized in an intensive care unit, and are immuno-compromised as a result of disease and treatment. Suspected P. aeruginosa require timely, adequate and empirical antibiotic therapy to ensure improved outcomes. The purpose of the study was to find the sensitivity and resistance pattern of P. aeruginosa to various groups of drugs, in clinical isolates collected from two major tertiary care hospitals of Peshawar. Methods: Different clinical isolate were taken from patients admitted in various wards of Khyber Teaching Hospital and Lady Reading Hospital Peshawar. Results: A total of 258 clinical isolates were positive for P. aeruginosa out of 2058 clinical isolates. Pseudomonas showed high degree of resistance to third generation Cephalosporins (Ceftazidime, and Ceftriaxone) and moderate degree of resistance to Quinolones and Aminoglycosides (Ofloxacin, Ciprofloxacin, Levofloxacin and Amikacin). Low resistance was observed to different combinations (Cefoperazone + Sulbactum, Piperacillin + Tazobactum). Meropenem and Imipenem had negligible resistance. Conclusion: There is growing resistance to different classes of antibiotics. Combination drugs are useful approach for empirical treatment in suspected Pseudomonas infection. Imipenem and Meropenem are extremely effective but should be in reserve. (author)

  14. [A selective nutrient medium for isolating clinical strains of Escherichia coli O157:H7].

    Science.gov (United States)

    Sultanov, Z Z; Stepanova, E D; Kakulina, E A

    2000-01-01

    A dried selective culture medium, electrolyte-deficient sorbitol agar (EDS agar), for the isolation and preliminary identification of E. coli O157:H7 from clinical material has been developed. The medium is not inferior in its quality to analogous foreign media and requires no scarce ingredients for its manufacture.

  15. Emergence of NDM-1 and OXA-72 producing Acinetobacter pittii clinical isolates in Lebanon

    Directory of Open Access Journals (Sweden)

    A. Al Atrouni

    2016-07-01

    Full Text Available Acinetobacter spp. have emerged as global opportunistic pathogen causing a wide range of infections. Emergence of carbapenem resistance in these organisms is a matter of great concern. We report here the first detection of Acinetobacter pittii clinical isolates in Lebanon carrying either the blaNDM-1 or the blaOXA-72 gene.

  16. Emergence of VIM-2 metallo-beta-lactamase producing Ralstonia pickettii clinical isolate in India

    Directory of Open Access Journals (Sweden)

    A Khajuria

    2014-01-01

    Full Text Available A multidrug-resistant clinical isolate of Ralstonia pickettii from a woman was analysed. Modified Hodge test was positive for carbapenemase production. Conjugation experiment revealed the presence of conjugative plasmid of >140 Kb size typed as IncN type. This is the first report of emergence blaVIM-2 in R. pickettii in India.

  17. Detection of Genes for Superantigen Toxins in Methicillin-Resistant Staphylococcus aureus Clinical Isolates in Karachi

    International Nuclear Information System (INIS)

    Objective: To detect genes for enterotoxins, exfoliative and toxic shock syndrome toxins in Staphylococcus aureus (S. aureus) strains isolated from clinical specimens. Study Design: Cross-sectional observational study. Place and Duration of Study: Department of Molecular Genetics, Dr. Ziauddin Hospital, Karachi, from January to December 2010. Methodology: Two hundred and ninety eight S. aureus clinical isolates were obtained from various clinical samples received at Dr. Ziauddin Hospital, Karachi. Out of these, 115 were detected as methicillin resistant (MRSA) by cefoxitin disk diffusion test showing a prevalence rate of 38.6%. Detection of individual toxin genes was performed by Polymerase Chain Reaction (PCR) by using only one primer pair for each tube. Uniplex primers were preferred as multiplex primers are longer in base pairs and have the potential for cross reaction due to non-specific binding and increase in optimization time. Results: The possession of a single gene or more than a single gene in MRSA isolates was found in 61.73% of clinical samples; the highest number was found in pus swab, followed by sputum, blood, urethral swab, and urine. The prevalence of toxin genes was higher in MRSA as compared to methicillin sensitive (MSSA) isolates (19.12%). Conclusion: PCR detects strains possessing toxin genes independent of their expression. The possession of genes for super-antigens seems to be a frequent and habitual trait of S. aureus more so in MRSA. (author)

  18. Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy

    DEFF Research Database (Denmark)

    Fredborg, M; Rosenvinge, F S; Spillum, E;

    2015-01-01

    (168 antimicrobial agent-organism combinations) demonstrated 3.6 % minor, no major and 1.2 % very major errors of the oCelloScope system compared to conventional susceptibility testing, as well as a rapid and correct phenotypic detection of strains with methicillin-resistant Staphylococcus aureus (MRSA......Rapid antimicrobial susceptibility testing (AST) is essential for early and appropriate therapy. Methods with short detection time enabling same-day treatment optimisation are highly favourable. In this study, we evaluated the potential of a digital time-lapse microscope system, the o......CelloScope system, to perform rapid AST. The oCelloScope system demonstrated a very high accuracy (96 % overall agreement) when determining the resistance profiles of four reference strains, nine clinical isolates, including multi-drug-resistant isolates, and three positive blood cultures. AST of clinical isolates...

  19. Species distribution and antibiotic sensitivity pattern of coagulase negative staphylococci isolated from various clinical specimens

    Directory of Open Access Journals (Sweden)

    Mohan U

    2002-01-01

    Full Text Available A study of 192 strains of Coagulase negative staphylococcus (CONS showed that Staphylococcus epidermidis was the most common species, 158 (82.29% isolated from all clinical specimens followed by S. saprophyticus (30, 15.62% isolated mainly from urine. Slime production was exhibited by 77 (48.7% strains of S. epidermidis and 8 (26.6% of S. saprophyticus and the difference in the slime producing activity was statistically significant (p<0.005. Antibiotic susceptibility testing against 15 commonly used antibiotics showed multidrug resistance with more than 90% resistance to penicillin, more than 50% to cephalexin and ciprofloxacin and more than 20% to methicillin, thus, highlighting the importance of species identification and antibiotic susceptibility testing for clinical isolates of CONS.

  20. Isolation of a mutant MDBK cell line resistant to bovine viral diarrhea virus infection due to a block in viral entry.

    Science.gov (United States)

    Flores, E F; Donis, R O

    1995-04-20

    A cell line, termed CRIB, resistant to infection with bovine viral diarrhea virus (BVDV) has been derived from the MDBK bovine kidney cell line. CRIB cells were obtained by selection and cloning of cells surviving infection with a highly cytolytic BVDV strain. CRIB cells contain no detectable infectious or defective BVDV as ascertained by cocultivation, animal inoculation, indirect immunofluorescence, Western immunoblot, Northern hybridization, and RNA PCR. Inoculation of CRIB cells with 24 cytopathic and noncytopathic BVDV strains does not result in expression of viral genes or amplification of input virus. Karyotype and isoenzyme analyses demonstrated that CRIB are genuine bovine cells. CRIB cells are as susceptible as the parental MDBK cells to 10 other bovine viruses, indicating that these cells do not have a broad defect blocking viral replication. Transfection of CRIB cells with BVDV RNA or virus inoculation in the presence of polyethylene-glycol results in productive infection, indicating that the defect of CRIB cells is at the level of virus entry. CRIB cells are the first bovine cells reported to be resistant to BVDV infection in vitro and may be a useful tool for studying the early interactions of pestiviruses with host cells.

  1. Molecular screening of 246 Portuguese Aspergillus isolates among different clinical and environmental sources.

    Science.gov (United States)

    Sabino, Raquel; Veríssimo, Cristina; Parada, Helena; Brandão, João; Viegas, Carla; Carolino, Elisabete; Clemons, Karl V; Stevens, David A

    2014-07-01

    Clinical and environmental samples from Portugal were screened for the presence of Aspergillus and the distributions of the species complexes were determined in order to understand how their distributions differ based on their source. Fifty-seven Aspergillus isolates from clinical samples were collected from 10 health institutions. Six species complexes were detected by internal transcribed spacer sequencing; Fumigati, Flavi, and Nigri were found most frequently (50.9%, 21.0%, and 15.8%, respectively). β-tubulin and calmodulin sequencing resulted in seven cryptic species (A. awamorii, A. brasiliensis, A. fructus, A. lentulus, A. sydowii, A. tubingensis, Emericella echinulata) being identified among the 57 isolates. Thirty-nine isolates of Aspergillus were recovered from beach sand and poultry farms, 31 from swine farms, and 80 from hospital environments, for a total 189 isolates. Eleven species complexes were found in these 189 isolates, and those belonging to the Versicolores species complex were found most frequently (23.8%). There was a significant association between the different environmental sources and distribution of the species complexes; the hospital environment had greater variability of species complexes than other environmental locations. A high prevalence of cryptic species within the Circumdati complex was detected in several environments; from the isolates analyzed, at least four cryptic species were identified, most of them growing at 37ºC. Because Aspergillus species complexes have different susceptibilities to antifungals, knowing the species-complex epidemiology for each setting, as well as the identification of cryptic species among the collected clinical isolates, is important. This may allow preventive and corrective measures to be taken, which may result in decreased exposure to those organisms and a better prognosis.

  2. The isolation of Candida rugosa and Candida mesorugosa from clinical samples in Ghana.

    Science.gov (United States)

    Adjapong, Gloria; Bartlett, Michael; Hale, Marie; Garrill, Ashley

    2016-03-01

    Members of the Candida rugosa species complex have been described as emerging fungal pathogens and are responsible for a growing number of Candida infections. In this communication we report the isolation of Candida rugosa and Candida mesorugosa in Ghana. To the best of our knowledge this is the first description of this species complex from a clinical setting in Africa.The isolates were identified on the basis of their rRNA gene internal transcribed spacer (ITS) sequences. For one isolate, obtained from sputum, the sequence grouped well with that of C. rugosa. Two other isolates from urine had sequences that grouped with Candida mesorugosa. Morphologically, C. rugosa formed white, wrinkled, and flat colonies on Sabouraud Dextrose Agar (SDA), whereas C. mesorugosa formed white, smooth colonies. On chromogenic medium, the isolates formed small, dry greenish-blue colonies with a pale or white border, similar to C. albicans. The C. rugosa isolate produced pseudohyphae in human serum and on CMA-Tween 80 agar. In contrast, the C. mesorugosa isolates did not generate pseudohyphae in human serum, but generated a few pseudohyphae with abundant blastoconidia on CMA-Tween 80 agar. Growth was observed at 37 °C and 42 °C but not at 45 °C.The two C. mesorugosa isolates had Minimum Inhibitory Concentrations (MICs) of 6 and 48 μg ml(-1) for fluconazole and are thus resistant. The C. rugosa isolate had an MIC of 24 μg ml(-1), indicative of resistance. All three isolates were susceptible to itraconazole and voriconazole (with respective MICs of < 0.125 μg ml(-1)).

  3. Analysis of pulsotypes of salmonella typhi isolates and their clinical profiles in typhoid fever patients

    Directory of Open Access Journals (Sweden)

    Retno K. Soemanto

    2003-03-01

    Full Text Available A study of genotyping (pulsotyping of Salmonella typhi (S. typhi isolates using pulse-field gel electrophoresis (PFGE methods was performed to examine their genetic diversity, and relationship between genetic characteristics and clinical outcomes.  Sixty-six S. typhi isolates obtained from sporadic hospitalized typhoid fever cases were used in this study. Four isolates were found identical and the dendogram constructed showed 33 pulsotypes in which 13 of them can be divided into 30 subtypes. Diversity among them were high as shown by the Dice coefficients that ranged from 0.486 to 1.000. Cluster analysis showed 2 main clusters with 65% degree of similarity, suggested that they were not originated from one clone. Further, at 90% degree of similarity, 9 clusters containing at least 3 isolates were determined to explore any possible existence of relationship between genetic profile and particular clinical outcomes. Clinical manifestations ranged from mild to severe were in fact distributed diversely among these clusters. Although the clinical data obtained were incomplete, 2 out of 4 patients infected by the S. typhi belonged to cluster 1 showed an elevation of total bilirubin, whereas it was not found in 19 other patients distributed in other 8 clusters. Even though specific clinical manifestations were apparently not found to relate with particular clusters of genotypes, S. typhi isolates grouped in cluster 1 seemed to show trophism to hepatobiliary system. (Med J Indones 2003; 12: 13-20 Keywords: S. typhi, typhoid fever, Pulsed-field Gel electrophoresis (PFGE

  4. Biofilm formation and genetic diversity of Salmonella isolates recovered from clinical, food, poultry and environmental sources.

    Science.gov (United States)

    Nair, Amruta; Rawool, Deepak B; Doijad, Swapnil; Poharkar, Krupali; Mohan, Vysakh; Barbuddhe, Sukhadeo B; Kolhe, Rahul; Kurkure, Nitin V; Kumar, Ashok; Malik, S V S; Balasaravanan, T

    2015-12-01

    In the present study, Salmonella isolates (n=40) recovered from clinical, food, poultry and environmental sources were characterized for serotype identification, genetic diversity and biofilm formation capability. Serotype identification using multiplex PCR assay revealed six isolates to be Salmonella Typhimurium, 14 as Salmonella Enteritidis, 11 as Salmonella Typhi, and the remaining nine isolates unidentified were considered as other Salmonella spp. Most of the Salmonella isolates (85%) produced biofilm on polystyrene surfaces as assessed by microtitre plate assay. About 67.5% isolates were weak biofilm producers and 17.5% were moderate biofilm producers. There was no significant difference in biofilm-forming ability among the Salmonella isolates recovered from different geographical regions or different sources. Among the genetic methods, Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR revealed greater discriminatory power (DI, 0.943) followed by pulsed field gel electrophoresis (PFGE) (DI, 0.899) and random amplification of polymorphic DNA (RAPD) PCR (DI, 0.873). However, composite analysis revealed the highest discrimination index (0.957). Greater discrimination of S. Typhimurium and S. Typhi was achieved using PFGE, while ERIC PCR was better for S. Enteritidis and other Salmonella serotypes. A strong positive correlation (r=0.992) was observed between biofilm formation trait and clustered Salmonella isolates in composite genetic analysis.

  5. Cdr2p contributes to fluconazole resistance in Candida dubliniensis clinical isolates.

    LENUS (Irish Health Repository)

    2011-05-01

    The development of resistance to azole antifungals used in the treatment of fungal infections can be a serious medical problem. Here, we investigate the molecular mechanisms associated with reduced susceptibility to fluconazole in clinical isolates of Candida dubliniensis , showing evidence of the trailing growth phenomenon. The changes in membrane sterol composition were studied in the presence of subinhibitory fluconazole concentrations. Despite lanosterol and eburicol accumulating as the most prevalent sterols after fluconazole treatment, these ergosterol precursors still support growth of Candida isolates. The overexpression of ABC transporters was demonstrated by immunoblotting employing specific antibodies against Cdr1p and Cdr2p. The presence of a full-length 170 kDa protein Cdr1p was detected in two isolates, while a truncated form of Cdr1p with the molecular mass of 85 kDa was observed in isolate 966\\/3(2). Notably, Cdr2p was detected in this isolate, and the expression of this transporter was modulated by subinhibitory concentrations of fluconazole. These results suggest that C. dubliniensis can display the trailing growth phenomenon, and such isolates express similar molecular mechanisms like that of fluconazole-resistant isolates and can therefore be associated with recurrent infections.

  6. SEQUENCE VARIABILITY OF HUMAN CYTOMEGALOVIRUS UL144 OPEN READING FRAME IN LOW-PASSAGE CLINICAL ISOLATES

    Institute of Scientific and Technical Information of China (English)

    Rong He; Yao-hua Ji; Qiang Ruan; Chang Xia; Lan-qing Liu; Sheng-min Lü; Ying Lu; Ying Qi; Yan-ping Ma; Qing Liu

    2004-01-01

    Objective To explore the relationship between human cytomegalovirus (HCMV) UL144 sequence variability and clinical disease.Methods HCMV UL144 open reading frame (ORF) was amplified by PCR assay in 72 lowpassage isolates [65 congenitally infective children and 7 healthy children who were HCMV-DNA positive by quantitative PCR (qPCR)]. All positive PCR products were analyzed by heteroduplex mobility assay and single-stranded conformation polymorphism (HMA-SSCP) and 32 of them were sequenced.Resuits Fifty-five patient isolates and five healthy children isolates were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in the UL144 ORF. Phylogenetic analysis indicated that the nucleotide sequences could be separated into 3 major genotypes. Comparing between UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. And genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively.Conclusions HCMV-UL144 existed in most of low passage isolates and sequences were hypervariable. The UL144ORF and its predicted product with the high level of sequence variability in different kinds of isolates suggest that UL144ORF might play a role in HCMV infectivity and subsequent diseases.

  7. Bovine immunoglobulin/protein isolate binds pro-inflammatory bacterial compounds and prevents immune activation in an intestinal co-culture model.

    Science.gov (United States)

    Detzel, Christopher J; Horgan, Alan; Henderson, Abigail L; Petschow, Bryon W; Warner, Christopher D; Maas, Kenneth J; Weaver, Eric M

    2015-01-01

    Intestinal barrier dysfunction is associated with chronic gastrointestinal tract inflammation and diseases such as IBD and IBS. Serum-derived bovine immunoglobulin/protein isolate (SBI) is a specially formulated protein preparation (>90%) for oral administration. The composition of SBI is greater than 60% immunoglobulin including contributions from IgG, IgA, and IgM. Immunoglobulin within the lumen of the gut has been recognized to have anti-inflammatory properties and is involved in maintaining gut homeostasis. The binding of common intestinal antigens (LPS and Lipid A) and the ligand Pam3CSK4, by IgG, IgA, and IgM in SBI was shown using a modified ELISA technique. Each of these antigens stimulated IL-8 and TNF-α cytokine production by THP-1 monocytes. Immune exclusion occurred as SBI (≤50 mg/mL) bound free antigen in a dose dependent manner that inhibited cytokine production by THP-1 monocytes in response to 10 ng/mL LPS or 200 ng/mL Lipid A. Conversely, Pam3CSK4 stimulation of THP-1 monocytes was unaffected by SBI/antigen binding. A co-culture model of the intestinal epithelium consisted of a C2BBe1 monolayer separating an apical compartment from a basal compartment containing THP-1 monocytes. The C2BBe1 monolayer was permeabilized with dimethyl palmitoyl ammonio propanesulfonate (PPS) to simulate a damaged epithelial barrier. Results indicate that Pam3CSK4 was able to translocate across the PPS-damaged C2BBe1 monolayer. However, binding of Pam3CSK4 by immunoglobulins in SBI prevented Pam3CSK4 translocation across the damaged C2BBe1 barrier. These results demonstrated steric exclusion of antigen by SBI which prevented apical to basal translocation of antigen due to changes in the physical properties of Pam3CSK4, most likely as a result of immunoglobulin binding. This study demonstrates that immunoglobulins in SBI can reduce antigen-associated inflammation through immune and steric exclusion mechanisms and furthers the mechanistic understanding of how SBI

  8. Biofilm formation in clinical isolates of nosocomial Acinetobacter baumannii and its relationship with multidrug resistance

    Institute of Scientific and Technical Information of China (English)

    Ebrahim Babapour; Azam Haddadi; Reza Mirnejad; Seyed-Abdolhamid Angaji; Nour Amirmozafari

    2016-01-01

    Objective: To check biofilm formation by Acinetobacter baumannii (A. baumannii) clinical isolates and show their susceptibility to different antibiotics and investigate a possible link between establishment of biofilm and multidrug resistance. Methods: This study was performed on clinical samples collected from patients with nosocomial infections in three hospitals of Tehran. Samples were initially screened by culture and biochemical tests for the presence of different species of Acinetobacter. Iden-tifications were further confirmed by PCR assays. Their susceptibilities to 11 antibiotics of different classes were determined by disc diffusion method according to Clinical and Laboratory Standards Institute guidelines. The ability to produce biofilm was investigated using methods:culture on Congo red agar, microtiter plate, and test tube method. Results: From the overall clinical samples, 156 specimens were confirmed to contain A. baumannii. The bacteria were highly resistant to most antibiotics except polymyxin B. Of these isolates, 10.26% were able to produce biofilms as shown on Congo red agar. However, the percentage of bacteria with positive biofilm in test tube, standard microtiter plate, and modified microtiter plate assays were 48.72%, 66.66%, and 73.72%, respec-tively. At least 92%of the biofilm forming isolates were multidrug resistant. Conclusions: Since most of the multidrug resistant strains produce biofilm, it seems necessary to provide continuous monitoring and determination of antibiotic susceptibility of clinical A. baumannii. This would help to select the most appropriate antibiotic for treatment.

  9. Application of MALDI-TOF MS for requalification of a Candida clinical isolates culture collection

    Directory of Open Access Journals (Sweden)

    Reginaldo Lima-Neto

    2014-06-01

    Full Text Available Microbial culture collections underpin biotechnology applications and are important resources for clinical microbiology by supplying reference strains and/or performing microbial identifications as a service. Proteomic profiles by MALDI-TOF MS have been used for Candida spp. identification in clinical laboratories and demonstrated to be a fast and reliable technique for the routine identification of pathogenic yeasts. The main aim of this study was to apply MALDI-TOF MS combined with classical phenotypic and molecular approaches to identify Candida clinical isolates preserved from 1 up to 52 years in a Brazilian culture collection and assess its value for the identification of yeasts preserved in this type of collections. Forty Candida spp. clinical isolates were identified by morphological and biochemical analyses. Identifications were also performed by the new proteomic approach based on MALDI-TOF MS. Results demonstrated 15% discordance when compared with morphological and biochemical analyses. Discordant isolates were analysed by ITS sequencing, which confirmed the MALDI-TOF MS identifications and these strains were renamed in the culture collection catalogue. In conclusion, proteomic profiles by MALDI-TOF MS represents a rapid and reliable method for identifying clinical Candida species preserved in culture collections and may present clear benefits when compared with the performance of existing daily routine methods applied at health centres and hospitals.

  10. Bacteremia with Streptococcus bovis and Streptococcus salivarius: clinical correlates of more accurate identification of isolates.

    Science.gov (United States)

    Ruoff, K L; Miller, S I; Garner, C V; Ferraro, M J; Calderwood, S B

    1989-01-01

    Two biotypes of Streptococcus bovis can be identified by laboratory testing and can be distinguished from the phenotypically similar organism Streptococcus salivarius. We assessed the clinical relevance of careful identification of these organisms in 68 patients with streptococcal bacteremia caused by these similar species. S. bovis was more likely to be clinically significant when isolated from blood (89%) than was S. salivarius (23%). There was a striking association between S. bovis I bacteremia and underlying endocarditis (94%) compared with that of S. bovis II bacteremia (18%). Bacteremia with S. bovis I was also highly correlated with an underlying colonic neoplasm (71% of patients overall, 100% of those with thorough colonic examinations) compared with bacteremia due to S. bovis II or S. salivarius (17% overall, 25% of patients with thorough colonic examinations). We conclude that careful identification of streptococcal bacteremic isolates as S. bovis biotype I provides clinically important information and should be more widely applied. PMID:2915024

  11. Mesenchymal Stem Cells Isolated from Adipose and Other Tissues: Basic Biological Properties and Clinical Applications

    Directory of Open Access Journals (Sweden)

    Hakan Orbay

    2012-01-01

    Full Text Available Mesenchymal stem cells (MSCs are adult stem cells that were initially isolated from bone marrow. However, subsequent research has shown that other adult tissues also contain MSCs. MSCs originate from mesenchyme, which is embryonic tissue derived from the mesoderm. These cells actively proliferate, giving rise to new cells in some tissues, but remain quiescent in others. MSCs are capable of differentiating into multiple cell types including adipocytes, chondrocytes, osteocytes, and cardiomyocytes. Isolation and induction of these cells could provide a new therapeutic tool for replacing damaged or lost adult tissues. However, the biological properties and use of stem cells in a clinical setting must be well established before significant clinical benefits are obtained. This paper summarizes data on the biological properties of MSCs and discusses current and potential clinical applications.

  12. STUDIES ON ANTIBACTERIAL EFFECT OF APAMARGA (ACHYRANTHES ASPERA ON MULTI-DRUG RESISTANT CLINICAL ISOLATES

    Directory of Open Access Journals (Sweden)

    Patil Usha

    2013-04-01

    Full Text Available Recent reports on emergence of multidrug resistant bacteria are cause of concern in medical world. Several ayurvedic drugs have been proved to contain the antimicrobial activity. Literature on effect of ayurvedic drugs on multidrug resistant bacterial pathogens is limited. Present study reports the antimicrobial effect of Achyranthes aspera (Apamarga crude extracts on the clinical isolates of multidrug resistant bacteria. The drug was evaluated by using phytochemical tests. Crude extracts of aqueous, methanol, ethanol and chloroform was prepared. Antibacterial activity against clinically isolated multidrug resistant bacteria belonging to groups of bacillus, citrobacter, E.coli, klebsiella, proteus and salmonella was tested. The drug showed highest efficacy against Bacillus organism while least effectiveness on Proteus spp bacteria. Results of the study conclude that the medicinal plant A. aspera might be useful against multidrug resistance in pathogens of clinical importance.

  13. Effects of early treatment with glatiramer acetate in patients with clinically isolated syndrome

    DEFF Research Database (Denmark)

    Comi, G.; Martinelli, V.; Rodegher, M.;

    2013-01-01

    Background: The placebo-controlled phase of the PreCISe study showed that glatiramer acetate delayed onset of clinically definite multiple sclerosis (CDMS) in patients with clinically isolated syndrome and brain lesions on MRI. Objective: To compare the effects of early versus delayed glatiramer...... acetate treatment in the open-label phase of PreCISe. Methods: Patients with a clinically isolated syndrome suggestive of MS with unifocal manifestation and 2 T2-weighted brain lesions were randomized to receive glatiramer acetate 20 mg/d (early-treatment, n=198) or placebo (delayed-treatment, n=211......) for 36 months or until conversion to CDMS, followed by open-label glatiramer acetate treatment for two years. Results: Early glatiramer acetate treatment reduced CDMS conversion risk by 41% (hazard ratio 0.59, 95% confidence interval 0.44-0.80; p=0.0005) versus delayed-treatment, and was associated...

  14. Isolation and identification of a bovine viral diarrhea virus isolate from yak in Qinghai Province%青海牦牛病毒性腹泻病毒的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    张芳芳; 王光华; 郑福英; 宫晓炜; 周继章; 吴润; 邱昌庆

    2011-01-01

    采集青海省泽库县某牧户疑似牦牛病毒性腹泻病牛的样品,将RT-PCR检测为阳性的样品处理后接种MDBK细胞,并盲传至第10代,检测每一代细胞中牛病毒性腹泻病毒的E0基因。通过分离株接种细胞后产生的病变效应观察、免疫荧光试验、电镜观察、RT-PCR扩增以及序列分析鉴定了该病毒。结果表明,盲传的每代细胞中均可检测到E0基因;接毒后的细胞在盲传至第7代时出现明显的细胞病变;免疫荧光试验中能观察到细胞内发出的特异性荧光;经浓缩、纯化的病毒在电镜下呈直径为40~60nm的病毒粒子,将其命名为QHZK株。对克隆的E0基因测序后提交至GenBank(登录号:JF927789),并将获得的序列与其他国内外分离株的E0序列比对,同源性为73.6%~98.2%;系统进化分析表明该分离株属于牛病毒性腹泻病毒1b亚型。%Samples were collected from yak with suspected bovine viral diarrhea on a yak farm in Zeku County,Qinghai Province.MDBK cell lines were inoculated with RT-PCR positive specimens,followed by blind passage culture for 10 generations,and the E0 gene of bovine viral diarrhea virus(BVDV) was detected from the cell cultures of each generation.The isolates were cultured in MDBK cell lines,and identified by cytopathic effect(CPE),direct immunofluorescence assay(DIFA),electron microscopy(EM),RT-PCR and sequence analysis. The results indicated that the E0 gene was detected from the cell cultures of each generation,and typical CPE was observed from cell cultures of the seventh generation.Specific fluorescence in cell cultures can be see in DIFA.Virions of 40-60nm were observed under EM,and was named QHZK strain.The obtained E0 gene sequence was submitted to GenBank with the accession number:JF927789.Alignment with other 9 strains of BVDV available in GenBank showed a homology of 73.6%-98.2% for nucleotide sequence.The phylogenetic analysis indicated that the isolated

  15. Molecular characterisation and antifungal susceptibility of clinical Trichosporon isolates in India.

    Science.gov (United States)

    Rastogi, Vijaylatha; Honnavar, Prasanna; Rudramurthy, Shivaprakash M; Pamidi, Umabala; Ghosh, Anup; Chakrabarti, Arunaloke

    2016-08-01

    In Asian countries, Trichosporon infection is a well-known disease in Japan. In India, the infection is increasingly recognised. The study was conducted to characterise the clinical Trichosporon isolates from India by phenotypic and molecular techniques. A total of 31 Trichosporon clinical isolates, recovered from patients of 14 hospitals across India were sequenced (ITS and IGS1 regions of rDNA). In vitro drug susceptibility testing of the isolates was performed against amphotericin-B, fluconazole, itraconazole, voriconazole and posaconazole. IGS1, rather than ITS sequences, correctly identified the isolates: Trichosporon asahii, 20; Trichosporon ovoides, 3; Trichosporon inkin, 2; Trichosporon asteroides, 1; Trichosporon mucoides, 1; Trichosporon loubieri, 1; Trichosporon debeurmannianum, 1; and Trichosporon dermatis, 1. Trichosporon asahii genotype III was the most common type, followed by genotype I and VII. Both these targets did not help to identify one Trichosporon to the species level. Trichosporon debeurmannianum, T. dermatis and T. asteroides were isolated for the first time from a human disease in India. The minimum inhibitory concentrations for voriconazole and posaconazole were within effective range. The study highlights the presence of wide range of Trichosporon species causing infection in India. Voriconazole or posaconazole may be the better drugs to treat such patients. PMID:27144725

  16. Antibacterial Activity of Rhodomyrtus tomentosa (Aiton Hassk. Leaf Extract against Clinical Isolates of Streptococcus pyogenes

    Directory of Open Access Journals (Sweden)

    Surasak Limsuwan

    2012-01-01

    Full Text Available Ethanol extract of Rhodomyrtus tomentosa (Aiton Hassk. leaf was evaluated for antibacterial activity against 47 clinical isolates of Streptococcus pyogenes. The extract exhibited good anti-S. pyogenes activity against all the tested isolates with similar minimum inhibitory concentration (MIC, 3.91–62.5 μg mL−1 and minimum bactericidal concentration (MBC, 3.91–62.5 μg mL−1 ranges. No surviving cells were detected at 16 h after treatment with 8 × MIC of the extract. The extract-treated cells demonstrated no lysis and cytoplasmic leakage through the bacterial membrane. Electron micrographs further revealed that the extract did not cause any dramatic changes on the treated cells. Rhodomyrtone, an isolated compound, exhibited good anti-S. pyogenes activity (14 isolates, expressed very low MIC (0.39–1.56 μg mL−1 and MBC (0.39-1.56 μg mL−1 values. Rhodomyrtus tomentosa leaf extract and rhodomyrtone displayed promising antibacterial activity against clinical isolates of S. pyogenes.

  17. Genotypic characterization, invasion index and antimicrobial resistance pattern in Listeria monocytogenes strains isolated from clinical samples

    Institute of Scientific and Technical Information of China (English)

    Behrooz Sadeghi Kalani; Abazar Pournajaf; Mansour Sedighi; Abbas Bahador; Gholamreza Irajian; Firuzeh Valian

    2015-01-01

    Objective: To evaluate antimicrobial resistance, invasion index and genetic profile in Listeriamonocytogenes isolated from clinical samples. Methods: At all, 170 clinical samples were collected from patients with spontaneous abortions hospitalized in Shariati hospital in Tehran during June 2010 to August 2013. Invasion index was determined using HeLa cells. The multiple-locus variable-number tandem-repeats analysis (MLVA) was used for evaluation of genetic relatedness. Results: Out of 14 L. monocytogenes isolates, 4 (28.57%), 2 (14.28%), 0 (0%), 5 (35.71%) and 3 (21.42%) were isolated from placental tissue, urine, blood, vaginal and rectal swabs, respectively. High resistance to penicillin and multidrug resistant were found amongst isolates. The invasion index was in the range of 0.001-0.007. Seven different types were obtained by MLVA assay and type 2 and 3 with 4 strains were the most frequent type. Strains isolated from the vagina and the placenta of the same type were also more resistant to penicillin. Conclusions: Since MLVA is a high-throughput screening method that is fairly inexpensive, easy to accomplish, rapid, and trustworthy, it is well suited to interlaboratory comparisons during epidemiological investigations. Also further studies of larger samples from a variety of sources such as food and animal specimens recommended comparing by MLVA method.

  18. Molecular typing of clinical isolates of Cryptococcus neoformans/Cryptococcus gattii species complex from Northeast Mexico.

    Science.gov (United States)

    González, Gloria M; Casillas-Vega, Néstor; Garza-González, Elvira; Hernández-Bello, Romel; Rivera, Gildardo; Rodríguez, Jesús Ancer; Bocanegra-Garcia, Virgilio

    2016-01-01

    Cryptococcosis is caused by members of the Cryptococcus neoformans/Cryptococcus gattii species complex. Based on molecular identification, these two species have been further differentiated into molecular types. The aim of this work was to characterize clinical cryptococcal isolates recovered from six hospitals in Northeast Mexico from 1995 to 2011. One hundred and sixty-six isolates, which were characterized by biochemical tests and in vitro susceptibility to amphotericin B, fluconazole, and voriconazole, and M13 PCR fingerprinting, were included in this study. Utilizing phenotypic tests, 153 isolates (92.16 %) were identified as C. neoformans and 13 (7.83 %) as C. gattii. All isolates were susceptible to all antifungals tested. Employing M13 PCR fingerprinting, eight molecular types were detected. VNI was the most common genotype (124 cases; 74.6 %), followed by VNII (15 cases; 9 %), VNIII (8 cases; 4.8 %), VNIV (6 cases; 3.6 %), VGI (6 cases; 3.6 %), VGII (3 cases; 1.8 %), and VGIII and VGIV (2 cases, 1.2 % each). We confirm the presence of C. gattii in clinical isolates in Northeast Mexico, and a high clonal diversity in the studied strains of C. neoformans/C. gattii species complex.

  19. In vitro drug susceptibility of 2275 clinical non-tuberculous Mycobacterium isolates of 49 species in The Netherlands.

    NARCIS (Netherlands)

    Ingen, J. van; Laan, T. van der; Dekhuijzen, R.; Boeree, M.J.; Soolingen, D. van

    2010-01-01

    In this study, 2275 clinical isolates of 49 species of non-tuberculous mycobacteria isolated in The Netherlands were subjected to standardised drug susceptibility testing using the Middlebrook 7H10 agar dilution method. Clarithromycin and rifabutin were most active, with 87% and 83% of all isolates,

  20. Exploring the contribution of efflux on the resistance to fluoroquinolones in clinical isolates of Staphylococcus aureus

    LENUS (Irish Health Repository)

    Costa, Sofia SANTOS

    2011-10-27

    Abstract Background Antimicrobial resistance mediated by efflux systems is still poorly characterized in Staphylococcus aureus, despite the description of several efflux pumps (EPs) for this bacterium. In this work we used several methodologies to characterize the efflux activity of 52 S. aureus isolates resistant to ciprofloxacin collected in a hospital in Lisbon, Portugal, in order to understand the role played by these systems in the resistance to fluoroquinolones. Results Augmented efflux activity was detected in 12 out of 52 isolates and correlated with increased resistance to fluoroquinolones. Addition of efflux inhibitors did not result in the full reversion of the fluoroquinolone resistance phenotype, yet it implied a significant decrease in the resistance levels, regardless of the type(s) of mutation(s) found in the quinolone-resistance determining region of grlA and gyrA genes, which accounted for the remaining resistance that was not efflux-mediated. Expression analysis of the genes coding for the main efflux pumps revealed increased expression only in the presence of inducing agents. Moreover, it showed that not only different substrates can trigger expression of different EP genes, but also that the same substrate can promote a variable response, according to its concentration. We also found isolates belonging to the same clonal type that showed different responses towards drug exposure, thus evidencing that highly related clinical isolates may diverge in the efflux-mediated response to noxious agents. The data gathered by real-time fluorometric and RT-qPCR assays suggest that S. aureus clinical isolates may be primed to efflux antimicrobial compounds. Conclusions The results obtained in this work do not exclude the importance of mutations in resistance to fluoroquinolones in S. aureus, yet they underline the contribution of efflux systems for the emergence of high-level resistance. All together, the results presented in this study show the potential

  1. Complete Genome Sequence of a Bovine Viral Diarrhea Virus 2 from Commercial Fetal Bovine Serum

    OpenAIRE

    Liu, Hua; Li, Yan; Gao, Mingchun; Wen, Kai; Jia, Ying; Liu, Xiaomei; Zhang, Wenlong; Ma, Bo; Wang, Junwei

    2012-01-01

    We isolated a bovine viral diarrhea virus (BVDV) from commercial fetal bovine serum and designated it HLJ-10. The complete genome is 12,284 nucleotides (nt); the open reading frame is 11,694 nt, coding 3,898 amino acids. Phylogenetic analysis indicated that this strain belongs to BVDV group 2.

  2. Antibacterial activity of mupirocin (pseudomonic Acid A) against, clinical isolates of methicillin-resistant staphylococcus aureus

    International Nuclear Information System (INIS)

    Colonized patients and health care workers are the main source of spread of methicillin resistant Staphylococcus aureus (MRSA) in hospitals. The elimination of nasal colonized MRSA plays a crucial role in infection control protocols. Mupirocin (pseudomonic acid A) is used for eradication of MRSA nasal carriage. Increasing use of pseudomonic acid A (mupirocin) has led to emergence of resistance. Objective To determine low and high level resistance of MRSA isolates from clinical specimens against mupirocin. Place and duration of study: Study was conducted at Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi from July 2006 to June 2007. Material and methods Three hundred methicillin-resistant Staphylococcus aureus isolates were studied. All clinical specimens were processed for culture and sensitivity. Staphylococcus aureus isolates were tested for methicillin resistance using 1 micro g oxacillin disk. The isolates were further tested by PCR for the presence of mecA gene. Minimum Inhibitory Concentration (MIC) of mupirocin against MRSA isolates was determined using agar dilution technique. Results Out of 300 MRSA isolates, 98% were found to have MlC against mupirocin as smaller than 4 micro g/mL. Remaining 2% isolates revealed low level resistance (MIC greater than 8 micro g/mL to 256 micro g/mL), no high level resistance (MIC greater than 512 micro g/mL) against mupirocin was detected. Conclusions: High level mupirocin resistance has not emerged so far in our setup. Due to increasing use of mupirocin, emergence of resistance against mupirocin among MRSA is a strong possibility. Strategy encompassing rational use of antimicrobials, hospital infection control, surveillance for the detection of mupirocin resistance and judicious use of this agent is required. (author)

  3. Variation of Slime Production and Ultrastructural Analysis in Clinical and Soil Isolates of Candida parapsilosis

    OpenAIRE

    Eugenio, Pontieri; Roberta, Sferra; Antonella, Vetuschi; Eugenio, Gaudio; Giuseppe, Carruba

    2011-01-01

    The slime production of 46 isolates of Candida parapsilosis from clinic (blood and vaginal) and soil sources was investigated. The cells were grown in liquid broth, testing several concentrations of glucose ranging from 0.9% (w/v) up to 20% in polystyrene tubes, and the presence of biofilm was checked by visual inspection of the internal wall of the tubes, scoring being: 0, no production; 1, weak; 2, moderate; 3, strong production. Each isolate had an identical slime value at the glucose conc...

  4. Helicobacter pylori HopE and HopV porins present scarce expression among clinical isolates

    Institute of Scientific and Technical Information of China (English)

    Maritza; Lienlaf; Juan; Pablo; Morales; María; Inés; Díaz; Rodrigo; Díaz; Elsa; Bruce; Freddy; Siegel; Gloria; León; Paul; R; Harris; Alejandro; Venegas

    2010-01-01

    AIM:To evaluate how widely Helicobacter pylori (H. pylori ) HopE and HopV porins are expressed among Chilean isolates and how seroprevalent they are among infected patients in Chile.METHODS: H. pylori hopE and hopV genes derived from strain CHCTX-1 were cloned by polymerase chain reaction (PCR), sequenced and expressed in Escherichia coli AD494 (DE3). Gel-purified porins were used to prepare polyclonal antibodies. The presence of both genes was tested by PCR in a collection of H. pylori clinical isolates an...

  5. Microbiologic characterization of isolates from a dalbavancin clinical trial for catheter-related bloodstream infections.

    Science.gov (United States)

    Goldstein, Beth P; Jones, Ronald N; Fritsche, Thomas R; Biedenbach, Douglas J

    2006-02-01

    Dalbavancin, a new-generation semisynthetic lipoglycopeptide in phase 3 clinical development, has been documented to be more active than vancomycin or teicoplanin against Gram-positive bacteria, including multidrug-resistant strains, by in vitro testing and in animal models. The human pharmacokinetics of dalbavancin predicts efficacy at weekly dosing intervals. In a phase 2 open-label clinical trial, dalbavancin exhibited superiority when compared with vancomycin against catheter-related bloodstream infection (CR-BSI). The majority of pathogens identified in this study as in clinical practice were coagulase-negative staphylococci (CoNS), necessitating rigorous characterization of duplicate isolates to rule out contaminants and to validate cases for study evaluations. At follow-up for the intent-to-treat population, overall pathogen eradication was 92.3% for dalbavancin and 75.9% for vancomycin. We describe the details of organisms isolated, their epidemiologic/genetic characterization, susceptibility patterns against glycopeptides, and the eradication rates by organism group. In conclusion, dalbavancin was active against all isolated pathogens associated with CR-BSI (CoNS, Staphylococcus aureus and Enterococcus faecalis; all MIC results, < or = 0.25 microg/mL) and achieved significant (P < 0.05) clinical success when compared with vancomycin. PMID:16458124

  6. Evaluation of Sialic Acid and Acute Phase Proteins (Haptoglobin and Serum Amyloid A) in Clinical and Subclinical Bovine Mastitis

    OpenAIRE

    S. Nazifi*, M. Haghkhah1, Z. Asadi, M. Ansari-Lari2, M. R. Tabandeh3, Z. Esmailnezhad and M. Aghamiri

    2011-01-01

    The present study was conducted to evaluate the concentrations of sialic acids (total, lipid bound and protein bound) and their correlation with acute phase proteins (haptoglobin and serum amyloid A) in clinical and subclinical mastitis of cattle. Thirty subclinical mastitic cows with positive California mastitis test (CMT) test and no clinical signs of mastitis, 10 clinical mastitic cows and 10 healthy cows with negative CMT test and normal somatic cell count were selected. Milk and blood sa...

  7. Prevalence of Virulence-Related Determinants in Clinical Isolates of Staphylococcus epidermidis

    Science.gov (United States)

    Najar Peerayeh, Shahin; Jazayeri Moghadas, Ali; Behmanesh, Mehrdad

    2016-01-01

    Background Staphylococcus epidermidis, a member of the human flora, is recognized as an opportunistic pathogen and cause of nosocomial infections. Staphylococcus epidermidis surface components are able to establish bacteria on the host surface, and cause infection. Objectives The frequency of icaA, IS256, aap, fbe and bhp in clinical isolates of S. epidermidis were investigated in this study. Materials and Methods Fifty-nine S. epidermidis isolates were collected from blood (50), wound (1), urine (4) and tracheal (4) samples (Tehran, Iran). Staphylococcus epidermidis isolates were identified with conventional bacteriological tests. Virulence-associated genes were detected by specific polymerase chain reactions (PCRs). Results Of the 59 S. epidermidis, fbe was found in 89.8%, while aap and bhp were observed in 64.4% and 15.3% of the samples, respectively. Coexistence of aap and fbe was found in 32 isolates, while coexistence of bhp and fbe was observed in five isolates. Two isolates were negative for the investigated genes. Conclusions Prevalence of fbe and aap was significantly different from similar studies, yet frequency of bhp was in accordance with other studies. Prevalence of icaA and IS256 was not significantly different from some studies while a significant difference was observed when results were compared with some other studies. PMID:27800129

  8. Macrolide-lincosamide-streptogramin B resistance phenotypes in clinical staphylococcal isolates.

    Science.gov (United States)

    Cetin, Emel Sesli; Gunes, Hayati; Kaya, Selcuk; Aridogan, Buket Cicioglu; Demirci, Mustafa

    2008-04-01

    The prevalence of macrolide-lincosamide-streptogramin B (MLSB) resistance as well as the MLSB resistance phenotypes were investigated by the double-disk diffusion test among 532 clinical staphylococci isolates in a Turkish university hospital. The activity of other antimicrobials, including trimethoprim/sulfamethoxazole, telithromycin, quinupristin/dalfopristin, linezolid, gentamicin, chloramphenicol, ciprofloxacin and vancomycin, was also evaluated. Of 532 isolates, 38.5% were resistant to MLSB antibiotics; 63.9% of the resistant isolates exhibited a constitutive phenotype (cMLSB) whereas 36.1% expressed an inducible resistance phenotype (iMLSB). MLSB resistance was more prevalent among coagulase-negative staphylococci (CoNS) strains. Oxacillin-resistant strains exhibited significantly higher MLSB resistance rates compared with oxacillin-susceptible strains (Pquinupristin/dalfopristin, the most effective antibiotics were telithromycin and chloramphenicol among all isolates. Susceptibility rates to other antibiotics tested were higher among isolates without MLS(B) resistance than isolates with MLSB resistance. The detection of a considerable rate (43.5%) of iMLSB resistance among erythromycin-resistant/clindamycin-susceptible strains suggests that the true percentage of clindamycin resistance may be underestimated if testing for inducible resistance is not performed. PMID:18206352

  9. Clinical significance, antimicrobial susceptibility and molecular identification of Nocardia species isolated from children with cystic fibrosis.

    Science.gov (United States)

    Betrán, Ana; Villuendas, M Cruz; Rezusta, Antonio; Pereira, Javier; Revillo, M José; Rodríguez-Nava, Verónica

    2016-01-01

    Nocardia is an opportunistic pathogen that causes respiratory infections in immunocompromised patients. The aim of this study was to analyze the epidemiology, clinical significance and antimicrobial susceptibility of Nocardia species isolated from eight children with cystic fibrosis. The isolated species were identified as Nocardia farcinica, Nocardia transvalensis, Nocardia pneumoniae, Nocardia veterana and Nocardia wallacei. N. farcinica was isolated in three patients and all of them presented lung affectation with a chronic colonization and pneumonia. N. farcinica showed resistance against gentamicin, tobramycin, cefotaxime, but was susceptible to trimethoprim-sulfamethoxazole and amikacin. N. transvalensis, which was isolated from two patients, showed an association with chronic colonization. N. transvalensis was resistant to tobramycin and amikacin, but susceptible to ciprofloxacin, trimethoprim-sulfamethoxazole and cefotaxime. N. veterana, N. pneumoniae and N. wallacei were isolated from three different patients and appeared in transitory lung colonization. N. veterana and N. pneumoniae were susceptible to imipenem, trimethoprim-sulfamethoxazole, amikacin, tobramycin, and cefotaxime. N. wallacei was resistant to amikacin, tobramycin, imipenem, and trimethoprim-sulfamethoxazole and susceptible to ciprofloxacin and cefotaxime. All the isolates were identified up to species level by 16S rRNA gene sequencing. The presence of Nocardia in the sputum of patients with cystic fibrosis is not always an indication of an active infection; therefore, the need for a treatment should be evaluated on an individual basis. The detection of multidrug-resistant species needs molecular identification and susceptibility testing, and should be performed for all Nocardia infections. PMID:27155949

  10. A method enabling high-throughput sequencing of human cytomegalovirus complete genomes from clinical isolates.

    Directory of Open Access Journals (Sweden)

    Steven Sijmons

    Full Text Available Human cytomegalovirus (HCMV is a ubiquitous virus that can cause serious sequelae in immunocompromised patients and in the developing fetus. The coding capacity of the 235 kbp genome is still incompletely understood, and there is a pressing need to characterize genomic contents in clinical isolates. In this study, a procedure for the high-throughput generation of full genome consensus sequences from clinical HCMV isolates is presented. This method relies on low number passaging of clinical isolates on human fibroblasts, followed by digestion of cellular DNA and purification of viral DNA. After multiple displacement amplification, highly pure viral DNA is generated. These extracts are suitable for high-throughput next-generation sequencing and assembly of consensus sequences. Throughout a series of validation experiments, we showed that the workflow reproducibly generated consensus sequences representative for the virus population present in the original clinical material. Additionally, the performance of 454 GS FLX and/or Illumina Genome Analyzer datasets in consensus sequence deduction was evaluated. Based on assembly performance data, the Illumina Genome Analyzer was the platform of choice in the presented workflow. Analysis of the consensus sequences derived in this study confirmed the presence of gene-disrupting mutations in clinical HCMV isolates independent from in vitro passaging. These mutations were identified in genes RL5A, UL1, UL9, UL111A and UL150. In conclusion, the presented workflow provides opportunities for high-throughput characterization of complete HCMV genomes that could deliver new insights into HCMV coding capacity and genetic determinants of viral tropism and pathogenicity.

  11. Clinical Grade Treg: GMP Isolation, Improvement of Purity by CD127pos Depletion, Treg Expansion, and Treg Cryopreservation

    OpenAIRE

    Peters, Jorieke H; Preijers, Frank W.; Rob Woestenenk; Hilbrands, Luuk B; Koenen, Hans J.P.M.; Irma Joosten

    2008-01-01

    BACKGROUND: Treg based immunotherapy is of great interest to facilitate tolerance in autoimmunity and transplantation. For clinical trials, it is essential to have a clinical grade Treg isolation protocol in accordance with Good Manufacturing Practice (GMP) guidelines. To obtain sufficient Treg for immunotherapy, subsequent ex vivo expansion might be needed. METHODOLOGY/PRINCIPAL FINDINGS: Treg were isolated from leukapheresis products by CliniMACS based GMP isolation strategies, using anti-C...

  12. Prevalence and Antigenic Differences Observed between Bovine Viral Diarrhea Virus Subgenotypes Isolated from Cattle in Australia and Feedlots in the Southwestern United States

    Science.gov (United States)

    Bovine viral diarrhea viruses (BVDV) are divided into two different species within the pestivirus genus, BVDV type 1 (BVDV1) and BVDV type 2 (BVDV2). Further phylogenetic analysis has revealed subgenotype groupings within the BVDV1 and BVDV2 species. Thus far twelve BVDV1 subgenotypes (BVDV1a throu...

  13. Mastitis Bovina: Resistencia a antibióticos de cepas de Staphylococcus aureus asiladas de leche (Bovine Mastitis: Antimicrobial resistance of Staphylococcus aureus strains isolated from milk

    Directory of Open Access Journals (Sweden)

    Pellegrino, MS

    2011-07-01

    Full Text Available ResumenLa mastitis bovina es considerada la enfermedad infecciosa del ganado lechero de mayor impacto económico mundial, siendo Staphylococcus aureus el principal agente patógeno en muchos países.SummaryBovine mastitis is a frequent cause of economic loss in worldwide dairy herds, being Staphylococcus aureus the main etiological agent in many countries.

  14. Mastitis Bovina: Resistencia a antibióticos de cepas de Staphylococcus aureus asiladas de leche (Bovine Mastitis: Antimicrobial resistance of Staphylococcus aureus strains isolated from milk)

    OpenAIRE

    Pellegrino, MS; Frola, ID; Odierno, LM; Bogni, CI

    2011-01-01

    ResumenLa mastitis bovina es considerada la enfermedad infecciosa del ganado lechero de mayor impacto económico mundial, siendo Staphylococcus aureus el principal agente patógeno en muchos países.SummaryBovine mastitis is a frequent cause of economic loss in worldwide dairy herds, being Staphylococcus aureus the main etiological agent in many countries.

  15. Microbiological and biochemical studies on certain antibiotic-resistant bacteria isolated from certain clinical specimens

    International Nuclear Information System (INIS)

    Infection is a dynamic process involving invasion of the body by pathogenic microorganisms and reactions of the tissues to microorganisms and their toxins. Pathogenic microorganisms isolated from clinical samples are of great threat to human health.The outcome of an infection depends on the virulence of the pathogen and the relative degree of resistance or susceptibility to antimicrobial chemotherapy. Antimicrobial agents interfere with specific processes that are essential for growth and division.Development of antibiotic resistance in bacteria is a problem of great concern. The high prevalence of resistant bacteria seems to be related to uncontrolled usage of antibiotics. B-lactamases are the most common cause of bacterial resistance to B-lactam antimicrobial agents, and it is one of the most important reason for increasing the resistance in pathogenic bacteria against some antibiotics especially those acting on inhibition of cell wall synthesis. One hundred and seven clinical samples and specimens were collected from public, private hospitals and National Cancer Institute (NCI) in Cairo, Egypt. Out of them 72 cases positive for microbial infection. Twelve cases were showed mixed infection. Eighty four isolates of pathogenic bacteria and yeast were collected from single and mixed culture. Susceptibilities of the isolates to 20 different antimicrobial agents were determined according to Kirby-Bauer method. Nine multi-drug resistant gram-negative bacterial strains were identified by (Micro Scan WalkAway 96 SI System). Six of them urine isolates, 2 wound (pus) isolates and one sputum isolate. The identified strains were exposed to in-vitro gamma irradiation at dose level of 24.4 Gy, which is biologically equivalent to the fractionated multiple therapeutic dose used in the protocol of cancer treatment of some patients. The antimicrobial susceptibility of the nine multi-drug resistant strains were carried out by disk diffusion method before and after irradiation

  16. Análise antigênica e molecular de amostras citopáticas do vírus da diarréia viral bovina Antigenic and molecular analysis of cytopathic isolates of bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    Fernando Luiz Tobias

    2000-03-01

    Mucosas.Seven cytopathic isolates of bovine viral diarrhea virus (BVDV isolated from clinical cases and blood of calves from herds with reproductive problems were analysed. All isolates contained a mixture of cytopathic (cp and noncytopathic (ncp viruses which were biologically cloned yielding pure populations of viruses of either biotype. The cp and ncp viruses obtained by cloning were characterized antigenically with a panel of monoclonal antibodies (MAbs and regarding to the expression of the non-structural polypeptide NS3. The analysis of MAb binding revealed two patterns of reactivity: 1. In five isolates, the cp and ncp viruses from the same isolate were antigenically very similar to each other, suggesting they represent true "pairs"of viruses. 2. Two isolates, however, yielded cp and ncp viruses antigenically different from each other. Western immunoblot analysis of non-structural polypeptides of ncp viruses revealed a unique band of reactivity, with a mass of approximately 125kDa, corresponding to the NS23 of the standard BVDV Singer strain. In addition to NS23, the cp viruses expressed a polypeptide of approximatelly 80kDa, corresponding to the NS3. The NS23 of two cp viruses displayed an altered migration in SDS-PAGE compared to the other viruses. In one virus, the NS23 had a molecular mass lower than expected whereas in other virus, two bands of reactivity were observed: one smaller and another bigger than the standard NS23, respectively. Our findings confirm previous results that cytopathic BVDV field isolates usually contain a mixture of viruses of both biotypes and that cytopathogenicity correlates with the expression of NS3. The isolation of cp viruses from the blood of clinically normal cattle, however, demonstrates that their occurrence is not restricted to cases of mucosal disease.

  17. Experimental infection of reindeer with bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    J.K. Morton

    1990-08-01

    Full Text Available Two 8-month reindeer (Rangifer tarandus and a 1-month-old Hereford-Holstein calf (Bos taurus were inoculated intranasally with the Singer (cytopathogenic strain of bovine viral diarrhea (BVD virus. Clinical signs in reindeer included loose stools containing blood and mucus, and transient laminitis or coronitis. Signs in the calf were limited to bloody mucus in the stool and lesions in the nasal mucosa. Antibody titers to BVD virus in the reindeer were intermittent, and titers in the calf persisted from days 14 to 63 post-inoculation (PI. Viremia was detected on PI day 4 in one reindeer, days 3-7 in the other, and days 2-7 in the calf. Bovine viral diarrhea virus was isolated from the lung of the calf at necropsy (PI day 63.

  18. [Isolation of Sporothrix pallida complex in clinical and environmental samples from Chile].

    Science.gov (United States)

    Cruz Choappa, Rodrigo M; Vieille Oyarzo, Peggy I; Carvajal Silva, Laura C

    2014-01-01

    The isolation of S. pallida complex from medical samples and home garden soil of a patient in Chile is here in reported. Fungi of the Sporothrix schenckii complex can cause various infections. In Chile, the medical and environmental isolates of these this complex are rare. The aim of this study was to identify an unusual agent in a case of onychomycosis and to detect its presence in the patient's home garden. For this purpose, clinical samples were obtained by scraping the patient's subungueal first right toe nail as well as by taking soil samples from different areas of her home garden. Species identification was performed by morphophysiology and one of the strains isolated from the patient's toe nail was sent to CBS for molecular confirmation (14.062). S. pallida complex was identified both from the patient's toe nail and samples taken from her home garden.

  19. PHENOTYPIC INVESTIGATION OF VIRULENCE PROFILES IN SOME CANDIDA SPP. STRAINS ISOLATED FROM DIFFERENT CLINICAL SPECIMENS

    Directory of Open Access Journals (Sweden)

    Alina Maria Holban

    2012-09-01

    Full Text Available Candida species great versatility may explain their wide range of infections occurred in a great variety of hosts. Even though many modern molecular techniques arise, phenotypic assays still provide valuable information and can guide the diagnosis and the treatment of infections. The phenotypic investigation of virulence determinants in some recent nosocomial isolates of Candida spp. revealed that the fungal clinical strains may develop specific virulence profiles depending on the taxonomic affiliation of the tested strains and the source of infection. Our data support the usefulness of phenotypic assays in predicting the clinical outcome of the fungal infections and in the adjustment of the antifungal treatment.

  20. Vancomycin modifies the expression of the agr system in multidrug-resistant Staphylococcus aureus clinical isolates

    Directory of Open Access Journals (Sweden)

    Vicenta eCázares-Domínguez

    2015-05-01

    Full Text Available Staphylococcus aureus is an opportunistic pathogen that colonizes human hosts and causes a wide variety of diseases. Two interacting regulatory systems called agr (accessory gene regulator and sar (staphylococcal accessory regulator are involved in the regulation of virulence factors. The aim of this study was to evaluate the effect of vancomycin on hld and spa gene expression during the exponential and post-exponential growth phases in multidrug resistant (MDR S. aureus. Methods. Antibiotic susceptibility was evaluated by the standard microdilution method. The phylogenetic profile was obtained by pulsed-field gel electrophoresis. Polymorphisms of agr and SCCmec were analyzed by multiplex polymerase chain reaction. The expression levels of hld and spa were analyzed by reverse transcription-PCR. An enzyme-linked immunosorbent assay assay was performed to detect protein A, and biofilm formation was analyzed via crystal violet staining. Results. In total, 60.60% (20/33 of S. aureus clinical isolates were MDR. Half (10/20 of the MDR S. aureus isolates were distributed in subcluster 10, with > 90% similarity among them. In the isolates of this subcluster, a high prevalence (100% for the agrII and the cassette SCCmec II polymorphisms was found. Our data showed significant increases in hld expression during the post-exponential phase in the presence and absence of vancomycin. Significant increases in spa expression, protein A production and biofilm formation were observed during the post-exponential phase when the MDR S. aureus isolates were challenged with vancomycin. Conclusion. The polymorphism agrII, which is associated with nosocomial isolates, was the most prevalent polymorphism in MDR S. aureus. Additionally, under our study conditions, vancomycin modified hld and spa expression in these clinical isolates. Therefore, vancomycin may regulate alternative systems that jointly participate in the regulation of these virulence factors.

  1. Prevalence of Aspergillus species in clinical samples isolated in an Indian tertiary care hospital

    Directory of Open Access Journals (Sweden)

    Xess Immaculata

    2004-12-01

    Full Text Available CONTEXT (BACKGROUND: In recent times, it has become important to determine the prevalence of different Aspergillus species in clinical samples in view of difference in antifungal susceptibility noted in some species. AIMS: To determine the species prevalence of Aspergillus isolates in various clinical samples received in the Mycology Laboratory at our institute. METHOD: Over a period of 4-years, a total of 18,731 samples were processed, and species identification carried out by standard microbiological methods. RESULTS: Four hundred and fifty six samples (2.43% were culture positive for Aspergillus species. A.flavus (46.93% was the most common isolate, followed by A.fumigatus (37.72% and A.niger (15.35%. It was observed that A.fumigatus was the predominant species isolated from blood and respiratory specimens, A.flavus was predominantly isolated from nasal polyps whereas A.niger predominated in nail specimens. Culture positivity was highest in the age group 12-65 years and in males. Sixty-nine patients (15.13% were admitted to the intensive care unit. CONCLUSIONS: The study highlights the diverse manifestations caused by Aspergillus species in human beings and also throws light on the different species prevalent locally. The knowledge would prove useful in selecting empirical antifungal therapy and formulating prophylactic and pre-emptive strategies.

  2. Phylogenetic assignment of Mycobacterium tuberculosis Beijing clinical isolates in Japan by maximum a posteriori estimation.

    Science.gov (United States)

    Seto, Junji; Wada, Takayuki; Iwamoto, Tomotada; Tamaru, Aki; Maeda, Shinji; Yamamoto, Kaori; Hase, Atsushi; Murakami, Koichi; Maeda, Eriko; Oishi, Akira; Migita, Yuji; Yamamoto, Taro; Ahiko, Tadayuki

    2015-10-01

    Intra-species phylogeny of Mycobacterium tuberculosis has been regarded as a clue to estimate its potential risk to develop drug-resistance and various epidemiological tendencies. Genotypic characterization of variable number of tandem repeats (VNTR), a standard tool to ascertain transmission routes, has been improving as a public health effort, but determining phylogenetic information from those efforts alone is difficult. We present a platform based on maximum a posteriori (MAP) estimation to estimate phylogenetic information for M. tuberculosis clinical isolates from individual profiles of VNTR types. This study used 1245 M. tuberculosis clinical isolates obtained throughout Japan for construction of an MAP estimation formula. Two MAP estimation formulae, classification of Beijing family and other lineages, and classification of five Beijing sublineages (ST11/26, STK, ST3, and ST25/19 belonging to the ancient Beijing subfamily and modern Beijing subfamily), were created based on 24 loci VNTR (24Beijing-VNTR) profiles and phylogenetic information of the isolates. Recursive estimation based on the formulae showed high concordance with their authentic phylogeny by multi-locus sequence typing (MLST) of the isolates. The formulae might further support phylogenetic estimation of the Beijing lineage M. tuberculosis from the VNTR genotype with various geographic backgrounds. These results suggest that MAP estimation can function as a reliable probabilistic process to append phylogenetic information to VNTR genotypes of M. tuberculosis independently, which might improve the usage of genotyping data for control, understanding, prevention, and treatment of TB.

  3. [Occurrence and antimicrobial susceptibility of Morganella morganii strains isolated from clinical samples].

    Science.gov (United States)

    Zalas-Wiecek, Patrycja; Gospodarek, Eugenia; Wróblewska, Joanna

    2012-01-01

    The aim of this study was the evaluation of occurrence and antimicrobial susceptibility of M morganii rods isolated from clinical samples. This study included 201 strains isolated in the Clinical Microbiology Department of Dr. A. Jurasz University Hospital in 2008-2010. Identification to species was carried out on the basis of the results of biochemical reactions included in the tests ID 32E and VITEK2 GN. Antimicrobial susceptibility of M. morganii rods was determined by the disk-diffusion method on Mueller-Hinton II Agar. Strains of M morganii most commonly isolated from skin and soft tissue, and material taken from the urinary tract, mainly from patients of Anesthesiology and Intensive Care Unit, Department of General and Vascular Surgery and Department of General Surgery and Endocrinology. All of M morganii strains isolated during the three years were susceptible to carbapenems. We reported decrease of strains susceptible to piperacillin and chloramphenicol. In 2010 we showed a higher percentage of strains intermediate to tigecycline, compared with 2009. We observed increase in the percentage of strains resistant to cefoperazone with sulbactam and reported decrease in the percentage of strains resistant and intermediate to aminoglycosides. Extended Spectrum Beta-Lactamases were produced by 13 (6,5%) of M morganii strains.

  4. Efficacy of amikacin and ciprofloxacin against clinical isolates of mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Background: Tuberculosis was a leading cause of death at the turn of the 20 century and continues to be one of the medical scourges of mankind. Before the availability of antimicrobial drugs the cornerstone of treatment was rest in the open air in sanatoria. The major breakthrough in treatment of tuberculosis came with the discovery of Streptomycin. Later, INH, Ethambutol, Pyrazinamide, Rifampicin were added to the arsenal. Objective of this study was to determine the sensitivity of clinical isolates of Mycobacterium tuberculosis against two second-line anti-tuberculosis drugs, Amikacin and Ciprofloxacin. Methods: This cross-sectional study was conducted at Department of Microbiology, Armed Forces Institute of Pathology (AFIP) Rawalpindi. All routine clinical samples received for acid fast bacilli (AFB) in the Department of Microbiology, AFIP, Rawalpindi were processed by modified Petroff's technique and inoculated on Lowenstein Jensen (LJ) medium and Bactec 460 Mycobacterium tuberculosis culture system. After identification of M. tuberculosis sensitivity was performed against first-line anti-tuberculosis drugs. Then susceptibility of M. tuberculosis isolates against Amikacin and Ciprofloxacin was performed on LJ medium. H37Rv was used as control strain. Results: Results were interpreted using resistance ratio method. Out of 100 M. tuberculosis isolates, 98% were sensitive to Amikacin and 97% to Ciprofloxacin. Conclusion: Amikacin and Ciprofloxacin are very effective second line anti-tuberculosis drugs against tuberculosis isolates in our set-up. (author)

  5. Slime Production and Expression of the Slime-Associated Antigen by Staphylococcal Clinical Isolates

    OpenAIRE

    Ammendolia, M. G.; R. Di Rosa; Montanaro, L; Arciola, C. R.; Baldassarri, L.

    1999-01-01

    The ability to produce slime and to express a slime-associated antigen was examined in a collection of staphylococcal clinical isolates. Slime-producing strains were found among coagulase-negative staphylococci in percentages comparable to those reported in other studies; surprisingly, a high percentage of Staphylococcus aureus strains also were able to produce this extracellular material. In the latter case, this ability was strongly dependent on the presence of an additional carbohydrate so...

  6. Candidiasis in Pediatrics; Identification and In vitro Antifungal Susceptibility of the Clinical Isolates

    OpenAIRE

    MOHAMMADI, R.; Ataei, B

    2016-01-01

    Background Candida species are normal microflora of oral cavity, vagina, and gastrointestinal tract. They are the third most prevalent cause of pediatric health care–associated bloodstream fungal infection. This study aimed to provide an epidemiological feature of candidiasis and also presents an antifungal susceptibility profile of clinical Candida isolates among children. Materials and Methods During July 2013 to February 2015, 105 patients from different hospitals of Isfahan, Iran, were ex...

  7. Chlamydia trachomatis Laboratory Strains versus Recent Clinical Isolates: Implications for Routine Microbicide Testing ▿

    OpenAIRE

    Skinner, M. C.; Stamm, W. E.; Lampe, M. L.

    2009-01-01

    A topical microbicide that women can use to prevent sexually transmitted diseases (STDs) is essential, and many microbicide candidates are being tested for activity against human immunodeficiency virus and other STDs, including Chlamydia trachomatis. Screening assays for assessing the activity of microbicides against C. trachomatis are typically done with laboratory-adapted strains, but it is possible that recent clinical isolates may have different susceptibilities to microbicides, as has be...

  8. Prevalence and role of efflux pump activity in ciprofloxacin resistance in clinical isolates of Klebsiella pneumoniae

    OpenAIRE

    Aathithan, S.; French, G L

    2011-01-01

    Abstract We investigated the prevalence and role of efflux pump activity and possible drug influx resistance in ciprofloxacin susceptibility amongst 26 distinct clinical isolates of Klebsiella pneumoniae of varying ciprofloxacin susceptibilities and known quinolone resistance-determining region (QRDR) genotypes. Cellular [14C]ciprofloxacin accumulation patterns and the amount of cell-associated [14C]ciprofloxacin of mid-logarithmic phase cells were determined before and after chall...

  9. Variation in germination of Clostridium difficile clinical isolates correlates to disease severity

    OpenAIRE

    Carlson, Paul E.; Kaiser, Alyssa M.; McColm, Sarah A.; Bauer, Jessica M.; Vincent B Young; Aronoff, David M.; Hanna, Philip C.

    2015-01-01

    Over the past two decades, Clostridium difficile infections have been increasing in both number and severity throughout the world. As with other spore forming bacteria, germination is a vital step in the life cycle of this pathogen. Studies have examined differences in sporulation and toxin production among a number of C. difficile clinical isolates; however, few have examined differences in germination and the relationship between this phenotype and disease severity. Here, over 100 C. diffic...

  10. Recent Trends in Clinically Significant Nontuberculous Mycobacteria Isolates at a Korean General Hospital

    OpenAIRE

    Kim, Hyun Soo; Lee, Yangsoon; Lee, Sangsun; Kim, Young Ah; Sun, Young-Kyu

    2013-01-01

    Lung disease caused by nontuberculous mycobacteria (NTM) represents an increasing proportion of all mycobacterial diseases. We investigated recent occurrences of NTM and evaluated the clinical significance of NTM isolates from 752 respiratory specimens collected from patients at National Health Insurance Service Ilsan Hospital between January 2007 and May 2011. Specimens were incubated on solid and liquid media (BACTEC MGIT 960, BD, USA) for 6-8 weeks, and PCR and reverse blot hybridization w...

  11. Virulence diversity among bacteremic Aeromonas isolates: ex vivo, animal, and clinical evidences.

    Directory of Open Access Journals (Sweden)

    Po-Lin Chen

    Full Text Available BACKGROUND: The objective of this study was to compare virulence among different Aeromonas species causing bloodstream infections. METHODOLOGY/PRINCIPAL FINDINGS: Nine of four species of Aeromonas blood isolates, including A. dhakensis, A. hydrophila, A. veronii and A. caviae were randomly selected for analysis. The species was identified by the DNA sequence matching of rpoD. Clinically, the patients with A. dhakensis bacteremia had a higher sepsis-related mortality rate than those with other species (37.5% vs. 0%, P = 0.028. Virulence of different Aeromonas species were tested in C. elegans, mouse fibroblast C2C12 cell line and BALB/c mice models. C. elegans fed with A. dhakensis and A. caviae had the lowest and highest survival rates compared with other species, respectively (all P values <0.0001. A. dhakensis isolates also exhibited more cytotoxicity in C2C12 cell line (all P values <0.0001. Fourteen-day survival rate of mice intramuscularly inoculated with A. dhakensis was lower than that of other species (all P values <0.0001. Hemolytic activity and several virulence factor genes were rarely detected in the A. caviae isolates. CONCLUSIONS/SIGNIFICANCE: Clinical data, ex vivo experiments, and animal studies suggest there is virulence variation among clinically important Aeromonas species.

  12. Characterization of Cell Wall Proteins in Saccharomyces cerevisiae Clinical Isolates Elucidates Hsp150p in Virulence.

    Directory of Open Access Journals (Sweden)

    Pang-Hung Hsu

    Full Text Available The budding yeast Saccharomyces cerevisiae has recently been described as an emerging opportunistic fungal pathogen. Fungal cell wall mannoproteins have been demonstrated to be involved in adhesion to inert surfaces and might be engaged in virulence. In this study, we observed four clinical isolates of S. cerevisiae with relatively hydrophobic cell surfaces. Yeast cell wall subproteome was evaluated quantitatively by liquid chromatography/tandem mass spectrometry. We identified totally 25 cell wall proteins (CWPs from log-phase cells, within which 15 CWPs were quantified. The abundance of Scw10p, Pst1p, and Hsp150p/Pir2p were at least 2 folds higher in the clinical isolates than in S288c lab strain. Hsp150p is one of the members in Pir family conserved in pathogenic fungi Candida glabrata and Candida albicans. Overexpression of Hsp150p in lab strain increased cell wall integrity and potentially enhanced the virulence of yeast. Altogether, these results demonstrated that quantitative cell wall subproteome was analyzed in clinical isolates of S. cerevisiae, and several CWPs, especially Hsp150p, were found to be expressed at higher levels which presumably contribute to strain virulence and fungal pathogenicity.

  13. [Performance evaluation of VITEK 2 system in meropenem susceptibility testing of clinical Pseudomonas aeruginosa isolates].

    Science.gov (United States)

    Acuner, Ibrahim Cağatay; Bayramoğlu, Gülçin; Birinci, Asuman; Cekiç Cihan, Ciğdem; Bek, Yüksel; Durupınar, Belma

    2011-07-01

    Pseudomonas aeruginosa is an important opportunistic pathogen associated with various community-acquired or nosocomial infections. Multi-drug resistant P.aeruginosa strains increasingly cause epidemics and spread in various hospital wards and geographic regions. Carbapenems are among the most effective antimicrobials in the treatment of multi-drug resistant P.aeruginosa infections, and meropenem is the most successful among alternatives in initial therapy. Particularly in severe infections, inappropriate or inadequate initial antimicrobial therapy is independently associated with adverse clinical and economic outcomes. Availability of accurate and rapid susceptibility testing is a priority. Most of the automated microbiology systems can provide rapid results within 8 to 12 hours. In comparison to standard methods, problems in the antimicrobial susceptibility testing of particular microorganisms and antimicrobial agents have been reported for automated microbiology systems. Failures have been reported previously especially in the susceptibility testing of P.aeruginosa versus carbapenem. Most of these studies are designed according to the Food and Drug Administration (FDA, USA) performance analysis scheme (Class II Special Controls Guidance Document: Antimicrobial Susceptibility Test Systems) in a simplified form. However, there are many lacking issues in the design of most of these studies. Among these, insufficient sample size, use of inappropriate reference method, lack of reproducibility testing, and inadequate distribution of study isolates in interpretative categories are of notice. There are only few studies in the literature that evaluate the performance of automated systems in antimicrobial susceptibility testing of carbapenems in clinical P.aeruginosa isolates with a sufficient sample size (n ? 100). However, most of these studies still have one or more major deficiencies in the study design. Furthermore, none of these studies evaluate the performance of

  14. Trends towards Lower Antimicrobial Susceptibility and Characterization of Acquired Resistance among Clinical Isolates of Brachyspira hyodysenteriae in Spain

    DEFF Research Database (Denmark)

    Hidalgo, Alvaro; Carvajal, Ana; Vester, Birte;

    2011-01-01

    The antimicrobial susceptibility of clinical isolates of Brachyspira hyodysenteriae in Spain was monitored, and the underlying molecular mechanisms of resistance were investigated. MICs of tylosin, tiamulin, valnemulin, lincomycin, and tylvalosin were determined for 87 B. hyodysenteriae isolates ...... G2032A and high levels of tiamulin and lincomycin MICs was found, suggesting an increasing importance of this mutation in antimicrobial resistance of clinical isolates of B. hyodysenteriae.......The antimicrobial susceptibility of clinical isolates of Brachyspira hyodysenteriae in Spain was monitored, and the underlying molecular mechanisms of resistance were investigated. MICs of tylosin, tiamulin, valnemulin, lincomycin, and tylvalosin were determined for 87 B. hyodysenteriae isolates...... by multiple-locus variable-number tandem repeats analysis. A comparison with antimicrobial susceptibility data from 2000 to 2007 showed an increase in pleuromutilin resistance over time, doubling the number of isolates with decreased susceptibility to tiamulin. No alteration in susceptibility was detected...

  15. Isolation and identification of pathogenic bacteria causing bovine subclinical mastitis in Eastern Hebei Province%冀东地区奶牛隐性乳房炎病原茵的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    高桂生; 高光平; 邵新华; 史秋梅; 张艳英; 梁银聚

    2012-01-01

    According to the method of LMT, pH detection and count of the somatic cells, the epidemiology of recessive mastitis, the isolation and identification of pathogenic bacteria were carried out on 1 021 subclinical bovine from 8 bovine farms and 3 large bovine factories in Eastern Hebei Province. The results showed that the incidence rates of recessive mastitis and isolalion of bacteria were 60.63% (619/1021) and 88.21%(546/619),respectively.82 strains of bacteria were isolated from 546 subclinical mastitis cows and identified as 4 classes and 14 species. The main bacteria included 36 strains of Staphylococcus, 33 strains of Streptococcus, 8 strains of Enterobtwteria and 5 strains of Microbacterium. with the proportion of 43.90% ,40.24% ,9.76% and 6.10% ,respectively.The mare pathogenic bacteria causing bovine subclinical mastitis in Eastern Hebei Province were Staphylococcus aureus and Streptococcus agalactiae.%通过LMT法、乳汁pH检验法和体细胞直接计数等方法相结合,对冀东地区3个大型奶牛场、8个奶牛养殖户选取的1021头奶牛进行隐性乳房炎流行病学调查与病菌分离鉴定。结果表明,隐性乳房炎发病率为60.63%(619/1021),乳房炎阳性乳样品中细菌分离率达88.21%(546/619)。从543头隐性乳房炎患牛的阳性乳区乳样中分得细菌共4类14种菌82株,其中葡萄球菌36株,占43.90%;链球菌33株,占40.24%;肠杆菌类8株,占9.76%;棒状杆菌5株。占6.10%。冀东地区奶牛隐性乳房炎主要病原菌为金黄色葡萄球菌和无乳链球菌。

  16. 内蒙古无乳链球菌耐药性及四环素耐药基因检测%Drug resistance and tetracycline resistance gene identification of Streptococcus agalactiae isolates from bovine in Inner Mongolia

    Institute of Scientific and Technical Information of China (English)

    杜琳; 郝永清

    2016-01-01

    为了解内蒙古地区隐性乳房炎无乳链球菌分离株耐药性及耐药基因,采集内蒙古不同地区的规模化养殖场隐性乳房炎乳样,采用 Kirby-Bauer(K-B)纸片扩散法测试分离株对16种抗菌药物的敏感性,PCR 方法检测其耐药基因.结果显示:无乳链球菌对大部分抗生素较敏感.对其有较强抑菌作用的药物为:青霉素 G、头孢噻肟、头孢唑啉、阿莫西林、红霉素、环丙沙星、恩诺沙星、氧氟沙星、林可霉素、呋喃妥因、万古霉素.其敏感性达到90%~100%.该菌株对四环素有较强的耐药性,耐药率达77.27%.PCR 扩增四环素耐药基因 tetM 、tetO 、tetK 、tetL ,结果显示22株无乳链球菌均含有 tetM 基因,其基因的检出率为100%.其中7株同时含有 tetK 基因,tetO 、tetL 基因未检出.%In order to understand drug resistance and tetracycline resistance gene of Streptococcus agalactiae epidemic strains,the prevalent strains of S .agalactiae from the main bovine culture area of Inner Mongolia Autonomous Region were detected through Kirby-B auer(K-B)method and PCR.Drug susceptibility testing indicated that the S .agalactiae isolated strains were sensitive to most antibiotics. The S .agalactiae isolates were susceptible to penicillin,cefotaxime,cefazolin,amoxicillin,erythromy-cin,ciprofloxacin,enrofloxacin,ofloxacin,clindamycin,nitrofurantoin,and vancomycin with susceptibili-ties rates of 90% to 100%.In contrast,the S .agalactiae isolates were resistant to tetracycline with re-sistance rates of 77.27%.Four resistance genes including tetM ,tet O ,tetK ,and tetL were characterized by PCR.All strains carried the resistance gene tetM and 7 strains also carried the resistance gene tetK . The resistance genes tetO and tetL were not detected.The results indicated that the resistance gene were related to the resistant type of antimicrobial agents and was important for further research and clinical therapy resistance mechanisms.

  17. DIFFERENTIAL EFFECT OF PAROMOMYCIN AND NITAZOXANIDE ON CLINICAL ISOLATES OF CRYPTOSPORIDIA IN VITRO

    Directory of Open Access Journals (Sweden)

    Poonam Sharma

    2014-11-01

    Full Text Available Background: Cryptosporidiosis is one of the most difficult protozoan infections to treat, with only two drugs i.e. nitazoxanide and paromomycin known for treatment with variable response in different patients. Human cryptosporidiosis is accounted mainly by C. hominis and C. parvum. These two species or their subtypes are known to differ in clinical manifestations, and may differ in their response to drugs. So, we planned the study to see the effect of nitazoxanide and paromomycin on different isolates of Cryptosporidium in vitro. Methods: MDCK cell lines were used for in vitro growth of parasite and cytotoxicity of drugs to MDCK cells was determined by MTT assay after 3, 12 and 24 hours of drug exposure. Efficacy of non-toxic drug concentrations (25% cytotoxic effect was seen with nitazoxanide and paromomycin at concentrations of more than, 25μg/ml and 6mg/ml, respectively. Nitazoxanide was more effective than paromomycin in decreasing in vitro growth, invasion inhibition and reducing oocyst viability of Cryptosporidium isolates. Drugs effect was higher on growth inhibition followed by invasion inhibition and least in decreasing oocyst viability. Different isolates had variable response to drugs; cumulatively C. parvum isolates were more susceptible at particular drug concentrations than C. hominis isolates.

  18. Characterization of persistent and transient Escherichia coli isolates recovered from clinical mastitis episodes in dairy cows.

    Science.gov (United States)

    Fairbrother, Julie-Hélène; Dufour, Simon; Fairbrother, John Morris; Francoz, David; Nadeau, Éric; Messier, Serge

    2015-03-23

    Escherichia coli usually cause transient intramammary infections in dairy cows, but persistent intramammary infections have been observed. The objective of the study was to compare antimicrobial resistance and virulence genes found in persistent and transient E. coli isolated from clinical mastitis cases in a cohort of 91 Canadian dairy herds monitored over a 2-year period. Antimicrobial susceptibility was determined by broth microdilution and the presence of 27 virulence genes associated with extra-intestinal E. coli infections was determined by colony hybridization. Proportion of resistance in persistent E. coli ranged from 0.0% (enrofloxacin) to 27.8% (ampicillin and tetracycline). Proportion of resistance in transient E. coli ranged from 0.0% (enrofloxacin) to 16.8% (tetracycline). Odds of being classified as a persistent isolate increased by a factor of 1.6 (95% CI: 1.1, 2.4) for each aditional resistance observed (e.g. isolates resistant to four antimicrobial agents had 1.6 times higher odds of belonging to the persistent groups compared to isolates demonstrating resistance to three agents). Persistency was associated with higher odds of resistance to ampicillin (OR: 9.8, Pcoli isolates is associated with certain traits. Findings concerning iron-acquisition shed new light on the mechanisms of intramammary survival.

  19. Impact of Plant Extracts and Antibiotics on Biofilm Formation of Clinical Isolates From Otitis Media

    Science.gov (United States)

    Rehman, Saba; Mujtaba Ghauri, Shahbaz; Sabri, Anjum Nasim

    2016-01-01

    Background: Otitis media can lead to severe health consequences, and is the most common reason for antibiotic prescriptions and biofilm-mediated infections. However, the increased pattern of drug resistance in biofilm forming bacteria complicates the treatment of such infections. Objectives: This study was aimed to estimate the biofilm formation potential of the clinical isolates of otitis media, and to evaluate the efficacy of antibiotics and plant extracts as alternative therapeutic agents in biofilm eradication. Materials and Methods: The ear swab samples collected from the otitis media patients visiting the Mayo Hospital in Lahore were processed to isolate the bacteria, which were characterized using morphological, biochemical, and molecular (16S rRNA ribotyping) techniques. Then, the minimum inhibitory concentrations (MICs) of the antibiotics and crude plant extracts were measured against the isolates. The cell surface hydrophobicity and biofilm formation potential were determined, both qualitatively and quantitatively, with and without antibiotics. Finally, the molecular characterization of the biofilm forming proteins was done by amplifying the ica operon. Results: Pseudomonas aeruginosa (KC417303-05), Staphylococcus hemolyticus (KC417306), and Staphylococcus hominis (KC417307) were isolated from the otitis media specimens. Among the crude plant extracts, Acacia arabica showed significant antibacterial characteristics (MIC up to 13 mg/ml), while these isolates exhibited sensitivity towards ciprofloxacin (MIC 0.2 µg/mL). All of the bacterial strains had hydrophobic cellular surfaces that helped in their adherence to abiotic surfaces, leading to strong biofilm formation potential (up to 7 days). Furthermore, the icaC gene encoding polysaccharide intercellular adhesion protein was amplified from S. hemolyticus. Conclusions: The bacterial isolates exhibited strong biofilm formation potential, while the extracts of Acacia arabica significantly inhibited biofilm

  20. Evaluation of Sialic Acid and Acute Phase Proteins (Haptoglobin and Serum Amyloid A in Clinical and Subclinical Bovine Mastitis

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    S. Nazifi*, M. Haghkhah1, Z. Asadi, M. Ansari-Lari2, M. R. Tabandeh3, Z. Esmailnezhad and M. Aghamiri

    2011-01-01

    Full Text Available The present study was conducted to evaluate the concentrations of sialic acids (total, lipid bound and protein bound and their correlation with acute phase proteins (haptoglobin and serum amyloid A in clinical and subclinical mastitis of cattle. Thirty subclinical mastitic cows with positive California mastitis test (CMT test and no clinical signs of mastitis, 10 clinical mastitic cows and 10 healthy cows with negative CMT test and normal somatic cell count were selected. Milk and blood samples were collected after confirmation of clinical and subclinical mastitis by somatic cell count and bacterial identification. Serum haptoglobin (Hp, serum amyloid A (SAA, total sialic acid (TSA, lipid bound sialic acid (LBSA and protein bound sialic acid (PBSA were measured by validated standard methods. Haptoglobin and SAA increased significantly in both types of mastitis compared with control group (P<0.001. However, the ratio of HP/SAA was significantly different from the control group only in clinical mastitis. The results showed that TSA and LBSA were significantly different in control group compared with clinical and subclinical mastitis (P<0.001. Protein bound sialic acid did not change in subclinical mastitis in comparison with control group (P=0.86. There was positive correlation between LBSA and PBSA in clinical mastitis (r=0.72, P=0.02 whereas significant negative correlation was observed between LBSA and PBSA in subclinical mastitis (r=-0.62, P<0.001. Results also showed no correlation between Hp and SAA with each other or with any other parameters in study groups.

  1. Incidence of virulence determinants in clinical Enterococcus faecalis and Enterococcus faecium isolates collected in Bulgaria

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    Tanya Strateva

    2016-04-01

    Full Text Available Abstract Objectives To evaluate the prevalence of some virulence genes among 510 clinical Enterococcus spp. isolates and to assess the association of those genes with the species, infection site, and patient group (inpatients/outpatients. Methods Adhesins genes (aggregation substances agg and asa1 of Enterococcus faecalis and Enterococcus faecium, respectively, enterococcal surface protein (esp, endocarditis-specific antigen A (efaA, collagen-binding proteins (ace/acm; invasins (hyaluronidase (hyl and gelatinase (gelE; cytotoxines (activation of cytolysin (cylA in E. faecalis; and modulators of the host immunity and inflammation (enhanced expression pheromone (eep in E. faecalis were detected by polymerase chain reaction. Results The overall prevalence was: esp – 44.3%, agg/asa1 – 38.4%, ace/acm – 64.3%, efaA – 85.9%, eep – 69.4%, gelE – 64.3%, hyl – 25.1%, and cylA – 47.1%. E. faecalis isolates had significantly higher frequency of adhesin genes (esp and agg/asa1 and gelatinase in comparison to E. faecium. Multiple virulence genes in E. faecalis were significantly more prevalent than in E. faecium isolates. Domination of E. faecium with or without only one gene compared to the isolates of E. faecalis were found. Enterococcus spp. isolates obtained from outpatients compared to inpatients isolates had significantly higher frequency of agg/asa1, eep, gelE and cylA. Some adhesins genes (esp, agg/asa1 and efaA had higher prevalence among the non-invasive Enterococcus spp. isolates compared to those causing invasive bacteremia, while ace/acm revealed higher dissemination in isolates causing invasive infections compared to non-invasive isolates. Conclusion Most E. faecalis attaches to abiotic surfaces in hospital environment, which correlates with higher prevalence of gene encoding for virulence factors involved in biofilm formation, such as enterococcal surface protein, aggregation substance, and gelatinase. The intestinal tract is an

  2. Survey on vertical infection of bovine viral diarrhea virus from fetal bovine sera in the field

    OpenAIRE

    NAGAYAMA, Kumiko; OGUMA, Keisuke; SENTSUI, Hiroshi

    2015-01-01

    Bovine viral diarrhea virus (BVDV) isolation and antibody survey were performed using 2,758 fetal bovine sera (FBS) collected from slaughterhouses in New Zealand, Australia and the Dominican Republic, and then sent to Japan to manufacture commercial serum for cell culture use. FBS in the Dominican Republic were pooled for each several individuals, and those collected in other countries were separated according to each individual and subjected to the tests. BVDV was isolated from 25 (0.91%) FB...

  3. Comparison of genomic and antimicrobial resistance features of latex agglutination test-positive and latex agglutination test-negative Staphylococcus aureus isolates causing bovine mastitis

    OpenAIRE

    Moser, A.; Stephan, R.; Corti, S; Johler, S.

    2013-01-01

    The dairy industry suffers massive economic losses due to staphylococcal mastitis in cattle. The Staphaureux latex agglutination test (Oxoid, Basel, Switzerland) was reported to lead to negative results in 54% of bovine Staphylococcus aureus strains, and latex-negative strains are thought to be less virulent than Staphaurex latex-positive strains. However, comparative information on virulence and resistance profiles of these 2 groups of Staph. aureus is scarce. Our objective was to associate ...

  4. Modulation of the kinetics of cholesterol side-chain cleavage by an activator and by an inhibitor isolated from the cytosol of the cortex of bovine adrenals.

    OpenAIRE

    Warne, P A; Greenfield, N J; Lieberman, S.

    1983-01-01

    Two modulators of sterol side-chain cleavage activity have been detected in the cytosol from the cortex of bovine adrenals. One is an inhibitor of side-chain cleavage which increases the Km of a purified and reconstituted mitochondrial side-chain cleavage system for both cholesterol and cholesterol sulfate. It also lowers the Vmax of cleavage when cholesterol sulfate is the substrate. The other modulator is a low molecular weight protein which in the reconstituted system increases the Vmax of...

  5. Molecular Characterization and In Vitro Antifungal Susceptibility of 316 Clinical Isolates of Dermatophytes in Iran.

    Science.gov (United States)

    Ansari, Saham; Hedayati, Mohammad T; Zomorodian, Kamiar; Pakshir, Keyvan; Badali, Hamid; Rafiei, Abdollah; Ravandeh, Mostafa; Seyedmousavi, Seyedmojtaba

    2016-02-01

    Dermatophytosis is a common mycotic infection of the skin, nail, and hair, associated with major public health concern worldwide. Various species of dermatophytes show significant differences in susceptibility to antifungals. Here, we present the antifungal susceptibility of a large collection of molecularly identified dermatophyte isolates obtained from tropical region of south of Iran. A total of 9485 patients clinically suspected to have cutaneous fungal infections were examined. Dermatophytosis was confirmed in 1502 cases by direct microscopy and culture. Three hundred and sixteen isolates recovered in culture were identified to species level using PCR sequencing of ITS region and RFLP. Tinea corporis was the most prevalent type of clinical manifestation (35.2 %), followed by tinea cruris (17 %), tinea capitis (12.8 %), tinea pedis (11.3 %), tinea manuum (11 %), tinea unguium (6.9 %), and tinea barbae (5.8 %). Trichophyton interdigitale was the most common isolate (49.36 %), followed by Trichophyton rubrum (18.98 %), Epidermophyton floccosum (13.29 %), Microsporum canis (9.17 %), Arthroderma benhamiae (T. anamorph of A. benhamiae; 5.38 %), and Trichophyton tonsurans (3.79 %). Overall, irrespective of the geographical region, terbinafine was the most potent antifungal against all isolates, with an MIC range of 0.002-0.25 μg/mL, followed by itraconazole (0.004-0.5 μg/mL), griseofulvin (0.125-8 μg/mL), and fluconazole (4-128 μg/mL). Analysis of our data revealed a significant increase in the frequency of A.benhamiae, which definitely warrants further investigation to explore source of this infection in south of Iran. Moreover, terbinafine was the most effective antifungal against all isolates, in vitro.

  6. Molecular Characterization and In Vitro Antifungal Susceptibility of 316 Clinical Isolates of Dermatophytes in Iran.

    Science.gov (United States)

    Ansari, Saham; Hedayati, Mohammad T; Zomorodian, Kamiar; Pakshir, Keyvan; Badali, Hamid; Rafiei, Abdollah; Ravandeh, Mostafa; Seyedmousavi, Seyedmojtaba

    2016-02-01

    Dermatophytosis is a common mycotic infection of the skin, nail, and hair, associated with major public health concern worldwide. Various species of dermatophytes show significant differences in susceptibility to antifungals. Here, we present the antifungal susceptibility of a large collection of molecularly identified dermatophyte isolates obtained from tropical region of south of Iran. A total of 9485 patients clinically suspected to have cutaneous fungal infections were examined. Dermatophytosis was confirmed in 1502 cases by direct microscopy and culture. Three hundred and sixteen isolates recovered in culture were identified to species level using PCR sequencing of ITS region and RFLP. Tinea corporis was the most prevalent type of clinical manifestation (35.2 %), followed by tinea cruris (17 %), tinea capitis (12.8 %), tinea pedis (11.3 %), tinea manuum (11 %), tinea unguium (6.9 %), and tinea barbae (5.8 %). Trichophyton interdigitale was the most common isolate (49.36 %), followed by Trichophyton rubrum (18.98 %), Epidermophyton floccosum (13.29 %), Microsporum canis (9.17 %), Arthroderma benhamiae (T. anamorph of A. benhamiae; 5.38 %), and Trichophyton tonsurans (3.79 %). Overall, irrespective of the geographical region, terbinafine was the most potent antifungal against all isolates, with an MIC range of 0.002-0.25 μg/mL, followed by itraconazole (0.004-0.5 μg/mL), griseofulvin (0.125-8 μg/mL), and fluconazole (4-128 μg/mL). Analysis of our data revealed a significant increase in the frequency of A.benhamiae, which definitely warrants further investigation to explore source of this infection in south of Iran. Moreover, terbinafine was the most effective antifungal against all isolates, in vitro. PMID:26369643

  7. Clinical illnesses associated with isolation of dysgonic fermenter 3 from stool samples.

    Science.gov (United States)

    Blum, R N; Berry, C D; Phillips, M G; Hamilos, D L; Koneman, E W

    1992-02-01

    The clinical significance of the fastidious organism DF-3 isolated from stool cultures is unclear. We sought to improve our understanding of this organism and to further define its association with human disease. Stool cultures for DF-3 were obtained from three sources: an ongoing study of enteric pathogens in patients infected with the human immunodeficiency virus, a screening procedure in which all stool samples submitted for Clostridium difficile toxin assay were cultured for DF-3, and stool samples submitted specifically for DF-3 culture. Retrospective clinical data were obtained from chart reviews of patients with positive cultures. Antimicrobial susceptibility testing and cell wall fatty acid analysis were performed for each DF-3 isolated. Eight isolates of DF-3 were obtained over a period of 8 months. All patients either had severe underlying disease or were immunocompromised, including three patients coinfected with human immunodeficiency virus and two patients with inflammatory bowel disease. The spectrum of clinical disease ranged from chronic diarrhea with a well-defined response to therapy for DF-3 to an asymptomatic carrier state. Cell wall fatty acid analysis of these isolates demonstrated a consistent pattern with a large peak of 12-methyltetradecanoate. DF-3, a fastidious gram-negative coccobacillus, can be recovered from stool cultures of immunocompromised patients by using selective media. The presence of 12-methyltetradecanoate in cell wall fatty acid analysis assists in identification. The increased use of a selective medium-(cefoperazone-vancomycin-amphotericin B) in the evaluation of diarrhea in immunocompromised hosts, including persons with inflammatory bowel disease, may better define the association of DF-3 with human gastrointestinal disease.

  8. Antifungal susceptibility and virulence factors of clinically isolated dermatophytes in Tehran, Iran

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    Mohammad Ali Afshari

    2016-03-01

    Full Text Available Background and Objectives: Dermatophytes possess a wide array of virulence factors and various antifungal susceptibility patterns which influence their pathogenesis in humans and animals. The aim of this study was to evaluate antifungal suscep- tibility and keratinase and proteinase activity of 49 dermatophyte strains from the genera Microsporum, Trichophyton and Epidermophyton which were isolated from human cases of dermatophytosis.Materials and Methods: Forty-nine dermatophyte strains isolated from clinical samples were cultured on general and spe- cific culture media. Keratinase and proteinase activity was screened on solid mineral media and confirmed in liquid cultures. Drug susceptibility toward azoles (fluconazole, ketoconazole and itraconazole, griseofulvin and terbinafine was evaluated using disk diffusion method on Mueller-Hinton agar and minimum inhibitory concentrations (MICs were determined using microbroth dilution assay according to the Clinical and Laboratory Standards Institute (CLSI guidelines.Results: Our results indicated that clinically isolated dermatophytes from 7 major species produced keratinase and protein- ase at different extents. The mean keratinase and proteinase activity was reported as 6.69 ± 0.31 (U/ml and 2.10 ± 0.22 (U/ ml respectively. Disk diffusion and microbroth dilution (MIC results of antifungal susceptibility testing showed that ke- toconazole was the most effective drug against Epidermophyton floccosum and Trichophyton mentagrophytes, itraconazole against T. rubrum and E. floccosum, and griseofulvin and terbinafine against Trichophyton verrucosum. Our results showed that all dermatophyte isolates were resistant to fluconazole. Overall, ketoconazole and itraconazole were the most effective drugs for all dermatophyte species tested.Conclusion: Our results showed that antifungal susceptibility testing is an urgent need to select drugs of choice for treatment of different types of dermatophytosis and

  9. Review of clinical studies of Polygonum multiflorum Thunb. and its isolated bioactive compounds

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    Guy-Armel Bounda

    2015-01-01

    Full Text Available Polygonum multiflorum Thunb. (PMT, officially listed in the Chinese Pharmacopoeia, is one of the most popular perennial Chinese traditional medicines known as He shou wu in China and East Asia, and as Fo-ti in North America. Mounting pharmacological studies have stressed out its key benefice for the treatment of various diseases and medical conditions such as liver injury, cancer, diabetes, alopecia, atherosclerosis, and neurodegenerative diseases as well. International databases such as PubMed/Medline, Science citation Index and Google Scholar were searched for clinical studies recently published on P. multiflorum. Various clinical studies published articles were retrieved, providing information relevant to pharmacokinetics-pharmacodynamics analysis, sleep disorders, dyslipidemia treatment, and neurodegenerative diseases. This review is an effort to update the clinical picture of investigations ever carried on PMT and/or its isolated bio-compounds and to enlighten its therapeutic assessment.

  10. Antiviral effects of bovine interferons on bovine respiratory tract viruses.

    OpenAIRE

    Fulton, R W; Downing, M M; Cummins, J M

    1984-01-01

    The antiviral effects of bovine interferons on the replication of bovine respiratory tract viruses were studied. Bovine turbinate monolayer cultures were treated with bovine interferons and challenged with several bovine herpesvirus 1 strains, bovine viral diarrhea virus, parainfluenza type 3 virus, goat respiratory syncytial virus, bovine respiratory syncytial virus, bovine adenovirus type 7, or vesicular stomatitis virus. Treatment with bovine interferons reduced viral yield for each of the...

  11. Enhancement of antibiotic activity by efflux inhibitors against multidrug resistant Mycobacterium tuberculosis clinical isolates from Brazil

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    Tatiane eCoelho

    2015-04-01

    Full Text Available Drug resistant tuberculosis continues to increase and new approaches for its treatment are necessary. The identification of M. tuberculosis clinical isolates presenting efflux as part of their resistant phenotype has a major impact in tuberculosis treatment. In this work, we used a checkerboard procedure combined with the tetrazolium microplate-based assay (TEMA to study single combinations between antituberculosis drugs and efflux inhibitors (EIs against multidrug resistant M. tuberculosis clinical isolates using the fully susceptible strain H37Rv as reference. Efflux activity was studied on a real-time basis by a fluorometric method that uses ethidium bromide as efflux substrate. Quantification of efflux pump genes mRNA transcriptional levels were performed by RT-qPCR. The fractional inhibitory concentrations (FIC indicated synergistic activity for the interactions between isoniazid, rifampicin, amikacin, ofloxacin, and ethidium bromide plus the EIs verapamil, thioridazine and chlorpromazine. The FICs ranged from 0.25, indicating a four-fold reduction on the MICs, to 0.015, 64-fold reduction. The detection of active efflux by real-time fluorometry showed that all strains presented intrinsic efflux activity that contributes to the overall resistance which can be inhibited in the presence of the EIs. The quantification of the mRNA levels of the most important efflux pump genes on these strains shows that they are intrinsically predisposed to expel toxic compounds as the exposure to subinhibitory concentrations of antibiotics were not necessary to increase the pump mRNA levels when compared with the non-exposed counterpart. The results obtained in this study confirm that the intrinsic efflux activity contributes to the overall resistance in multidrug resistant clinical isolates of M. tuberculosis and that the inhibition of efflux pumps by the EIs can enhance the clinical effect of antibiotics that are their substrates.

  12. Plasmodium knowlesi genome sequences from clinical isolates reveal extensive genomic dimorphism.

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    Miguel M Pinheiro

    Full Text Available Plasmodium knowlesi is a newly described zoonosis that causes malaria in the human population that can be severe and fatal. The study of P. knowlesi parasites from human clinical isolates is relatively new and, in order to obtain maximum information from patient sample collections, we explored the possibility of generating P. knowlesi genome sequences from archived clinical isolates. Our patient sample collection consisted of frozen whole blood samples that contained excessive human DNA contamination and, in that form, were not suitable for parasite genome sequencing. We developed a method to reduce the amount of human DNA in the thawed blood samples in preparation for high throughput parasite genome sequencing using Illumina HiSeq and MiSeq sequencing platforms. Seven of fifteen samples processed had sufficiently pure P. knowlesi DNA for whole genome sequencing. The reads were mapped to the P. knowlesi H strain reference genome and an average mapping of 90% was obtained. Genes with low coverage were removed leaving 4623 genes for subsequent analyses. Previously we identified a DNA sequence dimorphism on a small fragment of the P. knowlesi normocyte binding protein xa gene on chromosome 14. We used the genome data to assemble full-length Pknbpxa sequences and discovered that the dimorphism extended along the gene. An in-house algorithm was developed to detect SNP sites co-associating with the dimorphism. More than half of the P. knowlesi genome was dimorphic, involving genes on all chromosomes and suggesting that two distinct types of P. knowlesi infect the human population in Sarawak, Malaysian Borneo. We use P. knowlesi clinical samples to demonstrate that Plasmodium DNA from archived patient samples can produce high quality genome data. We show that analyses, of even small numbers of difficult clinical malaria isolates, can generate comprehensive genomic information that will improve our understanding of malaria parasite diversity and

  13. Sequence variability of human cytomegalovirus UL143 in low-passage clinical isolates

    Institute of Scientific and Technical Information of China (English)

    HE Rong; RUAN Qiang; QI Ying; MA Yan-ping; HUANG Yu-jing; SUN Zheng-rong; JI Yao-hua

    2006-01-01

    Background Human cytomegalovirus (HCMV) infects a number of organs and tissues in vivo. The different symptoms and tissue tropisms of HCMV infection perhaps result from genetic polymorphism. A new region of DNA containing at least 19 open reading frames(ORFs) (denoted UL133 to 151) was found in the low-passage HCMV clinical strain, Toledo, and several other low-passage clinical isolates, but not present in the HCMV laboratory strain, AD169. One of these genes, UL143, was studied to explore the sequence variability of UL143 ORF in HCMV clinical isolates and examine the possible association between gene variability and the outcome of HCMV infection.Methods The UL143 gene of the strains obtained from suspected congenitally HCMV-infected infants was amplified by polymerase chain reaction (PCR) and sequenced.Results Nineteen sequences of the strains were divided into 2 major groups, G1(n=16) and G2(n=3). All of the sequences had frame-shift mutation compared to Toledo. Nucleotide polymorphisms conferred substantial amino acid substitutions when compared with Toledo. All 16 UL143 putative proteins of the strains in G1 had a new myristylation site and loss of two PKC sites owing to missense mutations. No convincing relationships were observed between the presence of HCMV disease and the UL 143 sequence group.Conclusions HCMV-UL143 existed in low passage isolates. Sequence variability caused by frame-shift mutation was found in all HCMV clinical strains. No obvious linkage was observed between UL143 polymorphisms and the outcome of suspected congenital HCMV infection.

  14. The effect of environmental conditions on biofilm formation of Burkholderia pseudomallei clinical isolates.

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    Nur Siti K Ramli

    Full Text Available Burkholderia pseudomallei, a Gram-negative saprophytic bacterium, is the causative agent of the potentially fatal melioidosis disease in humans. In this study, environmental parameters including temperature, nutrient content, pH and the presence of glucose were shown to play a role in in vitro biofilm formation by 28 B. pseudomallei clinical isolates, including four isolates with large colony variants (LCVs and small colony variants (SCVs morphotypes. Enhanced biofilm formation was observed when the isolates were tested in LB medium, at 30 °C, at pH 7.2, and in the presence of as little as 2 mM glucose respectively. It was also shown that all SVCs displayed significantly greater capacity to form biofilms than the corresponding LCVs when cultured in LB at 37 °C. In addition, octanoyl-homoserine lactone (C(8-HSL, a quorum sensing molecule, was identified by mass spectrometry analysis in bacterial isolates referred to as LCV CTH, LCV VIT, SCV TOM, SCV CTH, 1 and 3, and the presence of other AHL's with higher masses; decanoyl-homoserine lactone (C(10-HSL and dodecanoyl-homoserine lactone (C(12-HSL were also found in all tested strain in this study. Last but not least, we had successfully acquired two Bacillus sp. soil isolates, termed KW and SA respectively, which possessed strong AHLs degradation activity. Biofilm formation of B. pseudomallei isolates was significantly decreased after treated with culture supernatants of KW and SA strains, demonstrating that AHLs may play a role in B. pseudomallei biofilm formation.

  15. Pathological studies on bovine viral diarrhea

    International Nuclear Information System (INIS)

    Bovine viral diarrhea virus (BVDV) is classified as an RNA virus in the family flavin viride and is a member of the genus pest virus (Collet et al 1989). BVDV has a worldwide distribution and infections in cattle populations (Kahrs et al 1970). It was recognized since 50 years ago, the initial description of an acute enteric disease of cattle in North America, which was characterized by outbreaks of diarrhea and erosive of digestive tract (Olafsonp et al 1946). The disease and causative agent were named bovine viral diarrhea (B V D ) and (B V DV), respectively. This virus was subsequently associated with a sporadically occurring and highly fatal enteric disease that was termed mucosal disease (M D), (Ramsey and Chivers 1953). The initial isolate of BVDV did not produce cytopathic effect in cell culture, whereas an isolate from MD did produce cytopathic effects (Lee et al 1957). In vitro characteristic of non cytopathic or sytopathic effects of BVDV is referred to as the biotype of the virus. It has now been established that MD occurs only when xattle that are born immuno tolerant to and persistently infected with a noncyropathic BVDV become super infected with a cytopathic BVDV. The knowledge of the molecular biology. Pathogenesis and epidemiology of BVDV has greatly evolved in the past 10-15 years and has provided a better understanding of this complex infectious agent. Infection with BVDV can result in a wide spectrum of diseases ranging from subclinical infection s to a highly fatal from known as mucosal disease (ND). The clinical response to infection depends on multiple interactive factors. Host factors that influence the clinical outcome of BVDV infection include whether the host is immunocompetent or immuno tolerant to BVDV, pregnancy status, gestational age of the fetus, immune status (passively derived or actively derived from previous infection or vaccination) and concurrent level of environmental stress

  16. Evaluation of different detection methods of biofilm formation in the clinical isolates

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    Afreenish Hassan

    2011-08-01

    Full Text Available BACKGROUND: Microorganisms growing in a biofilm are associated with chronic and recurrent human infections and are highly resistant to antimicrobial agents. There are various methods to detect biofilm production like Tissue Culture Plate (TCP, Tube method (TM, Congo Red Agar method (CRA, bioluminescent assay, piezoelectric sensors, and fluorescent microscopic examination. OBJECTIVE: This study was conducted to compare three methods for the detection of biofilms. METHOD: The study was carried out at the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Pakistan, from January 2010 to June 2010. A total of 110 clinical isolates were subjected to biofilm detection methods. Isolates were identified by standard microbiological procedures. Biofilm detection was tested by TCP, TM and CRA. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the Kirby-Bauer disc diffusion technique according to CLSI guidelines. RESULTS: The TCP method was considered to be superior to TM and CRA. From the total of 110 clinical isolates, TCP method detected 22.7% as high, 41% moderate and 36.3% as weak or non-biofilm producers. We have observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers. CONCLUSION: We can conclude from our study that the TCP method is a more quantitative and reliable method for the detection of biofilm forming microorganisms as compared to TM and CRA methods, and it can be recommended as a general screening method for detection of biofilm producing bacteria in laboratories.

  17. Nitrogen laser irradiation (337 nm) causes temporary inactivation of clinical isolates of Mycobacterium tuberculosis.

    Science.gov (United States)

    Dube, Alok; Jayasankar, K; Prabakaran, L; Kumar, V; Gupta, P K

    2004-01-01

    We have investigated the effect of nitrogen laser irradiation (337 nm) on viability of clinical isolates of Mycobacterium tuberculosis. Bacteria were exposed to a nitrogen laser (average power 2.0 mW) in vitro at power density of 70 +/- 0.7 W/m2 for 0-30 min, and the cell viability was determined by luciferase reporter phage (LRP) assay. Immediately after laser exposure, all the clinical isolates investigated showed a dose-dependent decrease in cell viability. However, when the laser-exposed isolates were incubated in broth medium for 3 days, most of these showed significant recovery from laser-induced damage. Addition of 5.0 microg/ml acriflavine (a DNA repair inhibitor) in the incubation medium had no significant effect on recovery. This suggests that DNA damage may not be involved in the cell inactivation. Electron paramagnetic resonance (EPR) studies using 5-doxyl strearic acid (5-DS) as a probe suggest alterations in lipid regions of the cell wall. Implications of these results for understanding therapeutic effect of nitrogen laser on drug-resistant tuberculosis are discussed. PMID:15278725

  18. Antibacterial Activity of Twenty Iranian Plant Extracts Against Clinical Isolates of Helicobacter pylori

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    Farahnaz Nariman

    2009-06-01

    Full Text Available Objective(sDue to increasing emergence of drug-resistance in Helicobacter pylori isolates, traditional plants arepotentially valuable sources of novel anti-H. pylori agents. In this research, anti-H. pylori activity of theorganic extracts of twenty native Iranian plants was determined against ten clinical isolates of H. pylori.Materials and MethodsDisc diffusion was used to determine the biological activity of 20 plant extracts as well as 8 antibioticscommonly used to treat H. pylori infections. Minimum inhibitory concentrations were also measured by tubeand agar dilution methods for the biologically active plant extracts.ResultsOf the twenty plant extracts analyzed, sixteen exhibited good anti-H. pylori activity, using disc diffusion.The ten most active extracts were Carum bulbocastanum, Carum carvi, Mentha longifolia, Saliva limbata,Saliva sclarea, Ziziphora clinopodioides, Thymus caramanicus, Glycyrrhiza glabra, Xanthium brasilicumand Trachyspermum copticum. Minimum inhibitory concentrations measured for the 10 biologically activeplant extracts were within the range of 31.25 to 500 μg/ml.ConclusionAmong the ten plant extracts effective against H. pylori clinical isolates, Carum carvi, Xanthium brasilicumand Trachyspermum copticum showed the highest activity.Keywords: Anti-Helicobacter pylori, Iranian plants, Organic extracts

  19. Effect of edible sesame oil on growth of clinical isolates of Candida albicans.

    Science.gov (United States)

    Ogawa, Toshiko; Nishio, Junko; Okada, Shinobu

    2014-07-01

    Elderly individuals are at increased risk of oral thrush (oral candidiasis) due to decreased saliva secretion. Due to their antimicrobial properties, edible oils can be effective natural agents for oral care. The objective of the present study was to compare the effects of sesame oil, which is widely used for cooking in Asian countries, and two other edible oils on the growth of both mycelial and yeast forms of five clinical isolates of Candida albicans, a causative microorganism of oral thrush. We assessed the effect of each oil in concentrations of 0.078%, 0.156%, and 0.313% on growth of the mycelial forms of the clinical isolates over 24 hr using the crystal violet method. We also evaluated the effect of each oil on growth of the yeast forms by counting the number of viable yeast cells after culturing in the oils for 24 hr. Sesame oil inhibited the growth of both mycelial and yeast forms. Safflower and olive oil also inhibited the growth of both forms of C. albicans but to a lesser extent than sesame oil. The ability to inhibit the growth of the mycelial form correlated with sesame oil concentration. Roasting influenced growth inhibition ability and high-roasted sesame oil most effectively inhibited the yeast form. The growth inhibitory effect differed among the five isolates. We hypothesize that the sesamin and fatty acid components of sesame oil are involved in its antifungal activity.

  20. Use of Trichomonas vaginalis clinical isolates to evaluate correlation of gene expression and metronidazole resistance.

    Science.gov (United States)

    Mead, J R; Fernadez, M; Romagnoli, P A; Secor, W E

    2006-02-01

    We investigated whether variations in gene expression of enzymes associated with anaerobic resistance of laboratory-derived strains of Trichomonas vaginalis could be detected in a group of 28 clinical isolates with variations in metronidazole sensitivity. We compared isolates by real-time PCR because this method allows for highly sensitive quantification of mRNA and for evaluation of several genes simultaneously. We found that PFOR gene A mRNA levels were highly correlated with PFOR gene B levels, as well as the D subunit of malic enzyme and ferrodoxin. Ferrodoxin mRNA expression was also significantly correlated with that of malic enzyme and hydrogenase. However, when we evaluated relationships between these enzymes and resistance to metronidazole, we found no significant correlations between aerobic or anaerobic in vitro sensitivity to drug and mRNA levels of any of the enzymes tested. Similarly, using a Student's t-test, no significant differences in enzyme mRNA levels were observed between isolates separated by metronidazole resistance or susceptibility. The lack of correlation between gene expression and resistance or susceptibility could be the result of differences in expression at the protein level or because other biochemical pathways or genes are involved in the resistance observed in clinical settings. PMID:16629339

  1. In vitro time-kill curves study of three antituberculous combinations against Mycobacterium tuberculosis clinical isolates.

    Science.gov (United States)

    López-Gavín, Alexandre; Tudó, Griselda; Rey-Jurado, Emma; Vergara, Andrea; Hurtado, Juan Carlos; Gonzalez-Martín, Julian

    2016-01-01

    The objective of this study was to examine the in vitro synergism of three-drug combinations against Mycobacterium tuberculosis (levofloxacin/linezolid/ethambutol, levofloxacin/amikacin/ethambutol and levofloxacin/linezolid/amikacin) using the time-kill curves method. In total, 8 multidrug-resistant and 12 drug-susceptible M. tuberculosis isolates were used. Minimum inhibitory concentrations (MICs) of the isolates for each drug were determined by the proportions method. Time-kill curves were studied for the three combinations proposed over 14 days using two different protocols. In protocol 1, 0.5× MIC for each drug was used. In protocol 2, 0.5× MIC for levofloxacin and linezolid and 0.25× MIC for amikacin and ethambutol were used. The MICs for all of the isolates studied were 0.5 mg/L for levofloxacin and linezolid and 2.5 mg/L for ethambutol and amikacin. All of the combinations displayed an additive activity compared with the most active individual drug. In conclusion, these results demonstrate that the three combinations tested were equally effective against M. tuberculosis isolates. The study of antituberculous combinations using in vitro methods is an excellent first step to predict their effect in clinical development phases as well as to test new regimens of the antituberculous drugs currently available. PMID:26691020

  2. Aminoglycosides resistance in clinical isolates of Staphylococcus aureus from a University Hospital in Bialystok, Poland.

    Directory of Open Access Journals (Sweden)

    Katarzyna Kaczyńska

    2008-06-01

    Full Text Available Staphylococcus aureus obtained from a University Hospital in Poland were characterized in relation to resistance to aminoglycoside antibiotics and the distribution of the genes encoding the most clinically relevant aminoglycoside modifying enzymes (AMEs. Of a total of 118 S. aureus, 45 (38.1% isolates were found to be resistant to at least one of the tested antibiotics. All aminoglycoside resistant isolates except one 44 (97.8% were resistant to kanamycin. The majority of strains 37 (82.2% and 32 (71.1% expressed resistance to neomycin and tobramycin, respectively. Eleven strains (24.4% were resistant to gentamicin or amikacin. All S. aureus strains were sensitive to netilmicin. The most prevalent resistance gene was aac(6'-Ie+aph(2' found in 13 (28.9% strains and 12 (26.7% isolates carried ant(4'-Ia gene, whilst aph(3'-IIIa gene was detected in only 7 (15.6% isolates. Additionally, the ant(6-Ia and str genes were detected in 14 (31.1% and 2 (4.4% strains, respectively. Ten (22.2% strains resistant to amikacin, tobramycin, kanamycin or neomycin did not harbor any of the above-noted genes.

  3. Relatedness of Mycobacterium avium subspecies hominissuis clinical isolates of human and porcine origins assessed by MLVA.

    Science.gov (United States)

    Leão, Célia; Canto, Ana; Machado, Diana; Sanches, Ilda Santos; Couto, Isabel; Viveiros, Miguel; Inácio, João; Botelho, Ana

    2014-09-17

    Mycobacterium avium subsp. hominissuis (MAH) is an important opportunistic pathogen, infecting humans and animals, notably pigs. Several methods have been used to characterize MAH strains. RFLP and PFGE typing techniques have been used as standard methods but are technically demanding. In contrast, the analysis of VNTR loci is a simpler, affordable and highly reliable PCR-based technique, allowing a numerical and reproductive digitalization of typing data. In this study, the analysis of Mycobacterium avium tandem repeats (MATRs) loci was adapted to evaluate the genetic diversity of epidemiological unrelated MAH clinical strains of human (n=28) and porcine (n=69) origins, collected from diverse geographical regions across mainland Portugal. These MAH isolates were found to be genetically diverse and genotypes are randomly distributed across the country. Some of the human strains shared identical VNTR profiles with porcine isolates. Our study shows that the VNTR genotyping using selected MATR loci is a useful analysis technique for assessing the genetic diversity of MAH isolates from Portugal. This typing method could be successfully applied in other countries toward the implementation of a worldwide open-access database of MATR-VNTR profiles of MAH isolates, allowing a better assessment of the global epidemiology traits of this important pathogenic species.

  4. Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP

    DEFF Research Database (Denmark)

    Taponen, S.; Simojoki, H.; Haveri, M.;

    2006-01-01

    The coagulase-negative staphylococcal species causing mastitis in lactating cattle were identified and possible differences in the clinical characteristics or persistence of mastitis caused by different CNS were evaluated. The effect of antimicrobial treatment was also assessed. In addition, AFLP...... of these species. Approximately half of the mastitis cases were clinical, and in the majority clinical signs were mild. The severity and persistence of intramammary infection were unaffected by CNS species. Fifty-nine percent of the quarter cases were treated with antimicrobials, and the rest were left without...... treatment. Mastitis due to P-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were P-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45...

  5. Comparison of lipopolysaccharide structures of Bordetella pertussis clinical isolates from pre- and post-vaccine era.

    Science.gov (United States)

    Albitar-Nehme, Sami; Basheer, Soorej M; Njamkepo, Elisabeth; Brisson, Jean-Robert; Guiso, Nicole; Caroff, Martine

    2013-08-30

    Endotoxins are lipopolysaccharides (LPS), and major constituents of the outer membrane of Gram-negative bacteria. Bordetella pertussis LPS were the only major antigens, of this agent of whooping-cough, that were not yet analyzed on isolates from the pre- and post-vaccination era. We compared here the LPS structures of four clinical isolates with that of the vaccine strain BP 1414. All physico-chemical analyses, including SDS-PAGE, TLC, and different MALDI mass spectrometry approaches were convergent. They helped demonstrating that, on the contrary to some other B. pertussis major antigens, no modification occurred in the dodecasaccharide core structure, as well as in the whole LPS molecules. These results are rendering these major antigens good potential vaccine components. Molecular modeling of this conserved LPS structure also confirmed the conclusions of previous experiments leading to the production of anti-LPS monoclonal antibodies and defining the main epitopes of these major antigens.

  6. In vitro activity of tigecycline and 10 other antimicrobials against clinical isolates of the genus Corynebacterium.

    Science.gov (United States)

    Fernandez-Roblas, R; Adames, H; Martín-de-Hijas, N Z; Almeida, D García; Gadea, I; Esteban, J

    2009-05-01

    We studied the in vitro activity of tigecycline and 10 other commonly used antibiotics against 135 clinical isolates of non-diphtheria Corynebacterium spp. using the Etest system. Tigecycline minimum inhibitory concentrations for 50% and 90% of the organisms (MIC(50) and MIC(90) values, respectively, in mg/L) were: Corynebacterium urealyticum, 0.094 and 0.125; Corynebacterium amycolatum, 0.125 and 2; Corynebacterium jeikeium, 0.094 and 0.75; Corynebacterium coyleae, 0.064 and 0.064; Corynebacterium striatum, 0.064 and 1; Corynebacterium aurimucosum, 0.094 and 0.125; and Corynebacterium afermentans, 0.064 and 0.094. The activities of all other antimicrobials were variable, with good activity of glycopeptides, linezolid, quinupristin/dalfopristin and daptomycin and with resistance to macrolides in a high number of isolates. Tigecycline is a good alternative for the therapy of infections caused by non-diphtheria corynebacteria. PMID:19153032

  7. Multiple sclerosis presented as clinically isolated syndrome: the need for early diagnosis and treatment

    Directory of Open Access Journals (Sweden)

    Sigliti-Henrietta Pelidou

    2008-06-01

    Full Text Available Sigliti-Henrietta Pelidou, Sotirios Giannopoulos, Sotiria Tzavidi, Georgios Lagos, Athanassios P KyritsisDepartment of Neurology, University of Ioannina School of Medicine, GreeceObjective: To aid in the timely diagnosis of patients who present with clinically isolated syndrome (CIS.Patients and methods: We studied 25 patients (18 women, 7 men, originally presented in our clinic with a CIS suggestive of multiple sclerosis (MS. All patients underwent the full investigation procedure including routine tests, serology, cerebrospinal fluid (CSF examinations, evoked potentials (EPs, and magnetic resonance imaging (MRI of brain and cervical spinal cord. Patients were imaged at baseline, and every three months thereafter up to a year.Results: The CIS was consisted of optic neuritis in 12 cases, incomplete transverse myelitis (ITM in 7 cases, Lhermitte sign in 2 cases, internuclear ophthalmoplegia (INO in 2 cases, mild brainstem syndrome in 1 case, and tonic-clonic seizures in 1 case. Using the baseline and three-month scans 18/25 (72% patients developed definite MS in one year of follow up while 7 (28% had no further findings during this observation period. Immunomodulatory treatments were applied to all definite MS patients.Conclusion: In light of new treatments available, MRIs at 3 month intervals are helpful to obtain the definite diagnosis of MS as early as possible.Keywords: multiple sclerosis, clinically isolated syndrome, optic neuritis, transverse myelitis

  8. Genomic characterization of isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis

    OpenAIRE

    Silva, E.; Gaivão, M.; Leitão, S.; Jost, B.H.; Carneiro, C.; Vilela, C.L.; Da Costa, L. Lopes; Mateus, L.

    2008-01-01

    Genomic characterization of Arcanobacterium pyogenes isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis PORTUGAL (Silva, E.) PORTUGAL Received: 2008-03-18 Revised: 2008-04-23 Accepted: 2008-04-25

  9. Molecular characterization of clinical isolates of Cyclospora cayetanensis from patients with diarrhea in India

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    P Yadav

    2015-01-01

    Full Text Available Purpose: Cyclospora cayetanensis is an intestinal coccidian protozoan that has emerged as an important cause of both epidemic and endemic protracted diarrhea worldwide. Though humans appear to be the only natural hosts; the role of animals as natural reservoir is uncertain but of increasing concern. The present study aimed to study the prevalence of coccidian in different groups such as immunocompromised, clinically apparent immunocompetent and healthy individuals. Also, the study isolates were assessed for heterogeneity among the sequences. Materials and Methods: Stool samples from different groups of patients were collected. The parasite was detected in stool by different diagnostic tools such as light microscopy and nested PCR-restriction fragment length polymorphism using 18S ribosomal RNA as the target gene. Results: The prevalence of C. cayetanensis was 2.4% (19/800 in the present study. The PCR assay amplified Cyclospora cayetanensis DNA in only 89% (17/19 isolates. Further, sequencing revealed no significant difference among the study isolates and the non-primates. Phylogenetic analysis of the study isolates however, formed two clusters. While one cluster showed close evolutionary association with the C. cayetanensis strains, the other cluster showed evolutionary association with the two non-primate species. Conclusion: The methods described here for detection of C. cayetanensis oocysts are simple, efficient, specific, and sensitive and therefore can be effectively applied for laboratory diagnosis and environmental assessment of fresh produce and water sources. Clinicians should include Cyclospora infection in the differential diagnosis of prolonged or relapsing diarrheal illness even in clinically apparent immunocompetent individuals.

  10. Speciation and Biofilm Production of Coagulase Negative Staphylococcal Isolates from Clinically Significant Specimens and their Antibiogram

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    S. S. Vijayasri Badampudi

    2016-04-01

    Full Text Available Background: Coagulase Negative Staphylococci (CONS are increasingly recognized as significant nosocomial pathogens. Their ability of biofilm formation and multiple drug resistance are causing serious human infections. Aim and Objectives: To isolate, identify, speciate clinically significant CONS from various specimens, to study antibiotic resistance pattern and biofilm production. Material and Methods: Specimens were collected aseptically, processed and identified upto the species level by a simple scheme of tests urease, novobiocin resistance, mannose and mannitol fermentation, ornithine decarboxylase. Antibiotic sensitivity was done with special reference to methicillin resistance. Biofilm formation was detected by Congo Red Agar (CRA method and Tube Method (TM. Results: Study groupOf 100 isolates majority were pus (40, followed by urine (28, blood (16, CSF (5, body fluids (4 and catheter tips and implants (7. The most common species isolated was S. epidermidis (40% followed by S. haemolyticus (26%, S. saprophyticus (15%, S. schleiferi (13%, S. simulans (2%, S. cohnii (2% and S. warneri and S. capitis each 1%. Resistance to penicillin was 91% followed by ampicillin (79%, cotrimoxazole (67%. Methicillin resistance was 72%. Biofilm producers were 69% by CRAmethod and 33% by TM with majority species S. epidermidis (82.5%- CRA and 55%-TM. Biofilm production was significantly associated with MRCONS (p value 0.0036. Control group-Of 30 isolates were S. epidermidis 66.6% followed by S. haemolyticus (16.66%. Biofilm producers were 53.33% by CRA method and 26.65% by TM with majority species S. epidermidis (65%-CRA and 30%-TM.Methicillin resistance was 26.6%. Conclusion: Clinical significance of CONS is increasing day by day, so there is a need for accurate identification to species level and their antibiogram to avoid multidrug resistance. Biofilm producing CONS species pose a risk and CRA method for screening biofilm can be used in all conventional

  11. Characterization of BKC-1 class A carbapenemase from Klebsiella pneumoniae clinical isolates in Brazil.

    Science.gov (United States)

    Nicoletti, Adriana Giannini; Marcondes, Marcelo F M; Martins, Willames M B S; Almeida, Luiz G P; Nicolás, Marisa F; Vasconcelos, Ana T R; Oliveira, Vitor; Gales, Ana Cristina

    2015-09-01

    Three Klebsiella pneumoniae clinical isolates demonstrating carbapenem resistance were recovered from different patients hospitalized at two medical centers in São Paulo, Brazil. Resistance to all β-lactams, quinolones, and some aminoglycosides was observed for these isolates that were susceptible to polymyxin B. Carbapenem hydrolysis, which was inhibited by clavulanic acid, was observed for all K. pneumoniae isolates that belonged to the same pulsed-field gel electrophoresis (PFGE) type and a novel sequence type (ST), ST1781 (clonal complex 442 [CC442]). A 10-kb nonconjugative incompatibility group Q (IncQ) plasmid, denominated p60136, was transferred to Escherichia coli strain TOP10 cells by electroporation. The full sequencing of p60136 showed that it was composed of a mobilization system, ISKpn23, the phosphotransferase aph3A-VI, and a 941-bp open reading frame (ORF) that codified a 313-amino acid protein. This ORF was named bla BKC-1. Brazilian Klebsiella carbapenemase-1 (BKC-1) showed a pI of 6.0 and possessed the highest identity (63%) with a β-lactamase of Sinorhizobium meliloti, an environmental bacterium. Hydrolysis studies demonstrated that purified BKC-1 not only hydrolyzed carbapenems but also penicillins, cephalosporins, and monobactams. However, the carbapenems were less efficiently hydrolyzed due to their very low kcat values (0.0016 to 0.031 s(-1)). In fact, oxacillin was the best substrate for BKC-1 (kcat /Km , 53,522.6 mM(-1) s(-1)). Here, we report a new class A carbapenemase, confirming the diversity and rapid evolution of β-lactamases in K. pneumoniae clinical isolates. PMID:26055384

  12. Antimicrobial resistance trends among canine Escherichia coli isolates obtained from clinical samples in the northeastern USA, 2004–2011

    OpenAIRE

    Cummings, Kevin J.; Aprea, Victor A.; Altier, Craig

    2015-01-01

    Our objectives were to describe the antimicrobial susceptibility of Escherichia coli isolates from dogs in the northeastern USA and to identify temporal trends in resistance to selected antimicrobial agents. Data were collected retrospectively for all canine E. coli isolates from clinical samples submitted to Cornell University’s Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Antimicrobial susceptibility testing was performed on 3519 canine E. coli isolates; fr...

  13. Genetics of oxacillin resistance in clinical isolates of Streptococcus pneumoniae that are oxacillin resistant and penicillin susceptible.

    OpenAIRE

    Dowson, C G; Johnson, A P; Cercenado, E.; George, R C

    1994-01-01

    It has recently been reported that penicillin-sensitive pneumococci may exhibit reduced susceptibility to oxacillin, resulting in their misclassification as being penicillin resistant by oxacillin disk testing. Intermediate oxacillin resistance (MIC, 1.0 microgram/ml) in three of these apparently unrelated penicillin-susceptible clinical isolates of Streptococcus pneumoniae isolated in the United Kingdom and in four Spanish isolates was shown to be solely due to the acquisition of a gene enco...

  14. Mutations in katG gene sequences in isoniazid-resistant clinical isolates of Mycobacterium tuberculosis are rare.

    OpenAIRE

    Pretorius, G.S.; van Helden, P. D.; Sirgel, F; Eisenach, K D; Victor, T C

    1995-01-01

    In this study, a battery of oligonucleotides was directed toward the katG gene and PCR-single-stranded conformation polymorphism (SSCP) analysis was used to search for katG gene deviations in clinical isolates of Mycobacterium tuberculosis from different geographical regions. Since a complete deletion of the katG gene was not found, it is suggested that deletion is not a major mechanism of isoniazid (isonicotinic acid hydrazide; INH) resistance in these isolates. However, 7 of 39 isolates (4 ...

  15. In vitro synergistic activities of tobramycin and selected beta-lactams against 75 gram-negative clinical isolates.

    OpenAIRE

    Owens, R C; Banevicius, M A; Nicolau, D. P.; Nightingale, C H; Quintiliani, R

    1997-01-01

    The microdilution checkerboard technique was utilized to distinguish synergistic activity between tobramycin and four beta-lactams: piperacillin-tazobactam, ticarcillin-clavulanate, ceftazidime, and ceftriaxone. Beta-lactam-aminoglycoside combinations were tested against 75 clinical isolates of Pseudomonas aeruginosa, Acinetobacter baumanii, Citrobacterfreundii, Serratia marcescens, and Enterobacter cloacae. Despite in vitro susceptibilities, all isolates demonstrated either synergism or indi...

  16. The pls Gene Found in Methicillin-Resistant Staphylococcus aureus Strains Is Common in Clinical Isolates of Staphylococcus sciuri

    OpenAIRE

    Juuti, Katri; Ibrahem, Salha; Virolainen-Julkunen, Anni; Vuopio-Varkila, Jaana; Kuusela, Pentti

    2005-01-01

    pls, a gene found in type I staphylococcal cassette chromosome mec (SCCmec) regions of methicillin-resistant Staphylococcus aureus strains, was present in 12 of the 15 human clinical Staphylococcus sciuri isolates studied. Pls was expressed in the S. sciuri isolates, although at a lower level than in S. aureus. Other parts of SCCmec could also be found in the S. sciuri genome.

  17. Antimicrobial susceptibility and clarithromycin resistance patterns of Helicobacter pylori clinical isolates in Vietnam.

    Science.gov (United States)

    Quek, Camelia; Pham, Son T; Tran, Kieu T; Pham, Binh T; Huynh, Loc V; Luu, Ngan B L; Le, Thao K T; Quek, Kelly; Pham, Van H

    2016-01-01

    Helicobacter pylori is a gastric pathogen that causes several gastroduodenal disorders such as peptic ulcer disease and gastric cancer.  Eradication efforts of H. pylori are often hampered by antimicrobial resistance in many countries, including Vietnam.  Here, the study aimed to investigate the occurrence of antimicrobial resistance among H. pylori clinical isolates across 13 hospitals in Vietnam.  The study further evaluated the clarithromycin resistance patterns of H. pylori strains.  In order to address the study interests, antimicrobial susceptibility testing, epsilometer test and PCR-based sequencing were performed on a total of 193 strains isolated from patients, including 136 children (3-15 years of age) and 57 adults (19-69 years of age).  Antimicrobial susceptibility testing showed that the overall resistance to amoxicillin, clarithromycin, levofloxacin, metronidazole, and tetracycline was 10.4%, 85.5%, 24.4%, 37.8%, and 23.8% respectively.  The distribution of minimum