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Sample records for streptoccocus agalactiae isolates

  1. [Isolation and characterization of siphovirus phages infecting bovine Streptococcus agalactiae].

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    Bai, Qinqin; Yang, Yongchun; Lu, Chengping

    2016-02-04

    To isolate and identify Streptococcus agalactiae phages and screen candidate phages to control infection caused by bovine S. agalactiae. We used two methods for isolation of S. agalactiae phages, namely (1) isolation of phages from milk and environmental samples, and (2) isolation of phages via induction of lysogens with Mitomycin C. Double-layer agar culture method was used to purify phages. Then the newly obtained phages, with S. agalactiae phage JX01 isolated from mastitis milk, were comparatively analyzed in the following aspects: morphology of phages by transmission electron microscopy, host range of phages to 55 S. agalactiae strains and other Streptococcus strains, phages DNA using EcoR I, Xba I, Pst I and Sal I, the optical multiplicity of infection, absorption curve and one step growth curve, and the stability of phages at different storage conditions. The comparative analysis of the 3 novel phages LYGO9, HZ04 and pA11 (induced from S. agalctiae bovine clinical isolate HAJL2011070601) with JX01 showed that the 4 phages were classified as the member of Siphovirdae family. EcoR I, Sal I, Xba I and Pst I separately digested the 4 phages DNA provided 4, 3, 3 and 2 profiles, respectively. This suggested that they were different strains. All the 4 phages specifically infected bovine S. agalactiae isolates. LYGO9, pA11, JX01 and HZ04 could lyse 12, 13, 20 and 23 of 42 tested bovine S. agalctiae isolates, respectively. This clearly indicated that these 4 phages are closely related. The 3 new phages which specifically lyse bovine S. agalactiae isolates are siphovirus phages. Phage LYGO9 was shown having a short latent period and a larger burst size.

  2. Emergence of respiratory Streptococcus agalactiae isolates in cystic fibrosis patients.

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    Vera Eickel

    Full Text Available Streptococcus agalactiae is a well-known pathogen for neonates and immunocompromized adults. Beyond the neonatal period, S. agalactiae is rarely found in the respiratory tract. During 2002-2008 we noticed S. agalactiae in respiratory secretions of 30/185 (16% of cystic fibrosis (CF patients. The median age of these patients was 3-6 years older than the median age CF patients not harboring S. agalactiae. To analyze, if the S. agalactiae isolates from CF patients were clonal, further characterization of the strains was achieved by capsular serotyping, surface protein determination and multilocus sequence typing (MLST. We found a variety of sequence types (ST among the isolates, which did not substantially differ from the MLST patterns of colonizing strains from Germany. However serotype III, which is often seen in colonizing strains and invasive infections was rare among CF patients. The emergence of S. agalactiae in the respiratory tract of CF patients may represent the adaptation to a novel host environment, supported by the altered surfactant composition in older CF patients.

  3. Antibiotic Susceptibilities and Serotyping of Clinical Streptococcus Agalactiae Isolates

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    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.

  4. Complete genome sequence of a virulent Streptococcus agalactiae strain 138P isolated from diseased Nile tilapia

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    Streptococcus agalactiae strain 138P was isolated from the kidney of diseased Nile tilapia in Idaho during a 2007 streptococcal disease outbreak. The full genome of S. agalactiae 138P is 1,838,716 bp. The availability of this genome will allow comparative genomics to identify genes for antigen disco...

  5. Short communication: In vitro antimicrobial susceptibility of Mycoplasma agalactiae strains isolated from dairy goats.

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    Paterna, A; Sánchez, A; Gómez-Martín, A; Corrales, J C; De la Fe, C; Contreras, A; Amores, J

    2013-01-01

    This study examined the susceptibility to several antimicrobials of 28 isolates of Mycoplasma agalactiae obtained from goats in a region (southeastern Spain) where contagious agalactia is endemic. For each isolate, the minimum inhibitory concentration (MIC) against 12 antimicrobials of the quinolone, macrolide, aminoglycoside, and tetracycline families was determined. The antimicrobials with the lowest MIC were enrofloxacin, ciprofloxacin, tylosin, and doxycycline, all with MIC90 (concentration at which growth of 90% of the isolates is inhibited) <1 µg/mL. Norfloxacin (a quinolone) showed a wide MIC range (0.1-12.8 µg/mL), suggesting a resistance mechanism toward this antimicrobial that was not elicited by enrofloxacin or ciprofloxacin (the other quinolones tested). Erythromycin showed the highest MIC90 such that its use against Mycoplasma agalactiae is not recommended. Finally, Mycoplasma agalactiae isolates obtained from goat herds with clinical symptoms of contagious agalactia featured higher MIC90 and MIC50 (concentration at which growth of 50% of the isolates is inhibited) values for many of the antimicrobials compared with isolates from asymptomatic animals. The relationship between the extensive use of antimicrobials in herds with clinical contagious agalactia and variations in MIC requires further study. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. Isolation and prevalence of Mycoplasma agalactiae in Kurdish sheep in Kurdistan, Iran

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    Mohammad Khezri

    Full Text Available Aim: Ruminant Mycoplasmosis are important diseases worldwide and several are listed by the World Organization for Animal Health (OIE to be of major economic significant. The aim of this study was to isolation mycoplasmas from sheep presenting contagious agalactiae (CA in Kurdistan in the West of Iran. Materials and Methods: Sixty-nine samples included (milk, conjuctiva swabs, synovial fluid and ear canal swabs were examined by PCR assay during 2011-2012. DNA was extracted from enriched samples. Two primers (forward and reverse amplify a 163bp region of 16S rRNA gene of Mycoplasma genus and two primers amplify 375bp region of 16S rRNA gene of Mycoplasma agalactiae (M. agalactiae species were used. Results: This proved that 46 samples (66.7% were infected with Mycoplasma in culture and PCR test, respectively. On the PCR test, 15 isolates (32.6% examined were positive for M. agalactiae that showed specific amplicon at 375bp. All Mycoplasma positive samples were analyzed for M. agalactiae infection by PCR method and 31 isolates (67.4% examined were negative for M. agalactiae. The finding of other mycoplasmas with significant epidemiology challenges existing plans for the control of CA in sheep population in Iran. Conclusion: The results of the present study show that M. agalactiae in CA disease in Kurdistan Province, 32.6% involved. In Iran, only M. agalactiae vaccines are commercially available, thus, the animals are not protected against these other species. [Vet World 2012; 5(12.000: 727-731

  7. Isolation, characterization, virulence and immunogenicity testing of field isolates of Pasteurella multocida, Staphylococcus aureus, and Streptococcus agalactiae in laboratory settings.

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    Qudratullah; Muhammad, G; Saqib, M; Bilal, M Qamar

    2017-08-01

    The present study was designed to investigate isolation, characterization, virulence and immunogenicity testing of field isolates of Pasteurella multocida, Staphylococcus aureus, and Streptococcus agalactiae in rabbits and mice. Isolates of P. multocida, S. aureus and Str. agalactiae recovered from field cases of Hemorragic septicemia and mastitis were scrutinized for virulence/pathogenicity and immunogenicity. Mouse LD 50 of P. multocida showed that P. multocida isolate No.1 was more virulent than isolates No. 2 and 3. Virulence of isolate No.1S. aureus and Str. agalactiae revealed that 100, 80% rabbits died within 18h of inoculation. Seven-digit numerical profiles of these 4 isolates with API ® Staph test strips isolates, No.1 (6736153) showed good identification (S. aureus id=90.3%). Indirect ELISA-based serum antibody titers to P. multocida isolate No.1, S. aureus No.1, Str. agalactiae, isolate No.1 elicited high antibody titers 1.9, 1.23, 1.12 respectively. All the pathogens of Isolate No. 1 (P. multocida, S. aureus Str. agalactiae), were high antibody than others isolates. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Identification of immunoreactive proteins of Streptococcus agalactiae isolated from cultured tilapia in China.

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    Liu, Guangjin; Zhang, Wei; Lu, Chengping

    2013-12-01

    Streptococcus agalactiae (Group B streptococcus, GBS) is an important zoonotic pathogen that can cause lethal infections in humans and animals, including aquatic species. Immunoreactive proteins of the S. agalactiae strain, GD201008-001, isolated from cultured tilapia in China, were screened by immunoproteomics using hyperimmune sera, convalescent guinea pig sera and GD201008-001-infected tilapia antisera as primary detection antibodies. A total of 16 different proteins were identified including 13 novel immunoreactive proteins of S. agalactiae. Four proteins, serine-rich repeat glycoprotein 1, branched-chain alpha-keto acid dehydrogenase (BKD) subunit E2, 5'-nucleotidase family protein and ornithine carbamoyltransferase, were shown to react with the three types of sera and thus were considered to represent novel S. agalactiae vaccine candidate antigens. Our findings represent the basis for vaccine development for piscine S. agalactiae and are necessary for understanding virulence factors and immunogenicity of S. agalactiae with different hosts. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

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

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    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.

  10. Comparison of virulence factors and capsular types of Streptococcus agalactiae isolated from human and bovine infections.

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    Emaneini, Mohammad; Khoramian, Babak; Jabalameli, Fereshteh; Abani, Samira; Dabiri, Hossein; Beigverdi, Reza

    2016-02-01

    Streptococcus agalactiae is a leading cause of human and bovine infections. A total of 194 S. agalactiae isolates, 55 isolates from bovines and 139 from humans, were analyzed for capsular types, virulence genes (scpB, hly, rib, bca and bac) and mobile genetic elements (IS1548 and GBSi1) using polymerase chain reaction (PCR) and multiplex PCR. Capsular type III was predominant (61%), followed by types V, II, Ib, and IV. The scpB, hly, bca and bac virulence genes were only found among human isolates. Twelve and 2 distinct virulence gene profiles were identified among human and bovine isolates respectively. The virulence gene profiles scpB- hly- IS1548- rib-bca (51%) and scpB- hly- IS1548- bca (19%) were only predominant among human isolates. The rib gene was the most common virulence gene in both human and bovine isolates. The study showed a high prevalence of virulence genes in S. agalactiae strains isolated from human infections, these result can support the idea that S. agalactiae isolated from humans and bovines are generally unrelated and probably belonged to separate populations. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

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

    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 clonal groups and were clustered in six different lineages, i.e. ST (sequence type) 103, ST568, ST67, ST301, ST313 and ST570, of which ST568 and ST570 were new genotypes. PFGE analysis revealed this isolates were clustered in 27 PFGE types, of which, types 7, 8, 14, 15, 16, 18, 23 and 25 were the eight major types, comprising close to 70% (71/102) of all the isolates. The most prevalent sequence types were ST103 (58% isolates) and ST568 (31% isolates), comprising capsular genotype Ia isolates without any of the detected Alp genes, suggesting the appearance of novel genomic backgrounds of prevalent strains of bovine S. agalactiae. All the strains possessed the pilus island 2b (PI-2b) gene and the prevalent capsular genotypes were types Ia (89% isolates) and II (11% isolates), the conserved pilus type providing suitable data for the development of vaccines against mastitis caused by S. agalactiae. PMID:23874442

  12. Molecular characterization of Streptococcus agalactiae strains isolated from fishes in Malaysia.

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    Amal, M N A; Zamri-Saad, M; Siti-Zahrah, A; Zulkafli, A R; Nur-Nazifah, M

    2013-07-01

    The aim of this study was to characterize Streptococcus agalactiae strains that were isolated from fishes in Malaysia using random amplified polymorphic DNA (RAPD) and repetitive extragenic palindromic PCR (REP-PCR) techniques. A total of 181 strains of Strep. agalactiae isolated from red hybrid tilapia (Oreochromis sp.) and golden pompano (Trachinotus blochii) were characterized using RAPD and REP-PCR techniques. Both the fingerprinting techniques generated reproducible band patterns, differing in the number and molecular mass amplicons. The RAPD technique displayed greater discriminatory power by its production of more complex binding pattern and divided all the strains into 13 groups, compared to 9 by REP-PCR technique. Both techniques showed the availability to differentiate the genetic profiles of the strains according to their geographical location of origin. Three strains of Strep. agalactiae that were recovered from golden pompano showed a genetic dissimilarity from the strains isolated from red hybrid tilapia, while the strain of ATCC 27956 that recovered from bovine displayed a unique profile for both methods. Both techniques possess excellent discriminative capabilities and can be used as a rapid means of comparing Strep. agalactiae strains for future epidemiological investigation. Framework as the guideline in traceability of this disease and in the search for potential local vaccine candidates for streptococcosis in this country. Journal of Applied Microbiology © 2013 The Society for Applied Microbiology.

  13. Streptococcus agalactiae in elephants - A comparative study with isolates from human and zoo animal and livestock origin.

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    Eisenberg, Tobias; Rau, Jörg; Westerhüs, Uta; Knauf-Witzens, Tobias; Fawzy, Ahmad; Schlez, Karen; Zschöck, Michael; Prenger-Berninghoff, Ellen; Heydel, Carsten; Sting, Reinhard; Glaeser, Stefanie P; Pulami, Dipen; van der Linden, Mark; Ewers, Christa

    2017-05-01

    Streptococcus (S.) agalactiae represents a significant pathogen for humans and animals. However, there are only a few elderly reports on S. agalactiae infections in wild and zoo elephants even though this pathogen has been isolated comparatively frequently in these endangered animal species. Consequently, between 2004 and 2015, we collected S. agalactiae isolates from African and Asian elephants (n=23) living in four different zoos in Germany. These isolates were characterised and compared with isolates from other animal species (n=20 isolates) and humans (n=3). We found that the isolates from elephants can be readily identified by classical biochemistry and MALDI-TOF mass spectrometry. Further characterisations for epidemiological issues were achieved using Fourier transform-infrared spectroscopy, capsule typing and molecular fingerprinting (PFGE, RAPD PCR). We could demonstrate that our elephant isolate collection contained at least six different lineages that were representative for their source of origin. Despite generally broad antimicrobial susceptibility of S. agalactiae, many showed tetracycline resistance in vitro. S. agalactiae plays an important role in bacterial infections not only in cattle and humans, but also in elephants. Comparative studies were able to differentiate S. agalactiae isolates from elephants into different infectious clusters based on their epidemiological background. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Serotypes, antibiotic susceptibilities, and multi-locus sequence type profiles of Streptococcus agalactiae isolates circulating in Beijing, China.

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    Wang, Ping; Tong, Jing-jing; Ma, Xiu-hua; Song, Feng-li; Fan, Ling; Guo, Cui-mei; Shi, Wei; Yu, Sang-jie; Yao, Kai-hu; Yang, Yong-hong

    2015-01-01

    To investigate the serotypes, antibiotic susceptibilities, and multi-locus sequence type (MLST) profiles of Streptococcus agalactiae (S. agalactiae) in Beijing to provide references for the prevention and treatment of S. agalactiae infections. All isolates were identified using the CAMP test and the latex-agglutination assay and serotyped using a Strep-B-Latex kit, after which they were assessed for antibiotic susceptibility, macrolide-resistance genes, and MLST profiles. In total, 56 S. agalactiae isolates were identified in 863 pregnant women (6.5%). Serotypes Ia, Ib, II, III, and V were identified, among which types III (32.1%), Ia (17.9%), Ib (16.1%), and V (14.3%) were the predominant serotypes. All isolates were susceptible to penicillin and ceftriaxone. The nonsusceptiblity rates measured for erythromycin, clarithromycin, azithromycin, telithromycin, clindamycin, tetracycline, and levofloxacin were 85.7%, 92.9%, 98.2%, 30.4%, 73.2%, 91%, and 39.3%, respectively. We identified 14 sequence types (STs) for the 56 isolates, among which ST19 (30.4%) was predominant. The rate of fluoroquinolone resistance was higher in serotype III than in the other serotypes. Among the 44 erythromycin-resistant isolates, 32 (72.7%) carried ermB. S. agalactiae isolates of the serotypes Ia, Ib, III, and V are common in Beijing. Among the S. agalactiae isolates, the macrolide and clindamycin resistance rates are extremely high. Most of the erythromycin-resistant isolates carry ermB.

  15. Isolation and identification of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR from sheep of Qom province, Iran

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    Abtin, A.R.

    2013-05-01

    Full Text Available Contagious agalactia (C.A is an infectious syndrome of sheep that is characterized by mastitis andsubsequent failure of milk production, arthritis, abortion and keratoconjunctivitis. Mycoplasma agalactiae(M. agalactiae is the main cause of the disease in sheep. The aim of this study was isolation andidentification of M. agalactiae with culture and polymerase chain reaction (PCR assay from sheep of Qomprovince in Iran. A total of 102 samples were collected from milk secretion, eye, ear and joint exudates ofsheep. All samples were cultured in PPLO broth supplemented for M. agalaciae isolation. The bacteriaDNAs were extracted by phenol/chloroform method and the PCR assay was applied for detecting ofMycoplasma genus in 163bp fragment of 16S rRNA gene and M. agalactiae in 375bp fragment oflipoprotein gene from culture as same as in clinical samples. Out of the 102 samples, 19(18.63% cultureswere shown positive and typical Mycoplasma colonies in PPLO agar culture diagnostic method and59(57.8% were scored positive by Mycoplasma genus PCR, 19(18.62% of the samples were scoredpositive by using M. agalactiae PCR as diagnostic method. Out of the 102 samples, 19 samples wereshown both positive in the culture and PCR, 42 samples were shown both negative in the culture and PCR.40 samples were negative in the culture and positive in PCR whereas only one sample was positive inculture and negative in PCR. The results showed that the more isolations of M. agalactiae were taken from milk and less in joint samples. M. agalactiae was one of the main factors of contagious agalactia that was detected for the first time from sheep in Qom province.

  16. The genetic diversity and phenotypic characterisation of Streptococcus agalactiae isolates from Rio de Janeiro, Brazil

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    Ana Beatriz de Almeida Corrêa

    2011-12-01

    Full Text Available Streptococcus agalactiae isolates are more common among pregnant women, neonates and nonpregnant adults with underlying diseases compared to other demographic groups. In this study, we evaluate the genetic and phenotypic diversity in S. agalactiae strains from Rio de Janeiro (RJ that were isolated from asymptomatic carriers. We analysed these S. agalactiae strains using pulsed-field gel electrophoresis (PFGE, serotyping and antimicrobial susceptibility testing, as well as by determining the macrolide resistance phenotype, and detecting the presence of the ermA/B, mefA/E and lnuB genes. The serotypes Ia, II, III and V were the most prevalent serotypes observed. The 60 strains analysed were susceptible to penicillin, vancomycin and levofloxacin. Resistance to clindamycin, chloramphenicol, erythromycin, rifampin and tetracycline was observed. Among the erythromycin and/or clindamycin resistant strains, the ermA, ermB and mefA/E genes were detected and the constitutive macrolides, lincosamides and streptogramin B-type resistance was the most prevalent phenotype observed. The lnuB gene was not detected in any of the strains studied. We found 56 PFGE electrophoretic profiles and only 22 of them were allocated in polymorphism patterns. This work presents data on the genetic diversity and prevalent capsular serotypes among RJ isolates. Approximately 85% of these strains came from pregnant women; therefore, these data may be helpful in developing future prophylaxis and treatment strategies for neonatal syndromes in RJ.

  17. Molecular epidemiology of Streptococcus agalactiae isolated from mastitis in Brazilian dairy herds.

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    Carvalho-Castro, Glei A; Silva, Juliana R; Paiva, Luciano V; Custódio, Dircéia A C; Moreira, Rafael O; Mian, Glaucia F; Prado, Ingrid A; Chalfun-Junior, Antônio; Costa, Geraldo M

    Streptococcus agalactiae is one of the most common pathogens leading to mastitis in dairy herds worldwide; consequently, the pathogen causes major economic losses for affected farmers. In this study, multilocus sequence typing (MLST), genotypic capsular typing by multiplex polymerase chain reaction (PCR), and virulence gene detection were performed to address the molecular epidemiology of 59 bovine (mastitis) S. agalactiae isolates from 36 dairy farms located in the largest milk-producing mesoregions in Brazil (Minas Gerais, São Paulo, Paraná, and Pernambuco). We screened for the virulence genes bac, bca, bibA, cfb, hylB, fbsA, fbsB, PI-1, PI-2a, and PI-2b, which are associated with adhesion, invasion, tissue damage, and/or immune evasion. Furthermore, five capsular types were identified (Ia, Ib, II, III, and IV), and a few isolates were classified as non-typeable (NT). MLST revealed the following eight sequence types (STs): ST-61, ST-67, ST-103, ST-146, ST-226, ST-314, and ST-570, which were clustered in five clonal complexes (CC64, CC67, CC103, CC17, and CC314), and one singleton, ST-91. Among the virulence genes screened in this study, PI-2b, fbsB, cfb, and hylB appear to be the most important during mastitis development in cattle. Collectively, these results establish the molecular epidemiology of S. agalactiae isolated from cows in Brazilian herds. We believe that the data presented here provide a foundation for future research aimed at developing and implementing new preventative and treatment options for mastitis caused by S. agalactiae. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  18. Molecular epidemiology of Streptococcus agalactiae isolated from mastitis in Brazilian dairy herds

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    Glei A. Carvalho-Castro

    Full Text Available Abstract Streptococcus agalactiae is one of the most common pathogens leading to mastitis in dairy herds worldwide; consequently, the pathogen causes major economic losses for affected farmers. In this study, multilocus sequence typing (MLST, genotypic capsular typing by multiplex polymerase chain reaction (PCR, and virulence gene detection were performed to address the molecular epidemiology of 59 bovine (mastitis S. agalactiae isolates from 36 dairy farms located in the largest milk-producing mesoregions in Brazil (Minas Gerais, São Paulo, Paraná, and Pernambuco. We screened for the virulence genes bac, bca, bibA, cfb, hylB, fbsA, fbsB, PI-1, PI-2a, and PI-2b, which are associated with adhesion, invasion, tissue damage, and/or immune evasion. Furthermore, five capsular types were identified (Ia, Ib, II, III, and IV, and a few isolates were classified as non-typeable (NT. MLST revealed the following eight sequence types (STs: ST-61, ST-67, ST-103, ST-146, ST-226, ST-314, and ST-570, which were clustered in five clonal complexes (CC64, CC67, CC103, CC17, and CC314, and one singleton, ST-91. Among the virulence genes screened in this study, PI-2b, fbsB, cfb, and hylB appear to be the most important during mastitis development in cattle. Collectively, these results establish the molecular epidemiology of S. agalactiae isolated from cows in Brazilian herds. We believe that the data presented here provide a foundation for future research aimed at developing and implementing new preventative and treatment options for mastitis caused by S. agalactiae.

  19. New Mutations of Penicillin-Binding Proteins in Streptococcus agalactiae Isolates from Cattle with Decreased Susceptibility to Penicillin.

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    Hu, Yun; Kan, Yunchao; Zhang, Zhengtian; Lu, Zhanning; Li, Yanqiu; Leng, Chaoliang; Ji, Jun; Song, Shiyang; Shi, Hongfei

    2018-02-23

    Streptococcus agalactiae is a causal agent of bovine mastitis and is treated by β-lactam antibiotics (BLAs). Compared to penicillin-resistant S. agalactiae from humans, resistant strains in bovine are rarely reported. In this study, we aimed to investigate BLA resistance and mutations in penicillin-binding proteins (PBPs) of S. agalactiae in central and northeast China. The minimum inhibitory concentrations (MICs) of 129 penicillin-resistant S. agalactiae isolates from cows with mastitis were determined, and the related PBP genes were detected and sequenced. All strains were unsusceptible to penicillin G and mostly resistant to ampicillin, cefalexin, and ceftiofur sodium. One hundred twenty-nine strains were divided into 4 clonal groups and 8 sequence types by multilocus sequence typing analysis. We found a set of new substitutions in PBP1B, PBP2B, and PBP2X from most strains isolated from three provinces. The strains with high PBP mutations showed a broader unsusceptible spectrum and higher MICs than those with few or single mutation. Our research indicates unpredicted mutations in the PBP genes of S. agalactiae isolated from cows with mastitis treated by BLAs. This screening is the first of S. agalactiae from cattle.

  20. Emerging trends in invasive and noninvasive isolates of Streptococcus agalactiae in a Latin American hospital: a 17-year study.

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    Crespo-Ortiz, Maria del Pilar; Castañeda-Ramirez, Claudia Rocio; Recalde-Bolaños, Monica; Vélez-Londoño, Juan Diego

    2014-08-03

    Streptococcus agalactiae or group B Streptococcus (GBS) has been recognized as a lethal pathogen in neonates worldwide. S. agalactiae infections also severely affect pregnant women and immunosuppressed adults with substantial attributable morbidity and mortality. However, in Latin America, studies on the epidemiology and behaviour of S. agalactiae infections remain limited. To better understand the behaviour of S. agalactiae infections in our region, we conducted a retrospective study to phenotypically describe S. agalactiae isolates collected in one of the largest hospitals in Colombia at two time periods: 1994-2001 and 2004-2012. The isolates were identified by biochemical analysis and tested for antimicrobial susceptibility. In 1994-2001 a total of 201 S. agalactiae isolates were found in urine 38.3%, vaginal exudates 27.8%, soft tissue 12.9%, and blood 8.5%. Susceptibility to ampicillin or penicillin was 94% whereas resistance to erythromycin and clindamycin were 2.8% and 5.2% respectively. In total 46 culture-positive cases of invasive infections were reported, 11 (24%) in neonates and 35 (76%) in adults. In 2004-2012 a total of 671 isolates were found in urine 47.8%, vaginal exudates 32.6%, soft tissue 2.7% and blood 9%. Susceptibility rates to ampicillin and penicillin were 98% whereas resistance to erythromycin and clindamycin were 12.5% and 9.4%. A total of 95 severe infections were reported: 12 (12.6%) were in neonates, 5 (5.3%) in children and 78 (82.1%) in adults. Over the 17-year study period the averaged prevalence of invasive S. agalactiae isolates was 17.4%. The estimated incidence for neonatal infections was 1.34 per 1000 livebirths (0.99 × 1000 livebirths for early- onset disease and 0.35 × 1000 livebirths for late- onset disease) whereas for non-pregnant adults the estimated incidence was 0.75 × 1000 admissions. A remarkable increase in bloodstream infections in immunosuppressed adults and a shift to early neonatal S. agalactiae infections

  1. Molecular characterization of Streptococcus agalactiae and Streptococcus uberis isolates from bovine milk.

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    Shome, Bibek Ranjan; Bhuvana, Mani; Mitra, Susweta Das; Krithiga, Natesan; Shome, Rajeswari; Velu, Dhanikachalam; Banerjee, Apala; Barbuddhe, Sukhadeo B; Prabhudas, Krishnamshetty; Rahman, Habibar

    2012-12-01

    Streptococci are one among the major mastitis pathogens which have a considerable impact on cow health, milk quality, and productivity. The aim of the present study was to investigate the occurrence and virulence characteristics of streptococci from bovine milk and to assess the molecular epidemiology and population structure of the Indian isolates using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Out of a total of 209 bovine composite milk samples screened from four herds (A-D), 30 Streptococcus spp. were isolated from 29 milk samples. Among the 30 isolates, species-specific PCR and partial 16S rRNA gene sequence analysis identified 17 Streptococcus agalactiae arising from herd A and 13 Streptococcus uberis comprising of 5, 7, and 1 isolates from herds B, C, and D respectively. PCR based screening for virulence genes revealed the presence of the cfb and the pavA genes in 17 and 1 S. agalactiae isolates, respectively. Similarly, in S. uberis isolates, cfu gene was present in six isolates from herd C, the pau A/skc gene in all the isolates from herds B, C, and D, whereas the sua gene was present in four isolates from herd B and the only isolate from herd D. On MLST analysis, all the S. agalactiae isolates were found to be of a novel sequence type (ST), ST-483, reported for the first time and is a single locus variant of the predicted subgroup founder ST-310, while the S. uberis isolates were found to be of three novel sequence types, namely ST-439, ST-474, and ST-475, all reported for the first time. ST-474 was a double locus variant of three different STs of global clonal complex ST-143 considered to be associated with clinical and subclinical mastitis, but ST-439 and ST-475 were singletons. Unique sequence types identified for both S. agalactiae and S. uberis were found to be herd specific. On PFGE analysis, identical or closely related restriction patterns for S. agalactiae ST-483 and S. uberis ST-439 in herds A and B

  2. First report of Streptococcus agalactiae isolated from Oreochromis niloticus in Piura, Peru: Molecular identification and histopathological lesions

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    Yessica Ortega Asencios

    2016-11-01

    Full Text Available The aim of this study was to identify the bacterium Streptococcus agalactiae isolated in farmed Nile tilapia (Oreochromis niloticus from Piura, Peru and to characterize the histopathological lesions caused by this pathogen. Sixteen tilapias were sampled with clinic signs of the disease such as erratic swimming, exophthalmia and haemorrhages on the body and fins. Qualitative PCR in real time and histopathological analysis were performed. Nine fishes positives to S. agalactiae were found. The main histopathological findings were fibrinosuppurative epicarditis, periesplenitis, meninigitis and panophtaltmitis with predominance of mononuclear infiltration in all tissues. The correlation between qPCR and histopathological findings demonstrated nine fish (prevalence of 56.25% with Cq lower than 30, associated to high degree of tissue injuries. This study reports the first isolation of S. agalactiae by PCR in real time in tilapia farmed in Peru and characterizes the major histopathological changes caused by this bacterium.

  3. Molecular serotyping, virulence gene profiling and pathogenicity of Streptococcus agalactiae isolated from tilapia farms in Thailand by multiplex PCR.

    Science.gov (United States)

    Kannika, K; Pisuttharachai, D; Srisapoome, P; Wongtavatchai, J; Kondo, H; Hirono, I; Unajak, S; Areechon, N

    2017-06-01

    This study aimed to biotype Streptococcus agalactiae isolated from tilapia farms in Thailand based on molecular biotyping methods and to determine the correlation between the serotype and virulence of bacteria. In addition to a biotyping (serotyping) technique based on multiplex PCR of cps genes, in this study, we developed multiplex PCR typing of Group B streptococcus (GBS) virulence genes to examine three clusters of virulence genes and their correlation with the pathogenicity of S. agalactiae. The epidemiology of S. agalactiae in Thailand was analysed to provide bacterial genetic information towards a future rational vaccine strategy for tilapia culture systems. Streptococcus agalactiae were isolated from diseased tilapia from different areas of Thailand. A total of 124 S. agalactiae isolates were identified by phenotypic analysis and confirmed by 16S rRNA PCR. Bacterial genotyping was conducted based on (i) molecular serotyping of the capsular polysaccharide (cps) gene cluster and (ii) virulence gene profiling using multiplex PCR analysis of 14 virulence genes (lmb, scpB, pavA, cspA, spb1, cyl, bca, rib, fbsA, fbsB, cfb, hylB, bac and pbp1A/ponA). Only serotypes Ia and III were found in this study; serotype Ia lacks the lmb, scpB and spb1 genes, whereas serotype III lacks only the bac gene. Virulence tests in juvenile Nile tilapia demonstrated a correlation between the pathogenicity of the bacteria and their virulence gene profile, with serotype III showing higher virulence than serotype Ia. Epidemiological analysis showed an almost equal distribution in all regions of Thailand, except serotype III was found predominantly in the southern areas. Only two serotypes of S. agalactiae were isolated from diseased tilapia in Thailand. Serotype Ia showed fewer virulence genes and lower virulence than serotype III. Both serotypes showed a similar distribution throughout Thailand. We identified two major serotypes of S. agalactiae isolates associated with the outbreak in

  4. Draft Genome Sequence of Streptococcus agalactiae Serotype Ia Strain M19, a Multidrug-Resistant Isolate from a Cow with Bovine Mastitis

    OpenAIRE

    Yang, Feng; Li, Hongsheng; Zhang, Shidong; Wang, Xurong

    2016-01-01

    Streptococcus agalactiae is a major contagious pathogen causing bovine mastitis worldwide. We report here the draft sequence of S.?agalactiae Ia strain M19, a multidrug-resistant isolate from a bovine mastitis case in Ningxia Hui autonomous region, China.

  5. Antimicrobial resistance and molecular characteristics of Streptococcus agalactiae isolated from women of reproductive age

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    Magdalena Musiorska

    2016-12-01

    Full Text Available Introduction. Streptococcus agalactiae infections are among the most significant causes of neonatal invasive diseases. Proper screening and detection of pregnant women carrying GBS allows intrapartum administration of antibiotic prophylaxis and is an effective measure in preventing transmission of bacteria from mother to newborns. Material and methods. Sixty three bacterial strains were isolated from vaginal swabs from pregnant and nonpregnant women of reproductive age. Species were identified by colony morphology, haemolysis type, Gram staining and SLIDEX® Strepto Plus latex test. Antimicrobial resistance of 56 strains was determined using disk-diffusion method. The presence of molecular resistance determinants was assessed using PCR with specific primers, and capsular types were identified using multiplex PCR. Results. None of the strains were resistant to the first drug of choice, penicillin. A large percentage of isolates (78.6% were resistant to doxycycline. The prevalence of resistance to macrolides and lincosamides, antibiotics used in women allergic to penicillin, was high. Those results corresponded with PCR tests, as tetM and ermA1 were most frequently detected genes (98.4 and 87.3%, respectively. 7.94% of strains possessed 7 different out of 13 tested genes determining resistance to different groups of antimicrobials. Among the capsular types, Ia, which proved to be associated with the most severe and invasive infections in mothers and neonates, was the most prevalent (65.08%. Conclusions. Even though they are susceptible to penicillin, multidrug resistance is common among S. agalactiae strains isolated from women of reproductive age and this resistance can be caused by more than one gene per single isolate

  6. Comparative genomics analysis of Streptococcus agalactiae reveals that isolates from cultured tilapia in China are closely related to the human strain A909.

    Science.gov (United States)

    Liu, Guangjin; Zhang, Wei; Lu, Chengping

    2013-11-11

    Streptococcus agalactiae, also referred to as Group B Streptococcus (GBS), is a frequent resident of the rectovaginal tract in humans, and a major cause of neonatal infection. In addition, S. agalactiae is a known fish pathogen, which compromises food safety and represents a zoonotic hazard. The complete genome sequence of the piscine S. agalactiae isolate GD201008-001 was compared with 14 other piscine, human and bovine strains to explore their virulence determinants, evolutionary relationships and the genetic basis of host tropism in S. agalactiae. The pan-genome of S. agalactiae is open and its size increases with the addition of newly sequenced genomes. The core genes shared by all isolates account for 50 ~ 70% of any single genome. The Chinese piscine isolates GD201008-001 and ZQ0910 are phylogenetically distinct from the Latin American piscine isolates SA20-06 and STIR-CD-17, but are closely related to the human strain A909, in the context of the clustered regularly interspaced short palindromic repeats (CRISPRs), prophage, virulence-associated genes and phylogenetic relationships. We identified a unique 10 kb gene locus in Chinese piscine strains. Isolates from cultured tilapia in China have a close genomic relationship with the human strain A909. Our findings provide insight into the pathogenesis and host-associated genome content of piscine S. agalactiae isolated in China.

  7. Genetic analysis of Streptococcus agalactiae strains isolated from neonates and their mothers.

    NARCIS (Netherlands)

    Melchers, W.J.G.; Bakkers, J.M.J.E.; Toonen, M.; Kuppeveld, F.J.M. van; Trijbels-Smeulders, M.J.A.M.; Hoogkamp-Korstanje, J.A.A.

    2003-01-01

    Streptococcus agalactiae or group B streptococcus (GBS) is the most common cause of neonatal sepsis and meningitis in neonates. One of the major questions is whether the GBS strains able to cause neonatal invasive disease have peculiar genetic features. A collection of S. agalactiae strains,

  8. Assignment of Streptococcus agalactiae isolates to clonal complexes using a small set of single nucleotide polymorphisms.

    Science.gov (United States)

    Honsa, Erin; Fricke, Thomas; Stephens, Alex J; Ko, Danny; Kong, Fanrong; Gilbert, Gwendolyn L; Huygens, Flavia; Giffard, Philip M

    2008-08-19

    Streptococcus agalactiae (Group B Streptococcus (GBS)) is an important human pathogen, particularly of newborns. Emerging evidence for a relationship between genotype and virulence has accentuated the need for efficient and well-defined typing methods. The objective of this study was to develop a single nucleotide polymorphism (SNP) based method for assigning GBS isolates to multilocus sequence typing (MLST)-defined clonal complexes. It was found that a SNP set derived from the MLST database on the basis of maximization of Simpsons Index of Diversity provided poor resolution and did not define groups concordant with the population structure as defined by eBURST analysis of the MLST database. This was interpreted as being a consequence of low diversity and high frequency horizontal gene transfer. Accordingly, a different approach to SNP identification was developed. This entailed use of the "Not-N" bioinformatic algorithm that identifies SNPs diagnostic for groups of known sequence variants, together with an empirical process of SNP testing. This yielded a four member SNP set that divides GBS into 10 groups that are concordant with the population structure. A fifth SNP was identified that increased the sensitivity for the clinically significant clonal complex 17 to 100%. Kinetic PCR methods for the interrogation of these SNPs were developed, and used to genotype 116 well characterized isolates. A five SNP method for dividing GBS into biologically valid groups has been developed. These SNPs are ideal for high throughput surveillance activities, and combining with more rapidly evolving loci when additional resolution is required.

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

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    Azizollah Ebrahimi

    2014-12-01

    Full Text Available 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 biofilm producers. This finding may indicate the high potential of pathogenicity in isolated strains. Sixteen (51.6% isolates were α hemolysin producers. Only 19.3%, 22.5% and 29.0% of the isolates were sensitive to streptomycin, flumequine and kanamycin, respectively. None of these three agents is recommended for treatment of mastitis cases.

  10. Assignment of Streptococcus agalactiae isolates to clonal complexes using a small set of single nucleotide polymorphisms

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    Gilbert Gwendolyn L

    2008-08-01

    Full Text Available Abstract Background Streptococcus agalactiae (Group B Streptococcus (GBS is an important human pathogen, particularly of newborns. Emerging evidence for a relationship between genotype and virulence has accentuated the need for efficient and well-defined typing methods. The objective of this study was to develop a single nucleotide polymorphism (SNP based method for assigning GBS isolates to multilocus sequence typing (MLST-defined clonal complexes. Results It was found that a SNP set derived from the MLST database on the basis of maximisation of Simpsons Index of Diversity provided poor resolution and did not define groups concordant with the population structure as defined by eBURST analysis of the MLST database. This was interpreted as being a consequence of low diversity and high frequency horizontal gene transfer. Accordingly, a different approach to SNP identification was developed. This entailed use of the "Not-N" bioinformatic algorithm that identifies SNPs diagnostic for groups of known sequence variants, together with an empirical process of SNP testing. This yielded a four member SNP set that divides GBS into 10 groups that are concordant with the population structure. A fifth SNP was identified that increased the sensitivity for the clinically significant clonal complex 17 to 100%. Kinetic PCR methods for the interrogation of these SNPs were developed, and used to genotype 116 well characterized isolates. Conclusion A five SNP method for dividing GBS into biologically valid groups has been developed. These SNPs are ideal for high throughput surveillance activities, and combining with more rapidly evolving loci when additional resolution is required.

  11. Isolation and identification of Mycoplasma agalactiae by culture and Polymerase Chain Reaction in Sheep and Goat Milk Samples in Kordestan province, Iran

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    Khaki, P.

    2011-06-01

    Full Text Available Contagious agalactiae (C.A. is one of the most common disease affecting small ruminants which is caused by Mycoplasma agalactiae. This disease is particularly widespread around the world and Iran is one of the countries that C.A. is present. The aim of this study was isolation and identification of M. agalactiae (MG with culture and PCR technique in milk samples in Kordestan province, Iran. A total of 367 milk samples were collected from sheep and goat. Specific published primers amplify a 375 bp gene of MG were used for PCR. Twenty (5.5% out of 367 were positive in PPLO agar and 5 (25% out of these isolates were positive with Mycoplasma agalactiae primers. Four (75% out of 5 isolates was from sheep and 1(25% from goat. Result of PCR with 367 milk samples showed that 11(3% of them were positive with these primers. The isolation of M. agalactiae showed that C.A is present in Kordestan province and our results suggested that PCR method because of reduces the time consuming could be an alternative method beside culture.

  12. In vitro antimicrobial susceptibility and genetic resistance determinants of Streptococcus agalactiae isolated from mastitic cows in Brazilian dairy herds

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    Juliana Rosa da Silva

    2017-08-01

    Full Text Available Streptococcus agalactiae is one of the main causative agents of bovine mastitis and is associated with several economic losses for producers. Few studies have evaluated antimicrobial susceptibility and the prevalence of genetic resistance determinants among isolates of this bacterium from Brazilian dairy cattle. This work aimed to evaluate the frequency of the antimicrobial resistance genes ermA, ermB, mefA, tetO, tetM, aphA3, and aad-6, and in vitro susceptibility to the antimicrobials amikacin, erythromycin, clindamycin, tetracycline, gentamicin, penicillin, ceftiofur, and cefalotin, and the associations between resistance genotypes and phenotypes among 118 S. agalactiae isolates obtained from mastitic cows in Brazilian dairy herds. Of the resistance genes examined, ermB was found in 19 isolates (16.1%, tetO in 23 (19.5%, and tetM in 24 (20.3%. The genes ermA, mefA, aphA3, and aad-6 were not identified. There was an association between the presence of genes ermB, tetM, and tetO and phenotypic resistance to erythromycin, clindamycin, and tetracycline. Rates of resistance to the tested antibiotics varied, as follows: erythromycin (19.5%, tetracycline (35.6%, gentamicin (9.3%, clindamycin (20.3%, penicillin (3.4%, and amikacin (38.1%; conversely, all isolates were susceptible to ceftiofur and cefalotin. Antimicrobial resistance testing facilitates the treatment decision process, allowing the most judicious choice of antibiotics. Moreover, it enables regional and temporal monitoring of the resistance dynamics of this pathogen of high importance to human and animal health.

  13. Streptococcus agalactiae: a vaginal pathogen?

    Science.gov (United States)

    Maniatis, A N; Palermos, J; Kantzanou, M; Maniatis, N A; Christodoulou, C; Legakis, N J

    1996-03-01

    The significance of Streptococcus agalactiae as an aetiological agent in vaginitis was evaluated. A total of 6226 samples from women who presented with vaginal symptoms was examined. The presence of >10 leucocytes/high-power field (h.p.f.) was taken to be the criterion of active infection. S. agalactiae was isolated from 10.1% of these samples. The isolation rates of other common pathogens such as Candida spp., Gardnerella vaginalis and Trichomonas spp. were 54.1%, 27.2% and 4.2%, respectively, in the same group of patients. In contrast, the isolation rates of these micro-organisms in the group of patients who had no infection (S. agalactiae was isolated, it was the sole pathogen isolated (83%) and its presence was associated with an inflammatory response in 80% of patients. Furthermore, the relative risk of vaginal infection with S. agalactiae (2.38) in patients with purulent vaginal discharge was greater than that of Candida spp. infection (1.41) and lower than that of Trichomonas spp. infection (8.32). These data suggest that S. agalactiae in symptomatic women with microscopic evidence of inflammation should be considered a causative agent of vaginitis.

  14. Pathogenicity of Human ST23 Streptococcus agalactiae to Fish and Genomic Comparison of Pathogenic and Non-pathogenic Isolates

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    Rui Wang

    2017-10-01

    Full Text Available Streptococcus agalactiae, or Group B Streptococcus (GBS, is a major pathogen causing neonatal sepsis and meningitis, bovine mastitis, and fish meningoencephalitis. CC23, including its namesake ST23, is not only the predominant GBS strain derived from human and cattle, but also can infect a variety of homeothermic and poikilothermic species. However, it has never been characterized in fish. This study aimed to determine the pathogenicity of ST23 GBS to fish and explore the mechanisms causing the difference in the pathogenicity of ST23 GBS based on the genome analysis. Infection of tilapia with 10 human-derived ST23 GBS isolates caused tissue damage and the distribution of pathogens within tissues. The mortality rate of infection was ranged from 76 to 100%, and it was shown that the mortality rate caused by only three human isolates had statistically significant difference compared with fish-derived ST7 strain (P < 0.05, whereas the mortality caused by other seven human isolates did not show significant difference compared with fish-derived ST7 strain. The genome comparison and prophage analysis showed that the major genome difference between virulent and non-virulent ST23 GBS was attributed to the different prophage sequences. The prophage in the P1 region contained about 43% GC and encoded 28–39 proteins, which can mediate the acquisition of YafQ/DinJ structure for GBS by phage recombination. YafQ/DinJ belongs to one of the bacterial toxin–antitoxin (TA systems and allows cells to cope with stress. The ST23 GBS strains carrying this prophage were not pathogenic to tilapia, but the strains without the prophage or carrying the pophage that had gene mutation or deletion, especially the deletion of YafQ/DinJ structure, were highly pathogenic to tilapia. In conclusion, human ST23 GBS is highly pathogenic to fish, which may be related to the phage recombination.

  15. Distribution of serotypes and evaluation of antimicrobial susceptibility among human and bovine Streptococcus agalactiae strains isolated in Brazil between 1980 and 2006

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    Tatiana Castro Abreu Pinto

    Full Text Available Streptococcus agalactiae is a common agent of clinical and subclinical bovine mastitis and an important cause of human infections, mainly among pregnant women, neonates and nonpregnant adults with underlying diseases. The present study describes the genetic and phenotypic diversity among 392 S. agalactiae human and bovine strains isolated between 1980 and 2006 in Brazil. The most prevalent serotypes were Ia, II, III and V and all the strains were susceptible to penicillin, vancomycin and levofloxacin. Resistance to clindamycin, chloramphenicol, erythromycin, rifampicin and tetracycline was observed. Among the erythromycin resistant strains, mefA/E, ermA and, mainly, ermB gene were detected, and a shift of prevalence from the macrolide resistance phenotype to the macrolidelincosamide- streptogramin B resistance phenotype over the years was observed. The 23 macrolide-resistant strains showed 19 different pulsed-field gel electrophoresis profiles. Regarding macrolide resistance, a major concern in S. agalactiae epidemiology, the present study describes an increase in erythromycin resistance from the 80s to the 90s followed by a decrease in the 2000-2006 period. Also, the genetic heterogeneity described points out that erythromycin resistance in Brazil is rather due to horizontal gene transmission than to spreading of specific macrolide-resistant clones.

  16. Field trial about occurrence and relevance of streptococcus agalactiae in Mauritanian camels

    OpenAIRE

    Ould Habiboullah, Habiboullah

    2010-01-01

    The objective of this study was to determine the presence and prevalence of Streptococcus (S.) agalactiae in Mauritanian camels. Twenty seven camel herds from 8 Mauritanian regions were included in the study, whereby a total of 276 milk samples and 104 udder skin swaps were investigated for S. agalactiae. S. agalactiae was isolated from 25 out of 27 investigated camel herds. A total of 41 S. agalactiae strains where isolated, 29 isolates from the milk samples and 12 isolates from the udd...

  17. Molecular Characterization of Streptococcus agalactiae Isolates From Pregnant and Non-Pregnant Women at Yazd University Hospital, Iran.

    Science.gov (United States)

    Sadeh, Maryam; Firouzi, Roya; Derakhshandeh, Abdollah; Bagher Khalili, Mohammad; Kong, Fanrong; Kudinha, Timothy

    2016-02-01

    Streptococcus agalactiae (Group B streptococcus, GBS) that colonize the vaginas of pregnant women may occasionally cause neonatal infections. It is one of the most common causes of sepsis and meningitis in neonates and of invasive diseases in pregnant women. It can also cause infectious disease among immunocompromised individuals. The distribution of capsular serotypes and genotypes varies over time and by geographic era. The serotyping and genotyping data of GBS in Iranian pregnant and non-pregnant women seems very limited. The aim of this study was to investigate the GBS ‎molecular capsular serotype ‎and genotype distribution of pregnant and non-pregnant carrier ‎women at Yazd university hospital, in Iran.‎. In this cross-sectional study, a total of 100 GBS strains isolated from 237 pregnant and 413 non-pregnant women were investigated for molecular capsular serotypes and surface protein genes using the multiplex PCR assay. The Chi-square method was used for statistical analysis. Out of 650 samples, 100 (15.4%) were identified as GBS, with a predominance of capsular serotypes III (50%) [III-1 (49), III-3 (1)], followed by II (25%), Ia (12%), V (11%), and Ib (2%), which was similar with another study conducted in Tehran, Iran, but they had no serotype Ia in their report. The surface protein antigen genes distribution was rib (53%), epsilon (38%), alp2/3 (6%), and alpha-c (3%). The determination of serotype and surface proteins of GBS strains distribution would ‎be ‎relevant ‎for the future possible formulation of a GBS vaccine.

  18. Molecular Characterization of Streptococcus agalactiae Isolates From Pregnant and Non-Pregnant Women at Yazd University Hospital, Iran

    OpenAIRE

    Sadeh, Maryam; Firouzi, Roya; Derakhshandeh, Abdollah; Bagher Khalili, Mohammad; Kong, Fanrong; Kudinha, Timothy

    2016-01-01

    Background: Streptococcus agalactiae (Group B streptococcus, GBS) that colonize the vaginas of pregnant women may occasionally cause neonatal infections. It is one of the most common causes of sepsis and meningitis in neonates and of invasive diseases in pregnant women. It can also cause infectious disease among immunocompromised individuals. The distribution of capsular serotypes and genotypes varies over time and by geographic era. The serotyping and genotyping data of GBS in Iranian pregna...

  19. First isolation of Mycoplasma capricolum subsp. capricolum, one of the causal agents of caprine contagious agalactia, on the island of Lanzarote (Spain).

    Science.gov (United States)

    De la Fe, C; Gutiérrez, A; Poveda, J B; Assunção, P; Ramírez, A S; Fabelo, F

    2007-03-01

    During an unusually long period of bad weather, several outbreaks of caprine contagious agalactia (CCA) were reported in a number of flocks on the island of Lanzarote (Canary Islands, Spain). Clinical and subclinical mastitis in lactating goats and some cases of arthritis and pneumonia in kids were observed in the affected flocks. Mycoplasma capricolum subsp. capricolum was isolated as the main causal agent of the outbreaks, associated with M. mycoides subsp. mycoides "large colony type" (Mmm LC) in two flocks. This is the first report of an isolation of M. capricolum subsp. capricolum on the island of Lanzarote. The finding is of epidemiological importance and could complicate plans to control the disease. The significance of this mycoplasma species in association with CCA must now be studied in detail.

  20. 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.

  1. Evaluation of the surveillance program of Streptococcus agalactiae in Danish dairy herds

    DEFF Research Database (Denmark)

    Andersen, H. J.; Pedersen, L. H.; Aarestrup, Frank Møller

    2003-01-01

    The aim of this study was to evaluate the Danish surveillance program of Streptococcus agalactiae in dairy herds with respect to 1) fluctuation over time of the presence of S. agalactiae in bulk tank milk, 2) sensitivity and specificity of the bacteriological method used, and 3) contamination...... the isolates. Streptococcus agalactiae was found in eight of 96 herds in which S. agalactiae had never previously been found during the surveillance program. Streptococcus agalactiae was not found in all seven sampling rounds in any of the eight herds. Comparing the approved method with supplemental findings...

  2. VNTR analysis reveals unexpected genetic diversity within Mycoplasma agalactiae, the main causative agent of contagious agalactia

    Directory of Open Access Journals (Sweden)

    Ayling Roger D

    2008-11-01

    Full Text Available Abstract Background Mycoplasma agalactiae is the main cause of contagious agalactia, a serious disease of sheep and goats, which has major clinical and economic impacts. Previous studies of M. agalactiae have shown it to be unusually homogeneous and there are currently no available epidemiological techniques which enable a high degree of strain differentiation. Results We have developed variable number tandem repeat (VNTR analysis using the sequenced genome of the M. agalactiae type strain PG2. The PG2 genome was found to be replete with tandem repeat sequences and 4 were chosen for further analysis. VNTR 5 was located within the hypothetical protein MAG6170 a predicted lipoprotein. VNTR 14 was intergenic between the hypothetical protein MAG3350 and the hypothetical protein MAG3340. VNTR 17 was intergenic between the hypothetical protein MAG4060 and the hypothetical protein MAG4070 and VNTR 19 spanned the 5' end of the pseudogene for a lipoprotein MAG4310 and the 3' end of the hypothetical lipoprotein MAG4320. We have investigated the genetic diversity of 88 M. agalactiae isolates of wide geographic origin using VNTR analysis and compared it with pulsed field gel electrophoresis (PFGE and random amplified polymorphic DNA (RAPD analysis. Simpson's index of diversity was calculated to be 0.324 for PFGE and 0.574 for VNTR analysis. VNTR analysis revealed unexpected diversity within M. agalactiae with 9 different VNTR types discovered. Some correlation was found between geographical origin and the VNTR type of the isolates. Conclusion VNTR analysis represents a useful, rapid first-line test for use in molecular epidemiological analysis of M. agalactiae for outbreak tracing and control.

  3. Characterization of Isolates of Streptococcus agalactiae from Diseased Farmed and Wild Marine Fish from the U.S. Gulf Coast, Latin America, and Thailand.

    Science.gov (United States)

    Soto, Esteban; Wang, Rui; Wiles, Judy; Baumgartner, Wes; Green, Christopher; Plumb, John; Hawke, John

    2015-06-01

    We examined Lancefield serogroup B Streptococcus isolates recovered from diseased, cultured hybrid Striped Bass (Striped Bass Morone saxatilis × White Bass M. chrysops) and wild and cultured Gulf Killifish Fundulus grandis from coastal waters of the U.S. Gulf of Mexico (Gulf coast) and compared those isolates to strains from tilapias Oreochromis spp. reared in Mississippi, Thailand, Ecuador, and Honduras and to the original Gulf coast strain identified by Plumb et al. ( 1974 ). The isolates were subjected to phylogenetic, biochemical, and antibiotic susceptibility analyses. Genetic analysis was performed using partial sequence comparison of (1) the 16S ribosomal RNA (rRNA) gene; (2) the sipA gene, which encodes a surface immunogenic protein; (3) the cspA gene, which encodes a cell surface-associated protein; and (4) the secY gene, which encodes components of a general protein secretion pathway. Phylogenies inferred from sipA, secY, and cspA gene sequence comparisons were more discriminating than that inferred from the 16S rRNA gene sequence comparison. The U.S. Gulf coast strains showed a high degree of similarity to strains from South America and Central America and belonged to a unique group that can be distinguished from other group B streptococci. In agreement with the molecular findings, biochemical and antimicrobial resistance analyses demonstrated that the isolates recovered from the U.S. Gulf coast and Latin America were more similar to each other than to isolates from Thailand. Three laboratory challenge methods for inducing streptococcosis in Gulf Killifish were evaluated-intraperitoneal (IP) injection, immersion (IMM), and immersion plus abrasion (IMMA)-using serial dilutions of S. agalactiae isolate LADL 97-151, a representative U.S. Gulf coast strain. The dose that was lethal to 50% of test fish by 14 d postchallenge was approximately 2 CFU/fish via IP injection. In contrast, the fish that were challenged via IMM or IMMA presented cumulative mortality

  4. Development of live attenuated Streptococcus agalactiae as potential vaccines by selecting for resistance to sparfloxacin.

    Science.gov (United States)

    Pridgeon, Julia W; Klesius, Phillip H

    2013-05-31

    To develop attenuated bacteria as potential live vaccines, sparfloxacin was used in this study to modify 40 isolates of Streptococcus agalactiae. Majority of S. agalactiae used in this study were able to develop at least 80-fold resistance to sparfloxacin. When the virulence of the sparfloxacin-resistant S. agalactiae isolates were tested in 10-12g Nile tilapia by intraperitoneal injection at dose of 2×10(7)CFU/fish, 31 were found to be avirulent to fish. Of the 31 avirulent sparfloxacin-resistant S. agalactiae isolates, 30 provided 75-100% protection to 10-12g Nile tilapia against challenges with a virulent S. agalactiae isolate Sag 50. When the virulence of the 30 sparfloxacin-resistant S. agalactiae isolates was tested in 3-5g Nile tilapia by intraperitoneal injection at dose of 2×10(7)CFU/fish, six were found to be avirulent to 3-5g Nile tilapia. Of the six avirulent sparfloxacin-resistant S. agalactiae isolates, four provided 3-5g Nile tilapia 100% protection against challenges with homologous isolates, including Sag 97-spar isolate that was non-hemolytic. However, Sag 97-spar failed to provide broad cross-protection against challenges with heterologous isolates. When Nile tilapia was vaccinated with a polyvalent vaccine consisting of 30 sparfloxacin-resistant S. agalactiae isolates at dose of 2×10(6)CFU/fish, the polyvalent vaccine provided significant (PS. agalactiae. Taken together, our results suggest that a polyvalent vaccine consisting of various strains of S. agalactiae might be essential to provide broader protection to Nile tilapia against infections caused by S. agalactiae. Published by Elsevier Ltd.

  5. Streptococcus agalactiae infection in domestic rabbits, Oryctolagus cuniculus.

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    Ren, S Y; Geng, Y; Wang, K Y; Zhou, Z Y; Liu, X X; He, M; Peng, X; Wu, C Y; Lai, W M

    2014-12-01

    Streptococcus agalactiae (Group B streptococcus, GBS) has emerged as an important pathogen that affects humans and animals, including aquatic species. In August 2011, a severe infectious disease affecting rabbits, which caused 42% mortality, occurred in Mianyang, Sichuan Province, China. The main clinical signs included acute respiratory distress syndrome, fever, paddling and convulsions. A Gram-positive, chain-forming coccus was isolated from the primary organs and tissues of diseased rabbits and then identified as S. agalactiae by morphology, biochemical and physiological characteristics, 16S rDNA and gyrB gene sequences analysis. All isolates of S. agalactiae showed a similar antibiotic susceptibility, which were sensitive to florfenicol, ampicillin,gentamicin and norfloxacin, as well as being resistant to penicillin, amoxicillin and tetracycline. To our knowledge, this is the first report on S. agalactiae natural infection in domestic rabbits. © 2013 Blackwell Verlag GmbH.

  6. Survey of strain distribution and antibiotic resistance pattern of group B streptococci (Streptococcus agalactiae isolated from clinical specimens

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    Mousavi, Seyed Masoud

    2016-09-01

    Full Text Available Aim: The aims of the present study were to determine the antibiotic susceptibility profils with particular emphasis on susceptible or resistant strains to macrolides and lincosamids antibiotics and to determine possible antibiotic resistance mechanisms occurring in group B streptococci (GBS strains using PCR assay and disk diffusion method.Methods: A total of 62 clinical GBS strains were investigated. Antibacterial susceptibility testing was performed using the disk diffusion method and inducible resistance test for clindamycin by standard double disk diffusion or D-zone test for all isolates to differentiate macrolide resistance phenotype (M, constitutive macrolide-lincosamide-streptogramin B phenotype (cMLS and induced macrolide-lincosamide-streptogramin B phenotype (iMLS. In addition, minimum inhibitory concentrations (MIC of penicillin were determined for all isolates. Finally, possible existence of antibiotic resistance genes for erythromycin , and and for clindamycin were examined among isolates using PCR assay.Results: All 62 isolates were susceptible to penicillin, ampicillin, linezolid, cefazoline and vancomycin. However, 93.5% (n=58 of isolates showed an increased MIC to penicillin. The overall rate of erythromycin resistance was 35.5% (n=22. All erythromycin-resistant isolates displayed the M phenotype (100%, n=22. All three erythromycin resistance genes (i.e. , and were found in erythromycin-resistant isolates.Conclusion: It was concluded that prescribing antibiotic without antibacterial susceptibility tests should be prevented because of the high prevalence of erythromycin-resistant GBS strains and the fact that erythromycin-resistant GBS strains has shown an increased MIC to penicillin, as the drug of choice for treating GBS infections.

  7. Streptococcus agalactiae isolates of serotypes Ia, III and V from human and cow are able to infect tilapia.

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    Chen, Ming; Wang, Rui; Luo, Fu-Guang; Huang, Yan; Liang, Wan-Wen; Huang, Ting; Lei, Ai-Ying; Gan, Xi; Li, Li-Ping

    2015-10-22

    Recent studies have shown that group B streptococcus (GBS) may be infectious across hosts. The purpose of this study is to investigate the pathogenicity of clinical GBS isolates with serotypes Ia, III and V from human and cow to tilapia and the evolutionary relationship among these GBS strains of different sources. A total of 27 clinical GBS isolates from human (n=10), cow (n=2) and tilapia (n=15) were analyzed using serotyping, multi-locus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Among them, 15 isolates were tested for their pathogenicity to tilapia. The results showed that five human GBS strains (2 serotype III, 2 serotype Ia and 1 serotype V) infected tilapia with mortality rate ranging from 56.67% to 100%, while the other five human GBS strains tested were unable to infect tilapia. In addition, two cow GBS strains C001 and C003 of serotype III infected tilapia. However, they had significantly lower pathogenicity than the five human strains. Furthermore, human GBS strains H005 and H008, which had very strong ability to infect tilapia, had the same PFGE pattern. MLST analysis showed that the five human and the two cow GBS strains that were able to infect tilapia belonged to clonal complexes CC19, CC23 and CC103. The study for the first time confirmed that human or cow GBS clonal complexes CC19, CC23 and CC103 containing strains with serotypes Ia, III and V could infect tilapia and induce clinical signs under experimental conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Development of live attenuated sparfloxacin-resistant Streptococcus agalactiae polyvalent vaccines to protect Nile tilapia

    Science.gov (United States)

    To develop attenuated bacteria as potential live vaccines, sparfloxacin was used in this study to modify 40 isolates of Streptococcus agalactiae. Majority of S. agalactiae used in this study were able to develop at least 80-fold resistance to sparfloxacin. When the virulence of the sparfloxacin-resi...

  9. Contagious agalactia by Mycoplasma agalactiae in small ruminants in Brazil: first report Agalaxia contagiosa por Mycoplasma agalactiae em pequenos ruminantes no Brasil: primeiro relato

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    Edisio Oliveira de Azevedo

    2006-12-01

    Full Text Available Two outbreaks of contagious agalactia by Mycoplasma agalactiae occurred in Paraíba State, Northeastern Region of Brazil are reported. The disease was characterized by mastitis, agalactia and polyarthritis in does and polyarthritis and conjunctivitis in kids and lambs. Fever and anorexia were also observed. Morbidy was from 26.1% to 100% in does, 36.5 to 100% in kids and 49% in lambs. In one farm 14.3% of the lactating goats and 6.4% of the kids died or were euthanized. In the other, 3.3% of the does, 36.5% of the kids and 22.9% of the lambs died and 84 affected goats were euthanized to control the disease. M. agalactiae was isolated from milk, joint exudates, nasal swabs and ear washings. The colonies were characteristic of Mycoplasma and the agent did not ferment both glucose and arginin. It was typed as Mycoplasma agalactiae by immunoperoxidase and PCR. This is the first report of M. agalactiae infection in Brazil, but the source of the infection remains unknown.Dois surtos de agalaxia contagiosa causada por Mycoplasma agalactiae são descritos no Estado da Paraíba, região Nordeste do Brasil. A doença caracterizou-se por mastite, agalaxia e poliartrite em cabras e poliartrite e cerato-conjuntivite em cabritos e cordeiros. Febre e anorexia também foram observadas. A morbidade variou de 26,1% a 100% nas cabras, 36,5% a 100% em cabritos e 49,0% em cordeiros. Na primeira fazenda, 14,3% das cabras em lactação e 6,4% dos cabritos morreram ou foram sacrificados. Na outra propriedade, 3,3% dos caprinos adultos, 36,5% dos cabritos e 22,9% dos cordeiros morreram e outros 84 caprinos foram sacrificados para controle da doença. M. agalactiae foi isolado a partir de leite, líquido articular, suabe nasal e lavado do conduto auditivo externo. Colônias características de Mycoplasma e que não fermentaram a glicose e arginina foram observadas. A identificação de M. agalactiae foi realizada por imunoperoxidase indireta e PCR. Sendo assim, M

  10. Human Streptococcus agalactiae strains in aquatic mammals and fish

    Science.gov (United States)

    2013-01-01

    Background In humans, Streptococcus agalactiae or group B streptococcus (GBS) is a frequent coloniser of the rectovaginal tract, a major cause of neonatal infectious disease and an emerging cause of disease in non-pregnant adults. In addition, Streptococcus agalactiae causes invasive disease in fish, compromising food security and posing a zoonotic hazard. We studied the molecular epidemiology of S. agalactiae in fish and other aquatic species to assess potential for pathogen transmission between aquatic species and humans. Methods Isolates from fish (n = 26), seals (n = 6), a dolphin and a frog were characterized by pulsed-field gel electrophoresis, multilocus sequence typing and standardized 3-set genotyping, i.e. molecular serotyping and profiling of surface protein genes and mobile genetic elements. Results Four subpopulations of S. agalactiae were identified among aquatic isolates. Sequence type (ST) 283 serotype III-4 and its novel single locus variant ST491 were detected in fish from Southeast Asia and shared a 3-set genotype identical to that of an emerging ST283 clone associated with invasive disease of adult humans in Asia. The human pathogenic strain ST7 serotype Ia was also detected in fish from Asia. ST23 serotype Ia, a subpopulation that is normally associated with human carriage, was found in all grey seals, suggesting that human effluent may contribute to microbial pollution of surface water and exposure of sea mammals to human pathogens. The final subpopulation consisted of non-haemolytic ST260 and ST261 serotype Ib isolates, which belong to a fish-associated clonal complex that has never been reported from humans. Conclusions The apparent association of the four subpopulations of S. agalactiae with specific groups of host species suggests that some strains of aquatic S. agalactiae may present a zoonotic or anthroponotic hazard. Furthermore, it provides a rational framework for exploration of pathogenesis and host-associated genome content of S

  11. Inapparent Streptococcus agalactiae infection in adult/commercial tilapia.

    Science.gov (United States)

    Sun, Jiufeng; Fang, Wei; Ke, Bixia; He, Dongmei; Liang, Yuheng; Ning, Dan; Tan, Hailing; Peng, Hualin; Wang, Yunxin; Ma, Yazhou; Ke, Changwen; Deng, Xiaoling

    2016-05-24

    We report on inapparent infections in adult/commercial tilapia in major tilapia fish farms in Guangdong. A total of 146 suspected isolates were confirmed to be S. agalactiae using an API 20 Strep system and specific PCR amplification. All isolates were identified as serotype Ia using multiplex serotyping PCR. An MLST assay showed single alleles of adhP (10), atr (2), glcK (2), glnA (1), pheS (1), sdhA (3) and tkt (2), and this profile was designated 'unique ST 7'. The analysis of virulence genes resulted in 10 clusters, of which dltr-bca-sodA-spb1-cfb-bac (62, 42.47%) was the predominant virulence gene profile. The PFGE analysis of S. agalactiae yielded 6 distinct PFGE types (A, B, C, D, F and G), of which Pattern C (103) was the predominant type, accounting for approximately 70.55% (103/146) of the total S. agalactiae strains. Therefore, unlike what has been found in juvenile tilapia, in which PFGE pattern D/F is the major prevalent pattern, we found that pattern C was the major prevalent pattern in inapparent infected adult/commercial tilapia in Guangdong, China. In conclusion, we close a gap in the current understanding of S. agalactiae epidemiology and propose that researchers should be alert for inapparent S. agalactiae infections in adult/commercial tilapia to prevent a potential threat to food safety.

  12. Streptococcus agalactiae infection in cancer patients: a five-year study.

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    Pimentel, B A S; Martins, C A S; Mendonça, J C; Miranda, P S D; Sanches, G F; Mattos-Guaraldi, A L; Nagao, P E

    2016-06-01

    Although the highest burden of Streptococcus agalactiae infections has been reported in industrialized countries, studies on the characterization and epidemiology are still limited in developing countries and implementation of control strategies remains undefined. The aim of this retrospective study was to assess the epidemiological, clinical, and microbiological aspects of S. agalactiae infections in cancer patients treated at a Reference Brazilian National Cancer Institute - INCA, Rio de Janeiro, Brazil. We reviewed the clinical and laboratory records of all cancer patients identified as having invasive S. agalactiae disease during 2010-2014. The isolates were identified by biochemical analysis and tested for antimicrobial susceptibility. A total of 263 strains of S. agalactiae were isolated from cancer patients who had been clinically and microbiologically classified as infected. S. agalactiae infections were mostly detected among adults with solid tumors (94 %) and/or patients who have used indwelling medical devices (77.2 %) or submitted to surgical procedures (71.5 %). Mortality rates (in-hospital mortality during 30 days after the identification of S. agalactiae) related to invasive S. agalactiae infections (n = 28; 31.1 %) for the specific category of neoplasic diseases were: gastrointestinal (46 %), head and neck (25 %), lung (11 %), hematologic (11 %), gynecologic (4 %), and genitourinary (3 %). We also found an increase in S. agalactiae resistance to erythromycin and clindamycin and the emergence of penicillin-less susceptible isolates. A remarkable number of cases of invasive infections due to S. agalactiae strains was identified, mostly in adult patients. Our findings reinforce the need for S. agalactiae control measures in Brazil, including cancer patients.

  13. Streptococcus iniae and Streptococcus agalactiae

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    Streptococcus iniae and S. agalactiae are economically important Gram positive bacterial pathogens of cultured and wild fish with a worldwide distribution. Both bacteria are potential zoonotic pathogens and have been associated most often with infections in immunocompromised people. Streptococcus in...

  14. Natural coinfection by Streptococcus agalactiae and Francisella noatunensis subsp. orientalis in farmed Nile tilapia (Oreochromis niloticus L.).

    Science.gov (United States)

    Assis, G B N; Tavares, G C; Pereira, F L; Figueiredo, H C P; Leal, C A G

    2017-01-01

    Streptococcus agalactiae and Francisella noatunensis subsp. orientalis (Fno) are important pathogens for farm-raised tilapia worldwide. There are no reports of coinfection caused by S. agalactiae and Fno in fish. This study aimed to determine the aetiology of atypical mortalities in a cage farm of Nile tilapia and to characterize the genetic diversity of the isolates. Fifty-two fish were sampled and subjected to parasitological and bacteriological examination. The S. agalactiae and Fno isolates were genotyped using MLST and REP-PCR, respectively. Whole-genome sequencing was performed to confirm the MLST results. Seven fish were shown coinfected by S. agalactiae and Fno. Chronic hypoxia and a reduction in the water temperature were determined as risk factors for coinfection. Fno isolates were shown clonally related in REP-PCR. The MLST analysis revealed that the S. agalactiae isolates from seven coinfected fish were negative for the glcK gene; however, these were determined to be members of clonal complex CC-552. This is the first description of coinfection by S. agalactiae and Fno in farm-raised Nile tilapia. The coinfection was predisposed by chronic hypoxia and was caused by the main genotypes of S. agalactiae and Fno reported in Brazil. Finally, a new S. agalactiae genotype with glcK gene partially deleted was described. © 2016 John Wiley & Sons Ltd.

  15. Comparative proteome analysis of two Streptococcus agalactiae strains from cultured tilapia with different virulence.

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    Li, Wei; Su, You-Lu; Mai, Yong-Zhan; Li, Yan-Wei; Mo, Ze-Quan; Li, An-Xing

    2014-05-14

    Streptococcus agalactiae is a major piscine pathogen, which causes significant morbidity and mortality among numerous fish species, and results in huge economic losses to aquaculture. Many S. agalactiae strains showing different virulence characteristics have been isolated from infected tilapia in different geographical regions throughout South China in the recent years, including natural attenuated S. agalactiae strain TFJ0901 and virulent S. agalactiae strain THN0901. In the present study, survival of tilapia challenged with S. agalactiae strain TFJ0901 and THN0901 (10(7)CFU/fish) were 93.3% and 13.3%, respectively. Moreover, there are severe lesions of the examined tissues in tilapia infected with strain THN0901, but no significant histopathological changes were observed in tilapia infected with the strain TFJ0901. In order to elucidate the factors responsible for the invasive potential of S. agalactiae between two strains TFJ0901 and THN0901, a comparative proteome analysis was applied to identify the different protein expression profiles between the two strains. 506 and 508 cellular protein spots of S. agalactiae TFJ0901 and THN0901 were separated by two dimensional electrophoresis, respectively. And 34 strain-specific spots, corresponding to 27 proteins, were identified successfully by MALDI-TOF mass spectrometry. Among them, 23 proteins presented exclusively in S. agalactiae TFJ0901 or THN0901, and the other 4 proteins presented in different isomeric forms between TFJ0901 and THN0901. Most of the strain-specific proteins were just involved in metabolic pathways, while 7 of them were presumed to be responsible for the virulence differences of S. agalactiae strain TFJ0901 and THN0901, including molecular chaperone DnaJ, dihydrolipoamide dehydrogenase, thioredoxin, manganese-dependent inorganic pyrophosphatase, elongation factor Tu, bleomycin resistance protein and cell division protein DivIVA. These virulence-associated proteins may contribute to identify new

  16. Central nervous system involvement in adult patients with invasive infection caused by Streptococcus agalactiae.

    Science.gov (United States)

    Oyanguren, B; Esteban, L; Guillán, M; de Felipe, A; Alonso Cánovas, A; Navas, E; Quereda, C; Corral, I

    2015-04-01

    Streptococcus agalactiae is frequently an asymptomatic coloniser and a cause of neonatal and puerperal sepsis. Infections in nonpregnant adults are uncommon. The frequency of neurological complications caused by invasive infection with this microorganism in adults remains unknown. Here, we study the frequency and characteristics of central nervous system (CNS) involvement in adults with invasive S. agalactiae infection. Review of all adults with invasive S. agalactiae infection between 2003 and 2011 in a tertiary hospital. S. agalactiae was isolated from blood, CSF or synovial fluid in 75 patients. Among them, 7 (9,3%) displayed neurological involvement: 5 men and 2 nonpregnant women, aged between 20 and 62 years. Diagnoses were spinal epidural abscess due to spondylodiscitis with spinal cord compression; acute bacterial meningitis; ischemic stroke as presentation of bacterial endocarditis (2 patients each); and meningoventriculitis after neurosurgery and ventricular shunting. One patient with endocarditis caused by S. agalactiae and S. aureus died in the acute phase, and another died 3 months later from metastatic cancer. The other patients recovered without sequelae. All patients had systemic predisposing factors for infection and 5 (71,4%) had experienced disruption of the mucocutaneous barrier as a possible origin of the infection. CNS involvement is not uncommon in adult patients with invasive infection caused by S. agalactiae. Isolating S. agalactiae, especially in cases of meningitis, should lead doctors to search for predisposing systemic disease and causes of mucocutaneous barrier disruption. Copyright © 2013 Sociedad Española de Neurología. Published by Elsevier Espana. All rights reserved.

  17. Streptococcus agalactiae: an emerging cause of septic arthritis.

    Science.gov (United States)

    Louthrenoo, Worawit; Kasitanon, Nuntana; Wangkaew, Suparaporn; Hongsongkiat, Sith; Sukitawut, Waraporn; Wichainun, Ramjai

    2014-03-01

    Invasive Streptococcus agalactiae infection in nonpregnant women has been reported increasingly worldwide. This study reports the clinical features and outcome of S. agalactiae septic arthritis in Thai patients. The medical records of cases with septic arthritis seen between July 1990 and December 2010 were reviewed. Only those with S. agalactiae were included in this study. From 244 cases of septic arthritis, 38 (15.57%, 13 men and 25 women) were caused by S. agalactiae, with 34 of them (89.48%) occurring between 2008 and 2010. Their mean age was 52.89 (SD, 18.95) years. Twenty-four of the 38 patients (63.16%) had 1 or more underlying disease that might predispose to joint infection. Fever and joint pain were the most common symptoms presented. Eleven cases (28.95%) presented monoarthritis, 15 (39.47%) oligoarthritis, and 12 (31.58%) polyarthritis, with a mean joint involvement of 3.34 (SD, 2.35) joints (range, 1-8). Cellulitis was seen in 27 cases (71.05%). Blood cultures were positive in 31 patients (81.58%). Thirty-five of the 38 synovial fluid specimens obtained were enough for cultures and stain smears, with 24 (68.57%) growing S. agalactiae and 19 (54.29%) showing gram-positive cocci. All isolates were sensitive to penicillin. Ten patients (26.31%) received arthroscopic drainage. The articular outcome was good in 11 patients, fair in 24, and poor in 3. There were no deaths. Streptococcus agalactiae is an emerging cause of septic arthritis in Thai patients. Physicians should be especially aware of this condition in patients presenting with acute oligopolyarthritis and prominent cellulitis.

  18. Streptococcus Agalactiae Research on Secretion Vaginal and Anal Pregnant Women of a City of Paulista Northwest

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    Cátia Rezende

    2015-12-01

    Full Text Available Streptococcus agalactiae has great medical importance in infections associated with severe neonatal morbidity and mortality. It is the most frequent bacterium isolated from the tables of septicemia, pneumonia and neonatal meningitis. This study aimed to determine the prevalence of anal and vaginal colonization of S. agalactiae in pregnant women at different gestational ages. We evaluated the incidence of colonization by S. agalactiae in 129 pregnant women of any age. Two samples of secretion were collected for culture: a perianal swab and a vaginal swab. Each of the two swabs were inoculated in test tubes containing Todd-Hewitt broth and subcultured on blood agar. After a period of 24 to 48 hours, the colonies suggestive of S. agalactiae were submitted to morfotintorial analysis and to biochemical tests for identification. Among 129 women studied, 3 (2.33% have tested positive for Streptococcus agalactiae and 126 (97.67% were negative. The results presented in this work are inferior to the data from other studies, however, the methodology used was compatible with most authors. Even so this rate is considered a relevant value taking into account the importance of Streptococcus agalactiae infections in pregnant women and newborns, emphasizing the importance of encouraging the culture of vaginal and anal secretion for the detection of Streptococcus agalactiae in surveys of prenatal care.

  19. Bacterial Protein Characterization of Streptococcus agalactiae by SDS-page Method for Subclinical Mastitis Irradiated Vaccine Materials in Dairy Cattle

    International Nuclear Information System (INIS)

    Tuasikal, B.J.; Wibawan, I.W.T.; Pasaribu, F.H; Estuningsih, S.

    2012-01-01

    A study have been conducted to isolate and characterize bacterial protein S. agalactiae, which is antigenic and can be used to test immunogenicity of vaccine in order to manufacture irradiated mastitis (inflammation of the udder) vaccine in ruminant. The study aims to determine the Molecular Weight (MW) bacterial protein S. agalactiae irradiation, which can be used to test the nature of its antigenic caharacteristic. The character of S. agalactiae antigenic stimulates antibody induction of the immune system, in which case is the body's defense system against mastitis disease in cattle. In this study, irradiation of gamma ray is used to attenuate the pathogenicity of bacteria by reducing S. agalactiae antigenic characteristic. Previous research, in irradiation dose orientation before antigenic protein isolation of S. agalactiae, indicated that irradiation lethal dose to 50% (LD 50 ) is 17 Gy. The characterization of S. agalactiae bacteria isolate using SDS-page method results in no significance different between irradiated and non-irradiated group, which indicated by MW range 75 - 100 kDa base on marker standard which used, or 99 kDa by the linier equation of Y = 11,60 - 0.05X (where Y = bands distance; X = MW standard protein); r 2 = 0.99. In conclusion, 17 Gy irradiation dose does not impair antigenic property of S. agalactiae and therefore, can be applied to produce base material of irradiated vaccine for mastitis. (author)

  20. Population structure and virulence gene profiles of Streptococcus agalactiae collected from different hosts worldwide.

    Science.gov (United States)

    Morach, Marina; Stephan, Roger; Schmitt, Sarah; Ewers, Christa; Zschöck, Michael; Reyes-Velez, Julian; Gilli, Urs; Del Pilar Crespo-Ortiz, María; Crumlish, Margaret; Gunturu, Revathi; Daubenberger, Claudia A; Ip, Margaret; Regli, Walter; Johler, Sophia

    2018-03-01

    Streptococcus agalactiae is a leading cause of morbidity and mortality among neonates and causes severe infections in pregnant women and nonpregnant predisposed adults, in addition to various animal species worldwide. Still, information on the population structure of S. agalactiae and the geographical distribution of different clones is limited. Further data are urgently needed to identify particularly successful clones and obtain insights into possible routes of transmission within one host species and across species borders. We aimed to determine the population structure and virulence gene profiles of S. agalactiae strains from a diverse set of sources and geographical origins. To this end, 373 S. agalactiae isolates obtained from humans and animals from five different continents were typed by DNA microarray profiling. A total of 242 different S. agalactiae strains were identified and further analyzed. Particularly successful clonal lineages, hybridization patterns, and strains were identified that were spread across different continents and/or were present in more than one host species. In particular, several strains were detected in both humans and cattle, and several canine strains were also detected in samples from human, bovine, and porcine hosts. The findings of our study suggest that although S. agalactiae is well adapted to various hosts including humans, cattle, dogs, rodents, and fish, interspecies transmission is possible and occurs between humans and cows, dogs, and rabbits. The virulence and resistance gene profiles presented enable new insights into interspecies transmission and make a crucial contribution to the identification of suitable targets for therapeutic agents and vaccines.

  1. Karakteristik dan Patogenisitas Streptococcus Agalactiae Tipe ?-hemolitik dan Non-hemolitik pada Ikan Nila

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    Esti Handayani Hardi

    2012-11-01

    Full Text Available Streptococcus agalactiae was isolated from cultured Nile tilapia (Oreochromis niloticus in Cirata gulfand Klaten. The isolates were Gram positive cocci, oxidative fermentative positive, motility, and catalasenegative, grown on media containing NaCl 6.5%, ?-haemolytic and non-haemolytic. Two types of S. agalactiae(?-haemolytic and non-haemolytic are different from their variety of sugars fermentation. Strains ?-haemolytic can ferment more sugars, including arabinose, sorbitol, lactose, and trehalose. Experimentalinfectivity trials on Nile tilapia (size 15 g, non-haemolytic type showed more virulent. This type causedfaster mortality, more severe behavior changes, and pathology changes than â-haemolytic type. NonhemoliticS. agalactiae caused 48% mortality 6-24 hours after injection, whereas â-haemolitic type caused17% mortality which it occured in 48 hours after injection (mortality of fish control 2,22%. Behaviordisease signs caused by non-haemolitic S. agalactiae started to happen 6 hours after injection whereas 12hours in ?-haemolytic type infection. Histopatological changes were observed on fish eye, spleen, andbrain. Hyperaemia, hyperthrophi, degeneration, and necrosis were also found on infected fish. Thisresearch was concluded that non-haemolytic of S. agalactiae was more virulent than ?-haemolytic.

  2. Presence and resistance of Streptococcus agalactiae in vaginal specimens of pregnant and adult non-pregnant women and association with other aerobic bacteria.

    Science.gov (United States)

    Numanović, Fatima; Smajlović, Jasmina; Gegić, Merima; Delibegović, Zineta; Bektaš, Sabaheta; Halilović, Emir; Nurkić, Jasmina

    2017-02-01

    Aim To determine the prevalence rate and resistance profile of Streptococcus agalactiae (S. agalactiae) in vaginal swabs of pregnant and adult non-pregnant women in the Tuzla region, Bosnia and Herzegovina (B&H), as well as its association with other aerobic bacteria. Methods This prospective study included 200 women, 100 pregnant and 100 adult non-pregnant. The research was conducted at the Institute of Microbiology, University Clinical Center Tuzla from October to December 2015. Standard aerobic microbiological techniques were used for isolation and identification of S. agalactiae and other aerobic bacteria. Antimicrobial susceptibility was determined by the disk diffusion and microdilution method(VITEK 2/AES instrument). Results Among 200 vaginal swabs, 17 (8.50%) were positive for S. agalactiae, e. g., 7% (7/100) of pregnant and 10% (10/100) of adult non-pregnant women. In the pregnant group, 71.4% (5/7) of S. agalactiae isolates were susceptible to clindamycin and 85.7%(6/7) to erythromycin. In the adult non-pregnant group, only resistance to clindamycin was observed in one patient (1/10; 10%). S. agalactiae as single pathogen was isolated in 57.14% (4/7) of pregnant and 60% (6/10) of adult non-pregnant S. agalactiae positive women. In mixed microbial cultures S. agalactiae was most frequently associated with Enterococcus faecalis and Escherichia coli. Conclusion The rate of S. agalactiae positive women in the population of pregnant and adult non-pregnant women of Tuzla Canton, B&H is comparable with other European countries. Large studies are needed to develop a common national strategy for the prevention of S. agalactiae infection in B&H, especially during pregnancy. Copyright© by the Medical Assotiation of Zenica-Doboj Canton.

  3. Incidence and serotype characterisation of Streptococcus agalactiae in a Portuguese hospital.

    Science.gov (United States)

    Pinto, Ana Mafalda; Pereira, Tamegão Aires; Alves, Valquíria; Araújo, António; Lage, Olga Maria

    2018-06-01

    Streptococcus agalactiae, commonly known as group B Streptococcus (GBS), has been recognised as a worldwide causative pathogenic agent of neonatal sepsis, meningitis and pneumonia. To better understand the behaviour of S. agalactiae in pregnant women from a hospital from the North of Portugal, retrospective analyses were performed to describe epidemiological, clinical and microbiological characteristics of the isolates obtained. Based on laboratorial records and the hospital's patient files, a 6-year retrospective study was performed to analyse S. agalactiae isolates from screened pregnant women between 35 and 37 weeks of gestation and hospitalised neonates from pregnant women between 24 and 41 weeks of gestation admitted in Hospital Pedro Hispano. Serotype characterisation was also performed in 67 GBS strains. In 6692 pregnant women between 35 and 37 weeks of gestation screened between 2011 and 2016, a total of 1377 S. agalactiae isolates (21%) were found. A high percentage (40%) of unknown colonisation status among hospitalised neonates from pregnant women between 24 and 41 weeks of gestations was also found. The incidence of neonatal sepsis was 8.7 (95% CI 7.0 to 10.8) cases per 1000 live births. Regarding serotype characterisation, serotype III (22.4%) was the most frequent, followed by serotype Ia (19.4%) and serotypes Ib and V (both with 17.9%). High epidemiological values of GBS colonisation and incidence were found in this study. In Portugal studies on the epidemiology and behaviour of S. agalactiae remain limited, reinforcing the importance and need for S. agalactiae screening across the country. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

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

    Science.gov (United States)

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

    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/absence of a subset of these loci in additional bovine and human strains revealed strong differentiation between the two groups (Fisher exact test: p S. agalactiae with Streptococcus uberis (nisin U operon) and Streptococcus dysgalactiae subsp. dysgalactiae (lactose operon). We also found evidence for LGT, involving the salivaricin operon, between the bovine S. agalactiae strain and either Streptococcus pyogenes or Streptococcus salivarius. Our findings provide insight intomechanismsfacilitatingenvironmentaladaptationandacquisitionofpotential virulence factors, while highlighting both the key role LGT has played in the recent evolution of the bovine S. agalactiae strain, and the importance of LGT among pathogens within a shared environment. PMID:21536150

  5. Acute Neonatal Parotitis with Late-Onset Septic Shock due to Streptococcus agalactiae

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    M. Boulyana

    2014-01-01

    Full Text Available Acute neonatal parotitis (ANP is a very rare disease. Most cases are managed conservatively; early antibiotics and adequate hydration may reduce the need for surgery. The most common cause of ANP is Staphylococcus aureus. We report a rare case of acute neonatal parotitis with late-onset septic shock due to Streptococcus agalactiae. The diagnosis was confirmed with ultrasound and isolation of Streptococcus agalactiae from blood culture. The patient was treated successfully with 10 days of intravenous antibiotics and supportive measures. Despite being rare, streptococcal ANP should be considered in the etiological diagnosis of neonatal sepsis. Early diagnosis and appropriate antibiotic might prevent serious complications.

  6. Discovery and Characterization of Human-Urine Utilization by Asymptomatic-Bacteriuria-Causing Streptococcus agalactiae.

    Science.gov (United States)

    Ipe, Deepak S; Ben Zakour, Nouri L; Sullivan, Matthew J; Beatson, Scott A; Ulett, Kimberly B; Benjamin, William H; Davies, Mark R; Dando, Samantha J; King, Nathan P; Cripps, Allan W; Schembri, Mark A; Dougan, Gordon; Ulett, Glen C

    2016-01-01

    Streptococcus agalactiae causes both symptomatic cystitis and asymptomatic bacteriuria (ABU); however, growth characteristics of S. agalactiae in human urine have not previously been reported. Here, we describe a phenotype of robust growth in human urine observed in ABU-causing S. agalactiae (ABSA) that was not seen among uropathogenic S. agalactiae (UPSA) strains isolated from patients with acute cystitis. In direct competition assays using pooled human urine inoculated with equal numbers of a prototype ABSA strain, designated ABSA 1014, and any one of several UPSA strains, measurement of the percentage of each strain recovered over time showed a markedly superior fitness of ABSA 1014 for urine growth. Comparative phenotype profiling of ABSA 1014 and UPSA strain 807, isolated from a patient with acute cystitis, using metabolic arrays of >2,500 substrates and conditions revealed unique and specific l-malic acid catabolism in ABSA 1014 that was absent in UPSA 807. Whole-genome sequencing also revealed divergence in malic enzyme-encoding genes between the strains predicted to impact the activity of the malate metabolic pathway. Comparative growth assays in urine comparing wild-type ABSA and gene-deficient mutants that were functionally inactivated for the malic enzyme metabolic pathway by targeted disruption of the maeE or maeK gene in ABSA demonstrated attenuated growth of the mutants in normal human urine as well as synthetic human urine containing malic acid. We conclude that some S. agalactiae strains can grow in human urine, and this relates in part to malic acid metabolism, which may affect the persistence or progression of S. agalactiae ABU. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Development of Streptococcus agalactiae vaccines for tilapia.

    Science.gov (United States)

    Liu, Guangjin; Zhu, Jielian; Chen, Kangming; Gao, Tingting; Yao, Huochun; Liu, Yongjie; Zhang, Wei; Lu, Chengping

    2016-12-21

    Vaccination is a widely accepted and effective method to prevent most pathogenic diseases in aquaculture. Various species of tilapia, especially Nile tilapia Oreochromis niloticus, are farmed worldwide because of their high consumer demand. Recently, the tilapia-breeding industry has been hampered by outbreaks of Streptococcus agalactiae infection, which cause high mortality and huge economic losses. Many researchers have attempted to develop effective S. agalactiae vaccines for tilapia. This review provides a summary of the different kinds of S. agalactiae vaccines for tilapia that have been developed recently. Among the various vaccine types, inactivated S. agalactiae vaccines showed superior protection efficiency when compared with live attenuated, recombinant and DNA vaccines. With respect to vaccination method, injecting the vaccine into tilapia provided the most effective immunoprotection. Freund's incomplete adjuvant appeared to be suitable for tilapia vaccines. Other factors, such as immunization duration and number, fish size and challenge dose, also influenced the vaccine efficacy.

  8. The Evaluation of Antibacterial Activity (Streptococcus agalactiae by Using Mahkota Dewa Extract (Phaleria macrocarpa L. with Diffirent Solvent

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    Lutfiana Safitri

    2017-07-01

    Full Text Available This research was aimed to determine the antibacterial activity of mahkota dewa extract and the effective concentration of mahkota dewa extract against Streptococcus agalactiae. This research was conducted from 2Mayth to 2Juneth 2016. The material ware Streptococcus agalactiae bacteria which isolated from mastitis milk, mahkota dewa which extracted using Aquades and ethanol. The method design was experiment research using Nested Design with 6 treatments and 5 replications. The concentrations of the treatments were P1 (10%, P2 (20%, P3 (30%, P4 (40% and P5 (50% and positive control P0 (iodips 10%. The collected data were analyzed using the analysis of variance indicated the significant effects, the least significant different test (LSD was then employed. Mahkota dewa extract using ethanol (50% result showed that significantly (P<0.01 on growth of Streptococcus agalactiae was 20.34± 0.92mm. Mahkota dewa extract using aquades (50% result showed that significantly (P<0.01 on growth of Streptococcus agalactiae was 10.0 ± 0.50 mm. The conclusion of this research is Mahkota dewa fruits extract with ethanol had inhibitory higher in habiting the Streptococcus agalactiae bacteria compared to aquades solvent. The best of treatment of Mahkota dewa fruits extract with ethanol and Aquades solvent in concentration 50% had a high ability to inhibit the growth of Streptococcus agalactiae.

  9. Major surfome and secretome profile of Streptococcus agalactiae from Nile tilapia (Oreochromis niloticus): Insight into vaccine development.

    Science.gov (United States)

    Li, Wei; Wang, Hai-Qing; He, Run-Zhen; Li, Yan-Wei; Su, You-Lu; Li, An-Xing

    2016-08-01

    Streptococcus agalactiae is a major piscine pathogen that is responsible for huge economic losses to the aquaculture industry. Safe recombinant vaccines, based on a small number of antigenic proteins, are emerging as the most attractive, cost-effective solution against S. agalactiae. The proteins of S. agalactiae exposed to the environment, including surface proteins and secretory proteins, are important targets for the immune system and they are likely to be good vaccine candidates. To obtain a precise profile of its surface proteins, S. agalactiae strain THN0901, which was isolated from tilapia (Oreochromis niloticus), was treated with proteinase K to cleave surface-exposed proteins, which were identified by liquid chromatography-tandem spectrometry (LC-MS/MS). Forty surface-associated proteins were identified, including ten proteins containing cell wall-anchoring motifs, eight lipoproteins, eleven membrane proteins, seven secretory proteins, three cytoplasmic proteins, and one unknown protein. In addition, culture supernatant proteins of S. agalactiae were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and all of the Coomassie-stained bands were subsequently identified by LC-MS/MS. A total of twenty-six extracellular proteins were identified, including eleven secretory proteins, seven cell wall proteins, three membrane proteins, two cytoplasmic proteins and three unknown proteins. Of these, six highly expressed surface-associated and secretory proteins are putative to be vaccine candidate of piscine S. agalactiae. Moreover, immunogenic secreted protein, a highly expressed protein screened from the secretome in the present study, was demonstrated to induce high antibody titer in tilapia, and it conferred protection against S. agalactiae, as evidenced by the relative percent survival (RPS) 48.61± 8.45%. The data reported here narrow the scope of screening protective antigens, and provide guidance in the development of a novel

  10. Streptococcus agalactiae in the environment of bovine dairy herds--rewriting the textbooks?

    Science.gov (United States)

    Jørgensen, H J; Nordstoga, A B; Sviland, S; Zadoks, R N; Sølverød, L; Kvitle, B; Mørk, T

    2016-02-29

    Many free-stall bovine dairy herds in Norway fail to eradicate Streptococcus agalactiae despite long-term control measures. In a longitudinal study of 4 free-stall herds with automatic milking systems (AMS), milk and extramammary sites were sampled 4 times with 1-2 month intervals. Composite milk, rectal- and vaginal swabs were collected from dairy cows; rectal swabs from heifers and young stock; rectal- and tonsillar swabs from calves; and environmental swabs from the AMS, the floors, cow beds, watering and feeding equipment. A cross sectional study of 37 herds was also conducted, with 1 visit for environmental sampling. Fifteen of the herds were known to be infected with S. agalactiae while the remaining 22 had not had evidence of S. agalactiae mastitis in the preceding 2 years. All samples were cultured for S. agalactiae, and selected isolates (n=54) from positive herds were genotyped by Multi Locus Sequence Typing (MLST). Results show that the bovine gastrointestinal tract and the dairy cow environment are reservoirs of S. agalactiae, and point to the existence of 2 transmission cycles; a contagious transmission cycle via the milking machine and an oro-fecal transmission cycle, with drinking water as the most likely vehicle for transmission. Ten sequence types were identified, and results suggest that strains differ in their ability to survive in the environment and transmit within dairy herds. Measures to eradicate S. agalactiae from bovine dairy herds should take into account the extra-mammary reservoirs and the potential for environmental transmission of this supposedly exclusively contagious pathogen. Copyright © 2016. Published by Elsevier B.V.

  11. Whole-cell vaccine of Streptococcus agalactiae in Oreochromis sp. with immersion method

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    , Sukenda

    2015-05-01

    Full Text Available ABSTRACT The study was aimed to evaluate the efficacy of formalin-killed non-hemolytic Streptococcus agalactiae N14G and NK1 isolates whole-killed vaccine to prevent streptococcosis in tilapia. Ten fishes were reared in a tank 60x30x35 cm3 with an average body weight at 10.79±0.99 g. Fish was vaccinated through bath immersion at a concentration of 109 cfu/mL. Fish was subsequently challenged by intraperitonial injection of Streptococcus agalactiae 105 cfu/mL at 11 days post-vaccination. Parameters observed were survival, relative percent survival (RPS, total leukocyte, phagocytic activity, antibody titer, total erythrocyte, haemoglobin level, haematocrit level, dan water quality. Samplings were performed in day-0, 20, and 30 after vaccination. Both vaccines have shown higher survival (60% and RPS (40% when challenged with pathogenic Streptococcus N14G isolates than other treatments. Based on RPS percentage observed, those vaccine were still not sufficiently effective to combat S. agalactiae infection. Keywords: tilapia, bath immersion, Streptococcus agalactiae, whole-cell vaccine ABSTRAK Penelitian ini bertujuan untuk mengevaluasi efikasi vaksin formalin-killed cell Streptococcus agalactiae tipe isolat nonhemolitik N14G dan NK1 se utuh yang diberikan melalui perendaman dalam mencegah penyakit streptococcosis pada ikan nila. Ikan nila yang digunakan memiliki bobot 10,79±0,99 g, dipelihara sebanyak sepuluh ekor dalam akuarium ukuran 60x30x35 cm3. Ikan divaksinasi dengan metode perendaman dengan dosis 109 cfu/mL. Uji tantang dilakukan pada hari ke-11 pascavaksinasi dengan dosis 105 cfu/mL. Parameter yang diamati meliputi sintasan (SR, sintasan relatif/relative percent survival (RPS, total leukosit, aktivitas fagositik, titer antibodi, total eritrosit, kadar hemoglobin, kadar hematokrit, dan kualitas air. Pengamatan parameter dilakukan pada hari ke-0, ke-10, ke-20, dan ke-30. Hasil penelitian menunjukkan perlakuan kedua vaksin yang diinfeksi

  12. Molecular identification and histopathological study of natural Streptococcus agalactiae infection in hybrid tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Laith, A A; Ambak, Mohd Azmi; Hassan, Marina; Sheriff, Shahreza Md; Nadirah, Musa; Draman, Ahmad Shuhaimi; Wahab, Wahidah; Ibrahim, Wan Nurhafizah Wan; Aznan, Alia Syafiqah; Jabar, Amina; Najiah, Musa

    2017-01-01

    The main objective of this study was to emphasize on histopathological examinations and molecular identification of Streptococcus agalactiae isolated from natural infections in hybrid tilapia ( Oreochromis niloticus ) in Temerloh Pahang, Malaysia, as well as to determine the susceptibility of the pathogen strains to various currently available antimicrobial agents. The diseased fishes were observed for variable clinical signs including fin hemorrhages, alterations in behavior associated with erratic swimming, exophthalmia, and mortality. Tissue samples from the eyes, brain, kidney, liver, and spleen were taken for bacterial isolation. Identification of S. agalactiae was screened by biochemical methods and confirmed by VITEK 2 and 16S rRNA gene sequencing. The antibiogram profiling of the isolate was tested against 18 standard antibiotics included nitrofurantoin, flumequine, florfenicol, amoxylin, doxycycline, oleandomycin, tetracycline, ampicillin, lincomycin, colistin sulfate, oxolinic acid, novobiocin, spiramycin, erythromycin, fosfomycin, neomycin, gentamycin, and polymyxin B. The histopathological analysis of eyes, brain, liver, kidney, and spleen was observed for abnormalities related to S. agalactiae infection. The suspected colonies of S. agalactiae identified by biochemical methods was observed as Gram-positive chained cocci, β-hemolytic, and non-motile. The isolate was confirmed as S. agalactiae by VITEK 2 (99% similarity), reconfirmed by 16S rRNA gene sequencing (99% similarity) and deposited in GenBank with accession no. KT869025. The isolate was observed to be resistance to neomycin and gentamicin. The most consistent gross findings were marked hemorrhages, erosions of caudal fin, and exophthalmos. Microscopic examination confirmed the presence of marked congestion and infiltration of inflammatory cell in the eye, brain, kidney, liver, and spleen. Eye samples showed damage of the lens capsule, hyperemic and hemorrhagic choroid tissue, and retina

  13. Molecular identification and histopathological study of natural Streptococcus agalactiae infection in hybrid tilapia (Oreochromis niloticus

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    Laith Abdul Razzak

    2017-01-01

    Full Text Available Aim: The main objective of this study was to emphasize on histopathological examinations and molecular identification of Streptococcus agalactiae isolated from natural infections in hybrid tilapia (Oreochromis niloticus in Temerloh Pahang, Malaysia, as well as to determine the susceptibility of the pathogen strains to various currently available antimicrobial agents. Materials and Methods: The diseased fishes were observed for variable clinical signs including fin hemorrhages, alterations in behavior associated with erratic swimming, exophthalmia, and mortality. Tissue samples from the eyes, brain, kidney, liver, and spleen were taken for bacterial isolation. Identification of S. agalactiae was screened by biochemical methods and confirmed by VITEK 2 and 16S rRNA gene sequencing. The antibiogram profiling of the isolate was tested against 18 standard antibiotics included nitrofurantoin, flumequine, florfenicol, amoxylin, doxycycline, oleandomycin, tetracycline, ampicillin, lincomycin, colistin sulfate, oxolinic acid, novobiocin, spiramycin, erythromycin, fosfomycin, neomycin, gentamycin, and polymyxin B. The histopathological analysis of eyes, brain, liver, kidney, and spleen was observed for abnormalities related to S. agalactiae infection. Results: The suspected colonies of S. agalactiae identified by biochemical methods was observed as Gram-positive chained cocci, β-hemolytic, and non-motile. The isolate was confirmed as S. agalactiae by VITEK 2 (99% similarity, reconfirmed by 16S rRNA gene sequencing (99% similarity and deposited in GenBank with accession no. KT869025. The isolate was observed to be resistance to neomycin and gentamicin. The most consistent gross findings were marked hemorrhages, erosions of caudal fin, and exophthalmos. Microscopic examination confirmed the presence of marked congestion and infiltration of inflammatory cell in the eye, brain, kidney, liver, and spleen. Eye samples showed damage of the lens capsule

  14. Natural outbreak of Streptococcus agalactiae (GBS) infection in wild giant Queensland grouper, Epinephelus lanceolatus (Bloch), and other wild fish in northern Queensland, Australia.

    Science.gov (United States)

    Bowater, R O; Forbes-Faulkner, J; Anderson, I G; Condon, K; Robinson, B; Kong, F; Gilbert, G L; Reynolds, A; Hyland, S; McPherson, G; Brien, J O'; Blyde, D

    2012-03-01

    Ninety-three giant Queensland grouper, Epinephelus lanceolatus (Bloch), were found dead in Queensland, Australia, from 2007 to 2011. Most dead fish occurred in northern Queensland, with a peak of mortalities in Cairns in June 2008. In 2009, sick wild fish including giant sea catfish, Arius thalassinus (Rüppell), and javelin grunter, Pomadasys kaakan (Cuvier), also occurred in Cairns. In 2009 and 2010, two disease epizootics involving wild stingrays occurred at Sea World marine aquarium. Necropsy, histopathology, bacteriology and PCR determined that the cause of deaths of 12 giant Queensland grouper, three wild fish, six estuary rays, Dasyatis fluviorum (Ogilby), one mangrove whipray, Himantura granulata (Macleay), and one eastern shovelnose ray, Aptychotrema rostrata (Shaw), was Streptococcus agalactiae septicaemia. Biochemical testing of 34 S. agalactiae isolates from giant Queensland grouper, wild fish and stingrays showed all had identical biochemical profiles. The 16S rRNA gene sequences of isolates confirmed all isolates were S. agalactiae; genotyping of selected S. agalactiae isolates showed the isolates from giant Queensland grouper were serotype Ib, whereas isolates from wild fish and stingrays closely resembled serotype II. This is the first report of S. agalactiae from wild giant Queensland grouper and other wild tropical fish and stingray species in Queensland, Australia. © 2012 Blackwell Publishing Ltd and State of Queensland.

  15. Identification of a virulence-related surface protein XF in piscine Streptococcus agalactiae by pre-absorbed immunoproteomics.

    Science.gov (United States)

    Liu, Guangjin; Zhang, Wei; Liu, Yongjie; Yao, Huochun; Lu, Chengping; Xu, Pao

    2014-10-26

    Since 2009, large-scale Streptococcus agalactiae infections have broken out in cultured tilapia farms in China, resulting in considerable economic losses. Screening of the surface proteins is required to identify virulence factors or protective antigens involved in piscine S.agalactiae infections in tilapia. Pre-absorbed immunoproteomics method (PAIM) is a useful method previously established in our laboratory for identifying bacterial surface proteins. A serine-rich repeat protein family 1 (Srr-1), designated XF, was identified by PAIM in piscine S. agalactiae isolate GD201008-001. To investigate the role of XF in the pathogenesis of piscine S. agalactiae, an isogenic xf mutant strain (Δxf) and a complemented strain (CΔxf) were successfully constructed. The Δxf mutant and CΔxf showed no significant differences in growth characteristics and adherence to HEp-2 cells compared with the wild-type strain. However the 50% lethal dose of Δxf was increased (4-fold) compared with that of the parental strain in a zebrafish infection model. The findings demonstrated that XF is a virulence-related, highly immunoreactive surface protein and is involved in the pathogenicity of S. agalactiae infections in fish.

  16. Prophagic DNA Fragments in Streptococcus agalactiae Strains and Association with Neonatal Meningitis

    Science.gov (United States)

    van der Mee-Marquet, Nathalie; Domelier, Anne-Sophie; Mereghetti, Laurent; Lanotte, Philippe; Rosenau, Agnès; van Leeuwen, Willem; Quentin, Roland

    2006-01-01

    We identified—by randomly amplified polymorphic DNA (RAPD) analysis at the population level followed by DNA differential display, cloning, and sequencing—three prophage DNA fragments (F5, F7, and F10) in Streptococcus agalactiae that displayed significant sequence similarity to the DNA of S. agalactiae and Streptococcus pyogenes. The F5 sequence aligned with a prophagic gene encoding the large subunit of a terminase, F7 aligned with a phage-associated cell wall hydrolase and a phage-associated lysin, and F10 aligned with a transcriptional regulator (ArpU family) and a phage-associated endonuclease. We first determined the prevalence of F5, F7, and F10 by PCR in a collection of 109 strains isolated in the 1980s and divided into two populations: one with a high risk of causing meningitis (HR group) and the other with a lower risk of causing meningitis (LR group). These fragments were significantly more prevalent in the HR group than in the LR group (P S. agalactiae strains to invade the neonatal brain endothelium. We then determined the prevalence of F5, F7, and F10 by PCR in a collection of 40 strains recently isolated from neonatal meningitis cases for comparison with the cerebrospinal fluid (CSF) strains isolated in the 1980s. The prevalence of the three prophage DNA fragments was similar in these two populations isolated 15 years apart. We suggest that the prophage DNA fragments identified have remained stable in many CSF S. agalactiae strains, possibly due to their importance in virulence or fitness. PMID:16517893

  17. Pathological analysis, detection of antigens, FasL expression analysis and leucocytes survival analysis in tilapia (Oreochromis niloticus) after infection with green fluorescent protein labeled Streptococcus agalactiae.

    Science.gov (United States)

    Wang, Jingyuan; Wu, Jinying; Yi, Liyuan; Hou, Zengxin; Li, Wensheng

    2017-03-01

    The pathogenesis of Streptococcus agalactiae infection in tilapia has not been fully described. To understand this, we investigated the clinic-pathological features of acute experimental septicemia in tilapia (Oreochromis niloticus) after receiving an intra-peritoneal injection with S. agalactiae THN-1901GFP. Immunohistochemistry and sections of pathological tissues were used to estimate the level of damage in the head-kidney, liver, spleen and trunk-kidney. The expression of FasL was analyzed by western blotting in these samples based on their damage levels. Leucocytes were isolated from the head-kidney and incubated with S. agalactiae THN-1901GFP. Then, phagocytosis, programmed cell death and the expression of FasL were analyzed. The infected tissues showed varying degrees of necrosis and histolysis. The serous membrane of the intestine was dissolved by S. agalactiae THN-1901GFP. Antigens of S. agalactiae THN-1901GFP accumulated in different parts of the infected organs. In the head-kidney and spleen, the expression of FasL was up-regulated in parallel with increased tissue damage. After being incubated with S. agalactiae THN-1901GFP, the phagocytic capacity and ability were both very high and the expression of FasL remained high in leucocytes. S. agalactiae THN-1901GFP was able to survive for a long period of time after being engulfed by phagocytic cells. These findings offer insight into the pathogenesis of S. agalactiae infection in tilapia. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Infecciones por Streptococcus agalactiae en un servicio de neonatología abierto Infections due to Streptococcus agalactiae at an open neonatology service

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    Manuel Díaz Álvarez

    2008-12-01

    infections due to Streptococcus agalactiae admitted in the Neonatology Service of «Juan M. Márquez» University Pediatric Hospital from February 1992 to December 2007 was conducted. Different clinical and epidemiological variables were processed and analyzed, with calculation of incidence and lethality rates and relation among the categoric variables. RESULTS. There were 76 newborns with infection due to Streptococcus agalactiae for an average annual rate of 1.9 x 100 admissions. It was observed a predominance of late onset infections and of those acquired in the community (89.5 % y 93.4 %, respectively. Meningitis was the most common clinical form followed by isolated bacteriemia. 56 of the 76 newborn infants had bacteriemia (73.7 %. Streptococcus agalactiae showed a high sensitivity to penicillin, erythromycine, vancomycin, cefotaxime and cloranphenicol. There were 7 deaths (9.2 %. All of them had infection of the CNS. CONCLUSIONS. Streptococcus agalactiae is an agent that causes infections affecting the newborn infant, both in the community and in the hospital. These infections may be lethal in some patients with infection of the CNS with o without bacteriemia, even when they maintain a pattern of elevated susceptibility to betalactamic antibiotics.

  19. Endocytosis‒Mediated Invasion and Pathogenicity of Streptococcus agalactiae in Rat Cardiomyocyte (H9C2)

    OpenAIRE

    Pooja, Sharma; Pushpanathan, Muthuirulan; Gunasekaran, Paramasamy; Rajendhran, Jeyaprakash

    2015-01-01

    Streptococcus agalactiae infection causes high mortality in cardiovascular disease (CVD) patients, especially in case of setting prosthetic valve during cardiac surgery. However, the pathogenesis mechanism of S. agalactiae associate with CVD has not been well studied. Here, we have demonstrated the pathogenicity of S. agalactiae in rat cardiomyocytes (H9C2). Interestingly, both live and dead cells of S. agalactiae were uptaken by H9C2 cells. To further dissect the process of S. agalactiae int...

  20. Genomic comparison of virulent and non-virulent Streptococcus agalactiae in fish.

    Science.gov (United States)

    Delannoy, C M J; Zadoks, R N; Crumlish, M; Rodgers, D; Lainson, F A; Ferguson, H W; Turnbull, J; Fontaine, M C

    2016-01-01

    Streptococcus agalactiae infections in fish are predominantly caused by beta-haemolytic strains of clonal complex (CC) 7, notably its namesake sequence type (ST) 7, or by non-haemolytic strains of CC552, including the globally distributed ST260. In contrast, CC23, including its namesake ST23, has been associated with a wide homeothermic and poikilothermic host range, but never with fish. The aim of this study was to determine whether ST23 is virulent in fish and to identify genomic markers of fish adaptation of S. agalactiae. Intraperitoneal challenge of Nile tilapia, Oreochromis niloticus (Linnaeus), showed that ST260 is lethal at doses down to 10(2) cfu per fish, whereas ST23 does not cause disease at 10(7) cfu per fish. Comparison of the genome sequence of ST260 and ST23 with those of strains derived from fish, cattle and humans revealed the presence of genomic elements that are unique to subpopulations of S. agalactiae that have the ability to infect fish (CC7 and CC552). These loci occurred in clusters exhibiting typical signatures of mobile genetic elements. PCR-based screening of a collection of isolates from multiple host species confirmed the association of selected genes with fish-derived strains. Several fish-associated genes encode proteins that potentially provide fitness in the aquatic environment. © 2014 John Wiley & Sons Ltd.

  1. Orientation of streptococcus agalactiae irradiation dose for subclinical mastitis vaccine in dairy cows

    International Nuclear Information System (INIS)

    Tuasikal, B.J.; Estuningsih, S.; Pasaribu, F.H.; Wibawan, I.W.T.

    2012-01-01

    An experiment to determine the effect of gamma-ray irradiation in debilitating Streptococcus agalactiae as a cause of subclinical mastitis (inflammation of the udder) in cows has been conducted. S. agalactiae bacteria was isolated from subclinical mastitis found in dairy cows in the field was then observed for its cell growth. The bacteria which have reached mid-log phase of growth, were divided into 5 treatment groups, of which each was irradiated at dose level of 0; 25; 50; 75; and 100Gy. Irradiated bacteria subsequently were grown on Brain Heart Infusion agar (BHI), and each of its colonies was counted for LD50 determination. The obtained curves from irradiated treatment bacterial shown viability percentage by the linier equation of Y = 95.414 e -0,0371X ; R 2 = 0.9979, while Y = % viability and X = dose of irradiation. The calculation done in this experiment shows that irradiation dose of 17.4 Gy weaken the bacteria pathogenicity of S. agalactiae to the level of LD 50 . (author)

  2. Identification of genes preferentially expressed by highly virulent piscine Streptococcus agalactiae upon interaction with macrophages.

    Directory of Open Access Journals (Sweden)

    Chang-Ming Guo

    Full Text Available Streptococcus agalactiae, long recognized as a mammalian pathogen, is an emerging concern with regard to fish. In this study, we used a mouse model and in vitro cell infection to evaluate the pathogenetic characteristics of S. agalactiae GD201008-001, isolated from tilapia in China. This bacterium was found to be highly virulent and capable of inducing brain damage by migrating into the brain by crossing the blood-brain barrier (BBB. The phagocytosis assays indicated that this bacterium could be internalized by murine macrophages and survive intracellularly for more than 24 h, inducing injury to macrophages. Further, selective capture of transcribed sequences (SCOTS was used to investigate microbial gene expression associated with intracellular survival. This positive cDNA selection technique identified 60 distinct genes that could be characterized into 6 functional categories. More than 50% of the differentially expressed genes were involved in metabolic adaptation. Some genes have previously been described as associated with virulence in other bacteria, and four showed no significant similarities to any other previously described genes. This study constitutes the first step in further gene expression analyses that will lead to a better understanding of the molecular mechanisms used by S. agalactiae to survive in macrophages and to cross the BBB.

  3. Identification of Genes Preferentially Expressed by Highly Virulent Piscine Streptococcus agalactiae upon Interaction with Macrophages

    Science.gov (United States)

    Guo, Chang-Ming; Chen, Rong-Rong; Kalhoro, Dildar Hussain; Wang, Zhao-Fei; Liu, Guang-Jin; Lu, Cheng-Ping; Liu, Yong-Jie

    2014-01-01

    Streptococcus agalactiae, long recognized as a mammalian pathogen, is an emerging concern with regard to fish. In this study, we used a mouse model and in vitro cell infection to evaluate the pathogenetic characteristics of S. agalactiae GD201008-001, isolated from tilapia in China. This bacterium was found to be highly virulent and capable of inducing brain damage by migrating into the brain by crossing the blood–brain barrier (BBB). The phagocytosis assays indicated that this bacterium could be internalized by murine macrophages and survive intracellularly for more than 24 h, inducing injury to macrophages. Further, selective capture of transcribed sequences (SCOTS) was used to investigate microbial gene expression associated with intracellular survival. This positive cDNA selection technique identified 60 distinct genes that could be characterized into 6 functional categories. More than 50% of the differentially expressed genes were involved in metabolic adaptation. Some genes have previously been described as associated with virulence in other bacteria, and four showed no significant similarities to any other previously described genes. This study constitutes the first step in further gene expression analyses that will lead to a better understanding of the molecular mechanisms used by S. agalactiae to survive in macrophages and to cross the BBB. PMID:24498419

  4. A multi locus variable number of tandem repeat analysis (MLVA scheme for Streptococcus agalactiae genotyping

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    Mereghetti Laurent

    2011-07-01

    Full Text Available Abstract Background Multilocus sequence typing (MLST is currently the reference method for genotyping Streptococcus agalactiae strains, the leading cause of infectious disease in newborns and a major cause of disease in immunocompromised children and adults. We describe here a genotyping method based on multiple locus variable number of tandem repeat (VNTR analysis (MLVA applied to a population of S. agalactiae strains of various origins characterized by MLST and serotyping. Results We studied a collection of 186 strains isolated from humans and cattle and three reference strains (A909, NEM316 and 2603 V/R. Among 34 VNTRs, 6 polymorphic VNTRs loci were selected for use in genotyping of the bacterial population. The MLVA profile consists of a series of allele numbers, corresponding to the number of repeats at each VNTR locus. 98 MLVA genotypes were obtained compared to 51 sequences types generated by MLST. The MLVA scheme generated clusters which corresponded well to the main clonal complexes obtained by MLST. However it provided a higher discriminatory power. The diversity index obtained with MLVA was 0.960 compared to 0.881 with MLST for this population of strains. Conclusions The MLVA scheme proposed here is a rapid, cheap and easy genotyping method generating results suitable for exchange and comparison between different laboratories and for the epidemiologic surveillance of S. agalactiae and analyses of outbreaks.

  5. ICESag37, a Novel Integrative and Conjugative Element Carrying Antimicrobial Resistance Genes and Potential Virulence Factors in Streptococcus agalactiae.

    Science.gov (United States)

    Zhou, Kaixin; Xie, Lianyan; Han, Lizhong; Guo, Xiaokui; Wang, Yong; Sun, Jingyong

    2017-01-01

    ICE Sag37 , a novel integrative and conjugative element carrying multidrug resistance and potential virulence factors, was characterized in a clinical isolate of Streptococcus agalactiae . Two clinical strains of S. agalactiae , Sag37 and Sag158, were isolated from blood samples of new-borns with bacteremia. Sag37 was highly resistant to erythromycin and tetracycline, and susceptible to levofloxacin and penicillin, while Sag158 was resistant to tetracycline and levofloxacin, and susceptible to erythromycin. Transfer experiments were performed and selection was carried out with suitable antibiotic concentrations. Through mating experiments, the erythromycin resistance gene was found to be transferable from Sag37 to Sag158. Sma I-PFGE revealed a new Sma I fragment, confirming the transfer of the fragment containing the erythromycin resistance gene. Whole genome sequencing and sequence analysis revealed a mobile element, ICE Sag37 , which was characterized using several molecular methods and in silico analyses. ICE Sag37 was excised to generate a covalent circular intermediate, which was transferable to S. agalactiae . Inverse PCR was performed to detect the circular form. A serine family integrase mediated its chromosomal integration into rumA , which is a known hotspot for the integration of streptococcal ICEs. The integration site was confirmed using PCR. ICE Sag37 carried genes for resistance to multiple antibiotics, including erythromycin [ erm(B) ], tetracycline [ tet(O) ], and aminoglycosides [ aadE, aphA , and ant(6) ]. Potential virulence factors, including a two-component signal transduction system ( nisK/nisR ), were also observed in ICE Sag37 . S1-PFGE analysis ruled out the existence of plasmids. ICE Sag37 is the first ICE Sa2603 family-like element identified in S. agalactiae carrying both resistance and potential virulence determinants. It might act as a vehicle for the dissemination of multidrug resistance and pathogenicity among S. agalactiae .

  6. Two novel functions of hyaluronidase from Streptococcus agalactiae are enhanced intracellular survival and inhibition of proinflammatory cytokine expression.

    Science.gov (United States)

    Wang, Zhaofei; Guo, Changming; Xu, Yannan; Liu, Guangjin; Lu, Chengping; Liu, Yongjie

    2014-06-01

    Streptococcus agalactiae is the causative agent of septicemia and meningitis in fish. Previous studies have shown that hyaluronidase (Hyl) is an important virulence factor in many Gram-positive bacteria. To investigate the role of S. agalactiae Hyl during interaction with macrophages, we inactivated the gene encoding extracellular hyaluronidase, hylB, in a clinical Hyl(+) isolate. The isogenic hylb mutant (Δhylb) displayed reduced survival in macrophages compared to the wild type and stimulated a significantly higher release of proinflammatory cytokines, such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor alpha (TNF-α), than the wild type in macrophages as well as in mice. Furthermore, only Hyl(+) strains could grow utilizing hyaluronic acid (HA) as the sole carbon source, suggesting that Hyl permits the organism to utilize host HA as an energy source. Fifty percent lethal dose (LD50) determinations in zebrafish demonstrated that the hylb mutant was highly attenuated relative to the wild-type strain. Experimental infection of BALB/c mice revealed that bacterial loads in the blood, spleen, and brain at 16 h postinfection were significantly reduced in the ΔhylB mutant compared to those in wild-type-infected mice. In conclusion, hyaluronidase has a strong influence on the intracellular survival of S. agalactiae and proinflammatory cytokine expression, suggesting that it plays a key role in S. agalactiae pathogenicity.

  7. Development of a loop-mediated isothermal amplification assay for the detection of Streptococcus agalactiae in bovine milk.

    Science.gov (United States)

    Bosward, Katrina L; House, John K; Deveridge, Amber; Mathews, Karen; Sheehy, Paul A

    2016-03-01

    Streptococcus agalactiae is a well-characterized bovine mastitis pathogen that is known to be highly contagious and capable of spreading rapidly in affected dairy herds. Loop-mediated isothermal amplification (LAMP) is a novel molecular diagnostic method that has the capability to provide rapid, cost-effective screening for pathogens to support on-farm disease control and eradication programs. In the current study, a LAMP test was developed to detect S. agalactiae in milk. The assay was validated on a bank of existing clinical mastitis milk samples that had previously been identified as S. agalactiae positive via traditional microbiological culture techniques and PCR. The LAMP assay was conducted on bacterial colonies and DNA extracted from milk in tube- and plate-based formats using multiple detection platforms. The 1-h assay conducted at 64 °C exhibited repeatability (coefficient of variation) of 2.07% (tube) and 8.3% (plate), sensitivity to ~20 pg of extracted DNA/reaction, and specificity against a panel of known bacterial mastitis pathogens. Of the 109 known S. agalactiae isolates assessed by LAMP directly from bacterial cells in culture, 108 were identified as positive, in accordance with PCR analysis. The LAMP analysis from the corresponding milk samples indicated that 104 of these milks exhibited a positive amplification curve. Although exhibiting some limitations, this assay provides an opportunity for rapid screening of milk samples to facilitate on-farm management of this pathogen. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Prevalence of the colonization by Streptococcus agalactiae in pregnant women from a public health system in Santo Ângelo – RS

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    Franciane Rios Senger

    2016-01-01

    Full Text Available ABSTRACT: Background and Objectives: Streptococcus agalactiae (GBS is part of the female genital tract microbiota, however, its clinical significance is related to infections in newborns and can cause severe cases of pneumonia, septicemia and meningitis. Thus, the aim of this study is to determine the prevalence of colonization by S. agalactiae in pregnant women and analyze the sensitivity profile of the isolated front of antimicrobials. Methods: Vaginal and anorectal samples of pregnant women over 30 weeks of gestation were collected from February to June 2013. The samples were stored in Stuart transport medium and then inoculated in Todd-Hewitt broth added gentamicin and acid nalidixic with subsequent subculture on blood agar plates. For identification were performed Gram test, catalase, CAMP and latex agglutination. Furthermore, the antimicrobial susceptibility tests and the test for the detection of resistance induced clindamycin in the samples resistant to erythromycin were conducted. They are also assessed, through interviews, demographic, socioeconomic and clinical and obstetric data. Results: It was observed colonization in 22.5% (18/80 of the pregnant women. The strains were sensitive to most of the antibiotics tested except erythromycin, showing a resistance of 22.2% (4/18 isolates. However, none of the samples erythromycin resistant had induced resistance to clindamycin. Conclusion: The high maternal colonization by S. agalactiae found emphasize the importance of isolation of this bacterium in late pregnancy, preventing the occurrence of neonatal infection. KEYWORDS: Streptococcus agalactiae. Pregnancy. Prevalence.

  9. Meningoencephalitis in farmed monosex Nile tilapia (Oreochromis niloticus L. caused by Streptococcus agalactiae

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    Adikesavalu Harresh

    2017-09-01

    Full Text Available Aquaculture of tilapia is a new research venture in India. With intensification in farming practices, tilapia are increasingly susceptible to bacterial infections. This article describes the isolation and identification of pathogenic bacteria from cultured monosex Nile tilapia, Oreochromis niloticus (L., that experienced moderate to severe mortalities in West Bengal, India between September and August 2014 and histopathological alterations in various organs. Gram-positive diplococci, identified as Streptococcus agalactiae with Streptococcus identification kits and 16S rDNA sequencing analysis, were isolated from the brain, operculum, and kidney. Other bacteria from the kidney were identified as Aeromonas sobria, A. caviae, Klebsiella pneumoniae ssp. pneumoniae, Escherichia coli, and Enterobacter cloacae. Staphylococcus epidermis was isolated from opercular hemorrhages. Histological sections of the infected tilapia brain revealed meningoencephalitis and granulomatous lesions. Sections from other organs indicated congestion, hemorrhagic and hyperplastic cells, necrosis, vacuolation, hemosiderin deposition, hypertrophic nuclei, melanomacrophage aggregation, and ruptured veins. This report is the first description of S. agalactiae as a primary pathogen causing meningoencephalitis in cultured tilapia in India.

  10. The homodimeric GBS1074 from Streptococcus agalactiae

    International Nuclear Information System (INIS)

    Shukla, Anshuman; Pallen, Mark; Anthony, Mark; White, Scott A.

    2010-01-01

    The homodimeric nature of the ESAT-6 homologue GBS1074 and the potential for fibre-like assemblies are revealed by the 2 Å resolution crystal structure. ESAT-6 is a well characterized secreted protein from Mycobacterium tuberculosis and represents the archetype of the WXG100 family of proteins. Genes encoding ESAT-6 homologues have been identified in the genome of the human pathogen Streptococcus agalactiae; one of these genes, esxA, has been cloned and the recombinant protein has been crystallized. In contrast to M. tuberculosis ESAT-6, the crystal structure of GBS1074 reveals a homodimeric structure similar to homologous structures from Staphylococcus aureus and Helicobacter pylori. Intriguingly, GBS1074 forms elongated fibre-like assemblies in the crystal structure

  11. Mixed infection zones may be important in the epidemiology of contagious agalactia

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    Jafarizadeh Amirreza

    2016-06-01

    Full Text Available Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma, Mycoplasma mycoides subsp. capri (Mmc, Mycoplasma capricolum subsp. capricolum (Mcc and Mycoplasma putrefaciens (Mp in sheep and goats with clinical signs consistent with contagious agalactia.

  12. Culture-Negative Neonatal Meningitis and Endocarditis Caused by Streptococcus agalactiae

    OpenAIRE

    Nègre, Valérie Lefranc; Colin-Gorski, Anne-Marie; Magnier, Suzel; Maisonneuve, Lydia; Aujard, Yannick; Bingen, Edouard; Bonacorsi, Stéphane

    2004-01-01

    We describe a case of culture-negative meningitis and endocarditis caused by Streptococcus agalactiae in a 27-day-old boy. S. agalactiae was detected in cerebrospinal fluid and serum by broad-spectrum PCR amplification.

  13. Relevance of Peptide Uptake Systems to the Physiology and Virulence of Streptococcus agalactiae

    OpenAIRE

    Samen, Ulrike; Gottschalk, Birgit; Eikmanns, Bernhard J.; Reinscheid, Dieter J.

    2004-01-01

    Streptococcus agalactiae is a major cause of invasive infections in human newborns. To satisfy its growth requirements, S. agalactiae takes up 9 of the 20 proteinogenic amino acids from the environment. Defined S. agalactiae mutants in one or several of four putative peptide permease systems were constructed and tested for peptide uptake, growth in various media, and expression of virulence traits. Oligopeptide uptake by S. agalactiae was shown to be mediated by the ABC transporter OppA1-F, w...

  14. Streptococcus agalactiae in pregnancies complicated by preterm prelabor rupture of membranes.

    Science.gov (United States)

    Musilova, Ivana; Pliskova, Lenka; Kutova, Radka; Jacobsson, Bo; Paterova, Pavla; Kacerovsky, Marian

    2016-01-01

    The main aim of this study was to evaluate the presence of Streptococcus agalactiae (S. agalactiae) in the vagina and the amniotic fluid in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). The next aim was to evaluate the incidence of S. agalactiae early onset sepsis in newborns from PPROM pregnancies, with respect to the presence of S. agalactiae in the vagina and the amniotic fluid. Singleton gestations with PPROM between 24 + 0 and 36 + 6 were included. A vaginal swab was obtained, and amniocentesis was performed at admission. The presence of S. agalactiae in the vagina and in the amniotic fluid was assessed by culture and by real-time polymerase chain reaction, respectively. In total, 336 women were included. The presence of S. agalactiae in the vaginal and amniotic fluid was found in 9% (31/336) and 1% (3/336) of women. One woman had S. agalactiae in the amniotic fluid but was negative for the presence of S. agalactiae in the vaginal fluid. Early onset neonatal sepsis developed in one newborn from pregnancies complicated by the presence of S. agalactiae in the amniotic fluid. The presence of S. agalactiae in the vagina and amniotic fluid complicated approximately each 10th and each 100th PPROM pregnancy. Cultivation-negative findings of S. agalactiae in the vagina did not exclude the positivity of the amniotic fluid for S. agalactiae and the development of early onset sepsis in newborns.

  15. Complete genome sequence of an attenuated Sparfloxacin resistant Streptococcus agalactiae strain 138spar

    Science.gov (United States)

    Through selection of resistance to sparfloxacin, an attenuated Streptococcus agalactiae strain 138spar was obtained from its virulent parent strain S. agalactiae 138P. The full genome of S. agalactiae 138spar is 1,838,126 bp. The availability of this genome will allow comparative genomics to identi...

  16. Molecular epidemiology and strain-specific characteristics of Streptococcus agalactiae at the herd and cow level.

    Science.gov (United States)

    Mahmmod, Y S; Klaas, I C; Katholm, J; Lutton, M; Zadoks, R N

    2015-10-01

    Host-adaptation of Streptococcus agalactiae subpopulations has been described whereby strains that are commonly associated with asymptomatic carriage or disease in people differ phenotypically and genotypically from those causing mastitis in dairy cattle. Based on multilocus sequence typing (MLST), the most common strains in dairy herds in Denmark belong to sequence types (ST) that are also frequently found in people. The aim of this study was to describe epidemiological and diagnostic characteristics of such strains in relation to bovine mastitis. Among 1,199 cattle from 6 herds, cow-level prevalence of S. agalactiae was estimated to be 27.4% based on PCR and 7.8% based on bacteriological culture. Quarter-level prevalence was estimated at 2.8% based on bacteriological culture. Per herd, between 2 and 26 isolates were characterized by pulsed-field gel electrophoresis (PFGE) and MLST. Within each herd, a single PFGE type and ST predominated, consistent with a contagious mode of transmission or point source infection within herds. Evidence of within-herd evolution of S. agalactiae was detected with both typing methods, although ST belonged to a single clonal complex (CC) per herd. Detection of CC23 (3 herds) was associated with significantly lower approximate count (colony-forming units) at the quarter level and significantly lower cycle threshold value at the cow level than detection of CC1 (2 herds) or CC19 (1 herd), indicating a lower bacterial load in CC23 infections. Median values for the number of infected quarters and somatic cell count (SCC) were numerically but not significantly lower for cows infected with CC23 than for cows with CC1 or CC19. For all CC, an SCC threshold of 200,000 cells/mL was an unreliable indicator of infection status, and prescreening of animals based on SCC as part of S. agalactiae detection and eradication campaigns should be discouraged. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights

  17. Molecular and bacteriological investigation of subclinical mastitis caused by Staphylococcus aureus and Streptococcus agalactiae in domestic bovids from Ismailia, Egypt.

    Science.gov (United States)

    Elhaig, Mahmoud Mohey; Selim, Abdelfattah

    2015-02-01

    A study was carried out to establish the prevalence of subclinical mastitis (SCM) in smallholder dairy farms in Ismailia, Egypt. A total of 340 milking cows and buffaloes were sampled from 60 farms, and 50 nasal swabs were collected from consenting farm workers. Milk samples were subjected to California mastitis test (CMT) and the positive samples were examined by bacterial culture and PCR to identify etiological agents. Based on CMT, the prevalence of SCM was 71.6 % in cattle and 43.5 % in buffaloes while the prevalence was 25.2 % at cow-quarter level and 21.7 % at buffaloes-quarter level. Bacteriological analysis showed that the most frequently identified bacteria were Staphylococcus (S.) aureus (38.3 %) and Streptococcus (Str.) agalactiae (20 %). The diagnostic sensitivity of PCR compared to bacterial culture was superior with S. aureus and Str. agalactiae detection being 41 and 22.6 %, respectively. Furthermore, methicillin-resistant S. aureus (MRSA) strains occurred in 52.2 and 45 % of isolates of animals and workers, respectively. Subclinical mastitis due to S. aureus and Str. agalactiae is endemic in smallholder dairy herds in Ismailia. The occurrence of MRSA in animals and workers highlights a need for wide epidemiological studies of MRSA and adopting control strategies.

  18. nalisis Keragaman Genetik Streptococcus agalactiae Penyebab Mastitis Subklinis Pada Sapi Perah Menggunakan pulsed field gel electrophoresis (PFGE) = Genetic Analysis of Streptococcus agalactiae Caused Subclinical ...

    OpenAIRE

    Trihastuti, Agnesia Endang

    2012-01-01

    Streptococcus agalactiae atau Streptokokus grup B (SGB) adalah salah satu bakteri utama penyebab mastitis subklinis pada sapi perch dan merupakan parasit obligat pada ambing. Karakterisasi S.agalactiae biasanya dilakukan secara konvensional menggunakan metode serotyping. Meslci metode ini sering digunakan namun masih mempunyai kelemahan apalagi masih adanya isolat S.agalactiae yang belum dapat dimasukkan ke dalam serotipe yang ada (nontypeable/NT), oleh karena itu pendekatan bare denga...

  19. An homolog of the Frz Phosphoenolpyruvate:carbohydrate phosphoTransferase System of extraintestinal pathogenic Escherichia coli is encoded on a genomic island in specific lineages of Streptococcus agalactiae.

    Science.gov (United States)

    Patron, Kévin; Gilot, Philippe; Camiade, Emilie; Mereghetti, Laurent

    2015-06-01

    We identified a Streptococcus agalactiae metabolic region (fru2) coding for a Phosphoenolpyruvate:carbohydrate phosphoTransferase System (PTS) homologous to the Frz system of extraintestinal pathogenic Escherichia coli strains. The Frz system is involved in environmental sensing and regulation of the expression of adaptation and virulence genes in E. coli. The S. agalactiae fru2 region codes three subunits of a PTS transporter of the fructose-mannitol family, a transcriptional activator of PTSs of the MtlR family, an allulose-6 phosphate-3-epimerase, a transaldolase and a transketolase. We demonstrated that all these genes form an operon. The fru2 operon is present in a 17494-bp genomic island. We analyzed by multilocus sequence typing a population of 492 strains representative of the S. agalactiae population and we showed that the presence of the fru2 operon is linked to the phylogeny of S. agalactiae. The fru2 operon is always present within strains of clonal complexes CC 1, CC 7, CC 10, CC 283 and singletons ST 130 and ST 288, but never found in other CCs and STs. Our results indicate that the fru2 operon was acquired during the evolution of the S. agalactiae species from a common ancestor before the divergence of CC 1, CC 7, CC 10, CC 283, ST 130 and ST 288. As S. agalactiae strains of CC 1 and CC 10 are frequently isolated from adults with invasive disease, we hypothesize that the S. agalactiae Fru2 system senses the environment to allow the bacterium to adapt to new conditions encountered during the infection of adults. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Diversity of prophage DNA regions of Streptococcus agalactiae clonal lineages from adults and neonates with invasive infectious disease.

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    Mazen Salloum

    Full Text Available The phylogenetic position and prophage DNA content of the genomes of 142 S. agalactiae (group-B streptococcus, GBS isolates responsible for bacteremia and meningitis in adults and neonates were studied and compared. The distribution of the invasive isolates between the various serotypes, sequence types (STs and clonal complexes (CCs differed significantly between adult and neonatal isolates. Use of the neighbor-net algorithm with the PHI test revealed evidence for recombination in the population studied (PHI, P = 2.01 × 10(-6, and the recombination-mutation ratio (R/M was 6:7. Nevertheless, the estimated R/M ratio differed between CCs. Analysis of the prophage DNA regions of the genomes of the isolates assigned 90% of the isolates to five major prophage DNA groups: A to E. The mean number of prophage DNA fragments amplified per isolate varied from 2.6 for the isolates of prophage DNA group E to 4.0 for the isolates of prophage DNA group C. The isolates from adults and neonates with invasive diseases were distributed differently between the various prophage DNA groups (P < 0.00001. Group C prophage DNA fragments were found in 52% of adult invasive isolates, whereas 74% of neonatal invasive isolates had prophage DNA fragments of groups A and B. Differences in prophage DNA content were also found between serotypes, STs and CCs (P < 0.00001. All the ST-1 and CC1 isolates, mostly of serotype V, belonged to the prophage DNA group C, whereas 84% of the ST-17 and CC17 isolates, all of serotype III, belonged to prophage DNA groups A and B. These data indicate that the transduction mechanisms, i.e., gene transfer from one bacterium to another by a bacteriophage, underlying genetic recombination in S. agalactiae species, are specific to each intraspecies lineage and population of strains responsible for invasive diseases in adults and neonates.

  1. Antibacterial activity and mechanism of berberine against Streptococcus agalactiae.

    Science.gov (United States)

    Peng, Lianci; Kang, Shuai; Yin, Zhongqiong; Jia, Renyong; Song, Xu; Li, Li; Li, Zhengwen; Zou, Yuanfeng; Liang, Xiaoxia; Li, Lixia; He, Changliang; Ye, Gang; Yin, Lizi; Shi, Fei; Lv, Cheng; Jing, Bo

    2015-01-01

    The antibacterial activity and mechanism of berberine against Streptococcus agalactiae were investigated in this study by analyzing the growth, morphology and protein of the S. agalactiae cells treated with berberine. The antibacterial susceptibility test result indicated minimum inhibition concentration (MIC) of berberine against Streptococcus agalactiae was 78 μg/mL and the time-kill curves showed the correlation of concentration-time. After the bacteria was exposed to 78 μg/mL berberine, the fragmentary cell membrane and cells unequal division were observed by the transmission electron microscopy (TEM), indicating the bacterial cells were severely damaged. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) study demonstrated that berberine could damage bacterial cells through destroying cellular proteins. Meanwhile, Fluorescence microscope revealed that berberine could affect the synthesis of DNA. In conclusion, these results strongly suggested that berberine may damage the structure of bacterial cell membrane and inhibit synthesis of protein and DNA, which cause Streptococcus agalactiae bacteria to die eventually.

  2. Molecular Characterization of Streptococcus agalactiae Causing Community- and Hospital-acquired Infections in Shanghai, China

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    Haoqin Jiang

    2016-08-01

    Full Text Available Streptococcus agalactiae, a colonizing agent in pregnant women and the main cause of neonatal sepsis and meningitis, has been increasingly associated with invasive disease in nonpregnant adults. We collected a total of 87 non-repetitive S. agalactiae isolates causing community-acquired (CA and hospital-acquired (HA infections in nonpregnant adults from a teaching hospital in Shanghai between 2009 and 2013. We identified and characterized their antibiotic resistance, sequence type (ST, serotype, virulence, and biofilm formation. The most frequent STs were ST19 (29.9%, ST23 (16.1%, ST12 (13.8%, and ST1 (12.6%. ST19 had significantly different distributions between CA- and HA-group B Streptococci (GBS isolates. The most frequent serotypes were III (32.2%, Ia (26.4%, V (14.9%, Ib (13.8%, and II (5.7%. Serotype III/ST19 was significantly associated with levofloxacin resistance in all isoates. The HA-GBS multidrug resistant rate was much higher than that of CA-GBS. Virulence genes pavA, cfb were found in all isolates. Strong correlations exist between serotype Ib (CA and HA and surface protein genes spb1 and bac, serotype III (HA and surface protein gene cps and GBS pilus cluster. The serotype, epidemic clone, PFGE-based genotype, and virulence gene are closely related between CA-GBS and HA-GBS, and certain serotypes and clone types were significantly associated with antibiotic resistance. However, CA-GBS and HA-GBS still had significant differences in their distribution of clone types, antibiotic resistance, and specific virulence genes, which may provide a basis for infection control.

  3. Endocytosis‒Mediated Invasion and Pathogenicity of Streptococcus agalactiae in Rat Cardiomyocyte (H9C2).

    Science.gov (United States)

    Pooja, Sharma; Pushpanathan, Muthuirulan; Gunasekaran, Paramasamy; Rajendhran, Jeyaprakash

    2015-01-01

    Streptococcus agalactiae infection causes high mortality in cardiovascular disease (CVD) patients, especially in case of setting prosthetic valve during cardiac surgery. However, the pathogenesis mechanism of S. agalactiae associate with CVD has not been well studied. Here, we have demonstrated the pathogenicity of S. agalactiae in rat cardiomyocytes (H9C2). Interestingly, both live and dead cells of S. agalactiae were uptaken by H9C2 cells. To further dissect the process of S. agalactiae internalization, we chemically inhibited discrete parts of cellular uptake system in H9C2 cells using genistein, chlorpromazine, nocodazole and cytochalasin B. Chemical inhibition of microtubule and actin formation by nocodazole and cytochalasin B impaired S. agalactiae internalization into H9C2 cells. Consistently, reverse‒ transcription PCR (RT‒PCR) and quantitative real time‒PCR (RT-qPCR) analyses also detected higher levels of transcripts for cytoskeleton forming genes, Acta1 and Tubb5 in S. agalactiae‒infected H9C2 cells, suggesting the requirement of functional cytoskeleton in pathogenesis. Host survival assay demonstrated that S. agalactiae internalization induced cytotoxicity in H9C2 cells. S. agalactiae cells grown with benzyl penicillin reduced its ability to internalize and induce cytotoxicity in H9C2 cells, which could be attributed with the removal of surface lipoteichoic acid (LTA) from S. agalactiae. Further, the LTA extracted from S. agalactiae also exhibited dose‒dependent cytotoxicity in H9C2 cells. Taken together, our data suggest that S. agalactiae cells internalized H9C2 cells through energy‒dependent endocytic processes and the LTA of S. agalactiae play major role in host cell internalization and cytotoxicity induction.

  4. Mycoplasma agalactiae Induces Cytopathic Effects in Infected Cells Cultured In Vitro.

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    Shrilakshmi Hegde

    Full Text Available Mycoplasma agalactiae is the etiological agent of the contagious agalactia syndrome in sheep and goats and causes significant economic losses worldwide. Yet the mechanism of pathogenesis is largely unknown. Even whole-genome sequence analysis of its pathogenic type strain did not lead to any conclusions regarding its virulence or pathogenicity factors. Although inflammation and tissue destruction at the local site of M. agalactiae infection are largely considered as effects of the host immune response, the direct effect of the agent on host cells is not completely understood. The aim of this study was to investigate the effect of M. agalactiae infection on the quality and viability of host cells in vitro. Changes in cell morphology including cell elongation, cytoplasm shrinkage and membrane blebbing were observed in infected HeLa cells. Chromatin condensation and increased caspase-3 cleavage in infected HeLa cells 48 h after infection suggests an apoptosis-like phenomenon in M. agalactiae-infected cells. In compliance with these results, decreased viability and cell lysis of M. agalactiae-infected HeLa cells was also observed. Measurement of the amount of LDH released after M. agalactiae infection revealed a time- and dose-dependent increase in HeLa cell lysis. A significant decrease in LDH released after gentamicin treatment of infected cells confirmed the major role of cytadherent M. agalactiae in inducing host cell lysis. This is the first study illustrating M. agalactiae's induction of cytopathic effects in infected HeLa cells. Further detailed investigation of infected host tissue for apoptotic markers might demonstrate the association between M. agalactiae-induced host cell lysis and the tissue destruction observed during M. agalactiae natural infection.

  5. Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus) and its transmission potential to cohabitated sheepshead minnows (Cyprinodon variegatus)

    Science.gov (United States)

    Streptococcus agalactiae has emerged as an economically important bacterial pathogen affecting global aquaculture. Worldwide aquaculture losses due to S. agalactiae are estimated around U.S. $1 billion, annually. Streptococcus agalactiae also known as a Lancefield Group B Streptococcus (GBS) is a Gr...

  6. Expression profiles of miRNAs from bovine mammary glands in response to Streptococcus agalactiae-induced mastitis.

    Science.gov (United States)

    Pu, Junhua; Li, Rui; Zhang, Chenglong; Chen, Dan; Liao, Xiangxiang; Zhu, Yihui; Geng, Xiaohan; Ji, Dejun; Mao, Yongjiang; Gong, Yunchen; Yang, Zhangping

    2017-08-01

    This study aimed to describe the expression profiles of microRNAs (miRNAs) from mammary gland tissues collected from dairy cows with Streptococcus agalactiae-induced mastitis and to identify differentially expressed miRNAs related to mastitis. The mammary glands of Chinese Holstein cows were challenged with Streptococcus agalactiae to induce mastitis. Small RNAs were isolated from the mammary tissues of the test and control groups and then sequenced using the Solexa sequencing technology to construct two small RNA libraries. Potential target genes of these differentially expressed miRNAs were predicted using the RNAhybrid software, and KEGG pathways associated with these genes were analysed. A total of 18 555 913 and 20 847 000 effective reads were obtained from the test and control groups, respectively. In total, 373 known and 399 novel miRNAs were detected in the test group, and 358 known and 232 novel miRNAs were uncovered in the control group. A total of 35 differentially expressed miRNAs were identified in the test group compared to the control group, including 10 up-regulated miRNAs and 25 down-regulated miRNAs. Of these miRNAs, miR-223 exhibited the highest degree of up-regulation with an approximately 3-fold increase in expression, whereas miR-26a exhibited the most decreased expression level (more than 2-fold). The RNAhybrid software predicted 18 801 genes as potential targets of these 35 miRNAs. Furthermore, several immune response and signal transduction pathways, including the RIG-I-like receptor signalling pathway, cytosolic DNA sensing pathway and Notch signal pathway, were enriched in these predicted targets. In summary, this study provided experimental evidence for the mechanism underlying the regulation of bovine mastitis by miRNAs and showed that miRNAs might be involved in signal pathways during S. agalactiae-induced mastitis.

  7. Severe infective endocarditis in a healthy adult due to Streptococcus agalactiae.

    Science.gov (United States)

    Fujita, Hiroaki; Nakamura, Itaru; Tsukimori, Ayaka; Sato, Akihiro; Ohkusu, Kiyofumi; Matsumoto, Tetsuya

    2015-09-01

    A case of severe endocarditis, with complications of multiple infarction, meningitis, and ruptured mitral chordae tendineae, caused by Streptococcus agalactiae in a healthy man, is reported. Emergency cardiovascular surgery was performed on the day of admission. Infective endocarditis caused by S. agalactiae is very rare, particularly in a healthy adult. In addition, microbiological analysis revealed that S. agalactiae of sequence type (ST) 19, which belongs to serotype III, was present in the patient's vegetation, mitral valve, and blood culture. It was therefore concluded that the endocarditis was caused by ST19, which has been reported as a non-invasive type of S. agalactiae. This was an extremely rare case in which S. agalactiae of ST19 caused very severe endocarditis in an adult patient with no underlying disease. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. Multiple Evolutionary Selections Involved in Synonymous Codon Usages in the Streptococcus agalactiae Genome.

    Science.gov (United States)

    Ma, Yan-Ping; Ke, Hao; Liang, Zhi-Ling; Liu, Zhen-Xing; Hao, Le; Ma, Jiang-Yao; Li, Yu-Gu

    2016-02-24

    Streptococcus agalactiae is an important human and animal pathogen. To better understand the genetic features and evolution of S. agalactiae, multiple factors influencing synonymous codon usage patterns in S. agalactiae were analyzed in this study. A- and U-ending rich codons were used in S. agalactiae function genes through the overall codon usage analysis, indicating that Adenine (A)/Thymine (T) compositional constraints might contribute an important role to the synonymous codon usage pattern. The GC3% against the effective number of codon (ENC) value suggested that translational selection was the important factor for codon bias in the microorganism. Principal component analysis (PCA) showed that (i) mutational pressure was the most important factor in shaping codon usage of all open reading frames (ORFs) in the S. agalactiae genome; (ii) strand specific mutational bias was not capable of influencing the codon usage bias in the leading and lagging strands; and (iii) gene length was not the important factor in synonymous codon usage pattern in this organism. Additionally, the high correlation between tRNA adaptation index (tAI) value and codon adaptation index (CAI), frequency of optimal codons (Fop) value, reinforced the role of natural selection for efficient translation in S. agalactiae. Comparison of synonymous codon usage pattern between S. agalactiae and susceptible hosts (human and tilapia) showed that synonymous codon usage of S. agalactiae was independent of the synonymous codon usage of susceptible hosts. The study of codon usage in S. agalactiae may provide evidence about the molecular evolution of the bacterium and a greater understanding of evolutionary relationships between S. agalactiae and its hosts.

  9. Comprehensive evaluation of immunomodulation by moderate hypoxia in S. agalactiae vaccinated Nile tilapia.

    Science.gov (United States)

    Gallage, Sanchala; Katagiri, Takayuki; Endo, Masato; Maita, Masashi

    2017-07-01

    Streptococcus agalactiae is a major bacterial pathogen in tilapia aquaculture. Vaccines are known to provide protection but S. agalactiae clearance in tilapia can be reduced by marginal environmental conditions. Therefore, the purpose of this study is to examine S. agalactiae clearance in vaccinated Nile tilapia under moderate hypoxic (55± 5% DO) and normoxic (85 ± 5%DO) conditions. Fish were acclimatized to either moderate hypoxia or normoxia and immunized with formalin-inactivated S. agalactiae. Fish were experimentally challenged with S. agalactiae at 30 days post-vaccination. Serum antibody titer was significantly higher in vaccinated fish kept under normoxic condition compared to the moderate hypoxic condition at fifteen and thirty days post-vaccination. The cumulative mortality following challenge was significantly reduced in vaccinated fish kept under normoxic condition compared to those in moderate hypoxic condition reflecting that pre-challenge antibody titer may correlate with survival of fish. Blood and tissue pathogen burden detection of S. agalactiae studies revealed that culturable S. agalactiae cells could not be detected in the blood of normoxic vaccinated fish at all the sampling points. In contrast, fish vaccinated in moderate hypoxic condition had considerable number of culturable S. agalactiae cells in their blood up to 5 days following challenge. Phagocytosis and intracellular reactive oxygen species (ROS) production were lowered by moderate hypoxia in vitro. Furthermore, presence of specific antibodies and higher specific antibody level in the serum increased phagocytosis, ROS production and lowered intracellular survival of S. agalactiae in head kidney leukocytes. Overall this study has highlighted that S. agalactiae clearance in vaccinated Nile tilapia is modulated by moderate hypoxia. One of the possible explanations for this might be less efficient phagocytic activities due to low oxygen availability and lower specific

  10. Effect Of Gamma Ray Irradiation On Streptococcus Agalactiae Growth For Vaccine Agent Of Mastitis Disease In Dairy Cattle

    International Nuclear Information System (INIS)

    Jeane Tuasikal, B; Sugoro, I; Tjiptosumirat, T; Lina, M

    2003-01-01

    A study has been conducted to determine the effect of gamma ray irradiation to attenuate infectivity of S. agalactiae as dominant bacteria causing mastitis in dairy cattle. The aim of the study is obtaining optimum irradiation dosage to provide radio vaccine for mastitis. S. agalactiae isolate bacteria of which has reach the mid log-phase was cultured and divided into 6 treatment groups of irradiation doses, i.e. 0, 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. Following irradiation, bacteria were then cultured in BHI agar media for colony counting to determine the LD 50 , resulting 7.5x10 8 ; 5.0x10 7 ; 7.0x10 6 ; 9.5x10 5 ; 1.5x10 4 ; and 3.5x10 3 cell/ml, respectively. Result of this study shows the higher irradiation doses the lower number of bacteria per ml, and LD 50 , which found to be under 0.2 kGy of irradiation dose

  11. Effects on goat milk quality of the presence of Mycoplasma spp. in herds without symptoms of contagious agalactia.

    Science.gov (United States)

    de la Fe, Christian; Sánchez, Antonio; Gutierrez, Aldo; Contreras, Antonio; Carlos Corrales, Juan; Assunçao, Patricia; Poveda, Carlos; Poveda, José B

    2009-02-01

    This study was designed to assess the possible effects of mycoplasmas on the quality of milk produced by goat herds in a contagious agalactia (CA) endemic area with absence of classical symptoms. Several factors related to milk quality (percentages of fat, total protein, lactose and total solids, standard plate counts (SPC) and presence of Staphylococcus aureus) were compared in mycoplasma-infected and non-infected herds. To define the CA status of 26 herds on the island of Lanzarote (Spain), where CA is endemic, 570 individual milk samples and 266 bulk tank milk (BTM) samples were microbiologically analysed for the presence of Mycoplasma spp. A herd was considered infected by mycoplasmas when at least a sample (individual or BTM) was positive. BTM samples were also used to determine milk quality parameters. Mycoplasma infection was confirmed in 13 herds. A total of 31, 10 and 11 strains of Mycoplasma mycoides subsp. mycoides LC (MmmLC), Mp. agalactiae and Mp. capricolum subsp. capricolum were isolated. No significant differences were observed between the least square means of the variables fat, total protein, lactose and total solids or SPC recorded for the infected v. non-infected herds. The Staph. aureus status of a herd was also found to be independent of the presence of Mycoplasma spp. Our findings indicate that neither the presence of mycoplasmas in a goat herd with absence of classical symptoms seem to compromise the quality of the BTM.

  12. Visualization of the role of host heme on the virulence of the heme auxotroph Streptococcus agalactiae.

    Science.gov (United States)

    Joubert, Laetitia; Dagieu, Jean-Baptiste; Fernandez, Annabelle; Derré-Bobillot, Aurélie; Borezée-Durant, Elise; Fleurot, Isabelle; Gruss, Alexandra; Lechardeur, Delphine

    2017-01-16

    Heme is essential for several cellular key functions but is also toxic. Whereas most bacterial pathogens utilize heme as a metabolic cofactor and iron source, the impact of host heme during bacterial infection remains elusive. The opportunist pathogen Streptococcus agalactiae does not synthesize heme but still uses it to activate a respiration metabolism. Concomitantly, heme toxicity is mainly controlled by the HrtBA efflux transporter. Here we investigate how S. agalactiae manages heme toxicity versus benefits in the living host. Using bioluminescent bacteria and heme-responsive reporters for in vivo imaging, we show that the capacity of S. agalactiae to overcome heme toxicity is required for successful infection, particularly in blood-rich organs. Host heme is simultaneously required, as visualized by a generalized infection defect of a respiration-negative mutant. In S. agalactiae, HrtBA expression responds to an intracellular heme signal via activation of the two-component system HssRS. A hssRS promoter-driven intracellular luminescent heme sensor was designed to identify host compartments that supply S. agalactiae with heme. S. agalactiae acquires heme in heart, kidneys, and liver, but not in the brain. We conclude that S. agalactiae response to heme is organ-dependent, and its efflux may be particularly relevant in late stages of infection.

  13. Detection and enumeration of Staphylococcus aureus and Streptococcus agalactiae in milk by Real-Time PCR

    OpenAIRE

    Aline Gerato Dibbern

    2015-01-01

    O presente trabalho foi organizado em dois estudos. O objetivo do primeiro estudo foi determinar o efeito da infecção intramamária (IIM) causada por S. aureus e S. agalactiae na contagem de células somáticas (CCS) e na composição do leite (gordura, proteína, lactose, sólidos totais e extrato seco desengordurado), o efeito da contagem de S. aureus e S. agalactiae sobre a composição do leite e a estimativa da frequência de vacas e de quartos infectados com S. aureus e S. agalactiae por meio da ...

  14. Poliartritis y tenosinovitis grave por Streptococcus agalactiae en un paciente con hipoesplenia funcional Severe polyarthritis and tenosynovitis caused by Streptococcus agalactiae in a patient with functional hyposplenia

    Directory of Open Access Journals (Sweden)

    Domingo C. Balderramo

    2002-08-01

    Full Text Available La artritis por Streptococcus agalactiae es infrecuente. No conocemos publicaciones de casos sobre la afección tendinosa por este microorganismo. Se presenta una mujer de 46 años que consultó por fiebre, poliartralgias, mialgias, diarrea y vómitos. Como antecedentes presentaba carcinoma papilar de tiroides e hipoesplenia funcional. Al examen se encontraba hemodinámicamente inestable, febril, con artritis de mano izquierda, muñecas, codos, hombro derecho y tobillo izquierdo. Presentaba tenosinovitis en ambos pies y en la mano izquierda. Los hemocultivos y el cultivo de la bursa olecraniana derecha fueron positivos para S. agalactiae. La ecografía mostró signos de tenosinovitis del tibial anterior izquierdo. Completó 20 días de tratamiento endovenoso con cefazolina y 12 días de cefuroxima oral. El cuadro articular revirtió completamente en 60 días. El Streptococcus agalactiae puede causar, en forma infrecuente, un síndrome de poliartritis, tenosinovitis y fiebre similar al producido por la infección gonocócica.Cases of arthritis caused by Streptococcus agalactiae are infrequent and in our knowledge there are no case reports of tenosynovitis caused by S. agalactiae. A 46-year-old woman presented with fever, polyarthralgia, myalgia, diarrhea and vomiting. She had a history of papillary thyroid carcinoma and functional hyposplenia. She was febrile, with arthritis in hands, wrists, elbows, right shoulder and left ankle joints, and presented tenosynovitis in both feet and left hand. Blood and right olecranon bursa sample cultures were positive for S. agalactiae. An ultrasound scan made at the musculus tibialis anterior of left foot revealed signs of tenosynovitis. She was treated with intravenous cefazolin for 20 days and oral cefuroxime for 12 days. The joint involvement completely subsided in 60 days. Streptococcus agalactiae can cause, infrequently, a polyarthritis and tenosynovitis syndrome similar to disseminated gonococcal

  15. Identification of Streptococcus agalactiae by fluorescent in situ hybridization compared to culturing and the determination of prevalence of Streptococcus agalactiae colonization among pregnant women in Bushehr, Iran.

    Science.gov (United States)

    Tajbakhsh, Saeed; Norouzi Esfahani, Marjan; Emaneini, Mohammad; Motamed, Niloofar; Rahmani, Elham; Gharibi, Somayyeh

    2013-09-08

    Pregnant women colonized by Streptococcus agalactiae (group B streptococci [GBS]) may transfer this microorganism to their newborns. S. agalactiae is an important cause of pneumonia, sepsis, and meningitis in newborns. Fluorescent in situ hybridization (FISH) is considered as a method of identification in the field of diagnostic microbiology. In this paper, we have designed a study to compare the DNA FISH after 7 h Lim broth enrichment and culturing for the identification of S. agalactiae and to determine the prevalence of vaginal colonization by S. agalactiae among pregnant women in Bushehr, Iran. Vaginal swab specimens were obtained from 285 pregnant women at 35 weeks or more than 35 weeks of gestation. The specimens were inoculated into Lim broth. In order to evaluate the sensitivity and specificity of GBS DNA FISH after 7 h Lim broth enrichment, the specimens were tested using both FISH and conventional culture methods. In addition, the prevalence of GBS colonization was determined. Based on the results of this study, both the sensitivity and specificity of FISH were 100%. S. agalactiae was detected by both culture and FISH in 27 of the 285 pregnant women. Thus, the prevalence of GBS colonization was 9.5%. Since short-term (7 h) Lim broth enrichment followed by FISH using oligonucleotide probes showed a high sensitivity and specificity, this protocol is therefore a highly accurate and relatively rapid method for the detection of S. agalactiae. Our analysis suggests that the use of DNA FISH to screen for S. agalactiae colonization in pregnant women may be considered in the absence of GBS culture availability.

  16. Streptococcus agalactiae impairs cerebral bioenergetics in experimentally infected silver catfish.

    Science.gov (United States)

    Baldissera, Matheus D; Souza, Carine F; Parmeggiani, Belisa S; Santos, Roberto C V; Leipnitz, Guilhian; Moreira, Karen L S; da Rocha, Maria Izabel U M; da Veiga, Marcelo L; Baldisserotto, Bernardo

    2017-10-01

    It is becoming evident that bacterial infectious diseases affect brain energy metabolism, where alterations of enzymatic complexes of the mitochondrial respiratory chain and creatine kinase (CK) lead to an impairment of cerebral bioenergetics which contribute to disease pathogenesis in the central nervous system (CNS). Based on this evidence, the aim of this study was to evaluate whether alterations in the activity of complex IV of the respiratory chain and CK contribute to impairment of cerebral bioenergetics during Streptococcus agalactiae infection in silver catfish (Rhamdia quelen). The activity of complex IV of the respiratory chain in brain increased, while the CK activity decreased in infected animals compared to uninfected animals. Brain histopathology revealed inflammatory demyelination, gliosis of the brain and intercellular edema in infected animals. Based on this evidence, S. agalactiae infection causes an impairment in cerebral bioenergetics through the augmentation of complex IV activity, which may be considered an adaptive response to maintain proper functioning of the electron respiratory chain, as well as to ensure ongoing electron flow through the electron transport chain. Moreover, inhibition of cerebral CK activity contributes to lower availability of ATP, contributing to impairment of cerebral energy homeostasis. In summary, these alterations contribute to disease pathogenesis linked to the CNS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Capsular typing of Streptococcus agalactiae (Lancefield group B streptococci) from fish using multiplex PCR and serotyping

    Science.gov (United States)

    Streptococcus spp. including Streptococcus agalactiae (Lancefield group B streptococci) are considered emerging pathogens responsible for approximately $1 billion USD in annual losses to the global tilapia (Oreochromis sp.) aquaculture industry. This study evaluated a published multiplex PCR capsul...

  18. Comparison of transmission dynamics between Streptococcus uberis and Streptococcus agalactiae intramammary infections.

    Science.gov (United States)

    Leelahapongsathon, Kansuda; Schukken, Ynte Hein; Pinyopummintr, Tanu; Suriyasathaporn, Witaya

    2016-02-01

    The objectives of study were to determine the transmission parameters (β), durations of infection, and basic reproductive numbers (R0) of both Streptococcus agalactiae and Streptococcus uberis as pathogens causing mastitis outbreaks in dairy herds. A 10-mo longitudinal study was performed using 2 smallholder dairy herds with mastitis outbreaks caused by Strep. agalactiae and Strep. uberis, respectively. Both herds had poor mastitis control management and did not change their milking management during the entire study period. Quarter milk samples were collected at monthly intervals from all lactating animals in each herd for bacteriological identification. The durations of infection for Strep. uberis intramammary infection (IMI) and Strep. agalactiae IMI were examined using Kaplan-Meier survival curves, and the Kaplan-Meier survival functions for Strep. uberis IMI and Strep. agalactiae IMI were compared using log rank survival-test. The spread of Strep. uberis and Strep. agalactiae through the population was determined by transmission parameter, β, the probability per unit of time that one infectious quarter will infect another quarter, assuming that all other quarters are susceptible. For the Strep. uberis outbreak herd (31 cows), 56 new infections and 28 quarters with spontaneous cure were observed. For the Strep. agalactiae outbreak herd (19 cows), 26 new infections and 9 quarters with spontaneous cure were observed. The duration of infection for Strep. agalactiae (mean=270.84 d) was significantly longer than the duration of infection for Strep. uberis (mean=187.88 d). The transmission parameters (β) estimated (including 95% confidence interval) for Strep. uberis IMI and Strep. agalactiae IMI were 0.0155 (0.0035-0.0693) and 0.0068 (0.0008-0.0606), respectively. The R0 (including 95% confidence interval) during the study were 2.91 (0.63-13.47) and 1.86 (0.21-16.61) for Strep. uberis IMI and Strep. agalactiae IMI, respectively. In conclusion, the transmission

  19. Transcriptomic response of goat mammary epithelial cells to Mycoplasma agalactiae challenge – a preliminary study

    DEFF Research Database (Denmark)

    Ogorevc, Jernej; Mihevc, Sonja Prpar; Hedegaard, Jakob

    2015-01-01

    Mycoplasma agalactiae (Ma) is one of the main aetiological agents of intramammary infections in small ruminants, causing contagious agalactia. To better understand the underlying disease patterns a primary goat mammary epithelial cell (pgMEC) culture was established from the mammary tissue and ch....... Additionally, the results represent comprehensive goat mammary transcriptome information and demonstrate the applicability of the comparative genomics approach for annotation of goat data, using transcriptome information of a closely related species (Bos taurus) as a reference....

  20. Transcriptomic response of goat mammary epithelial cells to Mycoplasma agalactiae challenge – a preliminary study

    DEFF Research Database (Denmark)

    Ogorevc, Jernej; Mihevc, Sonja Prpar; Hedegaard, Jakob

    2015-01-01

    Mycoplasma agalactiae (Ma) is one of the main aetiological agents of intramammary infections in small ruminants, causing contagious agalactia. To better understand the underlying disease patterns a primary goat mammary epithelial cell (pgMEC) culture was established from the mammary tissue......, steroid metabolism, fatty acid metabolism, apoptosis signalling, transcription regulation, and cell cycle regulation. Based on the results we suggest that mammary epithelial cells in vivo contribute to the immune system by the induced expression of cytokines and other chemotactic agents, activation...

  1. Streptococcus agalactiae Inhibits Candida albicans Hyphal Development and Diminishes Host Vaginal Mucosal TH17 Response

    OpenAIRE

    Xiao-Yu Yu; Fei Fu; Wen-Na Kong; Qian-Kun Xuan; Dong-Hua Wen; Xiao-Qing Chen; Yong-Ming He; Li-Hua He; Jian Guo; Ai-Ping Zhou; Yang-Hong Xi; Li-Jun Ni; Yu-Feng Yao; Wen-Juan Wu

    2018-01-01

    Streptococcus agalactiae and Candida albicans often co-colonize the female genital tract, and under certain conditions induce mucosal inflammation. The role of the interaction between the two organisms in candidal vaginitis is not known. In this study, we found that co-infection with S. agalactiae significantly attenuated the hyphal development of C. albicans, and that EFG1-Hwp1 signal pathway of C. albicans was involved in this process. In a mouse model of vulvovaginal candidiasis (VVC), the...

  2. Development of a colloidal gold immunochromatographic strip for rapid detection of Streptococcus agalactiae in tilapia.

    Science.gov (United States)

    Wen-de, Wu; Min, Li; Ming, Chen; Li-Ping, Li; Rui, Wang; Hai-Lan, Chen; Fu-Yan, Chen; Qiang, Mi; Wan-Wen, Liang; Han-Zhong, Chen

    2017-05-15

    A colloidal gold immunochromatographic strip was developed for rapid detection of Streptococcus agalactiae (S. agalactiae) infection in tilapia. The monoclonal antibodies (mAb) 4C12 and 3A9 were used to target S. agalactiae as colloidal gold-mAb conjugate and captured antibody, respectively. The colloidal gold immunochromatographic strip was assembled via routine procedures. Optimal pH and minimum antibody levels in the reaction system for gold colloidal-mAb 4C12 conjugation were pH 7.4 and 18μg/mL, respectively. Optimal concentrations of the captured antibody 3A9 and goat anti-mouse antibody were 0.6mg/mL and 2mg/mL, respectively. The sensitivity of the strip for detecting S. agalactiae was 1.5×10 5 colony forming units (CFU). No cross-reaction was observed with other commonly encountered bacteria, including Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum and Streptococcus iniae. The assay time for S. agalactiae was less than 15min. Tilapia samples artificially infected with S. agalactiae were tested using the newly developed strip. The results indicated that blood, brain, kidney, spleen, metanephros and intestine specimens of infected fish can be used for S. agalactiae detection. The validity of the strip was maintained for 6 months at 4°C. These findings suggested that the immunochromatographic strip was effective for spot and rapid detection of S. agalactiae infected tilapia. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Macrolide resistance gene erm(TR) and erm(TR)-carrying genetic elements in Streptococcus agalactiae: characterization of ICESagTR7, a new composite element containing IMESp2907.

    Science.gov (United States)

    Mingoia, Marina; Morici, Eleonora; Marini, Emanuela; Brenciani, Andrea; Giovanetti, Eleonora; Varaldo, Pietro E

    2016-03-01

    The objective of this study was to investigate macrolide-resistant Streptococcus agalactiae isolates harbouring erm(TR), an erm(A) gene subclass, with emphasis on their erm(TR)-carrying genetic elements. Four erm(TR)-carrying elements have been described to date: three closely related (ICE10750-RD.2, Tn1806 and ICESp1108) in Streptococcus pyogenes, Streptococcus pneumoniae and S. pyogenes, respectively; and one completely different (IMESp2907, embedded in ICESp2906 to form ICESp2905) in S. pyogenes. Seventeen macrolide-resistant erm(TR)-positive S. agalactiae isolates were phenotypically and genotypically characterized. Their erm(TR)-carrying elements were explored by analysing the distinctive recombination genes of known erm(TR)-carrying integrative and conjugative elements (ICEs) and by PCR mapping. The new genetic context and organization of IMESp2907 in S. agalactiae were explored using several experimental procedures and in silico analyses. Five isolates harboured ICE10750-RD.2/Tn1806, five isolates harboured ICESp1108 and five isolates bore unknown erm(TR)-carrying elements. The remaining two isolates, exhibiting identical serotypes and pulsotypes, harboured IMESp2907 in a new genetic environment, which was further investigated in one of the two isolates, SagTR7. IMESp2907 was circularizable in S. agalactiae, as described in S. pyogenes. The new IMESp2907 junctions were identified based on its site-specific integration; the att sites were almost identical to those in S. pyogenes. In strain SagTR7, erm(TR)-carrying IMESp2907 was embedded in an erm(TR)-less internal element related to ICE10750-RD.2/Tn1806, which, in turn, was embedded in an ICESde3396-like element. The resulting whole ICE, ICESagTR7 (∼129 kb), was integrated into the chromosome downstream of the rplL gene, and was excisable in circular form and transferable by conjugation. This is the first study exploring erm(TR)-carrying genetic elements in S. agalactiae. © The Author 2015. Published by

  4. Capsule impairs efficient adherence of Streptococcus agalactiae to intestinal epithelium in tilapias Oreochromis sp.

    Science.gov (United States)

    Barato, P; Martins, E R; Vasquez, G M; Ramirez, M; Melo-Cristino, J; Martínez, N; Iregui, C

    2016-11-01

    Streptococcosis caused by Streptococcus agalactiae is one of the most important diseases in the tilapia aquaculture industry. The role of the capsule of Streptococcus agalactiae in adherence to fish surfaces has not been evaluated and the mechanism of capsular regulation during adhesion has not been described. The aim of this study was to evaluate the role of the capsule of S. agalactiae during adhesion to intestinal epithelium of tilapia (Oreochromis sp.) in an ex vivo infection model. We show that the capsule impairs the adhesion of bacteria to host intestinal epithelium. Wild type (WT) strain SaTiBe08-18 (S. agalactiae recovered from tilapia) had reduced adhesion (P S. agalactiae to tilapia intestine and that the acidic milieu could regulate adherence of encapsulated strains. We found GlcNAc on the surface of adherent Δcps but not over the capsule in WT. This difference could be explained by the GlcNAc composition of Lancefield group B antigen and the peptidoglycan in GBS (Group B Streptococcus) and also may be related with better exposure of glycosylated adhesins in unencapsulated fish GBS. Understanding capsular regulation during adhesion of S. agalactiae may provide new leads to find a successful anti-adherence therapy to prevent streptococcosis in tilapia. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Phosphoglycerate kinase enhanced immunity of the whole cell of Streptococcus agalactiae in tilapia, Oreochromis niloticus.

    Science.gov (United States)

    Wang, Yi-Ting; Huang, Hsing-Yen; Tsai, Ming-An; Wang, Pei-Chi; Jiang, Bo-Huang; Chen, Shih-Chu

    2014-12-01

    Streptococcus agalactiae is a Gram-positive bacterium and a severe aquaculture pathogen that can infect a wide range of warmwater fish species. The outer-surface proteins in bacterial pathogens play an important role in pathogenesis. We evaluated the immunogenicity of two of the identified surface proteins namely phosphoglycerate kinase (PGK) and ornithine carbamoyl-transferase (OCT). PGK and OCT were over-expressed and purified from Escherichia coli and used as the subunit vaccines in tilapia. Tilapia immunized with the S. agalactiae modified bacteria vaccine (whole cell preparations with recombinant PGK and OCT proteins) individually were tested for the efficacy. OCT and PGK combined with WC had a higher survival rate. A high-level protection and significant specific antibody responses against S. agalactiae challenge was observed upon the vaccinated tilapia with the purified PGK protein and S. agalactiae whole cells. The specific antibody titer against S. agalactiae antigen suggested that increased antibody titers were correlated with post-challenge survival rate. Il-1β expression profile was higher in PGK + WC-treated group. Tnf-α expression in the PGK + WC group was significantly increased. Taken together, our results suggested the combinations of recombinant protein and whole cell may elicit immune responses that reach greater protection than that of individual S. agalactiae components. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. GC-MS-Based Metabolome and Metabolite Regulation in Serum-Resistant Streptococcus agalactiae.

    Science.gov (United States)

    Wang, Zhe; Li, Min-Yi; Peng, Bo; Cheng, Zhi-Xue; Li, Hui; Peng, Xuan-Xian

    2016-07-01

    Streptococcus agalactiae causes severe systemic infections in human and fish. In the present study, we established a pathogen-plasma interaction model by which we explored how S. agalactiae evaded serum-mediated killing. We found that S. agalactiae grew faster in the presence of yellow grouper plasma than in the absence of the plasma, indicating S. agalactiae evolved a way of evading the fish immune system. To determine the events underlying this phenotype, we applied GC-MS-based metabolomics approaches to identify differential metabolomes between S. agalactiae cultured with and without yellow grouper plasma. Through bioinformatics analysis, decreased malic acid and increased adenosine were identified as the most crucial metabolites that distinguish the two groups. Meanwhile, they presented with decreased TCA cycle and elevated purine metabolism, respectively. Finally, exogenous malic acid and adenosine were used to reprogram the plasma-resistant metabolome, leading to elevated and decreased susceptibility to the plasma, respectively. Therefore, our findings reveal for the first time that S. agalactiae utilizes a metabolic trick to respond to plasma killing as a result of serum resistance, which may be reverted or enhanced by exogenous malic acid and adenosine, respectively, suggesting that the metabolic trick can be regulated by metabolites.

  7. Novel substrate specificity of glutathione synthesis enzymes from Streptococcus agalactiae and Clostridium acetobutylicum

    International Nuclear Information System (INIS)

    Kino, Kuniki; Kuratsu, Shoko; Noguchi, Atsushi; Kokubo, Masahiro; Nakazawa, Yuji; Arai, Toshinobu; Yagasaki, Makoto; Kirimura, Kohtaro

    2007-01-01

    Glutathione (GSH) is synthesized by γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GS) in living organisms. Recently, bifunctional fusion protein, termed γ-GCS-GS catalyzing both γ-GCS and GS reactions from gram-positive firmicutes Streptococcus agalactiae, has been reported. We revealed that in the γ-GCS activity, S. agalactiae γ-GCS-GS had different substrate specificities from those of Escherichia coli γ-GCS. Furthermore, S. agalactiae γ-GCS-GS synthesized several kinds of γ-glutamyltripeptide, γ-Glu-X aa -Gly, from free three amino acids. In Clostridium acetobutylicum, the genes encoding γ-GCS and putative GS were found to be immediately adjacent by BLAST search, and had amino acid sequence homology with S. agalactiae γ-GCS-GS, respectively. We confirmed that the proteins expressed from each gene showed γ-GCS and GS activity, respectively. C. acetobutylicum GS had broad substrate specificities and synthesized several kinds of γ-glutamyltripeptide, γ-Glu-Cys-X aa . Whereas the substrate specificities of γ-GCS domain protein and GS domain protein of S. agalactiae γ-GCS-GS were the same as those of S. agalactiae γ-GCS-GS

  8. Regulation of PI-2b Pilus Expression in Hypervirulent Streptococcus agalactiae ST-17 BM110.

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    Bruno Périchon

    Full Text Available The widely spread Streptococcus agalactiae (also known as Group B Streptococcus, GBS "hypervirulent" ST17 clone is strongly associated with neonatal meningitis. The PI-2b locus is mainly found in ST17 strains but is also present in a few non ST17 human isolates such as the ST-7 prototype strain A909. Here, we analysed the expression of the PI-2b pilus in the ST17 strain BM110 as compared to the non ST17 A909. Comparative genome analyses revealed the presence of a 43-base pair (bp hairpin-like structure in the upstream region of PI-2b operon in all 26 ST17 genomes, which was absent in the 8 non-ST17 strains carrying the PI-2b locus. Deletion of this 43-bp sequence in strain BM110 resulted in a 3- to 5-fold increased transcription of PI-2b. Characterization of PI-2b promoter region in A909 and BM110 strains was carried out by RNAseq, primer extension, qRT-PCR and transcriptional fusions with gfp as reporter gene. Our results indicate the presence of a single promoter (Ppi2b with a transcriptional start site (TSS mapped 37 bases upstream of the start codon of the first PI-2b gene. The large operon of 16 genes located upstream of PI-2b codes for the group B carbohydrate (also known as antigen B, a major constituent of the bacterial cell wall. We showed that the hairpin sequence located between antigen B and PI-2b operons is a transcriptional terminator. In A909, increased expression of PI-2b probably results from read-through transcription from antigen B operon. In addition, we showed that an extended 5' promoter region is required for maximal transcription of gfp as a reporter gene in S. agalactiae from Ppi2b promoter. Gene reporter assays performed in Lactococcus lactis strain NZ9000, a related non-pathogenic Gram-positive species, revealed that GBS-specific regulatory factors are required to drive PI-2b transcription. PI-2b expression is up-regulated in the BM110ΔcovR mutant as compared to the parental BM110 strain, but this effect is probably

  9. Effect of Eugenol against Streptococcus agalactiae and Synergistic Interaction with Biologically Produced Silver Nanoparticles

    Directory of Open Access Journals (Sweden)

    Renata Perugini Biasi-Garbin

    2015-01-01

    Full Text Available Streptococcus agalactiae (group B streptococci (GBS is an important infections agent in newborns associated with maternal vaginal colonization. Intrapartum antibiotic prophylaxis in GBS-colonized pregnant women has led to a significant reduction in the incidence of early neonatal infection in various geographic regions. However, this strategy may lead to resistance selecting among GBS, indicating the need for new alternatives to prevent bacterial transmission and even to treat GBS infections. This study reported for the first time the effect of eugenol on GBS isolated from colonized women, alone and in combination with silver nanoparticles produced by Fusarium oxysporum (AgNPbio. Eugenol showed a bactericidal effect against planktonic cells of all GBS strains, and this effect appeared to be time-dependent as judged by the time-kill curves and viability analysis. Combination of eugenol with AgNPbio resulted in a strong synergistic activity, significantly reducing the minimum inhibitory concentration values of both compounds. Scanning and transmission electron microscopy revealed fragmented cells and changes in bacterial morphology after incubation with eugenol. In addition, eugenol inhibited the viability of sessile cells during biofilm formation and in mature biofilms. These results indicate the potential of eugenol as an alternative for controlling GBS infections.

  10. Streptococcus agalactiae em gestantes: prevalência de colonização e avaliação da suscetibilidade aos antimicrobianos Streptococcus agalactiae in pregnant women: prevalence of colonization and antimicrobial susceptibility evaluation

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    Irina Lermontov Borger

    2005-10-01

    Full Text Available OBJETIVOS: verificar a ocorrência de colonização por Streptococcus agalactiae em gestantes e avaliar a suscetibilidade das amostras isoladas aos antimicrobianos. MÉTODOS: foram avaliadas 167 grávidas entre a 32ª e a 41ª semana de gestação, independente da presença ou não de fatores de risco, atendidas no ambulatório de pré-natal entre fevereiro de 2003 e fevereiro de 2004. O material vaginal/anal, colhido com um único swab, foi inoculado em caldo Todd-Hewitt acrescido de ácido nalidíxico (15 µg/mL e gentamicina (8 µg/mL, com posterior subcultura no meio de ágar sangue. A identificação foi feita por meio da avaliação da morfologia e tipo de hemólise das colônias no meio de ágar sangue, teste da catalase, teste de cAMP e testes sorológicos. A avaliação da suscetibilidade aos antimicrobianos foi realizada pelos testes de difusão e de diluição em ágar. A análise estatística foi realizada por meio do teste de chi2; valores de p0,05. Todas as 32 amostras isoladas foram sensíveis a penicilina, cefotaxima, ofloxacina, cloranfenicol, vancomicina e meropenem. A resistência a eritromicina e clindamicina foi detectada em 9,4 e 6,2% das amostras, respectivamente. CONCLUSÕES: a incidência relativamente elevada (19,2% de colonização por S. agalactiae entre as gestantes avaliadas e o isolamento de amostras resistentes, especialmente aos antimicrobianos recomendados nos casos de alergia à penicilina, enfatizam a importância de detectar esta colonização no final da gravidez, associada à avaliação da suscetibilidade aos antimicrobianos, para uma prevenção eficaz da infecção neonatal.PURPOSE: to verify the occurrence of colonization by Streptococcus agalactiae in pregnant women attended at the prenatal outpatient clinic of the Teaching Maternity Hospital of Rio de Janeiro University (UFRJ and to evaluate the susceptibility of the isolates to antimicrobial agents. METHODS: a total of 167 pregnant women between the

  11. Infective Endocarditis Complicated by Intraventricular Abscesses, Pericarditis, and Mycotic Aneurysm Due to an Emerging Strain of Serotype VI Streptococcus agalactiae.

    Science.gov (United States)

    Hirai, Nobuyasu; Kasahara, Kei; Uno, Kenji; Ogawa, Yoshihiko; Ogawa, Taku; Yonekawa, Shinsuke; Nakano, Ryuichi; Yano, Hisakazu; Sakagami, Azusa; Uemura, Takayuki; Okura, Hiroyuki; Saito, Yoshihiko; Yoshikawa, Masahide; Mikasa, Keiichi

    2017-11-22

    An increasing number of invasive infections due to Streptococcus agalactiae in non-pregnant adults have been reported. We report a case of infective endocarditis complicated by intraventricular abscesses, pericarditis, and mycotic aneurysm due to S. agalactiae belonging to ST681 with a capsular serotype VI in a woman with diabetes. The patient also had a myocardial infarction and was treated with percutaneous coronary intervention, pericardiocentesis, and 6 weeks of antibiotic treatment. Invasive infections due to serotype VI S. agalactiae are common in Asian countries such as Taiwan and Japan, so continuous monitoring of invasive S. agalactiae strains is warranted.

  12. A new mosaic integrative and conjugative element from Streptococcus agalactiae carrying resistance genes for chloramphenicol (catQ) and macrolides [mef(I) and erm(TR)].

    Science.gov (United States)

    Morici, Eleonora; Simoni, Serena; Brenciani, Andrea; Giovanetti, Eleonora; Varaldo, Pietro E; Mingoia, Marina

    2017-01-01

    To investigate the genetic basis of catQ-mediated chloramphenicol resistance in Streptococcus agalactiae. Two clinical strains of catQ-positive chloramphenicol-resistant S. agalactiae (Sag236 and Sag403) were recently isolated, typed (MLST, PFGE pulsotypes, capsular types) and their antibiotic resistances investigated by phenotypic and genotypic approaches. Several molecular methods (PCR mapping, restriction assays, Southern blotting, sequencing and sequence analysis, conjugal transfer assays) were used to determine the genetic context of catQ and characterize a genetic element detected in the isolates. Sag236 and Sag403 shared the same ST (ST19), but exhibited a different capsular type (III and V, respectively) and pulsotype. Both harboured the macrolide resistance genes mef(I) and erm(TR) and the tetracycline resistance gene tet(M). Accordingly, they were resistant to chloramphenicol, erythromycin and tetracycline. catQ and mef(I) were associated in an IQ module that was indistinguishable in Sag236 and Sag403. In mating assays, chloramphenicol and erythromycin resistance proved transferable, at low frequency, only from Sag236. Transconjugants carried not only catQ and mef(I), but also erm(TR), suggesting a linkage of the three resistance genes in a mobile element, which, though seemingly non-mobile, was also detected in Sag403. The new element (designated ICESag236, ∼110 kb) results from recombination of two integrative and conjugative elements (ICEs) originally described in different streptococcal species: S. agalactiae ICESagTR7, carrying erm(TR); and Streptococcus pneumoniae ICESpn529IQ, carrying the prototype IQ module. These findings strengthen the notion that widespread streptococcal ICEs may form mosaics that enhance their diversity and spread, broaden their host range and carry new cargo genes. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions

  13. The β-hemolysin and intracellular survival of Streptococcus agalactiae in human macrophages.

    Science.gov (United States)

    Sagar, Anubha; Klemm, Carolin; Hartjes, Lara; Mauerer, Stefanie; van Zandbergen, Ger; Spellerberg, Barbara

    2013-01-01

    S. agalactiae (group B streptococci, GBS) is a major microbial pathogen in human neonates and causes invasive infections in pregnant women and immunocompromised individuals. The S. agalactiae β-hemolysin is regarded as an important virulence factor for the development of invasive disease. To examine the role of β-hemolysin in the interaction with professional phagocytes, the THP-1 monocytic cell line and human granulocytes were infected with a serotype Ia S. agalactiae wild type strain and its isogenic nonhemolytic mutant. We could show that the nonhemolytic mutants were able to survive in significantly higher numbers than the hemolytic wild type strain, in THP-1 macrophage-like cells and in assays with human granulocytes. Intracellular bacterial multiplication, however, could not be observed. The hemolytic wild type strain stimulated a significantly higher release of Tumor Necrosis Factor-α than the nonhemolytic mutant in THP-1 cells, while similar levels of the chemokine Interleukin-8 were induced. In order to investigate bacterial mediators of IL-8 release in this setting, purified cell wall preparations from both strains were tested and found to exert a potent proinflammatory stimulus on THP-1 cells. In conclusion, our results indicate that the β-hemolysin has a strong influence on the intracellular survival of S. agalactiae and that a tightly controlled regulation of β-hemolysin expression is required for the successful establishment of S. agalactiae in different host niches.

  14. The highly dynamic CRISPR1 system of Streptococcus agalactiae controls the diversity of its mobilome.

    Science.gov (United States)

    Lopez-Sanchez, Maria-José; Sauvage, Elisabeth; Da Cunha, Violette; Clermont, Dominique; Ratsima Hariniaina, Elisoa; Gonzalez-Zorn, Bruno; Poyart, Claire; Rosinski-Chupin, Isabelle; Glaser, Philippe

    2012-09-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) confer immunity against mobile genetic elements (MGEs) in prokaryotes. Streptococcus agalactiae, a leading cause of neonatal infections contains in its genome two CRISPR/Cas systems. We show that type 1-C CRISPR2 is present in few strains but type 2-A CRISPR1 is ubiquitous. Comparative sequence analysis of the CRISPR1 spacer content of 351 S. agalactiae strains revealed that it is extremely diverse due to the acquisition of new spacers, spacer duplications and spacer deletions that witness the dynamics of this system. The spacer content profile mirrors the S. agalactiae population structure. Transfer of a conjugative transposon targeted by CRISPR1 selected for spacer rearrangements, suggesting that deletions and duplications pre-exist in the population. The comparison of protospacers located within MGE or the core genome and protospacer-associated motif-shuffling demonstrated that the GG motif is sufficient to discriminate self and non-self and for spacer selection and integration. Strikingly more than 40% of the 949 different CRISPR1 spacers identified target MGEs found in S. agalactiae genomes. We thus propose that the S. agalactiae type II-A CRISPR1/Cas system modulates the cohabitation of the species with its mobilome, as such contributing to the diversity of MGEs in the population. © 2012 Blackwell Publishing Ltd.

  15. Streptococcus agalactiae Inhibits Candida albicans Hyphal Development and Diminishes Host Vaginal Mucosal TH17 Response

    Directory of Open Access Journals (Sweden)

    Xiao-Yu Yu

    2018-02-01

    Full Text Available Streptococcus agalactiae and Candida albicans often co-colonize the female genital tract, and under certain conditions induce mucosal inflammation. The role of the interaction between the two organisms in candidal vaginitis is not known. In this study, we found that co-infection with S. agalactiae significantly attenuated the hyphal development of C. albicans, and that EFG1-Hwp1 signal pathway of C. albicans was involved in this process. In a mouse model of vulvovaginal candidiasis (VVC, the fungal burden and the levels of pro-inflammatory cytokines, IL-1β, IL-6 and TNF-α showed a increase on co-infection with S. agalactiae, while the level of TH17 T cells and IL-17 in the cervicovaginal lavage fluid were significantly decreased. Our results indicate that S. agalactiae inhibits C. albicans hyphal development by downregulating the expression of EFG1-Hwp1. The interaction between S. agalactiae and C. albicans may attenuate host vaginal mucosal TH17 immunity and contribute to mucosal colonization by C. albicans.

  16. Streptococcus agalactiae Inhibits Candida albicans Hyphal Development and Diminishes Host Vaginal Mucosal TH17 Response.

    Science.gov (United States)

    Yu, Xiao-Yu; Fu, Fei; Kong, Wen-Na; Xuan, Qian-Kun; Wen, Dong-Hua; Chen, Xiao-Qing; He, Yong-Ming; He, Li-Hua; Guo, Jian; Zhou, Ai-Ping; Xi, Yang-Hong; Ni, Li-Jun; Yao, Yu-Feng; Wu, Wen-Juan

    2018-01-01

    Streptococcus agalactiae and Candida albicans often co-colonize the female genital tract, and under certain conditions induce mucosal inflammation. The role of the interaction between the two organisms in candidal vaginitis is not known. In this study, we found that co-infection with S. agalactiae significantly attenuated the hyphal development of C. albicans , and that EFG1 -Hwp1 signal pathway of C. albicans was involved in this process. In a mouse model of vulvovaginal candidiasis (VVC), the fungal burden and the levels of pro-inflammatory cytokines, IL-1β, IL-6 and TNF-α showed a increase on co-infection with S. agalactiae , while the level of TH17 T cells and IL-17 in the cervicovaginal lavage fluid were significantly decreased. Our results indicate that S. agalactiae inhibits C. albicans hyphal development by downregulating the expression of EFG1 -Hwp1. The interaction between S. agalactiae and C. albicans may attenuate host vaginal mucosal TH17 immunity and contribute to mucosal colonization by C. albicans .

  17. TOKSISITAS PROTEIN 89 kDa PRODUK EKSTRASELULER Streptococcus agalactiae PADA IKAN NILA (Oreochromis niloticus

    Directory of Open Access Journals (Sweden)

    Amrullah Amrullah

    2015-09-01

    Full Text Available Identifikasi protein toksin yang bersifat antigenik dari produk ekstraselular (ECP crude Streptococcus agalactiae penting dilakukan untuk pengembangan vaksin protein toksoid. Penelitian ini bertujuan untuk mengevaluasi toksisitas protein dengan berat molekul 89 kDa dari ECP Streptococcus agalactiae hasil fraksinasi dengan SDS-PAGE (ECPP89. Ikan nila dengan bobot sekitar 25 g masing-masing diinjeksi secara intraperitonial dengan protein ECPP89 dosis 1, 2, 4, 8, dan 16 μg/mL/ekor ikan (secara berturut-turut diberi kode ECPP89-1, ECPP89-2, ECPP89-4, ECPP89-8, dan ECPP89-16. Sebagai kontrol positif ikan diinjeksi dengan bakteri utuh S. agalactiae 1 x 104 cfu/mL (WCB dan ECP crude S. agalactiae (ECPC, sementara kontrol negatif ikan diinjeksi larutan PBS. Ikan dipelihara selama 15 hari dengan kepadatan 10 ekor (70 L air. Hasil penelitian menunjukkan bahwa ikan nila yang diinjeksi dengan ECPP89 mengalami gejala seperti perubahan morfologi, perubahan nafsu makan, dan perubahan renang ikan. Mortalitas pada perlakuan ECPP89 secara umum lebih tinggi dibandingkan dengan ECPC, namun lebih rendah (p<0,05 dibandingkan dengan WCB. Mortalitas ikan pada perlakuan ECPP89-4, ECPP89-8, dan ECPP89-16 tidak berbeda tetapi secara signifikan lebih tinggi dibandingkan dengan ECPC dan dua dosis perlakuan ECPP89 lainnya (p<0,05. Ikan kontrol negatif tidak mengalami kematian maupun perubahan klinis. Hasil ini menunjukkan bahwa ECPP89 merupakan protein toksin dari ECP S. agalactiae.

  18. The β-Hemolysin and Intracellular Survival of Streptococcus agalactiae in Human Macrophages

    Science.gov (United States)

    Sagar, Anubha; Klemm, Carolin; Hartjes, Lara; Mauerer, Stefanie; van Zandbergen, Ger; Spellerberg, Barbara

    2013-01-01

    S. agalactiae (group B streptococci, GBS) is a major microbial pathogen in human neonates and causes invasive infections in pregnant women and immunocompromised individuals. The S. agalactiae β-hemolysin is regarded as an important virulence factor for the development of invasive disease. To examine the role of β-hemolysin in the interaction with professional phagocytes, the THP-1 monocytic cell line and human granulocytes were infected with a serotype Ia S. agalactiae wild type strain and its isogenic nonhemolytic mutant. We could show that the nonhemolytic mutants were able to survive in significantly higher numbers than the hemolytic wild type strain, in THP-1 macrophage-like cells and in assays with human granulocytes. Intracellular bacterial multiplication, however, could not be observed. The hemolytic wild type strain stimulated a significantly higher release of Tumor Necrosis Factor-α than the nonhemolytic mutant in THP-1 cells, while similar levels of the chemokine Interleukin-8 were induced. In order to investigate bacterial mediators of IL-8 release in this setting, purified cell wall preparations from both strains were tested and found to exert a potent proinflammatory stimulus on THP-1 cells. In conclusion, our results indicate that the β-hemolysin has a strong influence on the intracellular survival of S. agalactiae and that a tightly controlled regulation of β-hemolysin expression is required for the successful establishment of S. agalactiae in different host niches. PMID:23593170

  19. Annual incidence, prevalence and transmission characteristics of Streptococcus agalactiae in Danish dairy herds

    DEFF Research Database (Denmark)

    Mweu, Marshal M.; Nielsen, Søren S.; Hisham Beshara Halasa, Tariq

    2012-01-01

    -level incidence rates and apparent prevalences of Streptococcus agalactiae (S. agalactiae) in the population of Danish dairy cattle herds over a 10-year period from 2000 to 2009 inclusive and (2) to estimate the herd-level entry and exit rates (demographic parameters), the transmission parameter, β, and recovery...... rate for S. agalactiae infection. Data covering the specified period, on bacteriological culture of all bulk tank milk samples collected annually as part of the mandatory Danish S. agalactiae surveillance scheme, were extracted from the Danish Cattle Database and subsequently analysed....... There was an increasing trend in both the incidence and prevalence of S. agalactiae over the study period. Per 100 herd-years the value of β was 54.1 (95% confidence interval [CI] 46.0–63.7); entry rate 0.3 (95% CI 0.2–0.4); infection-related exit rate 7.1 (95% CI 5.6–8.9); non-infection related exit rate 9.2 (95% CI 7...

  20. The β-hemolysin and intracellular survival of Streptococcus agalactiae in human macrophages.

    Directory of Open Access Journals (Sweden)

    Anubha Sagar

    Full Text Available S. agalactiae (group B streptococci, GBS is a major microbial pathogen in human neonates and causes invasive infections in pregnant women and immunocompromised individuals. The S. agalactiae β-hemolysin is regarded as an important virulence factor for the development of invasive disease. To examine the role of β-hemolysin in the interaction with professional phagocytes, the THP-1 monocytic cell line and human granulocytes were infected with a serotype Ia S. agalactiae wild type strain and its isogenic nonhemolytic mutant. We could show that the nonhemolytic mutants were able to survive in significantly higher numbers than the hemolytic wild type strain, in THP-1 macrophage-like cells and in assays with human granulocytes. Intracellular bacterial multiplication, however, could not be observed. The hemolytic wild type strain stimulated a significantly higher release of Tumor Necrosis Factor-α than the nonhemolytic mutant in THP-1 cells, while similar levels of the chemokine Interleukin-8 were induced. In order to investigate bacterial mediators of IL-8 release in this setting, purified cell wall preparations from both strains were tested and found to exert a potent proinflammatory stimulus on THP-1 cells. In conclusion, our results indicate that the β-hemolysin has a strong influence on the intracellular survival of S. agalactiae and that a tightly controlled regulation of β-hemolysin expression is required for the successful establishment of S. agalactiae in different host niches.

  1. Infecção congênita em cabritos por Mycoplasma agalactiae

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    N.S. Silva

    2014-04-01

    Full Text Available This study aimed to report three cases of contagious agalactia (CA by Mycoplasma agalactiae in goat kids born with polyarthritis. The nanny goats belonging to two different herds presented clinical signs of CA during pregnancy and in parturition they were apparently healthy. The carpal articulations of the three goat kids, the tarsus articulation in one, and thigh-femoral articulation in another showed swelling, pain and impairment of the flexion-extension movements. The articular liquid was collected from two goat kids at birth and revealed a content which varied from transparent to fibrinopurulent, presenting a yellow coloring. The samples were plated on modified Hayflick. The colonies had the appearance of "fried egg" and were confirmed as being M. agalactiae by biochemical tests and 16S rRNA PCR. Blood was collected from three animals soon after birth and submitted to the indirect ELISA test for the determination of the titration of the anti- M. agalactiae antibodies. The results confirmed that the goat kids were infected during pregnancy by M. agalactiae and resulted in the birth of an offspring with clinical signs of CA being immune tolerant.

  2. A case of infective endocarditis along with a ruptured valve caused by Streptococcus agalactiae in an immunocompetent man.

    Science.gov (United States)

    Suzuki, Kiyozumi; Hirai, Yuji; Morita, Fujiko; Uehara, Yuki; Oshima, Hiroko; Mitsuhashi, Kazunori; Amano, Atsushi; Naito, Toshio

    2016-01-01

    Streptococcus agalactiae ( S. agalactiae ) is a major cause of invasive disease in neonates and pregnant women, but has also recently been observed among non-pregnant adults, especially elderly persons or persons with underlying chronic disease. S. agalactiae is also a rare cause of infective endocarditis, and most cases require early surgery. We report the case of a 43-year-old previously healthy man who experienced rapid progressive culture-negative infective endocarditis with aortic valve vegetation and severe aortic regurgitation, which was complicated by lumbar spondylodiscitis. Emergency aortic valve replacement was performed on the day of his admission, which revealed a congenital bicuspid aortic valve was ruptured by the vegetation. The resected aortic valve specimen was submitted for 16S ribosomal RNA gene sequencing, which revealed that the pathogen was S. agalactiae . Therefore, S. agalactiae should be considered a potentially causative pathogen in cases of rapid progressive infective endocarditis, even if it occurs in a non-pregnant immunocompetent adult.

  3. Experimental early pathogenesis of Streptococcus agalactiae infection in red tilapia Oreochromis spp.

    Science.gov (United States)

    Iregui, C A; Comas, J; Vásquez, G M; Verján, N

    2016-02-01

    Streptococcus agalactiae causes a severe systemic disease in fish, and the routes of entry are still ill-defined. To address this issue, two groups of 33 red tilapia Oreochromis spp. each of 10 g were orally infected with S. agalactiae (n = 30), and by immersion (n = 30), six individuals were control-uninfected fish. Three tilapias were killed at each time point from 30 min to 96 h post-inoculation (pi); controls were killed at 96 h. Samples from most tissues were examined by haematoxylin-eosin (H&E), indirect immunoperoxidase (IPI) and periodic acid-Schiff; only intestine from fish infected by gavage was evaluated by transmission electron microscopy. The results of both experiments suggest that the main entry site of S. agalactiae in tilapia is the gastrointestinal epithelium; mucus seems to play an important defensive role, and environmental conditions may be an important predisposing factor for the infection. © 2015 John Wiley & Sons Ltd.

  4. Streptococcus agalactiae endocarditis presenting as acalculous cholecystitis in a previously well woman.

    LENUS (Irish Health Repository)

    Brewer, Linda

    2013-01-01

    This case report describes the unusual presentation of a previously very well woman with Streptococcus agalactiae endocarditis in the emergency department. History, examination and preliminary laboratory and radiological investigations supported a diagnosis of acalculous cholecystitis, for which she was given intravenous broad spectrum antimicrobial therapy. One day following admission, the patient deteriorated and became unresponsive. Subsequent MRI of the brain revealed multiple bihemispheric cerebral emboli and a large, mobile mitral valve thrombus was visualised on her transoesophageal echocardiogram. S agalactiae was cultured from venous blood samples and her antimicrobial cover was adjusted accordingly. Despite her presumed guarded prognosis, this patient made a remarkable recovery. To our knowledge, the association of S agalactiae endocarditis with acalculous cholecystitis has not been previously described.

  5. Epidemiology of Mycoplasma agalactiae and Mycoplasma mycoides cluster in flocks of northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Sandra Batista dos Santos

    2018-04-01

    Full Text Available ABSTRACT: The present study aimed to investigate contagious agalactia (CA in flocks from Pernambuco State. The study involved 225 goats and 63 ewes; 288 milk samples and 100 vaginal swabs were collected in total. The PCR assays were carried out using specific primers to Mycoplasma agalactiae and the Mycoplasma mycoides cluster. Among the goat’s milk samples,12.0% (27/225 were positive for Mycoplasma agalactiae DNA, while 5.3% (12/225 contained the Mycoplasma mycoides cluster. Of the vaginal swabs taken from goats, 15.4% (12/78 were positive for Mycoplasma agalactiae DNA and 3.8% (3/78 contained the Mycoplasma mycoides cluster. In the case of ewes, 4.3% (1/23 of the milk samples contained Mycoplasma agalactiae DNA, and 7.5% (3/40 were positive for the Mycoplasma mycoides cluster. Vaginal swabs taken from sheep´s were negative. Analysis of risk factors for mycoplasmosis, showed that goats and sheep flocks on the extensive breeding system are more likely to have mycoplasmosis than those on the intensive breeding system (odds ratio (OR 6.2; p=0.004; meat goat and sheep flocks are more likely to have infection compared to dairy flocks (OR 4.8; p=0.011; unclean animal housing increases the chances of infection (OR 5.0; p=0.031 and not performing quarantine increases the chances of mycoplasmosis (OR 4.6; p=0.042. Based on these findings we conclude that CA syndrome in the semiarid region of Pernambuco state can be associated with Mycoplasma agalactiae and Mycoplasma mycoides cluster.

  6. Development of live attenuated Streptococcus agalactiae vaccine for tilapia via continuous passage in vitro.

    Science.gov (United States)

    Li, L P; Wang, R; Liang, W W; Huang, T; Huang, Y; Luo, F G; Lei, A Y; Chen, M; Gan, X

    2015-08-01

    Fish Streptococcus agalactiae (S. agalactiae) seriously harms the world's aquaculture industry and causes huge economic losses. This study aimed to develop a potential live attenuated vaccine of S. agalactiae. Pre-screened vaccine candidate strain S. agalactiae HN016 was used as starting material to generate an attenuated strain S. agalactiae YM001 by continuous passage in vitro. The biological characteristics, virulence, and stability of YM001 were detected, and the protective efficacy of YM001 immunization in tilapia was also determined. Our results indicated that the growth, staining, characteristics of pulsed-field gel electrophoresis (PFGE) genotype, and virulence of YM001 were changed significantly as compared to the parental strain HN016. High doses of YM001 by intraperitoneal (IP) injection (1.0 × 10(9) CFU/fish) and oral gavage (1.0 × 10(10) CFU/fish) respectively did not cause any mortality and morbidity in tilapia. The relative percent survivals (RPSs) of fishes immunized with YM001 (1.0 × 10(8) CFU/fish, one time) via injection, immersion, and oral administration were 96.88, 67.22, and 71.81%, respectively, at 15 days, and 93.61, 60.56, and 53.16%, respectively, at 30 days. In all tests with 1-3 times of immunization in tilapia, the dosages at 1 × 10(8) and 1 × 10(9) CFU/fish displayed the similar best results, whereas the immunoprotection of the dosages at 1 × 10(6) and 1 × 10(7) CFU/fish declined significantly (P 0.05). The level of protective antibody elicited by oral immunization was significantly higher compared to that of the control group (P S. agalactiae strain YM001; oral immunization of tilapia with this strain produced a good immune protection. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. GapA, a potential vaccine candidate antigen against Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Zhang, Ze; Yu, Angen; Lan, Jiangfeng; Zhang, Hua; Hu, Minqiang; Cheng, Jiewei; Zhao, Lijuan; Lin, Li; Wei, Shun

    2017-04-01

    Streptococcosis due to the bacterium Streptococcus agalactiae (S. agalactiae) has resulted in enormous economic losses in aquaculture worldwide, especially in the tilapia culture industry. Previously, there were limited vaccines that could be employed against streptococcosis in tilapia. This study aimed to develop a vaccine candidate using the glyceraldehyde-phosphate dehydrogenase protein (GapA) of S. agalactiae encoded by the gapA gene. Tilapia were intraperitoneally injected with PBS, PBS + Freund's adjuvant, PBS + Montanide's adjuvant, GapA + Freund's adjuvant, GapA + Montanide's adjuvant, killed S. agalactiae whole cells (WC)+Freund's adjuvant, or killed S. agalactiae whole cells (WC)+ Montanide's adjuvant. They were then challenged with S. agalactiae, and the relative percentage survival (RPS) was monitored 14 days after the challenge. The highest RPSs were observed in the WC groups, with 76.7% in WC + Freund's adjuvant and 74.4% in WC + Montanide's adjuvant groups; these were followed by the GapA groups, with 63.3% in GapA + Freund's adjuvant and 45.6% in GapA + Montanide's adjuvant groups. The RPS of the PBS group was 0%, and those of PBS + Freund's adjuvant and PBS + Montanide's adjuvant groups were 6.7% and 3.3%, respectively. Additionally, the IgM antibody responses elicited in GapA groups and WC groups were significantly higher than those in PBS groups. Furthermore, the expressions of cytokine (IL-1β and TNF-α) mRNAs in the GapA groups and WC groups were significantly higher than those in the PBS groups. Taken together, these results reveal that the GapA protein is a promising vaccine candidate that could be used to prevent streptococcosis in tilapia. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Serine-Rich Repeat Proteins and Pili Promote Streptococcus agalactiae Colonization of the Vaginal Tract

    NARCIS (Netherlands)

    Sheen, Tamsin R.; Jimenez, Alyssa; Wang, Nai-Yu; Banerjee, Anirban; van Sorge, Nina M.; Doran, Kelly S.

    2011-01-01

    Streptococcus agalactiae (group B streptococcus [GBS]) is a Gram-positive bacterium found in the female rectovaginal tract and is capable of producing severe disease in susceptible hosts, including newborns and pregnant women. The vaginal tract is considered a major reservoir for GBS, and maternal

  9. Amperometric biosensor based on a single antibody of dual function for rapid detection of Streptococcus agalactiae.

    Science.gov (United States)

    Vásquez, Gersson; Rey, Alba; Rivera, Camilo; Iregui, Carlos; Orozco, Jahir

    2017-01-15

    Pathogenic bacteria are responsible for several diseases in humans and in a variety of hosts. Detection of pathogenic bacteria is imperative to avoid and/or fight their potential harmful effects. This work reports on the first amperometric biosensor for the rapid detection of Streptococcus agalactiae (S. agalactiae). The biosensor relies on a single biotinylated antibody that immobilizes the bacteria on a screen-printed carbon electrode while is further linked to a streptavidin-conjugated HRP reporter. The biotinylated antibody provides selectivity to the biosensor whereas serves as an anchoring point to the reporter for further amplification of the electrochemical signal. The resultant immunosensor is simple, responds rapidly, and allows for the selective and highly sensitive quantification of S. agalactiae cells in a concentration range of 10 1 -10 7 CFUml -1 , with a detection limit of 10CFUml -1 . The approach not only enables a rapid detection and quantification of S. agalactiae in environmental samples but also opens up new opportunities for the simple fabrication of electrochemical immunosensors for different target pathogens. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Physiological impact of transposable elements encoding DDE transposases in the environmental adaptation of Streptococcus agalactiae.

    Science.gov (United States)

    Fléchard, Maud; Gilot, Philippe

    2014-07-01

    We have referenced and described Streptococcus agalactiae transposable elements encoding DDE transposases. These elements belonged to nine families of insertion sequences (ISs) and to a family of conjugative transposons (TnGBSs). An overview of the physiological impact of the insertion of all these elements is provided. DDE-transposable elements affect S. agalactiae in a number of aspects of its capability to adapt to various environments and modulate the expression of several virulence genes, the scpB-lmB genomic region and the genes involved in capsule expression and haemolysin transport being the targets of several different mobile elements. The referenced mobile elements modify S. agalactiae behaviour by transferring new gene(s) to its genome, by modifying the expression of neighbouring genes at the integration site or by promoting genomic rearrangements. Transposition of some of these elements occurs in vivo, suggesting that by dynamically regulating some adaptation and/or virulence genes, they improve the ability of S. agalactiae to reach different niches within its host and ensure the 'success' of the infectious process. © 2014 The Authors.

  11. Complete genome sequence of an attenuated Sparfloxacin-resistant Streptococcus agalactiae strain 138spar

    Science.gov (United States)

    The complete genome of a sparfloxacin-resistant Streptococcus agalactiae vaccine strain 138spar is 1,838,126 bp in size. The genome has 1892 coding sequences and 82 RNAs. The annotation of the genome is added by the NCBI Prokaryotic Genome Annotation Pipeline. The publishing of this genome will allo...

  12. Spring forward with improved Nile tilapia Oreochromis niloticus resistant to Streptococcus iniae and Streptococcus agalactiae IB

    Science.gov (United States)

    Tilapia aquaculture worldwide is valued around US $ 7 billion. Tilapia are an important source of protein for domestic (top 5 most consumed seafoods) and global food security. Two gram postitive bacteria, Streptococcus iniae and S. agalactiae, are responsible for billion dollar losses annually. Gen...

  13. Single nucleotide resolution RNA-seq uncovers new regulatory mechanisms in the opportunistic pathogen Streptococcus agalactiae.

    Science.gov (United States)

    Rosinski-Chupin, Isabelle; Sauvage, Elisabeth; Sismeiro, Odile; Villain, Adrien; Da Cunha, Violette; Caliot, Marie-Elise; Dillies, Marie-Agnès; Trieu-Cuot, Patrick; Bouloc, Philippe; Lartigue, Marie-Frédérique; Glaser, Philippe

    2015-05-30

    Streptococcus agalactiae, or Group B Streptococcus, is a leading cause of neonatal infections and an increasing cause of infections in adults with underlying diseases. In an effort to reconstruct the transcriptional networks involved in S. agalactiae physiology and pathogenesis, we performed an extensive and robust characterization of its transcriptome through a combination of differential RNA-sequencing in eight different growth conditions or genetic backgrounds and strand-specific RNA-sequencing. Our study identified 1,210 transcription start sites (TSSs) and 655 transcript ends as well as 39 riboswitches and cis-regulatory regions, 39 cis-antisense non-coding RNAs and 47 small RNAs potentially acting in trans. Among these putative regulatory RNAs, ten were differentially expressed in response to an acid stress and two riboswitches sensed directly or indirectly the pH modification. Strikingly, 15% of the TSSs identified were associated with the incorporation of pseudo-templated nucleotides, showing that reiterative transcription is a pervasive process in S. agalactiae. In particular, 40% of the TSSs upstream genes involved in nucleotide metabolism show reiterative transcription potentially regulating gene expression, as exemplified for pyrG and thyA encoding the CTP synthase and the thymidylate synthase respectively. This comprehensive map of the transcriptome at the single nucleotide resolution led to the discovery of new regulatory mechanisms in S. agalactiae. It also provides the basis for in depth analyses of transcriptional networks in S. agalactiae and of the regulatory role of reiterative transcription following variations of intra-cellular nucleotide pools.

  14. Streptococcus agalactiae Serotype IV in Humans and Cattle, Northern Europe

    DEFF Research Database (Denmark)

    Lyhs, Ulrike; Kulkas, Laura; Katholm, Jorgen

    2016-01-01

    not differentiate between populations isolated from different host species. Isolates from humans and cattle differed in lactose fermentation, which is encoded on the accessory genome and represents an adaptation to the bovine mammary gland. Serotype IV-ST196 isolates were obtained from multiple dairy herds in both...

  15. Simultaneous Identification of Potential Pathogenicity Factors of Mycoplasma agalactiae in the Natural Ovine Host by Negative Selection

    OpenAIRE

    Hegde, Shivanand; Hegde, Shrilakshmi; Zimmermann, Martina; Flöck, Martina; Spergser, Joachim; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-01-01

    Mycoplasmas possess complex pathogenicity determinants that are largely unknown at the molecular level. Mycoplasma agalactiae serves as a useful model to study the molecular basis of mycoplasma pathogenicity. The generation and in vivo screening of a transposon mutant library of M. agalactiae were employed to unravel its host colonization factors. Tn4001mod mutants were sequenced using a novel sequencing method, and functionally heterogeneous pools containing 15 to 19 selected mutants were sc...

  16. Effect of the Streptococcus agalactiae Virulence Regulator CovR on the Pathogenesis of Urinary Tract Infection.

    Science.gov (United States)

    Sullivan, Matthew J; Leclercq, Sophie Y; Ipe, Deepak S; Carey, Alison J; Smith, Joshua P; Voller, Nathan; Cripps, Allan W; Ulett, Glen C

    2017-02-01

    Streptococcus agalactiae can cause urinary tract infection (UTI). The role of the S. agalactiae global virulence regulator, CovR, in UTI pathogenesis is unknown. We used murine and human bladder uroepithelial cell models of UTI and S. agalactiae mutants in covR and related factors, including β-hemolysin/cytolysin (β-h/c), surface-anchored adhesin HvgA, and capsule to study the role of CovR in UTI. We found that covR-deficient serotype III S. agalactiae 874391 was significantly attenuated for colonization in mice and adhesion to uroepithelial cells. Mice infected with covR-deficient S. agalactiae produced less proinflammatory cytokines than those infected with wild-type 874391. Acute cytotoxicity in uroepithelial cells triggered by covR-deficient but not wild-type 874391 was associated with significant caspase 3 activation. Mechanistically, covR mutation significantly altered the expression of several genes in S. agalactiae 874391 that encode key virulence factors, including β-h/c and HvgA, but not capsule. Subsequent mutational analyses revealed that HvgA and capsule, but not the β-h/c, exerted significant effects on colonization of the murine urinary tract in vivo. S. agalactiae CovR promotes bladder infection and inflammation, as well as adhesion to and viability of uroepithelial cells. The pathogenesis of S. agalactiae UTI is complex, multifactorial, and influenced by virulence effects of CovR, HvgA, and capsule. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

  17. The protective duration of Streptococcus agalactiae vaccine in Nile Tilapia for the prevention of streptococcosis

    Directory of Open Access Journals (Sweden)

    , Sukenda

    2016-02-01

    Full Text Available ABSTRACT The aim of this study was to assess the protective duration of Streptococcus agalactiae vaccine against streptococcosis in Nile tilapia. Fish were treated by the whole cell vaccine, ECP vaccine and mixture of whole cell and ECP vaccine. After 14, 28, 42, and 52 day post-vaccination (DPV, the fish were intraperitoneally challenged with 104  cfu/mL S. agalactiae. The results showed mortality rate of whole-cell vaccine (A, ECP vaccine (B and mix vaccine (C up to day 42 was significantly (P<0.05 lower than the control treatment, namely 73.33%; 80%; and 76%, respectively. The mortality rate of vaccine treatments A, B, and C on day 56 had no significant difference (P>0.05 with the control. The value of antibody titer vaccine treatments A, B, and C indicate that antigen-antibody reaction on day 28 after the vaccination was significantly (P <0.05 higher than the control that were 3.67; 3.33; and 3.67. Antigen-antibody reaction on day 42 after the vaccination was founded, but did not different significantly (P>0.05 with the control. Bacterial population in treatment A, B, and C in the organs of the fish until the 28th day was still under the control of 104  cfu/mL. S. agalactiae vaccine protection duration is 42 days after the vaccination. Keywords : nile tilapia, Streptococcus agalactiae, duration, vaccine, streptococcosis  ABSTRAK Penelitian ini bertujuan untuk menganalisis durasi proteksi dari vaksin Streptococcus agalactiae sebagai pencegahan terhadap streptococcosis pada ikan nila. Ikan divaksinasi dengan vaksin sel utuh, ECP dan gabungan sel utuh dan ECP dari S. agalactiae yang diinjeksi secara intrapetorineal. Ikan diuji tantang S. agalactiae 104  cfu/mL pada hari ke-14, ke-28, ke-42, dan ke-56 pascavaksinasi. Hasil penelitian menunjukkan tingkat mortalitas perlakuan vaksin sel utuh (A, vaksin ECP (B, dan gabungan vaksin sel utuh dengan ECP (C hingga hari ke-42 masih signifikan (P<0,05 lebih rendah dari perlakuan kontrol yaitu 73

  18. Protective efficacy of cationic-PLGA microspheres loaded with DNA vaccine encoding the sip gene of Streptococcus agalactiae in tilapia.

    Science.gov (United States)

    Ma, Yan-Ping; Ke, Hao; Liang, Zhi-Ling; Ma, Jiang-Yao; Hao, Le; Liu, Zhen-Xing

    2017-07-01

    Streptococcus agalactiae (S. agalactiae) is an important fish pathogen, which has received more attention in the past decade due to the increasing economic losses in the tilapia industry worldwide. As existing effective vaccines of S. agalactiae in fish have obvious disadvantage, to select immunoprotective antigens and package materials would undoubtedly contribute to the development of novel oral vaccines. In the present study, surface immunogenic protein (sip) was selected from the S. agalactiae serovar I a genomes as immunogenic protein in DNA vaccine form with cationic chitosan and biodegradable and biocompatible PLGA. The pcSip plasmid in cationic-PLGA was successfully expressed in tissues of immunized tilapia and the immunogenicity was assessed in tilapia challenge model. A significant increase was observed in the cytokine levels of IL-1β, TNF-α, CC1, CC2 in spleen and kidney tissues. Furthermore, immunized tilapia conferred different levels of protection against challenge with a lethal dose of highly virulent serovar I a S. agalactiae. Our results indicated that the pcSip plasmid in cationic-PLGA induced high level of antibodies and protection against S. agalactiae infection, could be effective oral DNA vaccine candidates. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Analysis of the type II-A CRISPR-Cas system of Streptococcus agalactiae reveals distinctive features according to genetic lineages

    Science.gov (United States)

    Lier, Clément; Baticle, Elodie; Horvath, Philippe; Haguenoer, Eve; Valentin, Anne-Sophie; Glaser, Philippe; Mereghetti, Laurent; Lanotte, Philippe

    2015-01-01

    CRISPR-Cas systems (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) are found in 90% of archaea and about 40% of bacteria. In this original system, CRISPR arrays comprise short, almost unique sequences called spacers that are interspersed with conserved palindromic repeats. These systems play a role in adaptive immunity and participate to fight non-self DNA such as integrative and conjugative elements, plasmids, and phages. In Streptococcus agalactiae, a bacterium implicated in colonization and infections in humans since the 1960s, two CRISPR-Cas systems have been described. A type II-A system, characterized by proteins Cas9, Cas1, Cas2, and Csn2, is ubiquitous, and a type I–C system, with the Cas8c signature protein, is present in about 20% of the isolates. Unlike type I–C, which appears to be non-functional, type II-A appears fully functional. Here we studied type II-A CRISPR-cas loci from 126 human isolates of S. agalactiae belonging to different clonal complexes that represent the diversity of the species and that have been implicated in colonization or infection. The CRISPR-cas locus was analyzed both at spacer and repeat levels. Major distinctive features were identified according to the phylogenetic lineages previously defined by multilocus sequence typing, especially for the sequence type (ST) 17, which is considered hypervirulent. Among other idiosyncrasies, ST-17 shows a significantly lower number of spacers in comparison with other lineages. This characteristic could reflect the peculiar virulence or colonization specificities of this lineage. PMID:26124774

  20. Streptococcus agalactiae vaginitis: nonhemolytic variant on the Liofilchem® Chromatic StreptoB.

    Science.gov (United States)

    Savini, Vincenzo; Marrollo, Roberta; D'Antonio, Marianna; D'Amario, Claudio; Fazii, Paolo; D'Antonio, Domenico

    2013-01-01

    Streptococcus agalactiae (group B Streptococcus, GBS) vaginal pathogenicity is not uniformly acknowledged throughout the literature; accordingly, in women, genital itching and burning, along with leukorrhea are commonly and almost exclusively referred to bacterial vaginosis, candidiasis and trichomoniasis. Conversely, GBS virulence for vagina was recognized in the past, as the organism has been observed to potentially cause local inflammation and discharge, as well as lactobacilli rarefaction. We depict here a case where a nonhemolytic (γ-hemolytic) GBS strain was found to be the etiologic agent of vaginal infection. Such uncommon S. agalactiae phenotypes are hard to be recognized and may be therefore responsible for misdiagnosing and underestimation of GBS vaginitis prevalence; here, we had the support of the Liofilchem(®) Chromatic StreptoB medium, that successfully detected such an atypical variant.

  1. Expression, purification, crystallization and preliminary crystallographic analysis of laminin-binding protein (Lmb) from Streptococcus agalactiae

    International Nuclear Information System (INIS)

    Ragunathan, Preethi; Spellerberg, Barbara; Ponnuraj, Karthe

    2009-01-01

    Laminin-binding protein from S. agalactiae was expressed, purified and crystallized and X-ray diffraction data were collected to 2.5 Å resolution. Laminin-binding protein (Lmb), a surface-exposed lipoprotein from Streptococcus agalactiae (group B streptococcus), mediates attachment to human laminin and plays a crucial role in the adhesion/invasion of eukaryotic host cells. However, the structural basis of laminin binding still remains unclear. In the context of detailed structural analysis, the lmb gene has been cloned, expressed in Escherichia coli, purified and crystallized. The crystals diffracted to a resolution of 2.5 Å and belonged to the monoclinic space group P2 1 , with unit-cell parameters a = 56.63, b = 70.60, c = 75.37 Å, β = 96.77°

  2. Development and host compatibility of plasmids for two important ruminant pathogens, Mycoplasma bovis and Mycoplasma agalactiae.

    Directory of Open Access Journals (Sweden)

    Shukriti Sharma

    Full Text Available Mycoplasma bovis is a cause of pneumonia, mastitis, arthritis and otitis media in cattle throughout the world. However, despite its clinical significance, there is a paucity of tools to genetically manipulate it, impeding our capacity to further explore the molecular basis of its virulence. To address this limitation, we developed a series of homologous and heterologous replicable plasmids from M. bovis and M. agalactiae. The shortest replicable oriC plasmid based on the region downstream of dnaA in M. bovis was 247 bp and contained two DnaA boxes, while oriC plasmids based on the region downstream of dnaA in M. agalactiae strains 5632 and PG2 were 219 bp and 217 bp in length, respectively, and contained only a single DnaA box. The efficiency of transformation in M. bovis and M. agalactiae was inversely correlated with the size of the oriC region in the construct, and, in general, homologous oriC plasmids had a higher transformation efficiency than heterologous oriC plasmids. The larger pWholeoriC45 and pMM21-7 plasmids integrated into the genomic oriC region of M. bovis, while the smaller oriC plasmids remained extrachromosomal for up to 20 serial passages in selective media. Although specific gene disruptions were not be achieved in M. bovis in this study, the oriC plasmids developed here could still be useful as tools in complementation studies and for expression of exogenous genes in both M. bovis and M. agalactiae.

  3. Streptococcus agalactiae vaginitis: nonhemolytic variant on the Liofilchem® Chromatic StreptoB

    OpenAIRE

    Savini, Vincenzo; Marrollo, Roberta; D’Antonio, Marianna; D’Amario, Claudio; Fazii, Paolo; D’Antonio, Domenico

    2013-01-01

    Streptococcus agalactiae (group B Streptococcus, GBS) vaginal pathogenicity is not uniformly acknowledged throughout the literature; accordingly, in women, genital itching and burning, along with leukorrhea are commonly and almost exclusively referred to bacterial vaginosis, candidiasis and trichomoniasis. Conversely, GBS virulence for vagina was recognized in the past, as the organism has been observed to potentially cause local inflammation and discharge, as well as lactobacilli rarefaction...

  4. Development and Host Compatibility of Plasmids for Two Important Ruminant Pathogens, Mycoplasma bovis and Mycoplasma agalactiae

    Science.gov (United States)

    Sharma, Shukriti; Citti, Chistine; Sagné, Eveline; Marenda, Marc S.

    2015-01-01

    Mycoplasma bovis is a cause of pneumonia, mastitis, arthritis and otitis media in cattle throughout the world. However, despite its clinical significance, there is a paucity of tools to genetically manipulate it, impeding our capacity to further explore the molecular basis of its virulence. To address this limitation, we developed a series of homologous and heterologous replicable plasmids from M. bovis and M. agalactiae. The shortest replicable oriC plasmid based on the region downstream of dnaA in M. bovis was 247 bp and contained two DnaA boxes, while oriC plasmids based on the region downstream of dnaA in M. agalactiae strains 5632 and PG2 were 219 bp and 217 bp in length, respectively, and contained only a single DnaA box. The efficiency of transformation in M. bovis and M. agalactiae was inversely correlated with the size of the oriC region in the construct, and, in general, homologous oriC plasmids had a higher transformation efficiency than heterologous oriC plasmids. The larger pWholeoriC45 and pMM21-7 plasmids integrated into the genomic oriC region of M. bovis, while the smaller oriC plasmids remained extrachromosomal for up to 20 serial passages in selective media. Although specific gene disruptions were not be achieved in M. bovis in this study, the oriC plasmids developed here could still be useful as tools in complementation studies and for expression of exogenous genes in both M. bovis and M. agalactiae. PMID:25746296

  5. Experimental infections with Mycoplasma agalactiae identify key factors involved in host-colonization.

    Directory of Open Access Journals (Sweden)

    Eric Baranowski

    Full Text Available Mechanisms underlying pathogenic processes in mycoplasma infections are poorly understood, mainly because of limited sequence similarities with classical, bacterial virulence factors. Recently, large-scale transposon mutagenesis in the ruminant pathogen Mycoplasma agalactiae identified the NIF locus, including nifS and nifU, as essential for mycoplasma growth in cell culture, while dispensable in axenic media. To evaluate the importance of this locus in vivo, the infectivity of two knock-out mutants was tested upon experimental infection in the natural host. In this model, the parental PG2 strain was able to establish a systemic infection in lactating ewes, colonizing various body sites such as lymph nodes and the mammary gland, even when inoculated at low doses. In these PG2-infected ewes, we observed over the course of infection (i the development of a specific antibody response and (ii dynamic changes in expression of M. agalactiae surface variable proteins (Vpma, with multiple Vpma profiles co-existing in the same animal. In contrast and despite a sensitive model, none of the knock-out mutants were able to survive and colonize the host. The extreme avirulent phenotype of the two mutants was further supported by the absence of an IgG response in inoculated animals. The exact role of the NIF locus remains to be elucidated but these data demonstrate that it plays a key role in the infectious process of M. agalactiae and most likely of other pathogenic mycoplasma species as many carry closely related homologs.

  6. Mycoplasma agalactiae in semen and milk of goat from Pernambuco state, Brazil

    Directory of Open Access Journals (Sweden)

    Bruno H.L.S. Alves

    2013-11-01

    Full Text Available In goat and sheep flocks, mycoplasmosis is a disease that may cause severe economical losses associated with polyarthritis, mastitis, agalactia, conjunctivitis, pneumonia and reproductive failure. The latter may involve repeat breeding, granular vulvovaginitis, infertility and abortions. The aim of the present study was to assess the occurrence of Mycoplasma agalactiae (Ma in semen and milk samples from naturally infected goat in the semiarid region from Pernambuco State, Northeast from Brazil. Thirty-nine semen samples and 81 milk samples were submitted to DNA extraction using a commercially available kit and following the manufacturer's instructions. The polymerase chain reaction (PCR was then performed in accordance with protocols described in the literature. The results of the present study revealed the presence of Ma in the DNA of 17.9% (7/39 of the semen samples and 3.7% (3/81 of the milk samples. The results obtained in the present study confirm the elimination of the DNA of Ma in the semen and milk samples. The presence of this agent in goat flocks is considered very risky in terms of reproductive disorders and contagious agalactia outbreaks in the Northeast region of Brazil.

  7. DETEKSI POLYMORPHISME DENGAN SUBSTITUSI NUKLEOTIDA TUNGGAL PADA Streptococcus agalactiae ISOLAT LOKAL INDONESIA

    Directory of Open Access Journals (Sweden)

    Angela Mariana Lusiastuti

    2015-12-01

    Full Text Available Kasus penyakit pada budidaya ikan nila di wilayah di Jawa Barat, Jawa Tengah, Jawa Timur, Sulawesi Utara dan Papua Barat, disebabkan Streptococcus yang menyebabkan penyakit Streptococcosis di mana 80% disebabkan oleh grup B S. agalactiae. Tujuan penelitian ini adalah melakukan deteksi pada nukleotida isolat S. agalactiae untuk mengetahui sampai sejauh mana terjadinya nukleotida polimorfisme tunggal (SNP pada isolat tersebut. Identifikasi menggunakan PCR dilakukan terhadap 16S rDNA dan primer spesifik spesies terhadap S. agalactiae yaitu agal I 5’-ATAAGAGTAATTAACACATGTTAG-3’ (forward dan agal II 5’-ACTTCGGGTGTTACAAAC-3’(reverse dengan target 1250 bp. Produk PCR diamplifikasi terlebih dahulu menggunakan tujuh pasangan primer oligonukleotida yang berbeda yang didesain dari sekuens genom NEM316 GBS. Sekuens yang diperoleh dibandingkan dengan sekuens di Gene Bank database menggunakan National Center for Biotechnology Information Blast search tool. Hasil yang diperoleh ternyata ada dua basa yang berubah yaitu pada basa 24 dan basa 167. Pada basa 24 jelas terjadi subtitusi basa baru yaitu G, yang seharusnya tidak ada basa tersebut pada gen adhP-54 dan adhP-49 standar. Sedangkan pada basa 167 terjadi perbedaan basa dari seharusnya A pada standar menjadi G pada isolat 2.

  8. Structural characterization of the virulence factor nuclease A from Streptococcus agalactiae.

    Science.gov (United States)

    Moon, Andrea F; Gaudu, Philippe; Pedersen, Lars C

    2014-11-01

    The group B pathogen Streptococcus agalactiae commonly populates the human gut and urogenital tract, and is a major cause of infection-based mortality in neonatal infants and in elderly or immunocompromised adults. Nuclease A (GBS_NucA), a secreted DNA/RNA nuclease, serves as a virulence factor for S. agalactiae, facilitating bacterial evasion of the human innate immune response. GBS_NucA efficiently degrades the DNA matrix component of neutrophil extracellular traps (NETs), which attempt to kill and clear invading bacteria during the early stages of infection. In order to better understand the mechanisms of DNA substrate binding and catalysis of GBS_NucA, the high-resolution structure of a catalytically inactive mutant (H148G) was solved by X-ray crystallography. Several mutants on the surface of GBS_NucA which might influence DNA substrate binding and catalysis were generated and evaluated using an imidazole chemical rescue technique. While several of these mutants severely inhibited nuclease activity, two mutants (K146R and Q183A) exhibited significantly increased activity. These structural and biochemical studies have greatly increased our understanding of the mechanism of action of GBS_NucA in bacterial virulence and may serve as a foundation for the structure-based drug design of antibacterial compounds targeted to S. agalactiae.

  9. KERENTANAN IKAN NILA SULTANA, RED NIFI, SRIKANDI DAN AUREUS TERHADAP INFEKSI BAKTERI Streptococcus agalactiae

    Directory of Open Access Journals (Sweden)

    Rita Febrianti

    2015-06-01

    Full Text Available Penelitian ini bertujuan untuk menguji kerentanan empat strain ikan nila, yaitu: Sultana, Red NIFI, Srikandi, dan Aureus terhadap infeksi bakteri Streptococcus agalactiae. Ikan uji berukuran 15-20 g/ekor dan berasal dari populasi, serta batch umur yang sama. Infeksi bakteri S. agalactiae dilakukan secara buatan melalui penyuntikan intra peritoneal (IP pada dosis 104 cfu/mL, sedangkan kelompok kontrol diinjeksi dengan larutan Phosphate Buffered Saline (PBS. Pengamatan dilakukan terhadap gejala klinis dan mortalitas ikan uji yang berlangsung selama 14 hari. Hasil penelitian menunjukkan bahwa seluruh strain ikan nila mengalami respons yang sama terhadap infeksi bakteri S. agalactie yang ditandai dengan munculnya gejala klinis seperti: warna gelap/menghitam, sirip geripis, nekrosa pada mulut, mata menonjol, opaque, ulcer, dan dropsy. Kerentanan tertinggi terhadap infeksi bakteri S. agalactiae yang dimanifestasikan dengan rataan persentase mortalitas ikan uji diperoleh pada ikan nila Aureus sebesar 72%, Sultana 50%, Srikandi 36%, Red NIFI sebesar 24%, dan kontrol tidak ada mortalitas. Setelah diuji tantang, kadar limfosit mengalami kenaikan, netrofil dan monosit mengalami penurunan.

  10. Natural Mutations in Streptococcus agalactiae Resulting in Abrogation of β Antigen Production.

    Science.gov (United States)

    Vasilyeva, Anastasia; Santos Sanches, Ilda; Florindo, Carlos; Dmitriev, Alexander

    2015-01-01

    Streptococcus agalactiae genome encodes 21 two-component systems (TCS) and a variety of regulatory proteins in order to control gene expression. One of the TCS, BgrRS, comprising the BgrR DNA-binding regulatory protein and BgrS sensor histidine kinase, was discovered within a putative virulence island. BgrRS influences cell metabolism and positively control the expression of bac gene, coding for β antigen at transcriptional level. Inactivation of bgrR abrogated bac gene expression and increased virulence properties of S. agalactiae. In this study, a total of 140 strains were screened for the presence of bac gene, and the TCS bgrR and bgrS genes. A total of 53 strains carried the bac, bgrR and bgrS genes. Most of them (48 strains) expressed β antigen, while five strains did not express β antigen. Three strains, in which bac gene sequence was intact, while bgrR and/or bgrS genes had mutations, and expression of β antigen was absent, were complemented with a constructed plasmid pBgrRS(P) encoding functionally active bgrR and bgrS gene alleles. This procedure restored expression of β antigen indicating the crucial regulatory role of TCS BgrRS. The complemented strain A49V/BgrRS demonstrated attenuated virulence in intraperitoneal mice model of S. agalactiae infection compared to parental strain A49V. In conclusion we showed that disruption of β antigen expression is associated with: i) insertion of ISSa4 upstream the bac gene just after the ribosomal binding site; ii) point mutation G342A resulting a stop codon TGA within the bac gene and a truncated form of β antigen; iii) single deletion (G) in position 439 of the bgrR gene resulting in a frameshift and the loss of DNA-binding domain of the BgrR protein, and iv) single base substitutions in bgrR and bgrS genes causing single amino acid substitutions in BgrR (Arg187Lys) and BgrS (Arg252Gln). The fact that BgrRS negatively controls virulent properties of S. agalactiae gives a novel clue for understanding of S

  11. Simultaneous Identification of Potential Pathogenicity Factors of Mycoplasma agalactiae in the Natural Ovine Host by Negative Selection.

    Science.gov (United States)

    Hegde, Shivanand; Hegde, Shrilakshmi; Zimmermann, Martina; Flöck, Martina; Spergser, Joachim; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-07-01

    Mycoplasmas possess complex pathogenicity determinants that are largely unknown at the molecular level. Mycoplasma agalactiae serves as a useful model to study the molecular basis of mycoplasma pathogenicity. The generation and in vivo screening of a transposon mutant library of M. agalactiae were employed to unravel its host colonization factors. Tn4001mod mutants were sequenced using a novel sequencing method, and functionally heterogeneous pools containing 15 to 19 selected mutants were screened simultaneously through two successive cycles of sheep intramammary infections. A PCR-based negative selection method was employed to identify mutants that failed to colonize the udders and draining lymph nodes in the animals. A total of 14 different mutants found to be absent from ≥ 95% of samples were identified and subsequently verified via a second round of stringent confirmatory screening where 100% absence was considered attenuation. Using this criterion, seven mutants with insertions in genes MAG1050, MAG2540, MAG3390, uhpT, eutD, adhT, and MAG4460 were not recovered from any of the infected animals. Among the attenuated mutants, many contain disruptions in hypothetical genes, implying their previously unknown role in M. agalactiae pathogenicity. These data indicate the putative role of functionally different genes, including hypothetical ones, in the pathogenesis of M. agalactiae. Defining the precise functions of the identified genes is anticipated to increase our understanding of M. agalactiae infections and to develop successful intervention strategies against it. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. Development of a quantitative PCR assay for monitoring Streptococcus agalactiae colonization and tissue tropism in experimentally infected tilapia.

    Science.gov (United States)

    Su, Y-L; Feng, J; Li, Y-W; Bai, J-S; Li, A-X

    2016-02-01

    Streptococcus agalactiae has become one of the most important emerging pathogens in the aquaculture industry and has resulted in large economic losses for tilapia farms in China. In this study, three pairs of specific primers were designed and tested for their specificities and sensitivities in quantitative real-time polymerase chain reactions (qPCRs) after optimization of the annealing temperature. The primer pair IGS-s/IGS-a, which targets the 16S-23S rRNA intergenic spacer region, was finally chosen, having a detection limit of 8.6 copies of S. agalactiae DNA in a 20 μL reaction mixture. Bacterial tissue tropism was demonstrated by qPCR in Oreochromis niloticus 5 days post-injection with a virulent S. agalactiae strain. Bacterial loads were detected at the highest level in brain, followed by moderately high levels in kidney, heart, spleen, intestines, and eye. Significantly lower bacterial loads were observed in muscle, gill and liver. In addition, significantly lower bacterial loads were observed in the brain of convalescent O. niloticus 14 days post-injection with several different S. agalactiae strains. The qPCR for the detection of S. agalactiae developed in this study provides a quantitative tool for investigating bacterial tissue tropism in infected fish, as well as for monitoring bacterial colonization in convalescent fish. © 2015 John Wiley & Sons Ltd.

  13. Investigation of the translation-initiation factor IF2 gene, infB, as a tool to study the population structure of Streptococcus agalactiae

    DEFF Research Database (Denmark)

    Hedegaard, J; Hauge, M; Fage-Larsen, J

    2000-01-01

    The sequence of infB, encoding the prokaryotic translation-initiation factor 2 (IF2), was determined in eight strains of Streptococcus agalactiae (group B streptococcus) and an alignment revealed limited intraspecies diversity within S. agalactiae. The amino acid sequence of IF2 from S. agalactiae...... and from related species were aligned and revealed an interspecies conserved central and C-terminal part, and an N-terminal part that is highly variable in length and amino acid sequence. The diversity and relationships in a collection of 58 genetically distinct strains of S. agalactiae were evaluated...... by comparing a partial sequence of infB. A total of six alleles were detected for the region of infB analysed. The alleles correlated with the separation of the same strains of S. agalactiae into major evolutionary lineages, as shown in previous work. The partial sequences of infB were furthermore used...

  14. Whole genome shotgun sequencing of Indian strains of Streptococcus agalactiae

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    Balaji Veeraraghavan

    2017-12-01

    Full Text Available Group B streptococcus is known as a leading cause of neonatal infections in developing countries. The present study describes the whole genome shotgun sequences of four Group B Streptococcus (GBS isolates. Molecular data on clonality is lacking for GBS in India. The present genome report will add important information on the scarce genome data of GBS and will help in deriving comparative genome studies of GBS isolates at global level. This Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession numbers NHPL00000000 – NHPO00000000.

  15. Reactive oxygen species generation mediated by NADPH oxidase and PI3K/Akt pathways contribute to invasion of Streptococcus agalactiae in human endothelial cells.

    Science.gov (United States)

    Oliveira, Jessica Silva Santos de; Santos, Gabriela da Silva; Moraes, João Alfredo; Saliba, Alessandra Mattos; Barja-Fidalgo, Thereza Christina; Mattos-Guaraldi, Ana Luíza; Nagao, Prescilla Emy

    2018-01-01

    BACKGROUND Streptococcus agalactiae can causes sepsis, pneumonia, and meningitis in neonates, the elderly, and immunocompromised patients. Although the virulence properties of S. agalactiae have been partially elucidated, the molecular mechanisms related to reactive oxygen species (ROS) generation in infected human endothelial cells need further investigation. OBJECTIVES This study aimed to evaluate the influence of oxidative stress in human umbilical vein endothelial cells (HUVECs) during S. agalactiae infection. METHODS ROS production during S. agalactiae-HUVEC infection was detected using the probe CM-H2DCFDA. Microfilaments labelled with phalloidin-FITC and p47phox-Alexa 546 conjugated were analysed by immunofluorescence. mRNA levels of p47phox (NADPH oxidase subunit) were assessed using Real Time qRT-PCR. The adherence and intracellular viability of S. agalactiae in HUVECs with or without pre-treatment of DPI, apocynin (NADPH oxidase inhibitors), and LY294002 (PI3K inhibitor) were evaluated by penicillin/gentamicin exclusion. Phosphorylation of p47phox and Akt activation by S. agalactiae were evaluated by immunoblotting analysis. FINDINGS Data showed increased ROS production 15 min after HUVEC infection. Real-Time qRT-PCR and western blotting performed in HUVEC infected with S. agalactiae detected alterations in mRNA levels and activation of p47phox. Pre-treatment of endothelial cells with NADPH oxidase (DPI and apocynin) and PI3K/Akt pathway (LY294002) inhibitors reduced ROS production, bacterial intracellular viability, and generation of actin stress fibres in HUVECs infected with S. agalactiae. CONCLUSIONS ROS generation via the NADPH oxidase pathway contributes to invasion of S. agalactiae in human endothelial cells accompanied by cytoskeletal reorganisation through the PI3K/Akt pathway, which provides novel evidence for the involvement of oxidative stress in S. agalactiae pathogenesis.

  16. Pattern of Infection and Antibiotic Activity among Streptococcus agalactiae Isolates from Adults in Mashhad, Iran

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    Masoumeh Malek-Jafarian

    2015-05-01

    Full Text Available Background: One of the main causes of sexually transmitted diseases is group B β- hemolytic streptococci (GBS multiplying in the genital tracts. Penicillin is the most common drug for the treatment of infections caused by these bacteria, but in patients suffering from Penicillin allergy, Erythromycin and Clindamycin are used as alternative therapeutic drugs against GBS. Recently, resistance to these drugs has been reported more often. In this study, efforts have been made to determine the prevalence and antibiotic resistance of GBS. Methods: Modified Christie Atkins Munch-Petersen (CAMP test was conducted on over 2400 samples of urine and discharge taken from vagina, urethra and prostate. The drug sensitivity was performed by double disk sensitivity tests to Bacitracin, Trimethoprim, and Sulfamethoxazole and then the resistant samples were investigated by E-test to determine the minimal inhibitory concentrations (MICs value. Results: Twenty-three vaginal and 10 urethral discharge, 27urine and 6 prostatic secretion samples were GBS positive. The most symbiotic microorganisms with GBS were strains of Enterococci (90%, Staphylococcus saprophyticus (25% and Candida albicans (6%. The disk diffusion method showed 18 cases with Penicillin resistance (MIC: 1.5 mg/ml. Conclusion: Taken together, GBS carriers’ rate in this study was found 20.65% (8.24% men and 12.4% women. Furthermore, findings showed high-level resistance to Erythromycin and Clindamycin.

  17. Genome analysis of multiple pathogenic isolates of Streptococcus agalactiae : Implications for the microbial "pan-genome"

    NARCIS (Netherlands)

    Tettelin, H; Masignani, [No Value; Cieslewicz, MJ; Donati, C; Medini, D; Ward, NL; Angiuoli, SV; Crabtree, J; Jones, AL; Durkin, AS; DeBoy, RT; Davidsen, TM; Mora, M; Scarselli, M; Ros, IMY; Peterson, JD; Hauser, CR; Sundaram, JP; Nelson, WC; Madupu, R; Brinkac, LM; Dodson, RJ; Rosovitz, MJ; Sullivan, SA; Daugherty, SC; Haft, DH; Selengut, J; Gwinn, ML; Zhou, LW; Zafar, N; Khouri, H; Radune, D; Dimitrov, G; Watkins, K; O'Connor, KJB; Smith, S; Utterback, TR; White, O; Rubens, CE; Grandi, G; Madoff, LC; Kasper, DL; Telford, JL; Wessels, MR; Rappuoli, R; Fraser, CM

    2005-01-01

    The development of efficient and inexpensive genome sequencing methods has revolutionized the study of human bacterial pathogens and improved vaccine design. Unfortunately, the sequence of a single genome does not reflect how genetic variability drives pathogenesis within a bacterial species and

  18. Spatiotemporal patterns, annual baseline and movement-related incidence of Streptococcus agalactiae infection in Danish dairy herds: 2000-2009.

    Science.gov (United States)

    Mweu, Marshal M; Nielsen, Søren S; Halasa, Tariq; Toft, Nils

    2014-02-01

    Several decades after the inception of the five-point plan for the control of contagious mastitis pathogens, Streptococcus agalactiae (S. agalactiae) persists as a fundamental threat to the dairy industry in many countries. A better understanding of the relative importance of within- and between-herd sources of new herd infections coupled with the spatiotemporal distribution of the infection, may aid in effective targeting of control efforts. Thus, the objectives of this study were: (1) to describe the spatiotemporal patterns of infection with S. agalactiae in the population of Danish dairy herds from 2000 to 2009 and (2) to estimate the annual herd-level baseline and movement-related incidence risks of S. agalactiae infection over the 10-year period. The analysis involved registry data on bacteriological culture of all bulk tank milk samples collected as part of the mandatory Danish S. agalactiae surveillance scheme as well as live cattle movements into dairy herds during the specified 10-year period. The results indicated that the predicted risk of a herd becoming infected with S. agalactiae varied spatiotemporally; the risk being more homogeneous and higher in the period after 2005. Additionally, the annual baseline risks yielded significant yet distinctive patterns before and after 2005 - the risk of infection being higher in the latter phase. On the contrary, the annual movement-related risks revealed a non-significant pattern over the 10-year period. There was neither evidence for spatial clustering of cases relative to the population of herds at risk nor spatial dependency between herds. Nevertheless, the results signal a need to beef up within-herd biosecurity in order to reduce the risk of new herd infections. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. The Efficacy of Nile Tilapia (Oreochromis niloticus Broodstock and Larval Immunization against Streptococcus agalactiae and Aeromonas hydrophila

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    Wesly Pasaribu

    2018-03-01

    Full Text Available Streptococcus agalactiae and Aeromonas hydrophila have been recognized as the causative agents of mortality in tilapia larvae with single infection and coinfection. The objective of this study was to evaluate the efficacy of maternal transfer and offspring protection from the immunization of monovalent and bivalent vaccines on Nile tilapia (Oreochromis niloticus broodstock and larval immunization. Four groups of broodstock were intraperitoneally injected with formalin killed whole-cells of S. agalactiae (Sa group, A. hydrophila (Ah group, the bivalent mixed vaccine of them (Biv group, and phosphate-buffered saline as a control (Pbs group. Immunization of the larvae produced from immunized broodstock with a bivalent vaccine (Biv1 group and Pbs (Pbs1 group was performed by immersion at 20 days after hatch. Larvae produced from the Pbs group were unvaccinated as the control (Pbs2 group. Changes in the specific antibody and relative percent survival were measured. The Sa and Ah groups that could increase specific antibodies and protection against pathogenic bacteria were challenged with the homologous bacteria. The Biv group stimulated and protected against both S. agalactiae and A. hydrophila. The specific antibody of the Biv1 group was higher than the Pbs1 and Pbs2 groups. The last observation in this study showed that the relative percent survival of the Biv group after challenged S. agalactiae, A. hydrophila, and coinfection were 74.74 ± 3.18%, 73.81 ± 8.58%, and 71.48 ± 5.70%, respectively. The use of bivalent vaccines on the broodstock and larvae may be a strategy to reduce mortality in Nile tilapia larvae caused by single pathogen infection of S. agalactiae and A. hydrophila, or coinfection with both S. agalactiae and A. hydrophila.

  20. Genital infections and reproductive complications associated with Trichomonas vaginalis, Neisseria gonorrhoeae, and Streptococcus agalactiae in women of Qom, central Iran

    Directory of Open Access Journals (Sweden)

    Mahmoud Nateghi Rostam

    2017-07-01

    Full Text Available Background: Trichomonas vaginalis (T.vaginalis and Neisseria gonorrhoeae (N.gonorrhoeae are two most common non-viral sexually transmitted infections in the world. No data are available regarding the epidemiology of genital infections in women of Qom, central Iran. Objective: Epidemiological investigation of sexually transmitted infections in genital specimens of women referred to the referral gynecology hospital in Qom, central Iran. Materials and Methods: Genital swab specimens were collected from women volunteers and used for identification of bacterial and protozoal infections by conventional microbial diagnostics, porA pseudo gene LightCycler® real-time PCR (for N.gonorrhoeae and ITS-PCR (for T.vaginalis. Results: Of 420 volunteers, 277 (65.9% had genital signs/symptoms, including 38.3% malodorous discharge, 37.9% dyspareunia, and 54.8% abdominal pain. Totally, 2 isolates of Streptococcus agalactiae were identified. Five specimens (1.2% in Thayer-Martin culture and 17 (4.1% in real-time PCR were identified as N.gonorrhoeae. Fifty-four specimens (12.9% in wet mount, 64 (15.2% in Dorset’s culture, and 81 (19.3% in ITS-PCR showed positive results for T.vaginalis. Five mixed infections of T.vaginalis+ N.gonorrhoeae were found. The risk of T.vaginalis infection was increased in women with low-birth-weight (p=0.00; OR=43.29, history of abortion (p=0.00; OR=91.84, and premature rupture of membranes (PROM (p=0.00; OR=21.75. The probability of finding nuclear leukocytes (p=0.00; OR=43.34 in vaginal smear was higher in T.vaginalis infection. Conclusion: The significant prevalence of trichomoniasis and gonorrhea emphasizes the need for accurate diagnosis and effective surveillance to prevent serious reproductive complications in women.

  1. Combination of garlic - shatterstone herb powder to control Streptococcus agalactiae infection in tilapia

    Directory of Open Access Journals (Sweden)

    Ririn Nurul Fauziah

    2015-10-01

    Full Text Available ABSTRACT This study was aimed at determining potential of combination powder of garlic Allium sativum-shatterstone herb Phyllanthus niruri supplemented in feed against S. agalactiae infection in tilapia. Four concentrations of combination powder of A. sativum-P. Niruri; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in in vitro test. In vivo test consisted of three treatments with three replications, namely positive control (K+, negative control (K- and the treatment of A. sativum-P. niruri suplemented in feed (BM.  The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL S. agalactiae at concentration of 105 cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM showed better growth rate, feed response, and survival (83.3% than positive control (36.7% at P<0.05. In addition, A. sativum-P. niruri suplemented in feed was also able to enhance the immune response by increasing phagocytic activity. Keywords: Streptococcus agalactiae, phytopharmacy, Allium sativum-Phyllanthus niruri, tilapia  ABSTRAK Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih Allium sativum-meniran Phyllanthus niruri dalam pakan terhadap pencegahan infeksi bakteri S. agalactiae pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam  menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji in vitro. Uji in vivo terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif

  2. Fulminant Necrotizing Fasciitis and Toxic Shock Syndrome Caused by Streptococcus agalactiae

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    Emin UYSAL

    2018-03-01

    Full Text Available Necrotizing fasciitis is a rare and life-threatening soft tissue infection that spreads rapidly and involves the skin, subcutaneous tissue, fascia, and muscle layer. The treatment is possible by initiating appropriate antibiotherapy for the clinically suspected cause and by performing surgical intervention quickly and aggressively. However, it should be known that necrotizing fasciitis is a disease that is difficult to manage despite all interventions, effective treatment protocols, and patient care. This article presents the case of a 60-year-old patient with diabetes mellitus who died of toxic shock syndrome with fulminant necrotizing fasciitis caused by Streptococcus agalactiae.

  3. Development of a single-dose recombinant CAMP factor entrapping poly(lactide-co-glycolide) microspheres-based vaccine against Streptococcus agalactiae.

    Science.gov (United States)

    Liu, Gang; Yin, Jinhua; Barkema, Herman W; Chen, Liben; Shahid, Muhammad; Szenci, Otto; De Buck, Jeroen; Kastelic, John P; Han, Bo

    2017-03-01

    Streptococcus agalactiae is an important contagious bovine mastitis pathogen. Although it is well controlled and even eradicated in most Northern European and North American dairy herds, the prevalence of this pathogen remains very high in China. However, research on development of a vaccine against S. agalactiae mastitis is scarce. The aims of the present study were to: (1) develop a single-dose vaccine against S. agalactiae based on poly(lactic-co-glycolic acid) (PLGA) microspheres (MS) encapsulated CAMP factor, a conserved virulent protein encoded by S. agalactiae's cfb gene; and (2) evaluate its immunogenicity and protective efficacy in a mouse model. The cfb gene was cloned and expressed in a recombinant Escherichia coli strain Trans1-T1. The CAMP factor was tested to determine a safe dose range and then encapsulated in MS of PLGA (50:50) to assess its release pattern in vitro and immune reaction in vivo. Furthermore, a mouse model and a histopathological assay were developed to evaluate bacterial burden and vaccine efficacy. In the low dosage range (S. agalactiae challenge. Additionally, no pathological lesions were detected in the vaccinated group. Therefore, PLGA-CAMP conferred protective efficacy against S. agalactiae in our mouse model, indicating its potential as a vaccine against S. agalactiae mastitis. Furthermore, the slow-release kinetics of PLGA MS warranted optimism for development of a single-dose vaccine. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  4. Comparaison de la souche 7784 hypovirulente de Mycoplasma agalactiae à la souche type virulente PG2 par la technique d'hybridation soustractive suppressive

    OpenAIRE

    Bonnefin, Nathalie

    2003-01-01

    Mycoplasma agalactiae est l'agent principal responsable de l'agalactie contagieuse des ovins et caprins, un syndrome largement répandu dans le monde et regroupant des atteintes mammaires, articulaires et oculaires. Afin de rechercher les bases moléculaires du pouvoir pathogène de M. agalactiae, la technique d'hybridation soustractive suppressive, qui permet l'identification de séquences présentes dans un génome mais absentes d'un autre, a été utilisée. Deux souches de M. agalactiae ont été co...

  5. Effect of carryover and presampling procedures on the results of real-time PCR used for diagnosis of bovine intramammary infections with Streptococcus agalactiae at routine milk recordings

    DEFF Research Database (Denmark)

    Mahmmod, Yasser; Mweu, Marshal Mutinda; Nielsen, Søren Saxmose

    2014-01-01

    with Streptococcus agalactiae (S. agalactiae) in dairy herds with conventional milking parlours. Misclassification may result in unnecessary costs for treatment and culling. The objectives of this study were to (1) determine the effect of carryover on PCR-positivity for S. agalactiae at different PCR cycle threshold...... (Ct) cut-offs by estimating the between-cow correlation while accounting for the milking order, and (2) evaluate the effect of aseptic presampling procedures (PSP) on PCR-positivity at the different Ct-value cut-offs. The study was conducted in four herds with conventional milking parlours at routine...

  6. Serotype markers in a Streptococcus agalactiae strain collection from Zimbabwe

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    Mavenyengwa R

    2010-01-01

    Full Text Available Objective: Group B streptococci (GBS from Southern African areas have been less well characterized. Our objective was to study serotype and serovariant distribution of carrier GBS strains as part of a study of the epidemiology of GBS carriage in pregnant women from Zimbabwe. Materials and Methods: We studied GBS isolated from 121 healthy pregnant women living in Harare and surrounding areas, Zimbabwe. Capsular polysaccharide (CPS testing for serotype determination and surface-anchored protein testing for serosubtype determination were done by gene-based serotyping (PCR, except for the proteins R3 and a novel protein called Z, which were detected by antibody-based methods. Results: Strains of the CPS types Ia (15.7%, Ib (11.6%, II (8.3%, III (38.8%, V (24.0% and NT (1.7% were detected along with the strain-variable proteins Cί (15.7% of isolates, Cα (19.8%, Alp1 (epsilon-22.3%, Alp3 (5.0%, R4/Rib (46.3%, R3 (27.3%, Z (27.3%, and SAR5 (28.9%, which encodes the R5 protein. Up to four of the protein genes could be possessed or the gene product expressed by one and the same isolate. A total of 32 serovariants were detected. The findings assessed by us as most important were the very low prevalence of the gene Alp3 (Alp3 - 4.9%, high prevalence of R4 (Rib - 46.2%, the proteins R3 (27.3%, Z (27.3%, and of SAR5 (R5 - 28.9%. The low prevalence of Alp3, notably in GBS type V strains, differed from findings with CPS type V GBS from non-African areas. Bacteria of the various CPS types showed distinct CPS/protein-marker associations. Conclusion: The results are of importance in relation to regional variations of GBS phenotypes and genotypes and thus, of importance in planning and research in the context of future vaccine formulations.

  7. Molecular and functional characterization of CD59 from Nile tilapia (Oreochromis niloticus) involved in the immune response to Streptococcus agalactiae.

    Science.gov (United States)

    Gan, Zhen; Wang, Bei; Zhou, Wei; Lu, Yishan; Zhu, Weiwei; Tang, Jufen; Jian, JiChang; Wu, Zaohe

    2015-05-01

    CD59, the major inhibitor of membrane attack complex, plays a crucial role in regulation of complement activation. In this paper, a CD59 gene of Nile tilapia, Oreochromis niloticus (designated as On-CD59) was cloned and its expression pattern under the stimulation of Streptococcus agalactiae was investigated. Sequence analysis showed main structural features required for complement-inhibitory activity were detected in the deduced amino acid sequence of On-CD59. In healthy Nile tilapia, the On-CD59 transcripts could be detected in all the examined tissues, with the most abundant expression in the brain. When immunized with inactivated S. agalactiae, there was a clear time-dependent expression pattern of On-CD59 in the skin, brain, head kidney, thymus and spleen, with quite different kinetic expressions. The assays for the complement-inhibitory activity suggested that recombinant On-CD59 protein had a species-selective inhibition of complement. Moreover, our works showed that recombinant On-CD59 protein may possess both binding activities to PGN and LTA and inhibiting activity of S. agalactiae. These findings indicated that On-CD59 may play important roles in the immune response to S. agalactiae in Nile tilapia. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Resistance of Nile tilapia (Oreochromis niloticus) to Streptococcus iniae and S. agalactiae Ib is heritable but not correlated

    Science.gov (United States)

    Tilapia (Oreochromis sp.) are an important source of protein with an economic value approaching US $8 billion yearly. Streptococcal disease, caused by Streptococcus iniae and S. agalactiae (both Gram positive bacteria), is an emerging or re-emerging disease negatively affecting tilapia aquaculture w...

  9. The Adc/Lmb System Mediates Zinc Acquisition in Streptococcus agalactiae and Contributes to Bacterial Growth and Survival.

    Science.gov (United States)

    Moulin, Pauline; Patron, Kévin; Cano, Camille; Zorgani, Mohamed Amine; Camiade, Emilie; Borezée-Durant, Elise; Rosenau, Agnès; Mereghetti, Laurent; Hiron, Aurélia

    2016-12-15

    The Lmb protein of Streptococcus agalactiae is described as an adhesin that binds laminin, a component of the human extracellular matrix. In this study, we revealed a new role for this protein in zinc uptake. We also identified two Lmb homologs, AdcA and AdcAII, redundant binding proteins that combine with the AdcCB translocon to form a zinc-ABC transporter. Expression of this transporter is controlled by the zinc concentration in the medium through the zinc-dependent regulator AdcR. Triple deletion of lmb, adcA, and adcAII, or that of the adcCB genes, impaired growth and cell separation in a zinc-restricted environment. Moreover, we found that this Adc zinc-ABC transporter promotes S. agalactiae growth and survival in some human biological fluids, suggesting that it contributes to the infection process. These results indicated that zinc has biologically vital functions in S. agalactiae and that, under the conditions tested, the Adc/Lmb transporter constitutes the main zinc acquisition system of the bacterium. A zinc transporter, composed of three redundant binding proteins (Lmb, AdcA, and AdcAII), was characterized in Streptococcus agalactiae This system was shown to be essential for bacterial growth and morphology in zinc-restricted environments, including human biological fluids. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  10. Role of the serine-rich surface glycoprotein Srr1 of Streptococcus agalactiae in the pathogenesis of infective endocarditis.

    Directory of Open Access Journals (Sweden)

    Ho Seong Seo

    Full Text Available The binding of bacteria to fibrinogen and platelets are important events in the pathogenesis of infective endocarditis. Srr1 is a serine-rich repeat glycoprotein of Streptococcus agalactiae that binds directly to the Aα chain of human fibrinogen. To assess the impact of Srr1 on the pathogenesis of endocarditis due to S. agalactiae, we first examined the binding of this organism to immobilized human platelets. Strains expressing Srr1 had significantly higher levels of binding to human platelets in vitro, as compared with isogenic Δsrr1 mutants. In addition, platelet binding was inhibited by pretreatment with anti-fibrinogen IgG or purified Srr1 binding region. To assess the contribution of Srr1 to pathogenicity, we compared the relative virulence of S. agalactiae NCTC 10/84 strain and its Δsrr1 mutant in a rat model of endocarditis, where animals were co-infected with the WT and the mutant strains at a 1:1 ratio. At 72 h post-infection, bacterial densities (CFU/g of the WT strain within vegetations, kidneys, and spleens were significantly higher, as compared with the Δsrr1 mutant. These results indicate that Srr1 contributes to the pathogenesis of endocarditis due to S. agalactiae, at least in part through its role in fibrinogen-mediated platelet binding.

  11. Evaluation of colorimetric loop-mediated isothermal amplification assay for visual detection of Streptococcus agalactiae and Streptococcus iniae in tilapia.

    Science.gov (United States)

    Suebsing, R; Kampeera, J; Tookdee, B; Withyachumnarnkul, B; Turner, W; Kiatpathomchai, W

    2013-10-01

    Streptococcus agalactiae and Strep. iniae are bacterial pathogens that cause streptococcosis in many fish species. An accelerated colorimetric loop-mediated isothermal amplification (LAMP) assay with pre-addition of calcein was established, and the transmission and detection of Strep. agalactiae and Strep. iniae in tilapia under natural aquatic environment were investigated. A positive reaction was observed by a colour change from orange to green through the naked eyes after completion at 63°C for 30 min with 10 times higher sensitivity than that of nested PCR assays and without cross-amplification with other fish bacterial pathogens. All sample types of Nile and red tilapia (broodstock, fertilized egg, fry) were Strep. agalactiae- and Strep. iniae positive by this new method, implying that they could be vertically transmitted. With its application for screening broodstock and fry before stocking and for monitoring fish health in grow-out ponds, the method would become very useful in fish farming industry. The application of colorimetric LAMP with pre-addition of calcein offers simple, rapid and sensitive technique with applicability for small field laboratories. This technique explored the possible vertical transmission mode of Strep. agalactiae and Strep. iniae under natural aquatic environment. It could be such preliminary data provided for the screening broodstock before breeding and/or the specific-pathogen-free production. © 2013 The Society for Applied Microbiology.

  12. The economic benefit of treating subclinical Streptococcus agalactiae mastitis in lactating cows.

    Science.gov (United States)

    Yamagata, M; Goodger, W J; Weaver, L; Franti, C

    1987-12-15

    The economic benefits of treating lactating cows for Streptococcus agalactiae mastitis were studied at a large (689 milking cows) central California dairy. Postcure milk production of case cows (infected, treated, and cured) was compared with production of paired control cows (uninfected) and was matched for yield, days in milk, days in gestation, and parity. A simulation was used to plot expected lactation curves for mastitic cows (infected, not treated) with characteristics similar to those of each control cow, and these curves were compared with actual case-cow lactation curves. The difference in actual and expected production was used to calculate net economic benefits of treatment. Comparison of expected with actual production indicated a net benefit from treatment of $396/cow for cows treated in early lactation and $237 for cows treated in midlactation, but a net loss of $55 for cows treated in late lactation. Lactation number did not have a significant impact on economic benefits of treatment. In contrast to other studies indicating no economic benefit from treating mastitis during lactation, this study's positive results may have been attributable to the high cure rate (98%) and the subclinical form of mastitis being treated. Streptococcus agalactiae mastitis treatment during early and midlactation would appear to be an economically justifiable option for dairy managers.

  13. [Effect of the 10 kb sequence of piscine Streptococcus agalactiae on bacterial virulence].

    Science.gov (United States)

    Liu, Guangjin; Zhu, Jielian; Shi, Ziwei; Ding, Ming; Wang, Ruyi; Yao, Huochun; Lu, Chengping; Xu, Pao

    2016-01-04

    From the previous comparative genomic analysis, we found a specific unknown 10 kb sequence (including 11 Open reading Frames) in Chinese piscine strain GD201008-001 genome. To study the role of 10 kb in the pathogenicity of piscine S. agalactiae, the 10 kb sequence was deleted from the GD201008-001 genome. The isogenic mutant Δ10 kb was constructed by using the temperature-sensitive Streptococcus-E. coli shuttle vector pSET4s. We compared the growth characteristics, adherence to HEp-2 cell and bacterial virulence in a zebrafish infection model between wild strain and mutant. Meanwhile the expressions of the known virulence genes from GD201008-001 and Δ10 kb were also quantified by real-time PCR. The Δ10 kb showed no significant differences in bacterial morphology and adherence to HEp-2 cells compared with the wild-type strain, but the speed of growth was slightly slower than the wild strain. Furthermore the 50% lethal dose of Δ10 kb was decreased up to 10-fold (P kb sequence of piscine Streptococcus agalactiae exerts a significant effect on bacterial virulence and probably regulates the virulence genes expression of GD20 1008-001.

  14. The effects of fish gender on susceptibility to acute Streptococcus agalactiae infection in Javanese medaka Oryzias javanicus.

    Science.gov (United States)

    Amal, M N A; Zarif, S T; Suhaiba, M S; Aidil, M R M; Shaqinah, N N; Zamri-Saad, M; Ismail, A

    2017-12-05

    This study describes the susceptibility of different fish gender following acute Streptococcus agalactiae infection by using Javanese medaka Oryzias javanicus as test fish. The fish were grouped into four groups, which were: (1) all-male; (2) all-female; (3) mixed-gender (1 male: 1 female ratio); and (4) control non-infected (1 male: 1 female ratio). The fish in group 1, 2 and 3 were intraperitoneally exposed to 5.4 × 10 8  CFU/mL of S. agalactiae, while for group 4, the fish were exposed using sterile broth. The main clinical signs and histopathological changes of infected Javanese medaka were commonly observed in S. agalactiae infected fishes. However, no difference on clinical signs and histopathological changes of fish in group 1, 2 and 3 were noticed. The Javanese medaka mortality in group 1, 2 and 3 were observed from 4 h post infection (hpi) to 6 hpi, with the cumulative mortality from 3% to 30%. Then, the mortality increased at 12 hpi, with the range from 53% to 80%. However, 100% of the infected fish dead at 24 hpi. No clinical sign, histopathological change and fish mortality recorded in group 4. Generally, the clinical signs, mortality patterns, cumulative mortality and histopathological changes of Javanese medaka infected by S. agalactiae did not show any difference between the all-male, all-female and mixed-gender groups. This indicates that the susceptibility of fish to S. agalactiae infection is not influenced by their gender. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. The effect of fish gender on susceptibility to acute Streptococcus agalactiae infection in Javanese medaka Oryzias javanicus.

    Science.gov (United States)

    Amal, M N A; Zarif, S T; Suhaiba, M; Aidil, M R M; Shaqinah, N N; Zamri-Saad, M; Ismail, A

    2017-12-04

    This study describes the susceptibility of different fish gender following acute Streptococcus agalactiae infection by using Javanese medaka Oryzias javanicus as test fish. The fish were grouped into four groups, which were: (1) all-male; (2) all-female; (3) mixed-gender (1 male: 1 female ratio); and (4) control non-infected (1 male: 1 female ratio). The fish in group 1, 2 and 3 were intraperitoneally exposed to 5.4 × 10 8  CFU/ml of S. agalactiae, while for group 4, the fish were exposed using sterile broth. The main clinical signs and histopathological changes of infected Javanese medaka were commonly observed in S. agalactiae infected fishes. However, no difference on clinical signs and histopathological changes of fish in group 1, 2 and 3 were noticed. The Javanese medaka mortality in group 1, 2 and 3 were observed from 4 h post infection (hpi) to 6 hpi, with the cumulative mortality from 3% to 30%. Then, the mortality increased at 12 hpi, with the range from 53% to 80%. However, 100% of the infected fish dead at 24 hpi. No clinical signs, histopathological changes and fish mortality recorded in group 4. Generally, the clinical signs, mortality patterns, cumulative mortality and histopathological changes of Javanese medaka infected by S. agalactiae did not show any difference between the all-male, all-female and mixed-gender groups. This indicates that the susceptibility of fish to S. agalactiae infection is not influenced by their gender. Copyright © 2017. Published by Elsevier Ltd.

  16. Latent class analysis of real time qPCR and bacteriological culturing for the diagnosis of Streptococcus agalactiae in cow composite milk samples

    DEFF Research Database (Denmark)

    Holmøy, Ingrid H.; Toft, Nils; Jørgensen, Hannah J.

    2018-01-01

    Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows....... The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare...... it to conventional bacteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging...

  17. Additive genetic variation in resistance of Nile tilapia (Oreochromis niloticus) to Streptococcus iniae and S. agalactiae capsular type Ib: is genetic resistance correlated?

    Science.gov (United States)

    Streptococcus (S.) iniae and S. agalactiae are both economically important Gram positive bacterial pathogens affecting the globally farmed tilapia (Oreochromis spp.). Historically control of these bacteria in tilapia culture has included biosecurity, therapeutants and vaccination strategies. Genet...

  18. Molecular characteristics of Streptococcus agalactiae in a mother-baby prospective cohort study: Implication for vaccine development and insights into vertical transmission.

    Science.gov (United States)

    Li, Shunming; Wen, Guoming; Cao, Xuelian; Guo, Dan; Yao, Zhenjiang; Wu, Chuan'an; Ye, Xiaohua

    2018-04-05

    Streptococcus agalactiae (GBS) is a leading cause of neonatal sepsis and meningitis in many countries. This study aimed to determine the molecular characteristics of GBS colonized in mothers and their infants so as to provide implication for vaccine strategies and confirm vertical transmission. A prospective cohort study was conducted to recruit 1815 mother-neonate pairs. All GBS isolates from pregnant women and her infants were tested for serotypes, multilocus sequence types and virulence genes. The relationship between multiple molecular characteristics of GBS isolates was tested by the correspondence analysis, and the agreement between mother-neonate paired data in molecular characteristics was analyzed using Kappa tests. The predominant serotypes were III, Ia and V, and the most prevalent sequence types (STs) were ST19, ST17, ST10, and ST12. All isolates carried at least one pilus island (PI). The most common combination of PIs was PI-2b alone, followed by PI-1+PI-2a and PI-2a alone, and the most prevalent alpha-like protein (alp) genes were rib, epsilon and alphaC. Moreover, a strong relationship was noted between STs, serotypes, alp genes and PIs, including ST17 associated with serotype-III/rib/PI-2b, ST19 with serotype-III/rib/PI-1+PI-2a, and ST485 with serotype-Ia/epsilon/PI-2b. The rate of GBS vertical transmission was 14.1%, and the kappa test revealed good agreement in multiple molecular characteristics among GBS-positive mother-neonate pairs. Notably, the switching of molecular characteristics was found during vertical transmission. Our findings underscore the value of monitoring multiple molecular characteristics so as to provide implication for multivalent strategies and gain insights into GBS vertical transmission and vertical characteristic switching. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Complete Atrioventricular Block Complicating Mitral Infective Endocarditis Caused by Streptococcus Agalactiae.

    Science.gov (United States)

    Arai, Masaru; Nagashima, Koichi; Kato, Mahoto; Akutsu, Naotaka; Hayase, Misa; Ogura, Kanako; Iwasawa, Yukino; Aizawa, Yoshihiro; Saito, Yuki; Okumura, Yasuo; Nishimaki, Haruna; Masuda, Shinobu; Hirayama, Astushi

    2016-09-08

    BACKGROUND Infective endocarditis (IE) involving the mitral valve can but rarely lead to complete atrioventricular block (CAVB). CASE REPORT A 74-year-old man with a history of infective endocarditis caused by Streptococcus gordonii (S. gordonii) presented to our emergency room with fever and loss of appetite, which had lasted for 5 days. On admission, results of serologic tests pointed to severe infection. Electrocardiography showed normal sinus rhythm with first-degree atrioventricular block and incomplete right bundle branch block, and transthoracic echocardiography and transesophageal echocardiography revealed severe mitral regurgitation caused by posterior leaflet perforation and 2 vegetations (5 mm and 6 mm) on the tricuspid valve. The patient was initially treated with ceftriaxone and gentamycin because blood and cutaneous ulcer cultures yielded S. agalactiae. On hospital day 2, however, sudden CAVB requiring transvenous pacing occurred, and the patient's heart failure and infection worsened. Although an emergent surgery is strongly recommended, even in patients with uncontrolled heart failure or infection, surgery was not performed because of the Child-Pugh class B liver cirrhosis. Despite intensive therapy, the patient's condition further deteriorated, and he died on hospital day 16. On postmortem examination, a 2×1-cm vegetation was seen on the perforated posterior mitral leaflet, and the infection had extended to the interventricular septum. Histologic examination revealed extensive necrosis of the AV node. CONCLUSIONS This rare case of CAVB resulting from S. agalactiae IE points to the fact that in monitoring patients with IE involving the mitral valve, clinicians should be aware of the potential for perivalvular extension of the infection, which can lead to fatal heart block.

  20. Molecular mapping of the cell wall polysaccharides of the human pathogen Streptococcus agalactiae

    Science.gov (United States)

    Beaussart, Audrey; Péchoux, Christine; Trieu-Cuot, Patrick; Hols, Pascal; Mistou, Michel-Yves; Dufrêne, Yves F.

    2014-11-01

    The surface of many bacterial pathogens is covered with polysaccharides that play important roles in mediating pathogen-host interactions. In Streptococcus agalactiae, the capsular polysaccharide (CPS) is recognized as a major virulence factor while the group B carbohydrate (GBC) is crucial for peptidoglycan biosynthesis and cell division. Despite the important roles of CPS and GBC, there is little information available on the molecular organization of these glycopolymers on the cell surface. Here, we use atomic force microscopy (AFM) and transmission electron microscopy (TEM) to analyze the nanoscale distribution of CPS and GBC in wild-type (WT) and mutant strains of S. agalactiae. TEM analyses reveal that in WT bacteria, peptidoglycan is covered with a very thin (few nm) layer of GBC (the ``pellicle'') overlaid by a 15-45 nm thick layer of CPS (the ``capsule''). AFM-based single-molecule mapping with specific antibody probes shows that CPS is exposed on WT cells, while it is hardly detected on mutant cells impaired in CPS production (ΔcpsE mutant). By contrast, both TEM and AFM show that CPS is over-expressed in mutant cells altered in GBC expression (ΔgbcO mutant), indicating that the production of the two surface glycopolymers is coordinated in WT cells. In addition, AFM topographic imaging and molecular mapping with specific lectin probes demonstrate that removal of CPS (ΔcpsE), but not of GBC (ΔgbcO), leads to the exposure of peptidoglycan, organized into 25 nm wide bands running parallel to the septum. These results indicate that CPS forms a homogeneous barrier protecting the underlying peptidoglycan from environmental exposure, while the presence of GBC does not prevent peptidoglycan detection. This work shows that single-molecule AFM, combined with high-resolution TEM, represents a powerful platform for analysing the molecular arrangement of the cell wall polymers of bacterial pathogens.

  1. Comparative genome analysis identifies two large deletions in the genome of highly-passaged attenuated Streptococcus agalactiae strain YM001 compared to the parental pathogenic strain HN016.

    Science.gov (United States)

    Wang, Rui; Li, Liping; Huang, Yan; Luo, Fuguang; Liang, Wanwen; Gan, Xi; Huang, Ting; Lei, Aiying; Chen, Ming; Chen, Lianfu

    2015-11-04

    Streptococcus agalactiae (S. agalactiae), also known as group B Streptococcus (GBS), is an important pathogen for neonatal pneumonia, meningitis, bovine mastitis, and fish meningoencephalitis. The global outbreaks of Streptococcus disease in tilapia cause huge economic losses and threaten human food hygiene safety as well. To investigate the mechanism of S. agalactiae pathogenesis in tilapia and develop attenuated S. agalactiae vaccine, this study sequenced and comparatively analyzed the whole genomes of virulent wild-type S. agalactiae strain HN016 and its highly-passaged attenuated strain YM001 derived from tilapia. We performed Illumina sequencing of DNA prepared from strain HN016 and YM001. Sequencedreads were assembled and nucleotide comparisons, single nucleotide polymorphism (SNP) , indels were analyzed between the draft genomes of HN016 and YM001. Clustered regularly interspaced short palindromic repeats (CRISPRs) and prophage were detected and analyzed in different S. agalactiae strains. The genome of S. agalactiae YM001 was 2,047,957 bp with a GC content of 35.61 %; it contained 2044 genes and 88 RNAs. Meanwhile, the genome of S. agalactiae HN016 was 2,064,722 bp with a GC content of 35.66 %; it had 2063 genes and 101 RNAs. Comparative genome analysis indicated that compared with HN016, YM001 genome had two significant large deletions, at the sizes of 5832 and 11,116 bp respectively, resulting in the deletion of three rRNA and ten tRNA genes, as well as the deletion and functional damage of ten genes related to metabolism, transport, growth, anti-stress, etc. Besides these two large deletions, other ten deletions and 28 single nucleotide variations (SNVs) were also identified, mainly affecting the metabolism- and growth-related genes. The genome of attenuated S. agalactiae YM001 showed significant variations, resulting in the deletion of 10 functional genes, compared to the parental pathogenic strain HN016. The deleted and mutated functional genes all

  2. Effect of Excoecaria agallocha on non-specific immune responses and disease resistance of Oreochromis niloticus against Streptococcus agalactiae.

    Science.gov (United States)

    Laith, A A; Mazlan, A G; Effendy, A W; Ambak, M A; Nurhafizah, W W I; Alia, A S; Jabar, A; Najiah, M

    2017-06-01

    The current study was designed to evaluate the effects of Excoecaria agallocha leaf extracts on immune mechanisms and resistance of tilapia, Oreochromis niloticus, after challenge with Streptococcus agalactiae. Fish were divided into 6 groups; groups 1-5 fed with E. agallocha leaf extracts at 10, 20, 30, 40 and 50mgkg -1 level, respectively. Group 6 were fed without extract addition and acted as control. E. agallocha extracts were administered as feed supplement in fish diet for 28days and the hematological, immunological, and growth performance studies were conducted. Fish were infected with S. agalactiae at a dose of 15×105CFUmL -1 and the total white blood cell (WBC), phagocytosis and respiratory burst activities of leukocytes, serum bactericidal activity, lysozyme, total protein, albumin, and globulin levels were monitored and mortalities recorded for 15days post infection. Results revealed that feeding O. niloticus with 50mgkg -1 of E. agallocha enhanced WBC, phagocytic, respiratory burst, serum bactericidal and lysozyme activities on day 28 pre-challenge and on 3rd, 6th, 9th, 12th and 15th day post-challenge as compared to control. Total protein and albumin were not enhanced by E. agallocha diet. E. agallocha increased the survival of fish after challenge with S. agalactiae. The highest mortality rate (97%) was observed in control fish and the lowest mortality (27%) was observed with group fed with 50mgkg -1 extract. The results indicate that dietary intake of E. agallocha methanolic leaf extract in O. niloticus enhances the non-specific immunity and disease resistance against S. agalactiae pathogen. Copyright © 2017. Published by Elsevier Ltd.

  3. An Overview of Vaccination Strategies and Antigen Delivery Systems for Streptococcus agalactiae Vaccines in Nile Tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Munang'andu, Hetron Mweemba; Paul, Joydeb; Evensen, Øystein

    2016-12-13

    Streptococcus agalactiae is an emerging infectious disease adversely affecting Nile tilapia ( Niloticus oreochromis ) production in aquaculture. Research carried out in the last decade has focused on developing protective vaccines using different strategies, although no review has been carried out to evaluate the efficacy of these strategies. The purpose of this review is to provide a synopsis of vaccination strategies and antigen delivery systems currently used for S. agalactiae vaccines in tilapia. Furthermore, as shown herein, current vaccine designs include the use of replicative antigen delivery systems, such as attenuated virulent strains, heterologous vectors and DNA vaccines, while non-replicative vaccines include the inactivated whole cell (IWC) and subunit vaccines encoding different S. agalactiae immunogenic proteins. Intraperitoneal vaccination is the most widely used immunization strategy, although immersion, spray and oral vaccines have also been tried with variable success. Vaccine efficacy is mostly evaluated by use of the intraperitoneal challenge model aimed at evaluating the relative percent survival (RPS) of vaccinated fish. The major limitation with this approach is that it lacks the ability to elucidate the mechanism of vaccine protection at portals of bacterial entry in mucosal organs and prevention of pathology in target organs. Despite this, indications are that the correlates of vaccine protection can be established based on antibody responses and antigen dose, although these parameters require optimization before they can become an integral part of routine vaccine production. Nevertheless, this review shows that different approaches can be used to produce protective vaccines against S. agalactiae in tilapia although there is a need to optimize the measures of vaccine efficacy.

  4. An Overview of Vaccination Strategies and Antigen Delivery Systems for Streptococcus agalactiae Vaccines in Nile Tilapia (Oreochromis niloticus)

    Science.gov (United States)

    Munang’andu, Hetron Mweemba; Paul, Joydeb; Evensen, Øystein

    2016-01-01

    Streptococcus agalactiae is an emerging infectious disease adversely affecting Nile tilapia (Niloticus oreochromis) production in aquaculture. Research carried out in the last decade has focused on developing protective vaccines using different strategies, although no review has been carried out to evaluate the efficacy of these strategies. The purpose of this review is to provide a synopsis of vaccination strategies and antigen delivery systems currently used for S. agalactiae vaccines in tilapia. Furthermore, as shown herein, current vaccine designs include the use of replicative antigen delivery systems, such as attenuated virulent strains, heterologous vectors and DNA vaccines, while non-replicative vaccines include the inactivated whole cell (IWC) and subunit vaccines encoding different S. agalactiae immunogenic proteins. Intraperitoneal vaccination is the most widely used immunization strategy, although immersion, spray and oral vaccines have also been tried with variable success. Vaccine efficacy is mostly evaluated by use of the intraperitoneal challenge model aimed at evaluating the relative percent survival (RPS) of vaccinated fish. The major limitation with this approach is that it lacks the ability to elucidate the mechanism of vaccine protection at portals of bacterial entry in mucosal organs and prevention of pathology in target organs. Despite this, indications are that the correlates of vaccine protection can be established based on antibody responses and antigen dose, although these parameters require optimization before they can become an integral part of routine vaccine production. Nevertheless, this review shows that different approaches can be used to produce protective vaccines against S. agalactiae in tilapia although there is a need to optimize the measures of vaccine efficacy. PMID:27983591

  5. INHIBITION OF Malus sylvestris Mill. PEELEXTRACT USING ETANOL SOLVENT ON THE GROWTH OF Streptococcus agalactiae AND Escherichia coli CAUSING MASTITIS

    Directory of Open Access Journals (Sweden)

    Kanzul Kamal Putra

    2017-03-01

    Full Text Available The purpose of this research was to find the resistibility of Manalagi apple peel extract, using etanol, to the growth of was to determine the antibacterial activity of Manalagi apple peel (Malus sylvestris Mill extract in various solvent using ethanol concentration against the growth of Streptococcus agalactiae and Escherichia coli bacteria that causing mastitis.The research methodwas experimental using Completely Randomized Design with 4 treatments and 6 replication. The treatments consisted of P1 (10%, P2 (20%, P3 (50% concentrations and P0 (10% iodips as the control. The variable measured was diameter of inhibition zone. The data were analyzed using ANOVA and continued by Duncan’s New Multiple Range Test (DMRT test if there was significantly difference result. The result of the inhibition zone of Manalagi apple peel extract using etanol in preventing the growth of Streptococcus agalactiae and Escherichia coli bacteria was different (P<0,01. In P2 (30% concentration, the extract resistibility to the growth of Streptococcus agalactiae bacteria was equivalent to P0 (iodips and in P3 (50% concentration, the extract resistibility to Escherichia coli bacteria was greater than P0 (iodips. Manalagi apple peel extract using etanol can be used as a natural antiseptic solution for teat dipping on dairy cows. The recommendation from the research was using extract Manalagi apple peel with etanol solvent concentration of 30% as a solution of teat dipping.   Keywords : Manalagi apple peel, Teat dipping, Mastitis, Streptococcus agalactiaeand Escherichia coli

  6. Quantitative assessment of the blood-brain barrier opening caused by Streptococcus agalactiae hyaluronidase in a BALB/c mouse model.

    Science.gov (United States)

    Luo, Su; Cao, Qing; Ma, Ke; Wang, Zhaofei; Liu, Guangjin; Lu, Chengping; Liu, Yongjie

    2017-10-19

    Streptococcus agalactiae is a pathogen causing meningitis in animals and humans. However, little is known about the entry of S. agalactiae into brain tissue. In this study, we developed a BALB/c mouse model based on the intravenous injection of β-galactosidase-positive Escherichia coli M5 as an indicator of blood-brain barrier (BBB) opening. Under physiological conditions, the BBB is impermeable to E. coli M5. In pathological conditions caused by S. agalactiae, E. coli M5 is capable of penetrating the brain through a disrupted BBB. The level of BBB opening can be assessed by quantitative measurement of E. coli M5 loads per gram of brain tissue. Further, we used the model to evaluate the role of S. agalactiae hyaluronidase in BBB opening. The inactivation of hylB gene encoding a hyaluronidase, HylB, resulted in significantly decreased E. coli M5 colonization, and the intravenous injection of purified HylB protein induced BBB opening in a dose-dependent manner. This finding verified the direct role of HylB in BBB invasion and traversal, and further demonstrated the practicability of the in vivo mouse model established in this study. This model will help to understand the S. agalactiae-host interactions that are involved in this bacterial traversal of the BBB and to develop efficacious strategies to prevent central nervous system infections.

  7. NATURAL MUTATION IN THE GENE OF RESPONSE REGULATOR BgrR RESULTING IN REPRESSION OF Bac PROTEIN SYNTHESIS, A PATHOGENICITY FACTOR OF STREPTOCOCCUS AGALACTIAE

    Directory of Open Access Journals (Sweden)

    A. S. Rozhdestvenskaya

    2013-01-01

    Full Text Available Abstract. Streptococcus agalactiae can cause variety of diseases of newborns and adults. For successful colonization of different human tissues and organs as well as for suppression of the host immune system S. agalactiae expresses numerous virulence factors. For coordinated expression of the virulence genes S. agalactiae employs regulatory molecules including regulatory proteins of two-component systems. Results of the present study demonstrated that in S. agalactiae strain A49V the natural mutation in the brgR gene encoding for BgrR regulatory protein, which is component of regulatory system BgrRS, resulted in the repression of Bac protein synthesis, a virulence factor of S. agalactiae. A single nucleotide deletion in the bgrR gene has caused a shift of the reading frame and the changes in the primary, secondary and tertiary structures of the BgrR protein. The loss of functional activity of BgrR protein in A49V strain and repression of Bac protein synthesis have increased virulence of the strain in experimental animal streptococcal infection.

  8. Survey of immunological features of the alpha-like proteins of Streptococcus agalactiae.

    Science.gov (United States)

    Maeland, Johan A; Afset, Jan E; Lyng, Randi V; Radtke, Andreas

    2015-02-01

    Nearly all Streptococcus agalactiae (group B streptococcus [GBS]) strains express a protein which belongs to the so-called alpha-like proteins (Alps), of which Cα, Alp1, Alp2, Alp3, Rib, and Alp4 are known to occur in GBS. The Alps are chimeras which form mosaic structures on the GBS surface. Both N- and C-terminal stretches of the Alps possess immunogenic sites of dissimilar immunological specificity. In this review, we have compiled data dealing with the specificity of the N- and C-terminal immunogenic sites of the Alps. The majority of N-terminal sites show protein specificity while the C-terminal sites show broader cross-reactivity. Molecular serotyping has revealed that antibody-based serotyping has often resulted in erroneous Alp identification, due to persistence of cross-reacting antibodies in antisera for serotyping. Retrospectively, this could be expected on the basis of sequence analysis results. Some of the historical R proteins are in fact Alps. The data included in the review may provide a basis for decisions regarding techniques for the preparation of specific antisera for serotyping of GBS, for use in other approaches in GBS research, and for decision making in the context of GBS vaccine developments. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Remodeling of the Streptococcus agalactiae transcriptome in response to growth temperature.

    Directory of Open Access Journals (Sweden)

    Laurent Mereghetti

    Full Text Available BACKGROUND: To act as a commensal bacterium and a pathogen in humans and animals, Streptococcus agalactiae (group B streptococcus, GBS must be able to monitor and adapt to different environmental conditions. Temperature variation is a one of the most commonly encountered variables. METHODOLOGY/PRINCIPAL FINDINGS: To understand the extent to which GBS modify gene expression in response to temperatures encountered in the various hosts, we conducted a whole genome transcriptome analysis of organisms grown at 30 degrees C and 40 degrees C. We identified extensive transcriptome remodeling at various stages of growth, especially in the stationary phase (significant transcript changes occurred for 25% of the genes. A large proportion of genes involved in metabolism was up-regulated at 30 degrees C in stationary phase. Conversely, genes up-regulated at 40 degrees C relative to 30 degrees C include those encoding virulence factors such as hemolysins and extracellular secreted proteins with LPXTG motifs. Over-expression of hemolysins was linked to larger zones of hemolysis and enhanced hemolytic activity at 40 degrees C. A key theme identified by our study was that genes involved in purine metabolism and iron acquisition were significantly up-regulated at 40 degrees C. CONCLUSION/SIGNIFICANCE: Growth of GBS in vitro at different temperatures resulted in extensive remodeling of the transcriptome, including genes encoding proven and putative virulence genes. The data provide extensive new leads for molecular pathogenesis research.

  10. In vitro characterization of PlySK1249, a novel phage lysin, and assessment of its antibacterial activity in a mouse model of Streptococcus agalactiae bacteremia.

    Science.gov (United States)

    Oechslin, Frank; Daraspe, Jean; Giddey, Marlyse; Moreillon, Philippe; Resch, Grégory

    2013-12-01

    Beta-hemolytic Streptococcus agalactiae is the leading cause of bacteremia and invasive infections. These diseases are treated with β-lactams or macrolides, but the emergence of less susceptible and even fully resistant strains is a cause for concern. New bacteriophage lysins could be promising alternatives against such organisms. They hydrolyze the bacterial peptidoglycan at the end of the phage cycle, in order to release the phage progeny. By using a bioinformatic approach to screen several beta-hemolytic streptococci, a gene coding for a lysin was identified on a prophage carried by Streptococcus dysgalactiae subsp. equisimilis SK1249. The gene product, named PlySK1249, harbored an original three-domain structure with a central cell wall-binding domain surrounded by an N-terminal amidase and a C-terminal CHAP domain. Purified PlySK1249 was highly lytic and bactericidal for S. dysgalactiae (2-log10 CFU/ml decrease within 15 min). Moreover, it also efficiently killed S. agalactiae (1.5-log10 CFU/ml decrease within 15 min) but not several streptococcal commensal species. We further investigated the activity of PlySK1249 in a mouse model of S. agalactiae bacteremia. Eighty percent of the animals (n = 10) challenged intraperitoneally with 10(6) CFU of S. agalactiae died within 72 h, whereas repeated injections of PlySK1249 (45 mg/kg 3 times within 24 h) significantly protected the mice (P S. dysgalactiae, demonstrated high cross-lytic activity against S. agalactiae both in vitro and in vivo. These encouraging results indicated that PlySK1249 might represent a good candidate to be developed as a new enzybiotic for the treatment of systemic S. agalactiae infections.

  11. Molecular Methods for Typing of Streptococcus agalactiae with Special Emphasis on the Development and Validation of a Multi-Locus Variable Number of Tandem Repeats Assay (MLVA)

    OpenAIRE

    Radtke, Andreas

    2012-01-01

    Molekylære metoder for typing av Streptococcus agalactiae med særlig vektlegging av utvikling og validering av et multi-locus variable number of tandem repeats assay (MLVA) Sammendraget: Streptococcus agalactiae eller gruppe B streptokokker (GBS) forårsaker livsfarlige infeksjoner hos nyfødte, gravide eller voksne med kroniske sykdommer. Den forårsaker også jurbetennelse i storfe. Typing av GBS gir innblikk i bakteriens epidemiologi og dens fylogenetiske slektskap. Ulike deler av bakterie...

  12. RovS and its associated signaling peptide form a cell-to-cell communication system required for Streptococcus agalactiae pathogenesis.

    Science.gov (United States)

    Pérez-Pascual, David; Gaudu, Philippe; Fleuchot, Betty; Besset, Colette; Rosinski-Chupin, Isabelle; Guillot, Alain; Monnet, Véronique; Gardan, Rozenn

    2015-01-20

    Bacteria can communicate with each other to coordinate their biological functions at the population level. In a previous study, we described a cell-to-cell communication system in streptococci that involves a transcriptional regulator belonging to the Rgg family and short hydrophobic peptides (SHPs) that act as signaling molecules. Streptococcus agalactiae, an opportunistic pathogenic bacterium responsible for fatal infections in neonates and immunocompromised adults, has one copy of the shp/rgg locus. The SHP-associated Rgg is called RovS in S. agalactiae. In this study, we found that the SHP/RovS cell-to-cell communication system is active in the strain NEM316 of S. agalactiae, and we identified different partners that are involved in this system, such as the Eep peptidase, the PptAB, and the OppA1-F oligopeptide transporters. We also identified a new target gene controlled by this system and reexamined the regulation of a previously proposed target gene, fbsA, in the context of the SHP-associated RovS system. Furthermore, our results are the first to indicate the SHP/RovS system specificity to host liver and spleen using a murine model, which demonstrates its implication in streptococci virulence. Finally, we observed that SHP/RovS regulation influences S. agalactiae's ability to adhere to and invade HepG2 hepatic cells. Hence, the SHP/RovS cell-to-cell communication system appears to be an essential mechanism that regulates pathogenicity in S. agalactiae and represents an attractive target for the development of new therapeutic strategies. Rgg regulators and their cognate pheromones, called small hydrophobic peptides (SHPs), are present in nearly all streptococcal species. The general pathways of the cell-to-cell communication system in which Rgg and SHP take part are well understood. However, many other players remain unidentified, and the direct targets of the system, as well as its link to virulence, remain unclear. Here, we identified the different players

  13. Evaluation of the efficacy of intramuscular versus intramammary treatment of subclinical Streptococcus agalactiae mastitis in dairy cows in Colombia.

    Science.gov (United States)

    Reyes, J; Chaffer, M; Sanchez, J; Torres, G; Macias, D; Jaramillo, M; Duque, P C; Ceballos, A; Keefe, G P

    2015-08-01

    A randomized controlled trial was performed in 17 Colombian dairy herds to determine the cure risk among cows subclinically infected with Streptococcus agalactiae exposed to 2 antibiotic therapies. Composite milk samples were collected before milking at the onset of the trial (pretreatment) and 2 subsequent times over a period of approximately 63 d. The intramammary application (IMM) of ampicillin-cloxacillin was compared with the intramuscular application (IM) of penethamate hydriodide, and cure risks after an initial and retreatment application were assessed. Cure risk after the initial treatment was higher (82.4%) for the IMM treatment than for IM therapy (65.8%). However, no difference was observed in the cure risk of refractory cases after retreatment (IMM=52.6% vs. IM=51.2%). The cumulative cure risk (both initial and retreatment) was 90.4 and 82.9% for the IMM and IM products, respectively. A 2-level random effects logistic model that controlled for pretreatment cow-level somatic cell count, indicated that IM treatment (odds ratio=0.37) had a lower cure risk than IMM and a tendency for a lower cure risk with increasing baseline somatic cell count. Our findings suggest that both products and administration routes can reduce the prevalence of S. agalactiae in affected herds, but the IMM product had a better efficacy in curing the infection. In addition to the treatment protocol, the cow somatic cell count should be considered when making management decisions for cows infected with S. agalactiae. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Role of the Group B antigen of Streptococcus agalactiae: a peptidoglycan-anchored polysaccharide involved in cell wall biogenesis.

    Directory of Open Access Journals (Sweden)

    Élise Caliot

    Full Text Available Streptococcus agalactiae (Group B streptococcus, GBS is a leading cause of infections in neonates and an emerging pathogen in adults. The Lancefield Group B carbohydrate (GBC is a peptidoglycan-anchored antigen that defines this species as a Group B Streptococcus. Despite earlier immunological and biochemical characterizations, the function of this abundant glycopolymer has never been addressed experimentally. Here, we inactivated the gene gbcO encoding a putative UDP-N-acetylglucosamine-1-phosphate:lipid phosphate transferase thought to catalyze the first step of GBC synthesis. Indeed, the gbcO mutant was unable to synthesize the GBC polymer, and displayed an important growth defect in vitro. Electron microscopy study of the GBC-depleted strain of S. agalactiae revealed a series of growth-related abnormalities: random placement of septa, defective cell division and separation processes, and aberrant cell morphology. Furthermore, vancomycin labeling and peptidoglycan structure analysis demonstrated that, in the absence of GBC, cells failed to initiate normal PG synthesis and cannot complete polymerization of the murein sacculus. Finally, the subcellular localization of the PG hydrolase PcsB, which has a critical role in cell division of streptococci, was altered in the gbcO mutant. Collectively, these findings show that GBC is an essential component of the cell wall of S. agalactiae whose function is reminiscent of that of conventional wall teichoic acids found in Staphylococcus aureus or Bacillus subtilis. Furthermore, our findings raise the possibility that GBC-like molecules play a major role in the growth of most if not all beta-hemolytic streptococci.

  15. In silico analysis of candidate proteins sharing homology with Streptococcus agalactiae proteins and their role in male infertility.

    Science.gov (United States)

    Parida, Rajeshwari; Samanta, Luna

    2017-02-01

    Leukocytospermia is a physiologic condition defined as human semen with a leukocyte count of >1 x 10 6 cells/ml that is often correlated with male infertility. Moreover, bacteriospermia has been associated with leukocytospermia ultimately leading to male infertility. We have found that semen samples with >1 x 10 6 /ml leukocytes and/or bacteriospermia have oxidative predominance as evidenced by augmented protein carbonyl and lipid peroxidation status of the semen which is implicated in sperm dysfunction. It has been reported that Streptococcus agalactiae is present in bacteriospermic samples. Previous research has shown that human leukocyte antigen beta chain paralog (HLA-DRB) alleles interact best with the infected sperm cells rather than the non-infected cells. Little is known about the interaction of major histocompatibility complex (MHC) present on leukocytes with the sperm upon bacterial infection and how it induces an immunological response which we have addressed by epitope mapping. Therefore, we examined MHC class II derived bacterial peptides which might have human sperm-related functional aspects. Twenty-two S. agalactiae proteins were obtained from PUBMED protein database for our study. Protein sequences with more than two accession numbers were aligned using CLUSTAL Omega to check their conservation pattern. Each protein sequence was then analyzed for T-cell epitope prediction against HLA-DRB alleles using the immune epitope database (IEDB) analysis tool. Out of a plethora of peptides obtained from this analysis, peptides corresponding to proteins of interest such as DNA binding response regulator, hyaluronate lyase and laminin binding protein were screened against the human proteome using Blastp. Interestingly, we have found bacterial peptides sharing homology with human peptides deciphering some of the important sperm functions. Antibodies raised against these probable bacterial antigens of fertility will not only help us understand the mechanism of

  16. Influencia de la composici??n del medio de cultivo sobre la producci??n de pigmento por Streptococcus agalactiae

    OpenAIRE

    Carazo Carazo, Concepci??n

    1995-01-01

    Se ha estudiado la influencia de la composici??n del medio de cultivo sobre la producci??n de pigmento por s. agalactiae partiendo de un medio de cultivo dise??ado para tal finalidad (new granada medium), eliminando uno a uno los componentes incluidos en su formulaci??n original. Almid??n, proteosa peptona (difco), suero, fosfatos y metrotexato influyen de forma importante en la formaci??n y/o visualizaci??n del pigmento producido por este microorganismo. Se ha determinado que la fracci??n de...

  17. Streptococcus agalactiae: HASTA AHORA EL ÚNICO Streptococcus PATÓGENO DE TILAPIAS CULTIVADAS EN COLOMBIA

    Directory of Open Access Journals (Sweden)

    AP Jiménez

    2007-01-01

    Full Text Available Las estreptococosis son un conjunto de enfermedades ocasionadas por un grupo de cocos Gram positivos con similares signologías que involucran distintos órganos en los individuos afectados. La identiÀcación precisa de cada uno de estos microorganismos no se logra de manera deÀnitiva por los métodos tradicionales microbiológicos, por lo que se debe acudir a otro tipo de metodologías como las técnicas de biología molecular. En 1999 se identiÀcó por primera vez en Colombia la estreptococosis en híbridos de tilapias de cultivo. La pos-terior secuenciación del ADN de distintos aislamientos obtenidos de varias regiones del país demostró un 98,8% de aÀnidad con el Streptococcus agalactiae. El presente estudio pretende deÀnir si hasta la fecha existe solo esta especie de Streptococcus en el país causan-do infección o enfermedad en tilapias de cultivo o, por el contrario, son varias las especies que intervienen en los cuadros infecciosos. Se evaluaron aislamientos de tejidos, de agua y fango de los sitios de cultivo, así como de lugares de expendio de tilapia roja (Oreochromissp., utilizando técnicas microbiológicas, inmunoperoxidasa indirecta (IPI y PCR, espe-cíÀcas para el aislamiento e identiÀcación del S. agalactiae. Los resultados del presente estudio demostraron que hasta la fecha en el país únicamente se ha identiÀcado la especie S. agalactiae causando infección o enfermedad en tilapias. No tenemos evidencia de que otros Streptococcus reportados internacionalmente como S. iniae y otros Gram positivos causen estreptococosis en Colombia.La tilapia parece ser el principal reservorio del S. agalactiae en el país, y el riesgo zoonótico, aunque existe, es mínimo si se toman las medidas apropia-das de bioseguridad.

  18. Interaction of Streptococcus agalactiae and cellular innate immunity in colonization and disease

    Directory of Open Access Journals (Sweden)

    Sybille eLandwehr-Kenzel

    2014-10-01

    Full Text Available Streptococcus agalactiae (Group B streptococcus, GBS is highly adapted to humans, where it is a normal constituent of the intestinal and vaginal flora. Yet, GBS has highly invasive potential and causes excessive inflammation, sepsis and death at the beginning of life, in the elderly and in diabetic patients. Thus GBS is a model pathobiont that thrives in the healthy host, but has not lost its potential virulence during coevolution with mankind. It remains incompletely understood how the innate immune system contains GBS in the natural niches, the intestinal and genital tracts, and which molecular events underlie breakdown of mucocutaneous resistance. Newborn infants between days seven and 90 of life are at risk of a particularly striking sepsis manifestation (late onset disease, LOD, where the transition from colonization to invasion and dissemination, and thus from health to severe sepsis is typically fulminant and not predictable. The great majority of late-onset sepsis cases is caused by one clone, GBS ST-17, which expresses HvgA as a signature virulence factor and adhesin. In mice, HvgA promotes the crossing of both the mucosal and the blood brain barrier. Expression levels of HvgA and other GBS virulence factors, such as pili and toxins, are regulated by the upstream two-component control system CovR/S. This in turn is modulated by acidic epithelial pH, high glucose levels and during the passage through the mouse intestine. After invasion, GBS has the ability to subvert innate immunity by mechanisms like GAPDH-dependent induction of IL-10 and β-protein binding to the inhibitory phagocyte receptors sialic acid binding immunoglobulin-like lectin 5 and 14. On the host side, sensing of GBS nucleic acids and lipopeptides by both Toll-like receptors (TLRs and the inflammasome appears to be critical for host resistance against GBS. Yet, comprehensive models on the interplay between GBS and human immune cells at the colonizing site are just

  19. Serine-rich repeat proteins and pili promote Streptococcus agalactiae colonization of the vaginal tract.

    Science.gov (United States)

    Sheen, Tamsin R; Jimenez, Alyssa; Wang, Nai-Yu; Banerjee, Anirban; van Sorge, Nina M; Doran, Kelly S

    2011-12-01

    Streptococcus agalactiae (group B streptococcus [GBS]) is a Gram-positive bacterium found in the female rectovaginal tract and is capable of producing severe disease in susceptible hosts, including newborns and pregnant women. The vaginal tract is considered a major reservoir for GBS, and maternal vaginal colonization poses a significant risk to the newborn; however, little is known about the specific bacterial factors that promote GBS colonization and persistence in the female reproductive tract. We have developed in vitro models of GBS interaction with the human female cervicovaginal tract using human vaginal and cervical epithelial cell lines. Analysis of isogenic mutant GBS strains deficient in cell surface organelles such as pili and serine-rich repeat (Srr) proteins shows that these factors contribute to host cell attachment. As Srr proteins are heavily glycosylated, we confirmed that carbohydrate moieties contribute to the effective interaction of Srr-1 with vaginal epithelial cells. Antibody inhibition assays identified keratin 4 as a possible host receptor for Srr-1. Our findings were further substantiated in an in vivo mouse model of GBS vaginal colonization, where mice inoculated with an Srr-1-deficient mutant exhibited decreased GBS vaginal persistence compared to those inoculated with the wild-type (WT) parental strain. Furthermore, competition experiments in mice showed that WT GBS exhibited a significant survival advantage over the ΔpilA or Δsrr-1 mutant in the vaginal tract. Our results suggest that these GBS surface proteins contribute to vaginal colonization and may offer new insights into the mechanisms of vaginal niche establishment.

  20. Serine-Rich Repeat Proteins and Pili Promote Streptococcus agalactiae Colonization of the Vaginal Tract ▿

    Science.gov (United States)

    Sheen, Tamsin R.; Jimenez, Alyssa; Wang, Nai-Yu; Banerjee, Anirban; van Sorge, Nina M.; Doran, Kelly S.

    2011-01-01

    Streptococcus agalactiae (group B streptococcus [GBS]) is a Gram-positive bacterium found in the female rectovaginal tract and is capable of producing severe disease in susceptible hosts, including newborns and pregnant women. The vaginal tract is considered a major reservoir for GBS, and maternal vaginal colonization poses a significant risk to the newborn; however, little is known about the specific bacterial factors that promote GBS colonization and persistence in the female reproductive tract. We have developed in vitro models of GBS interaction with the human female cervicovaginal tract using human vaginal and cervical epithelial cell lines. Analysis of isogenic mutant GBS strains deficient in cell surface organelles such as pili and serine-rich repeat (Srr) proteins shows that these factors contribute to host cell attachment. As Srr proteins are heavily glycosylated, we confirmed that carbohydrate moieties contribute to the effective interaction of Srr-1 with vaginal epithelial cells. Antibody inhibition assays identified keratin 4 as a possible host receptor for Srr-1. Our findings were further substantiated in an in vivo mouse model of GBS vaginal colonization, where mice inoculated with an Srr-1-deficient mutant exhibited decreased GBS vaginal persistence compared to those inoculated with the wild-type (WT) parental strain. Furthermore, competition experiments in mice showed that WT GBS exhibited a significant survival advantage over the ΔpilA or Δsrr-1 mutant in the vaginal tract. Our results suggest that these GBS surface proteins contribute to vaginal colonization and may offer new insights into the mechanisms of vaginal niche establishment. PMID:21984789

  1. Optimización de metodologías de cribaje para la búsqueda de Streptococcus agalactiae en embarazadas Optimization of screening methodologies for the detection of Streptococcus agalactiae in pregnant women

    Directory of Open Access Journals (Sweden)

    Silvia E. Montibello

    2011-03-01

    Full Text Available Streptococcus agalactiae es una causa importante de morbimortalidad en mujeres embarazadas y neonatos en todo el mundo. El objetivo del presente trabajo fue determinar la utilidad del medio cromogénico chromID Strepto B de bioMérieux para detectar S. agalactiae en embarazadas cuando la muestra es sembrada directamente en dicho medio o después del enriquecimiento en caldo de Todd Hewitt selectivo, opciones que se compararon con la metodología propuesta por el CDC . Se analizaron 1924 hisopados, 962 de introito vaginal y 962 rectales, correspondientes a 962 embarazadas entre la semana 35 y 37 de gestación, asistidas en distintos hospitales. Los hisopados se sembraron directamente en el medio chromID Strepto B (CR y luego se colocaron en un caldo de Todd Hewitt selectivo, suplementado con 15 µg/ml de ácido nalidíxico y 10 µg/ml de colistina (CTH-sel. Luego de 24 h de incubación, se realizaron subcultivos en el medio CR y en agar con 5% de sangre de carnero (ASO. La prevalencia global de S. agalactiae fue de 17,4%. La sensibilidad, la especificidad y los valores predictivos positivo y negativo del subcultivo en CR del material desarrollado en el CTH -sel fueron 98,8%, 100%, 100% y 99,7% respectivamente, con una incubación de 48 h. Los valores correspondientes de la siembra directa fueron 57,8%, 100%, 100% y 90%. La sensibilidad del subcultivo en ASO del material desarrollado en el CTH -sel fue del 85%. Se destaca el excelente rendimiento del subcultivo en CR luego del enriquecimiento en caldo de Todd Hewitt selectivo en comparación con el método propuesto por el CDC.Streptococcus agalactiae is a significant worldwide cause of morbidity and mortality in pregnant women and their newborn infants. The objective of this work was to determine the usefulness of bioMrieux chromogenic medium chromID Strepto B (CR for detecting S. agalactiae in pregnant women from the selective Todd-Hewitt broth (sel-THB against the methods proposed by the CDC

  2. Spatiotemporal patterns, annual baseline and movement-related incidence of Streptococcus agalactiae infection in Danish dairy herds: 2000–2009

    DEFF Research Database (Denmark)

    Mweu, Marshal M.; Nielsen, Søren S.; Hisham Beshara Halasa, Tariq

    2014-01-01

    -herd sources of new herd infections coupled with the spatiotemporal distribution of the infection, may aid in effective targeting of control efforts. Thus, the objectives of this study were: (1) to describe the spatiotemporal patterns of infection with S. agalactiae in the population of Danish dairy herds from...

  3. Crystallization and preliminary crystallographic analysis of two Streptococcus agalactiae proteins: the family II inorganic pyrophosphatase and the serine/threonine phosphatase

    International Nuclear Information System (INIS)

    Rantanen, Mika K.; Lehtiö, Lari; Rajagopal, Lakshmi; Rubens, Craig E.; Goldman, Adrian

    2006-01-01

    Two S. agalactiae proteins, the inorganic pyrophosphatase and the serine/threonine phosphatase, were crystallized and diffraction data were collected and processed from these crystals. The data from the two protein crystals extended to 2.80 and 2.65 Å, respectively. Streptococcus agalactiae, which infects human neonates and causes sepsis and meningitis, has recently been shown to possess a eukaryotic-like serine/threonine protein phosphorylation signalling cascade. Through their target proteins, the S. agalactiae Ser/Thr kinase and Ser/Thr phosphatase together control the growth as well as the morphology and virulence of this organism. One of the targets is the S. agalactiae family II inorganic pyrophosphatase. The inorganic pyrophosphatase and the serine/threonine phosphatase have therefore been purified and crystallized and diffraction data have been collected from their crystals. The data were processed using XDS. The inorganic pyrosphosphatase crystals diffracted to 2.80 Å and the Ser/Thr phosphatase crystals to 2.65 Å. Initial structure-solution experiments indicate that structure solution will be successful in both cases. Solving the structure of the proteins involved in this cascade is the first step towards understanding this phenomenon in atomic detail

  4. A novel one-step real-time multiplex PCR assay to detect Streptococcus agalactiae presence and serotypes Ia, Ib, and III.

    Science.gov (United States)

    Furfaro, Lucy L; Chang, Barbara J; Payne, Matthew S

    2017-09-01

    Streptococcus agalactiae is the leading cause of early-onset neonatal sepsis. Culture-based screening methods lack the sensitivity of molecular assays and do not indicate serotype; a potentially important virulence marker. We aimed to develop a multiplex PCR to detect S. agalactiae while simultaneously identifying serotypes Ia, Ib, and III; commonly associated with infant disease. Primers were designed to target S. agalactiae serotype-specific cps genes and the dltS gene. The assay was validated with 512 vaginal specimens from pregnant women. 112 (21.9%) were dltS positive, with 14.3%, 0.9%, and 6.3% of these identified as cps Ia, Ib, and III, respectively. Our assay is a specific and sensitive method to simultaneously detect S. agalactiae and serotypes Ia, Ib, and III in a single step. It is of high significance for clinical diagnostic applications and also provides epidemiological data on serotype, information that may be important for vaccine development and other targeted non-antibiotic therapies. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Crystallization and preliminary crystallographic analysis of two Streptococcus agalactiae proteins: the family II inorganic pyrophosphatase and the serine/threonine phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Rantanen, Mika K.; Lehtiö, Lari [Institute of Biotechnology, University of Helsinki, PO Box 65, FIN-00014, Helsinki (Finland); Rajagopal, Lakshmi; Rubens, Craig E. [Division of Infectious Disease, Children’s Hospital and Regional Medical Center, Seattle, Washington 98105 (United States); Goldman, Adrian, E-mail: adrian.goldman@helsinki.fi [Institute of Biotechnology, University of Helsinki, PO Box 65, FIN-00014, Helsinki (Finland)

    2006-09-01

    Two S. agalactiae proteins, the inorganic pyrophosphatase and the serine/threonine phosphatase, were crystallized and diffraction data were collected and processed from these crystals. The data from the two protein crystals extended to 2.80 and 2.65 Å, respectively. Streptococcus agalactiae, which infects human neonates and causes sepsis and meningitis, has recently been shown to possess a eukaryotic-like serine/threonine protein phosphorylation signalling cascade. Through their target proteins, the S. agalactiae Ser/Thr kinase and Ser/Thr phosphatase together control the growth as well as the morphology and virulence of this organism. One of the targets is the S. agalactiae family II inorganic pyrophosphatase. The inorganic pyrophosphatase and the serine/threonine phosphatase have therefore been purified and crystallized and diffraction data have been collected from their crystals. The data were processed using XDS. The inorganic pyrosphosphatase crystals diffracted to 2.80 Å and the Ser/Thr phosphatase crystals to 2.65 Å. Initial structure-solution experiments indicate that structure solution will be successful in both cases. Solving the structure of the proteins involved in this cascade is the first step towards understanding this phenomenon in atomic detail.

  6. Laboratory-controlled Challenges of Nile Tilapia (Oreochromis niloticus) with Streptococcus agalactiae: Comparisons between Immersion, Oral, Intracoelomic and Intramuscular Routes of Infection.

    Science.gov (United States)

    Soto, E; Zayas, M; Tobar, J; Illanes, O; Yount, S; Francis, S; Dennis, M M

    2016-11-01

    Streptococcus agalactiae, the aetiological agent of streptococcosis in fish, is an important pathogen of cultured and wild fish worldwide. To gain a better understanding of the pathogenesis of streptococcosis in Nile tilapia (Oreochromis niloticus), and to identify the experimental route of infection that most closely mimics natural disease, fingerlings were challenged with S. agalactiae utilizing different delivery methods. Fingerlings were challenged via intracoelomic injection (ICinj), intramuscular injection (IMinj), orally or by immersion with serial dilutions of S. agalactiae. The dose lethal to 50% of test fish 15 days post challenge was 120 colony forming units (CFU)/fish after ICinj, and 10 5  CFU/fish after IMinj. Acute mortalities were present in both groups, but were higher in the fish challenged by ICinj. Very low mortalities were observed in the fish challenged via oral or immersion routes. Post-mortem evaluation of survivors revealed classical lesions associated with fish streptococcosis, including granulomatous or lymphohistiocytic epicarditis, splenitis, meningitis, myocarditis, choroiditis and exophthalmia. The information obtained improves our understanding of the pathogenesis of streptococcosis in fish, and provides useful information regarding controlled experimental infections in tilapia challenged with S. agalactiae. Results from this study suggest that IMinj challenge methods are not only suitable to induce streptococcosis in tilapia, but they may be the preferred method to study the pathogenesis of the naturally-occurring disease in this species. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Molecular and functional characterization of peptidoglycan-recognition protein SC2 (PGRP-SC2) from Nile tilapia (Oreochromis niloticus) involved in the immune response to Streptococcus agalactiae.

    Science.gov (United States)

    Gan, Zhen; Chen, Shannan; Hou, Jing; Huo, Huijun; Zhang, Xiaolin; Ruan, Baiye; Laghari, Zubair Ahmed; Li, Li; Lu, Yishan; Nie, Pin

    2016-07-01

    PGRP-SC2, the member of PGRP family, plays an important role in regulation of innate immune response. In this paper, a PGRP-SC2 gene of Nile tilapia, Oreochromis niloticus (designated as On-PGRP-SC2) was cloned and its expression pattern under the infection of Streptococcus agalactiae was investigated. Sequence analysis showed main structural features required for amidase activity were detected in the deduced amino acid sequence of On-PGRP-SC2. In healthy tilapia, the On-PGRP-SC2 transcripts could be detected in all the examined tissues, with the most abundant expression in the muscle. When infected with S. agalactiae, there was a clear time-dependent expression pattern of On-PGRP-SC2 in the spleen, head kidney and brain. The assays for the amidase activity suggested that recombinant On-PGRP-SC2 protein had a Zn(2+)-dependent PGN-degrading activity. Moreover, our works showed that recombinant On-PGRP-SC2 protein could significantly reduce bacterial load in target organs attacked by S. agalactiae. These findings indicated that On-PGRP-SC2 may play important roles in the immune response to S. agalactiae in Nile tilapia. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Involvement of cholinergic and adenosinergic systems on the branchial immune response of experimentally infected silver catfish with Streptococcus agalactiae.

    Science.gov (United States)

    Baldissera, M D; Souza, C F; Doleski, P H; Moreira, K L S; da Veiga, M L; da Rocha, M I U M; Santos, R C V; Baldisserotto, B

    2018-01-01

    It has been recognized that the cholinergic and adenosinergic systems have an essential role in immune and inflammatory responses during bacterial fish pathogens, such as the enzymes acetylcholinesterase (AChE) and adenosine deaminase (ADA), which are responsible for catalysis of the anti-inflammatory molecules acetylcholine (ACh) and adenosine (Ado) respectively. Thus, the aim of this study was to investigate the involvement of the cholinergic and adenosinergic systems on the immune response and inflammatory process in gills of experimentally infected Rhamdia quelen with Streptococcus agalactiae. Acetylcholinesterase activity decreased, while ACh levels increased in gills of infected animals compared to uninfected animals. On the other hand, a significant increase in ADA activity with a concomitant decrease in Ado levels was observed in infected animals compared to uninfected animals. Based on this evidence, we concluded that infection by S. agalactiae in silver catfish alters the cholinergic and adenosinergic systems, suggesting the involvement of AChE and ADA activities on immune and inflammatory responses, regulating the ACh and Ado levels. In summary, the downregulation of AChE activity exerts an anti-inflammatory profile in an attempt to reduce or prevent the tissue damage, while the upregulation of ADA activity exerts a pro-inflammatory profile, contributing to disease pathophysiology. © 2017 John Wiley & Sons Ltd.

  9. Characterization of the duodenase-1 gene and its associations with resistance to Streptococuus agalactiae in hybrid tilapia (Oreochromis spp.).

    Science.gov (United States)

    Shen, Yubang; Fu, Gui Hong; Liu, Feng; Yue, Gen Hua

    2015-08-01

    Tilapia is a group of cultured teleost fishes whose production is threatened by some diseases. Identification of DNA markers associated with disease resistance in candidate genes may facilitate to accelerate the selection of disease resistance. The gene encoding a duodenase, which can trigger immune response, has not been studied in fish. We characterized the cDNA of duodenase-1 gene of hybrid tilapia. Its ORF is 759 bp, encoding a serine protease of 252 amino acids. This gene consisted of five exons and four introns. Its expression was detected in all 10 tissues examined, and it was highly expressed in the intestine and kidney. After a challenge with the bacterial pathogen, Streptococcus agalactiae, its expression was up-regulated significantly in the intestine, liver and spleen. We identified seven SNPs in the gene and found that four of them were significantly associated with the resistance to S. agalactiae (P tilapia. The SNP markers in the duodenase-1 gene associated with resistance to the bacterial pathogen, may facilitate the selection of tilapia resistant to the bacterial disease. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Growth, immune responses and protection of Nile tilapia Oreochromis niloticus immunized with formalin-killed Streptococcus agalactiae serotype Ia and III vaccines

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    Atchariya Suwannasang

    2017-08-01

    Full Text Available The protective efficacy of formalin-killed Streptococcus agalactiae (Group B Streptococcus, GBS serotype Ia (GBS-Ia and III (GBS-III vaccines were assessed in Nile tilapia (Oreochromis niloticus. The fish with an average weight of 34.45± 0.08 g were immunized by intraperitoneal (i.p. injection with 4 different formalin-killed vaccines prepared from GBS-Ia (1x1010 CFU/mL, GBS-III (1x1010 CFU/mL, and combined GBS-Ia and GBS-III in an equal volume at final concentrations 1x1010 CFU/mL and 2x1010 CFU/mL in comparison with the non-immunized control group. At 2 and 4 weeks post vaccination, no significant differences were observed (p>0.05 among treatments in growth performance or haemato-immunological parameters, except the increased red blood cell at 2 weeks. Significantly increased antibody titers (p<0.05 against GBS-Ia and GBS-III antigens were noted in the groups immunized with homologous GBS vaccines, whereas the group reacted with heterologous GBS antigen showed less antibody titer as compared with the control group. The vaccination experiment indicated that i.p. injection of Nile tilapia with formalin-killed cells prepared from GBS-Ia or GBS-III provides significant protection, with relative percent survival (RPS value of 52.17 to 71.42%, against a challenge with the homologous serotype isolate, whereas the RPS in fish challenged with a heterologous serotype isolate varied from 20.00 to 53.57%. These results suggested that vaccines from either GBS-Ia or GBS-III have insufficient cross-protective efficacy against the other serotypes. However, a mixed vaccine produced from both GBS serotypes Ia and III provided significant protection with 65.00 to 95.66% RPS which could be an excellent vaccine to protect fish against streptococcosis caused by both GBS serotypes Ia and III.

  11. Vpma phase variation is important for survival and persistence of Mycoplasma agalactiae in the immunocompetent host.

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    Rohini Chopra-Dewasthaly

    2017-09-01

    Full Text Available Despite very small genomes, mycoplasmas retain large multigene families encoding variable antigens whose exact role in pathogenesis needs to be proven. To understand their in vivo significance, we used Mycoplasma agalactiae as a model exhibiting high-frequency variations of a family of immunodominant Vpma lipoproteins via Xer1-mediated site-specific recombinations. Phase-Locked Mutants (PLMs expressing single stable Vpma products served as first breakthrough tools in mycoplasmology to study the role of such sophisticated antigenic variation systems. Comparing the general clinical features of sheep infected with a mixture of phase-invariable PLMs (PLMU and PLMY and the wild type strain, it was earlier concluded that Vpma phase variation is not necessary for infection. Conversely, the current study demonstrates the in vivo indispensability of Vpma switching as inferred from the Vpma phenotypic and genotypic analyses of reisolates obtained during sheep infection and necropsy. PLMY and PLMU stably expressing VpmaY and VpmaU, respectively, for numerous in vitro generations, switched to new Vpma phenotypes inside the sheep. Molecular genetic analysis of selected 'switchover' clones confirmed xer1 disruption and revealed complex new rearrangements like chimeras, deletions and duplications in the vpma loci that were previously unknown in type strain PG2. Another novel finding is the differential infection potential of Vpma variants, as local infection sites demonstrated an almost complete dominance of PLMY over PLMU especially during early stages of both conjunctival and intramammary co-challenge infections, indicating a comparatively better in vivo fitness of VpmaY expressors. The data suggest that Vpma antigenic variation is imperative for survival and persistence inside the immunocompetent host, and although Xer1 is necessary for causing Vpma variation in vitro, it is not a virulence factor because alternative Xer1-independent mechanisms operate in

  12. Multicentre epidemiological survey on the incidence of Streptococcus agalactiae in pregnancy

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    Leonardo Lodolo

    2010-03-01

    Full Text Available ß-hemolytic streptococci group B (GBS cause infections of the urinary tract, endometrium, amnion, and superinfection of wounds, with complications in 2% -4% of cases in pregnant woman.The infection transmitted to newborns is a cause of sepsis and meningitis with high incidence of mortality.A recent report on a multicentric study of the ISS, showed that detection of S. agalactiae in pregnant women in different realities of healthcare, ranging from 3.9% to 19.4%. This survey covers a sample of 29607 women screened in a total of 36852 childbirths, with a positivity of 4183 subjects equal to 14.1%. Thanks to the GBS Study Group of Piemonte and Valle d’Aosta microbiology laboratories, with the support of 42 study centres have been involved. Collection techniques, types of medium, environmental of incubation of the culture, possible enrichment in broth, of the swabs of 28491 women during 2006/2007 have been considered. 24.36% were vaginal swab while 75.64% vagino/rectal. According to data collected at 31 centers with eligible data (22,175 pregnant GBS positivity was 12.7% (CI [Confidence Interval] + / -95%: 13.1% -12.2% with wide variability between individual centers (2% -22.6%. The enrichment broth is used by 6 out of 9 centers with positivity> 15% (M: 20.05%, from 6 to 14 with positivity between 10-15% (M: 12.61%, from none to 8 centers with less than 10% positivity (M: 4.95%.This multicentre survey underlined the preference for vagino/rectal swab, as recommended by the literature. The frequency of GBS positivity of pregnant women on the whole samples is similar to that found in other national surveys, even if with large differences between the different centers.The sensitivity tends to increase with the use of enrichment techniques, whose use is uneven between the centers, perhaps indicating the need for greater uniformity of protocols based on the results of microbiological analysis EBM (Evidence Based Medicine.

  13. Estimation of test characteristics of real-time PCR and bacterial culture for diagnosis of subclinical intramammary infections with Streptococcus agalactiae in Danish dairy cattle in 2012 using latent class analysis.

    Science.gov (United States)

    Mahmmod, Yasser S; Toft, Nils; Katholm, Jørgen; Grønbæk, Carsten; Klaas, Ilka C

    2013-05-01

    The misdiagnosis of intramammary infections (IMI) with Streptococcus agalactiae (S. agalactiae) could lead farmers to treat or cull animals unnecessarily. The objective of this field study was to estimate the sensitivity (Se) and specificity (Sp) of real-time PCR at different cut-offs for cycle threshold (Ct) values against bacterial culture (BC) for diagnosis of S. agalactiae IMI using latent class analysis to avoid the assumption of a perfect reference test. A total of 614 dairy cows were randomly selected from 6 herds with bulk tank PCR Ct value ≤ 39 for S. agalactiae and S. aureus. At milk recording, 2456 quarter milk samples were taken aseptically for BC and the routinely taken cow level milk samples were analyzed by PCR. Results showed that 53 cows (8.6%) were positive for S. agalactiae IMI by BC. Sensitivity of PCR at cut-offs; ≤ 39, ≤ 37, ≤ 34, and ≤ 32, was 96.2%, 91.9%, 87.2% and 73.9%, while Se of BC was 25.7%, 29.9%, 59.9% and 72.1%. Specificity of PCR at cut-offs; ≤ 39, ≤ 37, ≤ 34, and ≤ 32, was 96.8%, 96.9%, 96.7%, and 97.22%, while Sp of BC was 99.7%, 99.5%, 99.2%, and 98.9%. The estimated prevalence of S. agalactiae IMI by PCR was higher than the apparent prevalence at the tested cut-offs, indicating under estimation of S. agalactiae IMI in the examined dairy cows. In conclusion, Se of PCR is always higher than Se of BC at all tested cut-offs. The lower cut-off, the more comparable becomes Se of PCR and Se of BC. The changes in Se in both PCR and BC at different Ct-value cut-offs may indicate a change in the definition of the latent infection. The similar Se of both tests at cut-off ≤ 32 may indicate high concentrations of S. agalactiae viable cells, representing a cow truly/heavily infected with S. agalactiae and thus easier to detect with BC. At cut-off ≤ 39 the latent definition of infection may reflect a more general condition of cows being positive for S. agalactiae. Our findings indicate that PCR Ct-value cut-offs should

  14. Structural Differences between the Streptococcus agalactiae Housekeeping and Pilus-Specific Sortases: SrtA and SrtC1

    Energy Technology Data Exchange (ETDEWEB)

    Khare, B.; Krishnan, V.; Rajashankar, K.R.; I-Hsiu, H.; Xin, M.; Ton-That, H.; Narayana, S.V. (Texas-HSC); (Cornell); (UAB)

    2011-10-21

    The assembly of pili on the cell wall of Gram-positive bacteria requires transpeptidase enzymes called sortases. In Streptococcus agalactiae, the PI-1 pilus island of strain 2603V/R encodes two pilus-specific sortases (SrtC1 and SrtC2) and three pilins (GBS80, GBS52 and GBS104). Although either pilus-specific sortase is sufficient for the polymerization of the major pilin, GBS80, incorporation of the minor pilins GBS52 and GBS104 into the pilus structure requires SrtC1 and SrtC2, respectively. The S. agalactiae housekeeping sortase, SrtA, whose gene is present at a different location and does not catalyze pilus polymerization, was shown to be involved in cell wall anchoring of pilus polymers. To understand the structural basis of sortases involved in such diverse functions, we determined the crystal structures of S. agalactiae SrtC1 and SrtA. Both enzymes are made of an eight-stranded beta-barrel core with variations in their active site architecture. SrtA exhibits a catalytic triad arrangement similar to that in Streptococcus pyogenes SrtA but different from that in Staphylococcus aureus SrtA. In contrast, the SrtC1 enzyme contains an N-terminal helical domain and a 'lid' in its putative active site, which is similar to that seen in Streptococcus pneumoniae pilus-specific sortases, although with subtle differences in positioning and composition. To understand the effect of such differences on substrate recognition, we have also determined the crystal structure of a SrtC1 mutant, in which the conserved DP(W/F/Y) motif was replaced with the sorting signal motif of GBS80, IPNTG. By comparing the structures of WT wild type SrtA and SrtC1 and the 'lid' mutant of SrtC1, we propose that structural elements within the active site and the lid may be important for defining the role of specific sortase in pili biogenesis.

  15. Structural differences between the Streptococcus agalactiae housekeeping and pilus-specific sortases: SrtA and SrtC1.

    Directory of Open Access Journals (Sweden)

    B Khare

    Full Text Available The assembly of pili on the cell wall of Gram-positive bacteria requires transpeptidase enzymes called sortases. In Streptococcus agalactiae, the PI-1 pilus island of strain 2603V/R encodes two pilus-specific sortases (SrtC1 and SrtC2 and three pilins (GBS80, GBS52 and GBS104. Although either pilus-specific sortase is sufficient for the polymerization of the major pilin, GBS80, incorporation of the minor pilins GBS52 and GBS104 into the pilus structure requires SrtC1 and SrtC2, respectively. The S. agalactiae housekeeping sortase, SrtA, whose gene is present at a different location and does not catalyze pilus polymerization, was shown to be involved in cell wall anchoring of pilus polymers. To understand the structural basis of sortases involved in such diverse functions, we determined the crystal structures of S. agalactiae SrtC1 and SrtA. Both enzymes are made of an eight-stranded beta-barrel core with variations in their active site architecture. SrtA exhibits a catalytic triad arrangement similar to that in Streptococcus pyogenes SrtA but different from that in Staphylococcus aureus SrtA. In contrast, the SrtC1 enzyme contains an N-terminal helical domain and a 'lid' in its putative active site, which is similar to that seen in Streptococcus pneumoniae pilus-specific sortases, although with subtle differences in positioning and composition. To understand the effect of such differences on substrate recognition, we have also determined the crystal structure of a SrtC1 mutant, in which the conserved DP(W/F/Y motif was replaced with the sorting signal motif of GBS80, IPNTG. By comparing the structures of WT wild type SrtA and SrtC1 and the 'lid' mutant of SrtC1, we propose that structural elements within the active site and the lid may be important for defining the role of specific sortase in pili biogenesis.

  16. Construction of a Streptococcus agalactiae phoB mutant and evaluation of its potential as an attenuated modified live vaccine in golden pompano, Trachinotus ovatus.

    Science.gov (United States)

    Cai, Xiaohui; Wang, Bei; Peng, Yinhui; Li, Yuan; Lu, Yishan; Huang, Yucong; Jian, Jichang; Wu, Zaohe

    2017-04-01

    Streptococcus agalactiae is a Gram-positive pathogen that can survive inside professional phagocytes and nonphagocytic cells to cause septicemia and meningoencephalitis in freshwater and marine fish. However, vaccines based on extracellular products (ECP) and formalin-killed whole S. agalactiae cells, as well as subunit vaccine are unable to protect fish from infection by variant serotypes S. agalactiae. The search for live attenuated vaccine with highly conserved and virulent-related genes is essential for producing a vaccine to help understand and control streptococcosis In this study, the phoB gene was cloned from pathogenic S. agalactiae TOS01 strain and the mutant strain SAΔphoB was constructed via allelic exchange mutagenesis. The results showed that the deduced amino acid of S. agalactiae TOS01 shares high similarities with other Streptococcus spp. and has high conserved response regulator receiver domain (REC) and DNA-binding effector domain of two-component system response regulators (Trans_reg_C). Cell adherence and invasion assays, challenge experiments and histopathological changes post-vaccination were performed and observed, the results showed that the mutant strain SAΔphoB has a lower adherence and invasion rate and less virulent than the wild type strain in golden pompano, and it doesn't induce clinical symptoms and obvious pathological changes in golden pompano, thereby indicating that the deletion of phoB affects the virulence and infectious capacity of S. agalactiae. Golden pompano vaccinated via intraperitoneal injection SAΔphoB had the relative percent survival value of 93.1% after challenge with TOS01, demonstrating its high potential as an effective attenuated live vaccine candidate. Real-time PCR assays showed that the SAΔphoB was able to enhance the expression of immune-related genes, including MHC-I, MyD88, IL-22 and IL-10 after vaccination, indicating that the SAΔphoB is able to induce humoral and cell-mediated immune response

  17. Structure of Streptococcus agalactiae tip pilin GBS104: a model for GBS pili assembly and host interactions

    Energy Technology Data Exchange (ETDEWEB)

    Krishnan, Vengadesan [UNESCO Regional Centre for Biotechnology (RCB), Gurgaon 122 016, Haryana (India); Dwivedi, Prabhat [University of Texas Health Science Center, Houston, TX 77030 (United States); Kim, Brandon J. [San Diego State University, 5500 Campanile Drive, San Diego, CA 92182 (United States); Samal, Alexandra; Macon, Kevin [University of Alabama at Birmingham, Birmingham, AL 35294 (United States); Ma, Xin; Mishra, Arunima [University of Texas Health Science Center, Houston, TX 77030 (United States); Doran, Kelly S. [San Diego State University, 5500 Campanile Drive, San Diego, CA 92182 (United States); Ton-That, Hung [University of Texas Health Science Center, Houston, TX 77030 (United States); Narayana, Sthanam V. L., E-mail: narayana@uab.edu [University of Alabama at Birmingham, Birmingham, AL 35294 (United States); UNESCO Regional Centre for Biotechnology (RCB), Gurgaon 122 016, Haryana (India)

    2013-06-01

    The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. In addition, a homology model of the remaining two domains of GBS104 was built and a model of full-length GBS104 was generated by combining the homology model (the N1 and N4 domains) and the crystal structure of the 75 kDa fragment (the N2 and N3 domains). This rod-shaped GBS104 model is constructed of three IgG-like domains (the N1, N2 and N4 domains) and one vWFA-like domain (the N3 domain). The N1 and N2 domains of GBS104 are assembled with distinct and remote segments contributed by the N- and C-termini. The metal-binding site in the N3 domain of GBS104 is in the closed/low-affinity conformation. Interestingly, this domain hosts two long arms that project away from the metal-binding site. Using site-directed mutagenesis, two cysteine residues that lock the N3 domain of GBS104 into the open/high-affinity conformation were introduced. Both wild-type and disulfide-locked recombinant proteins were tested for binding to extracellular matrix proteins such as collagen, fibronectin, fibrinogen and laminin, and an increase in fibronectin binding affinity was identified for the disulfide-locked N3 domain, suggesting that induced conformational changes may play a possible role in receptor binding.

  18. Characterization of a new CAMP factor carried by an integrative and conjugative element in Streptococcus agalactiae and spreading in Streptococci.

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    Sarah Chuzeville

    Full Text Available Genetic exchanges between Streptococci occur frequently and contribute to their genome diversification. Most of sequenced streptococcal genomes carry multiple mobile genetic elements including Integrative and Conjugative Elements (ICEs that play a major role in these horizontal gene transfers. In addition to genes involved in their mobility and regulation, ICEs also carry genes that can confer selective advantages to bacteria. Numerous elements have been described in S. agalactiae especially those integrated at the 3' end of a tRNA(Lys encoding gene. In strain 515 of S. agalactiae, an invasive neonate human pathogen, the ICE (called 515_tRNA(Lys is functional and carries different putative virulence genes including one encoding a putative new CAMP factor in addition to the one previously described. This work demonstrated the functionality of this CAMP factor (CAMP factor II in Lactococcus lactis but also in pathogenic strains of veterinary origin. The search for co-hemolytic factors in a collection of field strains revealed their presence in S. uberis, S. dysgalactiae, but also for the first time in S. equisimilis and S. bovis. Sequencing of these genes revealed the prevalence of a species-specific factor in S. uberis strains (Uberis factor and the presence of a CAMP factor II encoding gene in S. bovis and S. equisimilis. Furthermore, most of the CAMP factor II positive strains also carried an element integrated in the tRNA(Lys gene. This work thus describes a CAMP factor that is carried by a mobile genetic element and has spread to different streptococcal species.

  19. Structure of Streptococcus agalactiae tip pilin GBS104: a model for GBS pili assembly and host interactions

    International Nuclear Information System (INIS)

    Krishnan, Vengadesan; Dwivedi, Prabhat; Kim, Brandon J.; Samal, Alexandra; Macon, Kevin; Ma, Xin; Mishra, Arunima; Doran, Kelly S.; Ton-That, Hung; Narayana, Sthanam V. L.

    2013-01-01

    The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. In addition, a homology model of the remaining two domains of GBS104 was built and a model of full-length GBS104 was generated by combining the homology model (the N1 and N4 domains) and the crystal structure of the 75 kDa fragment (the N2 and N3 domains). This rod-shaped GBS104 model is constructed of three IgG-like domains (the N1, N2 and N4 domains) and one vWFA-like domain (the N3 domain). The N1 and N2 domains of GBS104 are assembled with distinct and remote segments contributed by the N- and C-termini. The metal-binding site in the N3 domain of GBS104 is in the closed/low-affinity conformation. Interestingly, this domain hosts two long arms that project away from the metal-binding site. Using site-directed mutagenesis, two cysteine residues that lock the N3 domain of GBS104 into the open/high-affinity conformation were introduced. Both wild-type and disulfide-locked recombinant proteins were tested for binding to extracellular matrix proteins such as collagen, fibronectin, fibrinogen and laminin, and an increase in fibronectin binding affinity was identified for the disulfide-locked N3 domain, suggesting that induced conformational changes may play a possible role in receptor binding

  20. Desempenho e hematologia de tilápias-do-nilo alimentadas com Saccharomyces cerevisiae e vacinadas contra Streptococcus agalactiae

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    Rogério Salvador

    2013-08-01

    Full Text Available O objetivo deste trabalho foi avaliar a inter-relação entre a suplementação alimentar com parede celular de Saccharomyces cerevisae e a vacinação contra Streptococcus agalactiae e seu efeito sobre o desempenho produtivo e as variáveis hematológicas de tilápia-do-nilo (Oreochromis niloticus. Oitenta e quatro tilápias-do-nilo foram distribuídas em 12 caixas de fibra (n=7, em arranjo fatorial 2x2x3, correspondente a dois níveis de suplementação com parede celular de levedura, dois tipos de inoculação e três tempos de avaliação. Os peixes foram alimentados durante 77 dias. A vacinação dos peixes foi realizada 60 dias após o início da alimentação. Quinze dias após a vacinação, todos os peixes foram submetidos ao desafio com cepa viva de S. agalactiae, e 6, 24 e 48 horas após o desafio, o sangue foi colhido da veia caudal para avaliações. Peixes alimentados com ração suplementada apresentam maior ganho de peso e taxa de crescimento específico, e a interação entre os efeitos da dieta e da vacinação resulta em maiores taxas de hematócrito, hemoglobina e leucócitos.

  1. Development of Primer Sets for Loop-Mediated Isothermal Amplification that Enables Rapid and Specific Detection of Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae

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    Deguo Wang

    2015-05-01

    Full Text Available Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae are the three main pathogens causing bovine mastitis, with great losses to the dairy industry. Rapid and specific loop-mediated isothermal amplification methods (LAMP for identification and differentiation of these three pathogens are not available. With the 16S rRNA gene and 16S-23S rRNA intergenic spacers as targets, four sets of LAMP primers were designed for identification and differentiation of S. dysgalactiae, S. uberis and S. agalactiae. The detection limit of all four LAMP primer sets were 0.1 pg DNA template per reaction, the LAMP method with 16S rRNA gene and 16S-23S rRNA intergenic spacers as the targets can differentiate the three pathogens, which is potentially useful in epidemiological studies.

  2. Development of Primer Sets for Loop-Mediated Isothermal Amplification that Enables Rapid and Specific Detection of Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae.

    Science.gov (United States)

    Wang, Deguo; Liu, Yanhong

    2015-05-26

    Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae are the three main pathogens causing bovine mastitis, with great losses to the dairy industry. Rapid and specific loop-mediated isothermal amplification methods (LAMP) for identification and differentiation of these three pathogens are not available. With the 16S rRNA gene and 16S-23S rRNA intergenic spacers as targets, four sets of LAMP primers were designed for identification and differentiation of S. dysgalactiae, S. uberis and S. agalactiae. The detection limit of all four LAMP primer sets were 0.1 pg DNA template per reaction, the LAMP method with 16S rRNA gene and 16S-23S rRNA intergenic spacers as the targets can differentiate the three pathogens, which is potentially useful in epidemiological studies.

  3. PENGARUH LAMA DAN SUHU PENYIMPANAN EKSTRAK DAUN SIRIH HIJAU (Piper betle linn) DENGAN AQUADES TERHADAP DAYA HAMBAT BAKTERI Streptococcus agalactiae PENYEBAB MASTITIS PADA SAPI PERAH

    OpenAIRE

    Muhammad Sanjaya Kusuma; Tri Eko Susilorini; Puguh Surjowardojo

    2017-01-01

    Green betle leaf (Piper betle L.) is one of the plants used by the people of Indonesia for tradisional medicine. Green betle leaf contains antibacterial compounds consisting of phenol and its derivatives. This study aims to determine the antibacterial activity of green betle leaf (Piper betle L.) againt the bacteria Streptococcus agalactiae caused mastitis in dairy cows. Bacterial inhibition test by paper disc method. Data analysis using ANOVA by Nested design with 6 treatment and 6 repetitio...

  4. Safety and immunogenicity of an oral DNA vaccine encoding Sip of Streptococcus agalactiae from Nile tilapia Oreochromis niloticus delivered by live attenuated Salmonella typhimurium.

    Science.gov (United States)

    Huang, L Y; Wang, K Y; Xiao, D; Chen, D F; Geng, Y; Wang, J; He, Y; Wang, E L; Huang, J L; Xiao, G Y

    2014-05-01

    Attenuated Salmonella typhimurium SL7207 was used as a carrier for a reconstructed DNA vaccine against Streptococcus agalactiae. A 1.02 kb DNA fragment, encoding for a portion of the surface immunogenic protein (Sip) of S. agalactiae was inserted into pVAX1. The recombinant plasmid pVAX1-sip was transfected in EPC cells to detect the transient expression by an indirect immunofluorescence assay, together with Western blot analysis. The pVAX1-sip was transformed by electroporation into SL7207. The stability of pVAX1-sip into Salmonella was over 90% after 50 generations with antibiotic selection in vitro while remained stable over 80% during 35 generations under antibiotic-free conditions. The LD50 of SL/pVAX1-sip was 1.7 × 10(11) CFU/fish by intragastric administration which indicated a quite low virulence. Tilapias were inoculated orally at 10(8) CFU/fish, the recombinant bacteria were found present in intestinal tract, spleens and livers and eventually eliminated from the tissues 4 weeks after immunization. Fish immunized at 10(7), 10(8) and 10(9) CFU/fish with different immunization times caused various levels of serum antibody and an effective protection against lethal challenge with the wild-type strain S. agalactiae. Integration studies showed that the pVAX1-sip did not integrate with tilapia chromosomes. The DNA vaccine SL/pVAX1-sip was proved to be safe and effective in protecting tilapias against S. agalactiae infection. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. A metabolomic investigation into the effects of temperature on Streptococcus agalactiae from Nile tilapia (Oreochromis niloticus) based on UPLC-MS/MS.

    Science.gov (United States)

    Hu, Wen-Ting; Guo, Wei-Liang; Meng, Ai-Yun; Sun, Yun; Wang, Shi-Feng; Xie, Zhen-Yu; Zhou, Yong-Can; He, Chaozu

    2017-10-01

    Streptococcosis caused by Streptococcus agalactiae is one of the most serious diseases in farmed tilapia, and temperature is one of the most important environmental factors related to its outbreak. To elucidate the influence of temperature variation on the pathogen from a metabolic perspective, the global metabolomics of 2 pathogenic strains of S. agalactiae from sick tilapia were analyzed at 35°C and 25°C using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with pattern recognition approaches and pathway analysis. The result showed that the metabolic status of S. agalactiae was extensively affected by its culture temperature. Based on the results of metabolites contributing to these differences, a large number of nucleotides and their ramifications were markedly elevated at 35°C. Various energy substances, components of the cell wall and substances associated with stress regulation such as glyceraldehyde 3-phosphate, pyroglutamic acid, glutamate, d-Alanyl-d-alanine, glycerophosphocholine, dephospho-CoA, and oxidized glutathione increased when the strains were cultured at 35°C. Additionally, a general decrease in various precursors of capsule, antigen, and virulence protein formation were detected including mannose, maltotriose, N-acetyl-d-glucosamine 6-phosphate, uracil, proline, and citrulline. These metabolic changes indicated that metabolic activity decreased, while adaptive ability to environment and pathogenicity to host increased at high temperature. This study is the first to determine the metabolomic responses of S. agalactiae to temperature, and the results are useful to reveal its pathogenic mechanism and find effective disease control. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Disruption of the pdhB pyruvate dehydrogenase [corrected] gene affects colony morphology, in vitro growth and cell invasiveness of Mycoplasma agalactiae.

    Directory of Open Access Journals (Sweden)

    Shivanand Hegde

    Full Text Available The utilization of available substrates, the metabolic potential and the growth rates of bacteria can play significant roles in their pathogenicity. This study concentrates on Mycoplasma agalactiae, which causes significant economic losses through its contribution to contagious agalactia in small ruminants by as yet unknown mechanisms. This lack of knowledge is primarily due to its fastidious growth requirements and the scarcity of genetic tools available for its manipulation and analysis. Transposon mutagenesis of M. agalactiae type strain PG2 resulted in several disruptions throughout the genome. A mutant defective in growth in vitro was found to have a transposon insertion in the pdhB gene, which encodes a component of the pyruvate dehydrogenase complex. This growth difference was quite significant during the actively dividing logarithmic phase but a gradual recovery was observed as the cells approached stationary phase. The mutant also exhibited a different and smaller colony morphology compared to the wild type strain PG2. For complementation, pdhAB was cloned downstream of a strong vpma promoter and upstream of a lacZ reporter gene in a newly constructed complementation vector. When transformed with this vector the pdhB mutant recovered its normal growth and colony morphology. Interestingly, the pdhB mutant also had significantly reduced invasiveness in HeLa cells, as revealed by double immunofluorescence staining. This deficiency was recovered in the complemented strain, which had invasiveness comparable to that of PG2. Taken together, these data indicate that pyruvate dehydrogenase might be an important player in infection with and colonization by M. agalactiae.

  7. PENGARUH LAMA DAN SUHU PENYIMPANAN EKSTRAK DAUN SIRIH HIJAU (Piper betle linn DENGAN AQUADES TERHADAP DAYA HAMBAT BAKTERI Streptococcus agalactiae PENYEBAB MASTITIS PADA SAPI PERAH

    Directory of Open Access Journals (Sweden)

    Muhammad Sanjaya Kusuma

    2017-11-01

    Full Text Available Green betle leaf (Piper betle L. is one of the plants used by the people of Indonesia for tradisional medicine. Green betle leaf contains antibacterial compounds consisting of phenol and its derivatives. This study aims to determine the antibacterial activity of green betle leaf (Piper betle L. againt the bacteria Streptococcus agalactiae caused mastitis in dairy cows. Bacterial inhibition test by paper disc method. Data analysis using ANOVA by Nested design with 6 treatment and 6 repetitions. The results of this study green betle leaf (Piper betle L. extract inhibiting the growth of bacteria Streptococcus agalactiae was significantly different (P<0,01. The conclusion were the extract of green betel leaf (Piper betle L. can inhibit the growth of Streptococcus agalactiae and storage temperature has no effect, but storage periode gives effect the quality of green betel leaf extract (Piper betle L., so that the leaf extract storage green betel with distilled solvent recommended on 2nd days at refrigerator.

  8. Antimicrobial Resistance Profiles of Bacteria Isolated from the Nasal Cavity of Camels in Samburu, Nakuru, and Isiolo Counties of Kenya

    Directory of Open Access Journals (Sweden)

    J. M. Mutua

    2017-01-01

    Full Text Available This study was designed to determine antimicrobial resistance profiles of bacteria isolated from the nasal cavity of healthy camels. A total of 255 nasal samples (swabs were collected in Isiolo, Samburu, and Nakuru counties, Kenya, from which 404 bacterial isolates belonging to various genera and species were recovered. The bacterial isolates included Bacillus (39.60%, coagulase-negative Staphylococcus (29.95%, Streptococcus species other than Streptococcus agalactiae (25.74%, coagulase-positive Staphylococcus (3.96%, and Streptococcus agalactiae (0.74%. Isolates were most susceptible to Gentamicin (95.8%, followed by Tetracycline (90.5%, Kanamycin and Chloramphenicol (each at 85.3%, Sulphamethoxazole (84.2%, Co-trimoxazole (82.1%, Ampicillin (78.9%, and finally Streptomycin (76.8%. This translated to low resistance levels. Multidrug resistance was also reported in 30.5% of the isolates tested. Even though the antibiotic resistance demonstrated in this study is low, the observation is significant, since the few resistant normal flora could be harboring resistance genes which can be transferred to pathogenic bacteria within the animal, to other animals’ bacteria and, most seriously, to human pathogens.

  9. Latent class analysis of real time qPCR and bacteriological culturing for the diagnosis of Streptococcus agalactiae in cow composite milk samples.

    Science.gov (United States)

    Holmøy, Ingrid H; Toft, Nils; Jørgensen, Hannah J; Mørk, Tormod; Sølverød, Liv; Nødtvedt, Ane

    2018-06-01

    Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows. The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare it to conventional bacteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging to 6 herds were cultured and tested by qPCR. While 37 (6.4%) samples were positive for S. agalactiae by bacteriological culture, 66 (11.4%) samples were positive by qPCR. The within-herd prevalence in the six herds, as estimated by the latent class models ranged from 7.7 to 50.8%. At the recommended cut-off (cycle threshold 37), the sensitivity of the qPCR was significantly higher at 95.3 (95% posterior probability interval [PPI] [84.2; 99.6]) than that of bacteriological culture at 58.2 (95% PPI [43.8; 74.4]). However, bacterial culture had a higher specificity of 99.7 (95% PPI [98.5; 100.0]) compared to the qPCR at 98.5 (95% PPI [94.6; 99.9]). The median estimated negative predictive values of qPCR was consistently higher than those of the BC at all estimated prevalences, and the superiority of the qPCR increased with increasing within-herd prevalence. The median positive predictive values of BC was in general higher than the estimates for the qPCR, however, at the highest prevalence the predictive ability of both tests were similar. Copyright © 2018 Elsevier B.V. All

  10. Carriage and serotype distribution of Streptococcus agalactiae in third trimester pregnancy in southern Ghana

    DEFF Research Database (Denmark)

    Slotved, Hans-Christian; Dayie, Nicholas T K D; Banini, Josephine A N

    2017-01-01

    was performed on a single subcultured colony. Gram staining was performed, and isolates were evaluated for beta-haemolytic reactions. Furthermore, the isolates were serotyped using the GBS latex serotyping kit. RESULTS: The carriage rates were found to be 25.5% (95% CI: 19.6-32.1) to 28.0% (95% CI: 21...... of this study revealed that prevalence of GBS colonization in pregnant women in Greater Accra region is high and comparable to rates observed in South Africa and Western countries. The most prevalent serotypes were serotypes VII and IX, which have not been observed before in West Africa....

  11. 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.

  12. Isolation and identification of bacterial pathogen from mastitis milk in Central Java Indonesia

    Science.gov (United States)

    Harjanti, D. W.; Ciptaningtyas, R.; Wahyono, F.; Setiatin, ET

    2018-01-01

    Mastitis is a multi-etiologic disease of the mammary gland characterized mainly by reduction in milk production and milk quality due to intramammary infection by pathogenic bacteria. Nearly 83% of lactating dairy cows in Indonesia are infected with mastitis in various inflammation degrees. This study was conducted to isolate and identify the pathogen in milk collected from mastitis-infected dairy cows. The study was carried out in ten smallholder dairy farms in Central Java Indonesia based on animal examination, California mastitis test, isolation bacterial pathogens, Gram staining, Catalase and Coagulase test, and identification of bacteria species using Vitek. Bacteriological examination of milk samples revealed 15 isolates where Streptococcus was predominant species (73.3%) and the coagulase negative Staphylococcus species was identified at the least bacteria (26.7%). The Streptococcus bacteria found were Streptococcus uberis (2 isolates), Streptococcus sanguinis(6 isolates), Streptococcus dysgalactiaessp dysgalactiae(1 isolate) , Streptococcus mitis (1 isolate) and Streptococcus agalactiae (1 isolate). The Staphylococcus isolates comprising of Staphylococcus simulans (1 isolate) and Staphylococcus chromogens (3 isolates). Contamination of raw milkwith pathogenic bacteria can cause outbreaks of human disease (milk borne disease). Thus, proper milk processing method that couldinhibit the growth or kill these pathogenic bacteria is important to ensure the safety of milk and milk products.

  13. Identificação dos sorotipos de Streptococcus agalactiae pela técnica de PCR de amostras isoladas em pacientes colonizados e infectados na cidade de Campinas e região

    OpenAIRE

    Katelí Fiolo

    2011-01-01

    Resumo: Streptococcus agalactiae, conhecido como Estreptococo beta-hemolítico do grupo B (EGB), é classificado por diferenças capsulares que podem variar em dez sorotipos, alguns responsáveis por infecções materno-infantis sérias e debilitantes ou podendo ainda levar ao óbito. O EGB pode ocasionar também, infecções graves em adultos e idosos. OBJETIVO: Descrever e analisar o perfil epidemiológico dos sorotipos prevalentes de Streptococcus agalactiae, provenientes de infecção em recém-nascidos...

  14. Influência da temperatura na infecção de tilápias do Nilo por Streptococcus agalactiae

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    Paulo Fernandes Marcusso

    2015-04-01

    Full Text Available Mudanças ambientais afetam a homeostase dos peixes, tornando-os mais suscetíveis a doenças. No Brasil, tem sido relatados surtos de infecção por Streptococcus agalactiae em tilápia do Nilo, principalmente quando se encontram fora da zona de conforto térmico. No presente trabalho, foi avaliada a taxa de mortalidade e determinado quais foram os órgãos mais afetados por essa bactéria em temperaturas que ocorrem naturalmente no Sul do Brasil. Quarenta tilápias-do-nilo (Oreochromis niloticus foram infectadas por Streptococcus agalactiae e distribuídas em quatro grupos (n = 10, cada um deles submetidos a diferentes temperaturas: G1: 24°C, G2: 26°C, G3: 28°C e G4: 32°C. Os peixes foram monitorados durante 10 dias. Os peixes com sinais clínicos irreversíveis foram sacrificados e coletadas amostras de cérebro, fígado e rins para analise bacteriológica e molecular. Foram observados sinais compatíveis com infecção estreptocócica em todos os grupos. A taxa de mortalidade mais elevada ocorreu nos grupos mantidos nas temperaturas de 24°C e 32°C. O cérebro foi o órgão mais afetado, com a maior percentagem de isolamento de S. agalactiae pelos dois métodos de diagnostico. Os resultados sugerem que, tal como nos mamíferos, temperaturas que estão mais afastadas da zona de conforto afetam significativamente a homeostase dos peixes, aumentando a sua susceptibilidade para infecções bacterianas.

  15. Hibridación in situ para la detección de streptococcus agalactiae en tejidos de tilapia (oreochromis sp.)

    OpenAIRE

    Pulido, E. A.; Iregui, C. A.

    2010-01-01

    La estreptococosis es uno de los problemas sanitarios más serios en la acuicultura mundial. En Colombia la enfermedad afecta de manera importante las explotaciones de tilapia. se estandarizó la técnica de hibridación in situ (HIS ) en tejidos de tilapia previamente identificados como positivos a la presencia de S. agalactiae por la técnica de inmunoperoxidasa indirecta (IPI) y microbiología. Se obtuvo señal positiva en el interior de los granulomas con u...

  16. Caracterização Genética da Resistência à Eritromicina em Streptococcus agalactia e Degestantes saudáveis

    OpenAIRE

    Pinheiro, Sandra Marisa de Oliveira Sequeira

    2009-01-01

    Dissertação de Mestrado em Biologia Clínica Laboratorial Streptococcus agalactiae (Grupo B de Lancefield, EGB), um microrganismo comensal do homem, reconhecido em 1920 como o agente etiológico da mastite bovina, tem sido associado a infecções em parturientes e recém-nascidos, sendo o principal agente de septicemia e meningite neonatal. Para evitar a infecção perinatal recomenda-se a pesquisa da bactéria na região vagino-perianal durante o terceiro semestre de gravidez, indicando o tratamen...

  17. Influencia de la composición del medio de cultivo sobre la producción de pigmento por Streptococcus agalactiae

    OpenAIRE

    Carazo Carazo, Concepción

    2011-01-01

    Se ha estudiado la influencia de la composición del medio de cultivo sobre la producción de pigmento por s. agalactiae partiendo de un medio de cultivo diseñado para tal finalidad (new granada medium), eliminando uno a uno los componentes incluidos en su formulación original. Almidón, proteosa peptona (difco), suero, fosfatos y metrotexato influyen de forma importante en la formación y/o visualización del pigmento producido por este microorganismo. Se ha determinado que la fracción de proteos...

  18. Binding of glycoprotein Srr1 of Streptococcus agalactiae to fibrinogen promotes attachment to brain endothelium and the development of meningitis.

    Directory of Open Access Journals (Sweden)

    Ho Seong Seo

    Full Text Available The serine-rich repeat glycoprotein Srr1 of Streptococcus agalactiae (GBS is thought to be an important adhesin for the pathogenesis of meningitis. Although expression of Srr1 is associated with increased binding to human brain microvascular endothelial cells (hBMEC, the molecular basis for this interaction is not well defined. We now demonstrate that Srr1 contributes to GBS attachment to hBMEC via the direct interaction of its binding region (BR with human fibrinogen. When assessed by Far Western blotting, Srr1 was the only protein in GBS extracts that bound fibrinogen. Studies using recombinant Srr1-BR and purified fibrinogen in vitro confirmed a direct protein-protein interaction. Srr1-BR binding was localized to amino acids 283-410 of the fibrinogen Aα chain. Structural predictions indicated that the conformation of Srr1-BR is likely to resemble that of SdrG and other related staphylococcal proteins that bind to fibrinogen through a "dock, lock, and latch" mechanism (DLL. Deletion of the predicted latch domain of Srr1-BR abolished the interaction of the BR with fibrinogen. In addition, a mutant GBS strain lacking the latch domain exhibited reduced binding to hBMEC, and was significantly attenuated in an in vivo model of meningitis. These results indicate that Srr1 can bind fibrinogen directly likely through a DLL mechanism, which has not been described for other streptococcal adhesins. This interaction was important for the pathogenesis of GBS central nervous system invasion and subsequent disease progression.

  19. Estimation of test characteristics of real-time PCR and bacterial culture for diagnosis of subclinical intramammary infections with Streptococcus agalactiae in Danish dairy cattle in 2012 using latent class analysis

    DEFF Research Database (Denmark)

    Mahmmod, Yasser; Toft, Nils; Katholm, Jørgen

    2013-01-01

    threshold (Ct) values against bacterial culture (BC) for diagnosis of S. agalactiae IMI using latent class analysis to avoid the assumption of a perfect reference test. A total of 614 dairy cows were randomly selected from 6 herds with bulk tank PCR Ct value ≤ 39 for S. agalactiae and S. aureus. At milk...... cut-offs, indicating under estimation of S. agalactiae IMI in the examined dairy cows. In conclusion, Se of PCR is always higher than Se of BC at all tested cut-offs. The lower cut-off, the more comparable becomes Se of PCR and Se of BC. The changes in Se in both PCR and BC at different Ct-value cut...... definition of infection may reflect a more general condition of cows being positive for S. agalactiae. Our findings indicate that PCR Ct-value cut-offs should be chosen according to the underlying latent infection definition of interest. Latent class analysis proposes a useful alternative to classic test...

  20. Expression, purification, crystallization and preliminary X-ray diffraction studies of the human keratin 4-binding domain of serine-rich repeat protein 1 from Streptococcus agalactiae

    International Nuclear Information System (INIS)

    Sundaresan, Ramya; Samen, Ulrike; Ponnuraj, Karthe

    2011-01-01

    Expression, purification and crystallization of Srr-1-K4BD, a human keratin 4-binding domain of serine-rich repeat protein 1 from S. agalactiae, was carried out. Native crystals of Srr-1-K4BD diffracted to 3.8 Å resolution using synchrotron radiation. Serine-rich repeat protein 1 (Srr-1) is a surface protein from Streptococcus agalactiae. A 17 kDa region of this protein has been identified to bind to human keratin 4 (K4) and is termed the Srr-1 K4-binding domain (Srr-1-K4BD). Recombinant Srr-1-K4BD was overexpressed in Escherichia coli BL21 (DE3) cells. Native and selenomethionine-substituted proteins were prepared using Luria–Bertani (LB) and M9 minimal media, respectively. A two-step purification protocol was carried out to obtain a final homogenous sample of Srr-1-K4BD. Crystals of native Srr-1-K4BD were obtained using PEG 3350 as a precipitant. The crystals diffracted to 3.8 Å resolution using synchrotron radiation and belonged to space group P2 1 , with unit-cell parameters a = 47.56, b = 59.48, c = 94.71 Å, β = 93.95°

  1. Cinética de fermentación y acción antimicrobiana de Weissella confusa contra Staphylococcus aureus y Streptococcus agalactiae

    Directory of Open Access Journals (Sweden)

    Liliana Serna Cock

    2010-01-01

    Full Text Available Se evaluaron las cinéticas de producción de biomasa, producción de ácido láctico, consumo de sustrato y actividad antimicrobiana de Weissella confusa, una bacteria acido láctica con actividad antimicrobiana frente a Streptococcus agalactiae y Staphylococcus aureus, microorganismos productores de mastitis bovina. Las fermentaciones se llevaron a cabo anaeróbicamente en discontinuo utilizando como sustratos, sustrato comercial (SC, leche suplementada con extracto levadura (LEL y leche suplementada con extracto de levadura y glucosa (LELG y se compararon los parámetros cinéticos. La mayor inhibición de los patógenos, la mayor producción de ácido láctico y el mayor rendimiento de biomasa se presentó en sustrato LELG. En éste último sustrato al cabo de la cuarta hora de fermentación, se presentó un diámetro de inhibición de 36,33 mm para Staphylococcus aureus y a la octava hora de fermentación 39 mm de diámetro para Streptococcus agalactiae; la producción máxima de ácido láctico fue 13,12 gL-1 (a las 48 h y la máxima concentración de biomasa fue 3,07 gL-1 (a las 48 h. Estos resultados fueron superiores a los obtenidos en SC donde, para el mismo tiempo de fermentación, se obtuvieron 24,38 mm para Staphylococcus aureus y 30,58 mm de diámetro de inhibición para Streptococcus agalactiae; la mayor producción de ácido láctico fue 11,6 gL-1 (a las 12 h y la mayor concentración de biomasa fue 1,18 gL-1 (a las 24 h. Los resultados sugieren que el LELG puede convertirse en una alternativa a bajo costo para la producción de Weissella confusa, microorganismo con gran potencial para el control y el tratamiento de mastitis bovina.

  2. Isolated galaxies

    International Nuclear Information System (INIS)

    Einasto, Maret

    1990-01-01

    To test for the possible presence of really isolated galaxies, which form a randomly distributed population in voids, we compare the distribution of most isolated galaxies in an observed sample with distributions of the same number of random points using the nearest neighbour test. The results show that the random population of really isolated galaxies does not exist - even the most isolated galaxies are connected with systems of galaxies, forming their outlying parts. (author)

  3. [Neonatal bacterial infection by maternal-fetal contamination: for a change in approach? 1. Detection of Streptococcus agalactiae infection: methods and evaluation of results].

    Science.gov (United States)

    Blond, M H; Gold, F; Pierre, F; Quentin, R; Aujard, Y

    2001-10-01

    Perinatal group B streptococcal infection has been the subject of numerous studies and despite guidelines established during the last decade remains a frequent disease with high mortality. The basic aim of the guidelines is to screen for Streptococcus agalactiae during the antepartum period in order to institute antibiotic therapy during delivery. A critical review of the literature highlights the real impact and adverse effect of these guidelines: difficult application (only two-thirds of all maternity units have a protocol and compliance is only 75%), maternal risks of antibiotic therapy (especially the emergence of resistant Gram negative bacteria), fetal risks (accentuation of neonatal sepsis with resistant strains, retarded neonatal infections, frequent use of antibiotics with a broader spectrum, higher frequency of nosocomial sepsis).

  4. The Effect of Decoction Leaves from Green Leaf (piper betle l. to Inhibition Activity of Streptococcus Agalactiae Cause of Mastitis in Dairy Cow

    Directory of Open Access Journals (Sweden)

    Oki Selfiana Marsono

    2017-07-01

    Full Text Available ABSTRACT This research was conducted from January to February 2017 at Microbiology Laboratory, Faculty Science and Technology, Islamic Maulana Maliki Ibrahim University, Malang and at Microbiology Laboratory, Faculty of Science, Muhammadiyah University, Malang. The purpose of this research was to determine the effectiveness of Piper betle L. leave extract on antibacterial activity the growth of Streptococcus agalactiae. The method used was Complete Randomized Design (CRD with 8 treatments and 6 replications. Piper betle L. leaves extract was used 20% for P0 (0-day, P1 (first day, P2 (second day, P3 (third day, P4 (fourth day, P5 (fiveth day, P6 (sixth days and P7 (seventh day. The results showed that Piper betle L. leaves extract had a highly significant effect (P<0.01 on inhibiting the growth of the bacteria so the results were continued with LSD test analysys because there were differences among variables. The results showed that P0 was the highest (3.93±0.50; then followed by P1 (3.83±0.69; P2 (3.47±0.21; P3 (3.13±0.22; P4 (3.12±0.58; P5 (3.04±0.24; P6 (3.00±0.44 include at medium categories, and P7 (2.97±0.19 include at low category, where inhibition zone which formed on each treatment have non significant decreased. In conclusion, the storage of Piper betle L. extract have effect on antibacterial activities (Streptococcus agalactiae, which the highest at day-0.

  5. Purinergic signaling modulates the cerebral inflammatory response in experimentally infected fish with Streptococcus agalactiae: an attempt to improve the immune response.

    Science.gov (United States)

    Souza, Carine F; Baldissera, Matheus D; Bottari, Nathiele B; Moreira, Karen L S; da Rocha, Maria Izabel U M; da Veiga, Marcelo L; Santos, Roberto C V; Baldisserotto, Bernardo

    2018-06-01

    Appropriate control of the immune response is a critical determinant of fish health, and the purinergic cascade has an important role in the immune and inflammatory responses. This cascade regulates the levels of adenosine triphosphate (ATP), adenosine diphosphate, adenosine monophosphate and adenosine (Ado), molecules involved in physiological or pathological events as inflammatory and anti-inflammatory mediators. Thus, the aim of this study was to evaluate whether purinergic signaling, through the activities of nucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase, and adenosine deaminase (ADA), is capable of modulating the cerebral immune and inflammatory responses in silver catfish that is experimentally infected with Streptococcus agalactiae. Cerebral NTPDase (with ATP as substrate) and 5'-nucleotidase activities increased, while ADA activity decreased in silver catfish that is experimentally infected with S. agalactiae, compared to the control group. Moreover, the cerebral levels of ATP and Ado increased in infected animals compared to the uninfected control group. Brain histopathology in infected animals revealed inflammatory demyelination (the presence of occasional bubbly collections), increased cellular density in the area near to pia-mater and intercellular edema. Based on this evidence, the modulation of the purinergic cascade by the enzymes NTPDase, 5'-nucleotidase, and ADA exerts an anti-inflammatory profile due to the regulation of ATP and Ado levels. This suggests involvement of purinergic enzymes on streptococcosis pathogenesis, through regulating cerebral ATP and Ado levels, molecules known to participate in physiological or pathological events as inflammatory and anti-inflammatory mediators, respectively. In summary, the modulation of the cerebral purinergic cascade exerts an anti-inflammatory profile in an attempt to reduce inflammatory damage.

  6. Indirect enzyme-linked immunosorbent assay method based on Streptococcus agalactiae rSip-Pgk-FbsA fusion protein for detection of bovine mastitis.

    Science.gov (United States)

    Bu, Ri-E; Wang, Jin-Liang; Wu, Jin-Hua; Xilin, Gao-Wa; Chen, Jin-Long; Wang, Hua

    2017-03-01

    The aim of this study was to establish a rapid and accurate method for the detection of the Streptococcus agalactiae antibody (SA-Ab) to determine the presence of the bovine mastitis (BM)-causative pathogen. The multi-subunit fusion protein rSip-Pgk-FbsA was prokaryotically expressed and purified. The triple activities of the membrane surface-associated proteins Sip, phosphoglycerate kinase (Pgk), and fibronectin (FbsA) were used as the diagnostic antigens to establish an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of SA-Ab in BM. The optimal antigen coating concentration was 2 μg/mL, the optimal serum dilution was 1:160, and the optimal dilution of the enzyme-labeled secondary antibody was 1:6000. The sensitivity, specificity, and repeatability tests showed that the method established in this study had no cross-reaction with antibodies to Streptococcus pyogenes, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis in the sera. The results of the sensitivity test showed that a positive result could be obtained even if the serum dilution reached 1:12,800, indicating the high sensitivity and good repeatability of the method. The positive coincidence rate of this method was 98.6%, which is higher than that of previous tests established with the Sip or Pgk mono-antigen fusion protein, respectively, demonstrating the relatively higher sensitivity of this newly established method. The detection rate for 389 clinical samples was 46.53%. The indirect ELISA method established in this study could provide a more accurate and reliable serological method for the rapid detection of S. agalactiae in cases of BM.

  7. Investigation of the Antimicrobial Activity of Bacillus licheniformis Strains Isolated from Retail Powdered Infant Milk Formulae.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Begley, Máire; Clifford, Tanya; Deasy, Thérèse; Considine, Kiera; O'Connor, Paula; Ross, R Paul; Hill, Colin

    2014-03-01

    This study investigated the potential antimicrobial activity of ten Bacillus licheniformis strains isolated from retail infant milk formulae against a range of indicator (Lactococcus lactis, Lactobacillus bulgaricus and Listeria innocua) and clinically relevant (Listeria monocytogenes, Staphylococcus aureus, Streptococcus agalactiae, Salmonella Typhimurium and Escherichia coli) microorganisms. Deferred antagonism assays confirmed that all B. licheniformis isolates show antimicrobial activity against the Gram-positive target organisms. PCR and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses indicated that four of the B. licheniformis isolates produce the bacteriocin lichenicidin. The remaining six isolates demonstrated a higher antimicrobial potency than lichenicidin-producing strains. Further analyses identified a peptide of ~1,422 Da as the most likely bioactive responsible for the antibacterial activity of these six isolates. N-terminal sequencing of the ~1,422 Da peptide from one strain identified it as ILPEITXIFHD. This peptide shows a high homology to the non-ribosomal peptides bacitracin and subpeptin, known to be produced by Bacillus spp. Subsequent PCR analyses demonstrated that the six B. licheniformis isolates may harbor the genetic machinery needed for the synthesis of a non-ribosomal peptide synthetase similar to those involved in production of subpeptin and bacitracin, which suggests that the ~1,422 Da peptide might be a variant of subpeptin and bacitracin.

  8. Avaliação da sensibilidade da cultura de leite do tanque para isolamento de agentes contagiosos da mastite bovina Evaluation of the sensitivity of bulk tank milk cultures for the isolation of contagious bovine mastitis pathogens

    Directory of Open Access Journals (Sweden)

    Maria Aparecida V. P. Brito

    1998-01-01

    herds had SCC levels below 400,000 ml-1. S. aureus and S. agalactiae were isolated from five of the nine herds with high SCC, S. agalactiae from one and S. aureus from three. Six herds had SSC below 200,000 ml-1. S. aureus and S. agalactiae were isolated from one, S. aureus from three, while the other two were negative for both pathogens. The results of herds A, B, C and D sampled at the farms showed that S. aureus was isolated from 1.8%, 19.2%, 17.0% and 8.4% of the animals and 0.9%, 5.9%, 5.4% and 2.2% of the mammary quarters, respectively. S. agalactiae was isolated from herds A, C and D. Within these herds the percentages of isolation were, respectively, 1.8%, 10.6% and 8.4% for the cows and 0.46%, 3.8% and 3.7% for the mammary quarters. S. aureus was recovered from all three bulk tank cultures from herds A, B and D. Only the third sample from herd C was positive for S. aureus. S. agalactiae was recovered from all samples collected from herd D, two samples from herd C and one sample from herd A. Coliforms were isolated from all tank samples from herds A, B, C and D and from all but one sample collected in the processing plant. Yeasts were recovered from 16 herds sampled at the processing plant and from all tank samples from herds A, B, C, and D. Neither coliforms or yeasts were isolated from the individual animals of herds A, B, C and D. These findings indicate that the milk was contaminated during or after milking, probably due to deficient hygiene and cleaning procedures. The analysis of the bulk tank milk cultures showed that the test was sensitive enough to detect contagious mastitis pathogens. The sensitivity of the test increased when more than two consecutive samples were examined.

  9. Rapid identification of pathogenic streptococci isolated from moribund red tilapia (Oreochromis spp.).

    Science.gov (United States)

    Abdelsalam, Mohamed; Elgendy, Mamdouh Y; Shaalan, Mohamed; Moustafa, Mohamed; Fujino, Masayuki

    2017-03-01

    Accurate and rapid identification of bacterial pathogens of fish is essential for the effective treatment and speedy control of infections. Massive mortalities in market-sized red tilapia (Oreochromis spp.) were noticed in mariculture concrete ponds in northern Egypt. Histopathological examination revealed marked congestion in the central vein of the liver with the presence of bacterial aggregates inside the lumen and in the vicinity of the central vein. A total of 12 isolates of streptococci were obtained from the moribund fish. This study documented the ability of the MicroSeq 500 16S bacterial sequencing method to accurately identify Streptococcus agalactiae and S. dysgalactiae mixed infections from moribund red tilapia that were difficult to be recognised by the commercial biochemical systems. The continuously decreasing cost of the sequencing technique should encourage its application in routine diagnostic procedures.

  10. Antibacterial Effects of Citrus aurantium on Bacteria Isolated from Urinary Tract Infection

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    Masoud Dadashi

    2015-10-01

    Full Text Available Background :  Emerging antibacterial resistance rates and beta-lactamase producing bacteria recovered from UTI is an increasing problem in different regions, limiting therapeutic options. Therefore, this survey consider to use the extract and essence of the citrus aurantium (which have a so many rate of planting in Iran and also survey on extract on bacteria whose cause urinary tract infections, and compare this with common antibiotics. Methods and Materials: This study was experimental design.We have been isolate the E.coli,Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae and Enterococcus faecalis from UTI and then determine of antibacterial effect of Citrus aurantium against this bacteria with subculture and put the exact diagnosis on them. Antibacterial effects of the herb extract by well diffusion assay and  nalidixic acid and Co-trimoxazol were evaluated by method of agar disc diffusion. Results:Enterococcus faecalis had 100% sensitivity against of extract,essence and Co-trimoxazole , and 80% against nalidixic acid . E.coli had 100% sensitivity against Co-trimoxazol, nalidixic acid and it was totally resistance to extract and essence.Klebsiella Pneumonie had 80% to Co-trimoxazol, 75% to nalidixic acid and resistance against extract and essence.Streptococcus agalactiae was 100% sensitivity to essence and Co-trimoxazol and 90% against nalidixic acid and shown 80% sensitivity against extract.Staphylococcus aureus MRSA shown 100% sensitivity against Co-trimoxazol and 70% sensitivity against essence, extract and nalidixic acid. Conclusion: Detection of antibiotic resistance among isolates is important in prevention and control of infections. In this study, it was shown that extracts of citrus aurantium have high antibacterial effects on gram positive bacteria compare to gram negative bacteria.

  11. Mastitogenic bacteria isolated from dairy cows in Kenya and their antimicrobial sensitivity

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    George K. Gitau

    2014-02-01

    Full Text Available There is limited epidemiological knowledge on udder health in Kenyan dairy cattle that would aid in a pro-active approach towards mastitis prevention. The study objectives were: (1 to investigate the prevalence and distribution of clinical and subclinical mastitis in dairy cattle in Mukurwe-ini and Nakuru Districts, Kenya, and (2 to determine the antibacterial sensitivity of the organisms causing bovine mastitis in these districts. The study involved field-screening of milk samples from 241 dairy cows on 128 farms by use of the California Mastitis Test (CMT and, if CMT-positive, followed by bacteriological culture of the major causative agents and their respective antibiotic sensitivity to eight commonly used antibiotics. All participating farms were visited twice during the study period. The results obtained during the first and second visits showed the prevalence of clinical mastitis to be very low: 0.9% and 0.5%, respectively; 56.0% and 65.0% of cows were CMT-positive on at least one quarter and 49.6% and 58.7% of cows were culture-positive, respectively. There was no significant difference in mastitis prevalence between Nakuru and Mukurwe-ini districts (p > 0.10. Staphylococcus aureus was isolated in 68.0% and 77.0% of samples during the first and second visits, respectively. Other frequently isolated agents included Streptococcus agalactiae, and other Streptococcus spp., S. aureus and S. agalactiae were most sensitive to gentamycin and norfloxacin, and least sensitive to cotrimazole and ampicillin. Knowing the prevalence of mastitogenic organisms and their antibiotic sensitivities could improve treatment efficacy and cow longevity.

  12. Prevalencia de Mycoplasma agalactiae y Toxoplasma gondii en caprinos y ovinos de los municipios del sur de la Guajira y el Norte del Cesar

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    Oscar Fabián Porras Pinto

    2016-06-01

    Full Text Available Introducción: Los departamentos de la Guajira y Cesar son considerados los principales productores de ovinos y caprinos del país, según censo del Instituto Colombiano Agropecuario (ICA en el año 2015. Estas especies animales revisten interés desde el punto de vista sanitario, no solo por los productos obtenidos de ellos, sino también por la capacidad que tienen de ser reservorio de diferentes agentes infecciosos. A la fecha en Colombia no se encuentran reportes relacionados con la prevalencia de agentes infecciosos, como es el caso de Mycoplasma agalactiae y Toxoplasma gondii en estas poblaciones animales, razón por la cual, en 2015 se comienza a desarrollar un trabajo inter-institucional titulado: “Proyecto Piloto de Excelencia Sanitaria en Ganadería para la Guajira y el Cesar”, proyecto en el que participan entidades públicas y privadas, el cual, anida el presente trabajo. Objetivo: Determinar la prevalencia de M. agalactiae y T. gondii en caprinos y ovinos, basada en la determinación de IgG dirigida a estos agentes infecciosos, para la construcción de un perfil sanitario en los municipios del sur de la Guajira y el Norte del Cesar. Materiales y métodos: El presente trabajo corresponde a un estudio descriptivo de corte transversal, que tuvo como universo de estudio una población de 15.300 ovinos y caprinos del sur de la Guajira y norte del Cesar definida por Censo realizado en 2015 por el ICA. Tomado como base esta población, se calculó un tamaño muestral de 1039 individuos mediante la herramienta WinEpi versión 2.0 (Zaragoza, España. Resultados: La determinación de la prevalencia de M. agalactiae y T. gondii se realizará mediante la evaluación de la presencia de IgG sérica específica para estos agentes infecciosos mediante pruebas comerciales de ELISA indirecta, según las recomendaciones de los fabricantes. Las pruebas serán realizadas a sueros provenientes de muestras sanguíneas de animales incluidos en la

  13. Hibridación in situ para la detección de Streptococcus agalactiae en tejidos de tilapia (Oreochromis sp.

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    E. A. Pulido

    2010-04-01

    Full Text Available La estreptococosis es uno de los problemas sanitarios más serios en la acuicultura mundial.En Colombia la enfermedad afecta de manera importante las explotaciones detilapia. Se estandarizó la técnica de hibridación in situ (HIS en tejidos de tilapia previamenteidentificados como positivos a la presencia de S. agalactiae por la técnicade inmunoperoxidasa indirecta (IPI y microbiología. Se obtuvo señal positiva en elinterior de los granulomas con una especificidad del 100%. Se logró amplificar significativamentela señal mediante el uso de la tiramida. La HIS en tejidos de tilapia puedeser usada para el diagnóstico y estudios de patogenésis y epidemiológicos con este microorganismo.Se requieren futuras investigaciones para optimizar la marcación de lasbacterias libres en los tejidos y evaluar la sensibilidad de la técnica.

  14. Hibridación in situ para la detección de streptococcus agalactiae en tejidos de tilapia (oreochromis sp.

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    E. A. Pulido

    2010-01-01

    Full Text Available La estreptococosis es uno de los problemas sanitarios más serios en la acuicultura mundial. En Colombia la enfermedad afecta de manera importante las explotaciones de tilapia. se estandarizó la técnica de hibridación in situ (HIS en tejidos de tilapia previamente identificados como positivos a la presencia de S. agalactiae por la técnica de inmunoperoxidasa indirecta (IPI y microbiología. Se obtuvo señal positiva en el interior de los granulomas con una especificidad del 100%. Se logró amplificar significativamente la señal mediante el uso de la tiramida. La HIS en tejidos de tilapia puede ser usada para el diagnóstico y estudios de patogenésis y epidemiológicos con este microorganismo. Se requieren futuras investigaciones para optimizar la marcación de las bacterias libres en los tejidos y evaluar la sensibilidad de la técnica.

  15. RNA-Seq revealed the impairment of immune defence of tilapia against the infection of Streptococcus agalactiae with simulated climate warming.

    Science.gov (United States)

    Wang, Le; Liu, Peng; Wan, Zi Yi; Huang, Shu Qing; Wen, Yan Fei; Lin, Grace; Yue, Gen Hua

    2016-08-01

    Global warming is one of the causes of disease outbreaks in fishes. Understanding its mechanisms is critical in aquaculture and fisheries. We used tilapia to study the effects of a high temperature on the infection of a bacterial pathogen Streptococcus agalactiae using RNA-Seq. We found that the dissolved oxygen level in water at 32 °C is lower than at 22 °C, and tilapia infected with the pathogen died more rapidly at 32 °C. The gene expression profiles showed significant differences in fish raised under different conditions. We identified 126 and 576 differentially expressed genes (DEGs) at 4 and 24 h post infection at 22 °C, respectively, whereas at 32 °C, the data were 312 and 1670, respectively. Almost all responding pathways at 22 °C were involved in the immune responses, whereas at 32 °C, the enriched pathways were not only involved in immune responses but also involved in oxygen and energy metabolisms. We identified significant signals of immunosuppression of immune responses at 32 °C. In addition, many of the enriched transcription factors and DEGs under positive selection were involved in immune responses, oxygen and/or energy metabolisms. Our results suggest that global warming could reduce the oxygen level in water and impair the defence of tilapia against bacterial infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Antimicrobial Sensitivity Pattern of Microorganisms Isolated from Vaginal Infections at a Tertiary Hospital in Bangalore, India

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    Nagalakshmi Narayana-Swamy

    2015-03-01

    Full Text Available Background: The vagina contains dozens of microbiological species in variable quantities and is, therefore, considered a complex environment. Among the microorganisms, bacteria have important repercussions on women’s health. The present study was conducted especially to elucidate this type of vaginal isolates and their sensitivity towards currently used antibiotics. Methods: This was a retrospective study conducted at the Department of Obstetrics and Gynaecology, Sapthagiri Hospital, Bangalore, India from January 2012 to December 2013. All symptomatic women who had a high vaginal swab taken for culture and sensitivity testing were included in this study. Antibiotic susceptibility was tested using disc diffusion method (modified Kirby-Bauer method. The antibiotic sensitivity patterns of isolated microorganisms were studied. Results: Out of 200 patients, 95% had positive vaginal cultures. Fifteen types of microorganisms were isolated. The highest frequency of infection was observed at the age of 20-30 years, followed by 41-50 years and 31-40 years, and a low frequency of infection was observed above 50 years of age. The most prevalent pathogen was Escherichia coli, followed by Streptococcus agalactiae and diphtheroids with equal incidence. Among the antibiotics tested, isolated pathogens were completely resistant to nalidixic acid and highly sensitive to meropenem and imepenem. Conclusion: The high prevalence of gynaecological infections demands that patients with symptoms undergo thorough investigation with cultures and sensitivity essays. Changes in treatment protocols are required to treat vaginal infections effectively.

  17. Isolation World

    OpenAIRE

    Núñez Martín, Eugeni

    2012-01-01

    El trabajo de fin de grado tiene como nombre “Isolation World”, que en su traducción literal significa “Aislamiento del mundo”, un videojuego diseñado y creado desde cero en su totalidad, utilizando herramientas y conocimiento de lógica en programación que se han ido aprendiendo y desarrollando a lo largo de la carrera.

  18. Bacteriocinogenic potential and virulence traits of Enterococcus faecium and E. faecalis isolated from human milk

    Science.gov (United States)

    Khalkhali, Soodabeh; Mojgani, Naheed

    2017-01-01

    Background and Objectives: Human milk is a continuous supply of Lactic Acid bacteria (LAB), including enterococci with probiotic potentials. The aim of this study was to analyze two Enterococcus species, isolated from human milk for their probiotic potential, bacteriocin producing ability and virulence traits. Materials and Methods: Enterococcus faecium TA0033 and E. faecalis TA102 were tested for acid and bile tolerance, survival in simulated gastric and intestinal conditions. The antibacterial spectrum of the isolates was tested by agar well diffusion assay. The antagonistic agent was characterized by physico-chemical methods. The enterocin structural genes, virulence determinants, vancomycin resistance and biogenic amine genes, such as hdc1, hdc2, tdc, ldc and odc were also determined. Results: The tested isolates survived acidic conditions, high bile salt (1%), simulated gastric and intestinal conditions. The culture supernatant fluids of the two isolates inhibited the growth of Escherichia coli, Listeria monocytogenes, Salmonella typhi, Staphylococcus aureus, Shigella dysenteriae and Streptococcus agalactiae. The antagonistic activity was lost in the presence of proteolytic enzymes but tolerated the action of catalase, lysozyme and lipase. In contrast to enterocin TA102, enterocin TA0033 possessed bactericidal mode of action. Bacteriocin structural genes, entA and entB were present in the genome of the two isolates, while E. faecalis TA102 additionally harboured entP and bac31 genes. The phenotypic and genotypic virulence assessment studies indicated hyaluronidase (hyl) production and vancomycin resistance in E. faecalis TA102 while, none of the isolates harboured the biogenic amine genes. Conclusion: The presence of virulence genes in E. faecalis TA102 calls for careful monitoring of Enterococcus isolates for their safety parameters. PMID:29238458

  19. [Evaluation of Prolex for the rapid identification of streptococci isolated in medical microbiology].

    Science.gov (United States)

    Loubinoux, J; Mihaila-Amrouche, L; Bouvet, A

    2004-10-01

    The need to rapidly identify streptococci responsible for acute infectious diseases has led to the development of agglutination techniques that are able to identify streptococcal group antigens (A, B, C, D, F, and G) directly from primoculture colonies on blood agar. The Prolex agglutination tests (Pro-Lab Diagnostics, Richmond Hill, Ontario, Canada), distributed in France by i2a, have been used for the determination of group antigens of 166 isolates of streptococci and enterococci previously identified in the National Reference Center for Streptococci. The results obtained with the Prolex reagents have permitted to correctly identify all pyogenic beta-hemolytic streptococci (23 Streptococcus pyogenes, 21 Streptococcus agalactiae, 33 Streptococcus dysgalactiae subsp. equisimilis including 6 group C and 27 group G, and 5 Streptococcus porcinus including 4 group B). Four differences between unexpected agglutinations (A or F) and species identifications have been obtained. These differences were observed for four non-hemolytic isolates of Streptococcus mutans, Streptococcus gordonii, Streptococcus infantarius, and Streptococcus suis. The anti-D reagent has been of value as a marker for isolates of enterococci. Thus, these results confirm the abilities of these agglutination tests for the grouping of beta-hemolytic streptococci. Moreover, the use of Prolex has the advantage to be rapid because of the non-enzymatic but chemical extraction of streptococcal antigens.

  20. Loop-mediated isothermal amplification assay for rapid detection of Streptococcus agalactiae (group B streptococcus) in vaginal swabs - a proof of concept study.

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    McKenna, James Patrick; Cox, Ciara; Fairley, Derek John; Burke, Rachael; Shields, Michael D; Watt, Alison; Coyle, Peter Valentine

    2017-03-01

    Neonatal sepsis caused by Streptococcus agalactiae [group B streptococcus (GBS)] is a life-threatening condition, which is preventable if colonized mothers are identified and given antibiotic prophylaxis during labour. Conventional culture is time consuming and unreliable, and many available non-culture diagnostics are too complex to implement routinely at point of care. Loop-mediated isothermal amplification (LAMP) is a method that, enables the rapid and specific detection of target nucleic acid sequences in clinical materials without the requirement for extensive sample preparation. A prototype LAMP assay targeting GBS sip gene is described. The assay was 100 % specific for GBS, with a limit of detection of 14 genome copies per reaction. The clinical utility of the LAMP assay for rapid direct molecular detection of GBS was determined by testing a total of 157 vaginal swabs with minimal sample processing using a rapid lysis solution. Compared to a reference quantitative real-time PCR assay, the direct LAMP protocol had a sensitivity and specificity of 95.4 and 100 %, respectively, with positive and negative predictive values of 100 and 98.3 %, respectively. Positive and negative likelihood ratios were infinity and 0.05, respectively. The direct LAMP method required a mean time of 45 min from the receipt of a swab to generation of a confirmed result, compared to 2 h 30 min for the reference quantitative real-time PCR test. The direct LAMP protocol described is easy to perform, facilitating rapid and accurate detection of GBS in vaginal swabs. This test has a potential for use at point of care.

  1. The effects of dietary kefir and low molecular weight sodium alginate on serum immune parameters, resistance against Streptococcus agalactiae and growth performance in Nile tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Van Doan, Hien; Hoseinifar, Seyed Hossein; Tapingkae, Wanaporn; Khamtavee, Pimporn

    2017-03-01

    The present study evaluates the effects of dietary kefir and low molecular weight sodium alginate (LWMSA) (singular or combined) on non-specific immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Fish with average weight of 18.60 ± 0.04 g were supplied and randomly stocked in sixteen glass tanks (150 L) at density of 20 fish per tank. Fish were fed experimental diets as follows: 0 g kg -1 LMWSA (Control, Diet 1), 10 g kg -1 LMWSA (Diet 2), 40 g kg -1 kefir (Diet 3), and 10 g kg -1 LMWSA + 40 g kg -1 kefir (Diet 4) for 50 days. At the end of the feeding trial, serum lysozyme (SL), phagocytosis (PI), respiratory burst (RB), and alternative complement (ACH50) activities as well as growth performance were measured. Singular and combined administration of kefir and low molecular weight sodium alginate (LMWSA) significantly increased serum SL, PI, RB, and ACH50 activities compared control group (P < 0.05); the highest innate immune responses were observed in fish fed combinational diet (kefir + LMWSA) (P < 0.05). The results of experimental challenge revealed significantly higher resistance against Streptococcus agalactiae in fish fed supplemented diets and the highest post challenge survival rate was observed in synbiotic diet (P < 0.05). Similar results obtained in case of growth parameters. Feeding on supplemented diet significantly improved SGR and FCR and the highest growth parameters was observed in fish fed synbiotic diet (P < 0.05). These finding revealed that combined administration of dietary kefir and LMWSA can be considered for improving immune response, disease resistance and growth performance of Nile tilapia. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Isolation of Arcobacter spp from the milk of dairy cows in Brazil Isolamento de Arcobacter spp do leite de vacas leiteiras no Brasil

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    Celso Pianta

    2007-02-01

    Full Text Available Bacteriologic examinations were performed on 188 milk samples collected from cows from 11 farms for diagnosis of mastitis in three cities of Rio Grande do Sul, Brazil. Among the common causes of mastitis, the most frequent isolates were Staphylococcus aureus, followed by Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae and Streptococcus agalactiae. Bacteriologic examination of 32 milk samples from one farm didn't show bacteria known as common etiologic agent of mastitis. Six samples of Arcobacter spp typed by genotypic tests as Arcobacter cryaerophilus (five strains and Arcobacter butzleri (one strain were isolated from cows' milk of that farm. It is reported the isolation of Arcobacter species from the milk of cows in absence of clinical signs of mastitis. This is the first report of the detection of the microorganisms in the milk of dairy cows in Brazil. No previous reports are known from other countries.Foram realizados exames bacteriológicos em 188 amostras de leite colhidas de vacas de 11 propriedades leiteiras para diagnóstico de mastite, em três municípios no Rio Grande do Sul, Brasil. Entre as causas comuns de mastite, os isolados mais freqüentes foram Staphylococcus aureus, seguido de Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae e Streptococcus agalactiae. O exame bacteriológico realizado em 32 amostras de leite de vacas de uma propriedade não demonstrou a presença de bactérias conhecidas como causadoras de mastite. Foram isoladas do leite de vacas desta propriedade seis amostras de Arcobacter spp, classificadas por testes moleculares como Arcobacter cryaerophilus (cinco amostras e Arcobacter butzleri (uma amostra. É relatado o isolamento de espécies de Arcobacter do leite de vacas na ausência de sinais clínicos de mastite. Este é o primeiro relato da detecção dos microorganismos no leite de vacas leiteiras no Brasil.

  3. Susceptibility to antimicrobial agents among bovine mastitis pathogens isolated from North American dairy cattle, 2002-2010.

    Science.gov (United States)

    Lindeman, Cynthia J; Portis, Ellen; Johansen, Lacie; Mullins, Lisa M; Stoltman, Gillian A

    2013-09-01

    Approximately 8,000 isolates of Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli, isolated by 25 veterinary laboratories across North America between 2002 and 2010, were tested for in vitro susceptibility to beta-lactam, macrolide, and lincosamide drugs. The minimal inhibitory concentrations (MICs) of the beta-lactam drugs remained low against most of the Gram-positive strains tested, and no substantial changes in the MIC distributions were seen over time. Of the beta-lactam antimicrobial agents tested, only ceftiofur showed good in vitro activity against E. coli. The MICs of the macrolides and lincosamides also remained low against Gram-positive mastitis pathogens. While the MIC values given by 50% of isolates (MIC50) for erythromycin and pirlimycin and the streptococci were all low (≤0.5 µg/ml), the MIC values given by 90% of isolates (MIC90) were higher and more variable, but with no apparent increase over time. Staphylococcus aureus showed little change in erythromycin susceptibility over time, but there may be a small, numerical increase in pirlimycin MIC50 and MIC90 values. Overall, the results suggest that mastitis pathogens in the United States and Canada have not shown any substantial changes in the in vitro susceptibility to beta-lactam, macrolide, and lincosamide drugs tested over the 9 years of the study.

  4. Benih Keturunan Induk Ikan Nila yang Divaksinasi pada Tingkat Kematangan Gonad-2 Lebih Tahan Terhadap Infeksi Streptococcus agalactiae (RESISTANCE OF TILAPIA (OREOCHRIMIS NILOTICUS FRY VACCINATED AT DIFFERENT GONADAL DEVELOPMENTAL STAGES TOWARD STREPTOCO

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    Khairun Nissa

    2016-11-01

    Full Text Available The aim of this study was to evaluate the effectiveness of vaccination based on gonad maturationstages on tilapia brood stocks in which the released antibodies was able to be transferred to the seed.Vaccine composed with whole cells and extracellular product (ECP was injected at stage 2 and stage 3 ofthe gonad development stages at concentration of 109 CFU mL1 as much as 4 mL to 1 kg of brood fish.Control fish was unvaccinated treatment. Challenge study at seed was conducted by immersing S. agalactiaefor 30 minutes at 7, 14, 21, and 28 days post hatching (DPH in 107 CFU/mL. Antibody levels on broodstocks, eggs, and body fluids of seed, and relative percentage survival (RPS of seed post challenge studywere evaluated. The results showed that stage 2 of gonad developmental stages was found on 7 days postinitial spawning and stage 3 found on 14 days post initial spawning of brood fish. Vaccinated done in stage 2 of gonad developmental stages gave immunoglobulin serum in brood (0,166±0,001, egg (0,165±0,002,and seed aged 7, 14, 21, and 28 days post hatching (0,164±0,002, 0,162±0,005, 0,155±0,006, and 0,14±0,008respectively were significantly higher (P<0,05 compared to other treatment. Challenged test that doneby immersing with S. agalactiae suspension on larval aged 7, 14, 21, and 28 days had highest RPS(95,24%, 83,33%, 72,22%, and 56,02% respectively formed on seed from brood stock vaccination in gonaddevelopment stage 2. Vaccination in tilapia brood stocks at stage 2 of gonad developmental stages gavehighest protection by maternal immunity to the seed against S. agalactiae.

  5. Prevalence and Antimicrobial Susceptibility Profiles of Pathogen Isolated from Bovine Mastitis Milk in Transylvania, Romania

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    Cosmina Bouari

    2016-11-01

    Full Text Available Mastitis in cows, one of the most common and economically important infectious diseases of dairy cattle, all over the world, with significant impact due to economic losses, occurs when the udder becomes inflamed because the leukocytes are released into the mammary gland usually in response to bacteria invasion of the teat canal. The main objective of this study was to evaluate the in vitro antimicrobial susceptibility of bacteria isolated from milk in order to design specific control programs for bovine mastitis in this area. A total of 204 milk samples aseptically collected both from farms and private owners were processed during May 2014 and March 2016 within the Microbiology Laboratory of the Faculty of Veterinary Cluj-Napoca, Romania. The microbiological examination was carried out by inoculation on blood agar and MacConkey medium. After the overnight incubation in aerobic conditions, the identification of the isolates was performed using microscopic, cultural and biochemical methods. Biochemical identification was based on API 20 Biomerieux system. Susceptibility to antibiotics was evaluated using Kirby Bauer disk diffusion method on Mueller-Hinton agar; the antibiotics were represented by Amoxicillin and Clavulanic Acid, Ceftiofur, Florfenicol, Mastidiscs, Enrofloxacin, Penicillin and Tetracycline. Staphylococcus spp. was the most common isolated pathogen, in 54.9% of the specimens, followed by Streptococcus spp. in 20.1%, Escherichia coli in 10.78%, Klebsiella spp. in 8.34%, Bacillus spp. in 5.88%. The most frequent associations were represented by staphylococci-streptococci in 62.7% of the samples, followed by streptococci-bacillus in 19.8% of the samples. The most important etiological agents identified were Staphylococcus aureus, S uberis, Streptococcus agalactiae, and Escherichia coli. Antimicrobial susceptibility test for the total isolates revealed good sensitivity to Enrofloxacin, Mastidiscs and Amoxicillin and Clavulanic Acid

  6. Prevalência de colonização por Streptococcus agalactiae em gestantes atendidas em maternidade do Ceará, no Brasil, correlacionando com os resultados perinatais Prevalence of the colonization by Streptococcus agalactiae in pregnant women from a maternity in Ceará, Brazil, correlating with perinatal outcomes

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    José Juvenal Linhares

    2011-12-01

    Full Text Available OBJETIVO: Analisar a prevalência de Streptococcus agalactiae, um estreptococo do Grupo B, em gestantes e seus possíveis fatores de risco, bem como o impacto perinatal e a suscetibilidade antimicrobiana das colonizadas. MÉTODOS: Foram avaliadas 213 gestantes a partir de 20 semanas de gestação, independente dos fatores de risco, atendidas em um hospital-escola terciário da zona Norte do Estado de Ceará, no Brasil. O cálculo do tamanho amostral ocorreu por conveniência. Foi utilizada técnica do swab estéril único para coleta de secreção das regiões vaginal e perianal. As amostras recém-obtidas eram armazenadas em meio de transporte Stuart e, no laboratório, inoculadas em meio seletivo Todd-Hewitt adicionado de gentamicina (8 ug/mL e ácido nalidíxico (15 ug/mL, com posterior subcultivo em placas em ágar-sangue. Nos materiais eram realizados teste de Gram, catalase com peróxido de oxigênio e CAMP (Christie, Atkins, Munch-Petersen, sendo confirmados sorologicamente com Streptococcal Grouping Kit, Oxoid®. As positivas foram submetidas a testes de suscetibilidade antimicrobiana. Foram também avaliadas variáveis socioeconômicas, reprodutivas, clínico-obstétricas e neonatais. Os dados foram analisados utilizando o programa Epi-Info 6.04. RESULTADOS: A prevalência de colonização encontrada foi de 9,8% pelo teste de CAMP, embora apenas 4,2% pelo sorológico. O único fator de proteção observado foi cor da pele branca (p=0,01, 0.45>OR>0.94, IC95%. Não foi observada diferença de prevalência do estreptococo do Grupo B com outras variáveis reprodutivas ou obstétricas. Ocorreu infecção em apenas um dos recém-nascidos de mães colonizadas, entretanto revelou-se infecção por Pseudomonas spp. Foi encontrada resistência para ampicilina (4/9 e cefalotina (4/9, penicilina (4/9 casos, eritromicina (3/9, clindamicina (7/9 e cloranfenicol (1/9. CONCLUSÕES: A taxa de infecção foi inferior à encontrada em outros estudos

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

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    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

  8. Optimum Shock Isolation

    National Research Council Canada - National Science Library

    Bolotnik, Nikolai

    2001-01-01

    .... Several types of performance criteria for isolation are considered, the most important of which are the peak force transmitted to the body to be isolated and the maximum displacement of the body relative to the base...

  9. Isolated patellofemoral osteoarthritis.

    NARCIS (Netherlands)

    Jonbergen, J.P.W. van; Poolman, R.W.; Kampen, A. van

    2010-01-01

    BACKGROUND AND PURPOSE: The optimal treatment for isolated patellofemoral osteoarthritis is unclear at present. We systematically reviewed the highest level of available evidence on the nonoperative and operative treatment of isolated patellofemoral osteoarthritis to develop an evidenced-based

  10. Isolation and characterization of Streptococcus spp. group B in Nile tilapias (Oreochromis niloticus reared in hapas nets and earth nurseries in the northern region of Parana State, Brazil

    Directory of Open Access Journals (Sweden)

    Salvador Rogério

    2005-01-01

    Full Text Available The objective of this study was to isolate and characterize Streptococcus spp. in Nile tilapias (Oreochromis niloticus reared in net-pens and earth nurseries. Eight intensive tilapia-rearing farms were investigated in north Paraná, Brazil from April 1st 2001 to April 30th 2002. The fish were reared in a system of hapas nets on four farms and in earth nurseries on other four farms. A total of 370 samples were analyzed of material collected from 120 fish (brain, liver, kidney, skin scrapes, ascites liquid and eye that were sown on BHI agar (Brain Heart Infusion supplemented with 1% yeast extract and sheep blood. Streptococcus spp. was isolated in 36 of the samples (18 brain, eight liver, eight kidney and two ascites liquid from 25 fish. Streptococci were isolated in both systems, almost in the same proportion. First the streptococci were characterized by the catalase and esculin test, growth in methylene blue and sodium chloride at 6.5%. They were classified in groups by the Slidex Strepto-Kit (BioMerieux, France. The phenotypic characteristics were determined by the Api 20 Strep microtest system (BioMerieux, France. The 36 Streptococcus spp. samples did not present hemolysis and were classified as Lancefield group B. Further 16 samples were identified as Streptococcus agalactiae and 20 were not identified by the Api 20 Strep, but presented the same biochemical profile described for the reference strain of Streptococcus difficile (ND-2-22.

  11. Mutation and premating isolation

    Science.gov (United States)

    Woodruff, R. C.; Thompson, J. N. Jr

    2002-01-01

    While premating isolation might be traceable to different genetic mechanisms in different species, evidence supports the idea that as few as one or two genes may often be sufficient to initiate isolation. Thus, new mutation can theoretically play a key role in the process. But it has long been thought that a new isolation mutation would fail, because there would be no other individuals for the isolation-mutation-carrier to mate with. We now realize that premeiotic mutations are very common and will yield a cluster of progeny carrying the same new mutant allele. In this paper, we discuss the evidence for genetically simple premating isolation barriers and the role that clusters of an isolation mutation may play in initiating allopatric, and even sympatric, species divisions.

  12. Physics in isolation

    International Nuclear Information System (INIS)

    Anon.

    1986-01-01

    In late May, about 330 physicists made their way up to isolated and beautiful Lake Louise high in the Canadian Rockies about 100 miles west of Calgary in a second effort to increase interactions between particle and nuclear physicists. The conference series aims to foster exciting and diverse physics by bringing the different physicists together somewhere which is so isolated that they must interact with each other. The formula worked very well and isolated Lake Louise was a huge success

  13. Physics in isolation

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    1986-09-15

    In late May, about 330 physicists made their way up to isolated and beautiful Lake Louise high in the Canadian Rockies about 100 miles west of Calgary in a second effort to increase interactions between particle and nuclear physicists. The conference series aims to foster exciting and diverse physics by bringing the different physicists together somewhere which is so isolated that they must interact with each other. The formula worked very well and isolated Lake Louise was a huge success.

  14. Isolated optic nerve pseudotumour

    International Nuclear Information System (INIS)

    Patankar, T.; Prasad, S.; Krishnan, A.; Laxminarayan, R.

    2000-01-01

    Isolated optic nerve involvement by the idiopathic inflammatory process is a rare finding and very few reports are available. Here a case of an isolated optic nerve inflammatory pseudotumour presenting with gradually progressive unilateral loss of vision is described. It showed dramatic response to a trial of steroids and its differential diagnoses are discussed. Copyright (1999) Blackwell Science Pty Ltd

  15. Antibacterial screening of Rumex species native to the Carpathian Basin and bioactivity-guided isolation of compounds from Rumex aquaticus.

    Science.gov (United States)

    Orbán-Gyapai, Orsolya; Liktor-Busa, Erika; Kúsz, Norbert; Stefkó, Dóra; Urbán, Edit; Hohmann, Judit; Vasas, Andrea

    2017-04-01

    Plants belonging to the genus Rumex (family Polygonaceae) are used worldwide in traditional medicine for the treatment of various diseases caused by different microorganisms (e.g. bacteria-related dermatologic conditions, dysentery and enteritis). The present study focused on the antibacterial screening of Rumex species native to the Carpathian Basin, and isolation of compounds from one of the most efficient species, Rumex aquaticus. The antibacterial effects of n-hexane, chloroform and aqueous fractions of methanol extracts prepared from different parts of 14 Rumex species (R. acetosella, R. acetosa, R. alpinus, R. aquaticus, R. conglomeratus, R. crispus, R. hydrolapathum, R. obtusifolius subsp. obtusifolius, R. obtusifolius subsp. subalpinus, R. patientia, R. pulcher, R. scutatus, R. stenophyllus and R. thyrsiflorus) were investigated against Staphylococcus epidermidis, S. aureus, MRSA, Bacillus subtilis, Moraxella catarrhalis, Streptococcus pyogenes, S. pneumoniae, S. agalactiae, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae using the disc diffusion method. Mainly the n-hexane and chloroform extracts prepared from the roots of the plants displayed high antibacterial activity (inhibition zones>15mm) against one or more bacterial strains. The highly active extracts of the aerial part and root of R. aquaticus were subjected to a multistep separation procedure. 19 Compounds, among them naphthalenes (musizin, and its glucoside, torachrysone-glucoside, 2-methoxystypandrone), anthraquinones (emodin, chrysophanol, physcion, citreorosein, chrysophanol-8-O-glucoside), flavonoids (quercetin, quercetin-3,3'-dimethylether, isokaempferide, quercetin 3-O-arabinoside, quercetin 3-O-galactoside, catechin), stilbenes (resveratrol, piceid), and 1-stearoylglycerol were isolated from the plant. The antibacterial activities of isolated compounds were determined, and it was observed that especially naphthalenes exerted remarkable antibacterial effects against

  16. Multiple-locus variant-repeat assay (MLVA) is a useful tool for molecular epidemiologic analysis of Streptococcus agalactiae strains causing bovine mastitis.

    Science.gov (United States)

    Radtke, Andreas; Bruheim, Torkjel; Afset, Jan Egil; Bergh, Kåre

    2012-06-15

    Group B streptococci (GBS) were considered a major cause of mastitis in cattle until preventive measures succeeded in controlling the disease in the 1970s and 1980s. During the last 5-6 years an increasing number of cases have been observed in some Scandinavian countries. A total of 187 GBS isolates from mastitis cases were collected from 119 animals in 34 Norwegian farms in the period from April 2007 to November 2010. 133 (71%) of the isolates were from farms with automated milking systems. The strains underwent typing of capsular polysaccharides (CPS) and surface proteins, and were analyzed by multi-locus variable repeat assay (MLVA) to investigate the epidemiological relationship of strains within and between farms. The GBS strains were differentiated into 12 types by CPS and surface protein analysis, with CPS types V (54%) and IV (34%) predominating. MLVA was superior to CPS and protein typing for strain differentiation, resolving the 187 strains into 37 types. In 29 of 34 farms all GBS strains had identical MLVA profiles specific for each farm. However, in one farm represented with 48 isolates, four MLVA variants with differences in one repeat locus were observed during the almost 3-year long collection period. Similar variations were observed at four other farms. This might reflect the stability of repeat loci under in vivo conditions. Farms with automated milking systems were overrepresented in this material. In conclusion, the five-loci MLVA allowed rapid high-resolution genotyping of the bovine GBS strains within and between farms. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. MOX Fabrication Isolation Considerations

    Energy Technology Data Exchange (ETDEWEB)

    Eric L. Shaber; Bradley J Schrader

    2005-08-01

    This document provides a technical position on the preferred level of isolation to fabricate demonstration quantities of mixed oxide transmutation fuels. The Advanced Fuel Cycle Initiative should design and construct automated glovebox fabrication lines for this purpose. This level of isolation adequately protects the health and safety of workers and the general public for all mixed oxide (and other transmutation fuel) manufacturing efforts while retaining flexibility, allowing parallel development and setup, and minimizing capital expense. The basis regulations, issues, and advantages/disadvantages of five potential forms of isolation are summarized here as justification for selection of the preferred technical position.

  18. isolated from Trichoderma harzianum

    African Journals Online (AJOL)

    SAM

    2014-05-21

    May 21, 2014 ... Chitinase gene from Trichoderma harzianum was cloned and hetrologously over expressed in ... used by Trichoderma to inhibit the growth of other fungi. ..... actinomycete isolates from niche habitats in Manipur for antibiotic.

  19. Inhibitory Effect of Lactobacillus reuteri on Some Pathogenic Bacteria Isolated From Women With Bacterial Vaginosis

    Directory of Open Access Journals (Sweden)

    Eslami

    2014-08-01

    Full Text Available Background Considering the high prevalence of bacterial vaginosis and its association with urinary tract infection in women and treatment of gynecologic problems occur when a high recurrence of bacterial vaginosis is often treated with antibiotics. Objectives The purpose of this study was to investigate the inhibitory effect of Lactobacillus reuteri on pathogenic bacteria isolated from women with bacterial vaginosis. Materials and Methods Ninety-six samples were obtained from vaginal discharge of women with bacterial vaginosis by a gynecologist with a Dacron swab and put in sterile tubes containing TSB broth and Thioglycollate broth. Then were immediately sent to the laboratory in cold chain for further assessment. Afterward, culture was transferred on blood agar, EMB, Palcam and differential diagnosis environments. Then cultures were incubated for 24 hours at 37 °C. Lactobacillus reuteri strains were cultured in MRS environment and transferred to laboratory. After purification of pathogenic bacteria, Lactobacillus reuteri inhibitory effect on pathogenic bacteria was evaluated by minimum inhibitory concentration (MIC and antibiogram. Statistical analysis was performed using SPSS software v.16. Results The results of this study demonstrated the inhibitory effect of Lactobacillus reuteri on some pathogenic bacteria that cause bacterial, including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Enterococcus, Listeria monocytogenes and E. coli. Microscopic examination of stained smears of most Lactobacillus and pathogenic bacteria showed reduced. The prevalence of abnormal vaginal discharge, history of drug use, contraceptive methods and douching were 61%, 55%, 42% and 13%, respectively. Significant difference was observed between the use and non-use of IUD in women with bacterial. Conclusions Our findings indicated the inhibitory effect of Lactobacillus reuteri on pathogenic bacteria that

  20. Seismic isolation structure for pool-type LMFBR - isolation building with vertically isolated floor for NSSS

    International Nuclear Information System (INIS)

    Sakurai, A.; Shiojiri, H.; Aoyagi, S.; Matsuda, T.; Fujimoto, S.; Sasaki, Y.; Hirayama, H.

    1987-01-01

    The NSSS isolation floor vibration characteristics were made clear. Especially, the side support bearing (rubber bearing) is effective for horizontal floor motion restraint and rocking motion control. Seismic isolation effects for responses of the reactor components can be sufficiently expected, using the vertical seismic isolation floor. From the analytical and experimental studies, the following has been concluded: (1) Seismic isolation structure, which is suitable for large pool-type LMFBR, were proposed. (2) Seismic response characteristics of the seismic isolation structure were investigated. It was made clear that the proposed seismic isolation (Combination of the isolated building and the isolated NSSS floor) was effective. (orig./HP)

  1. [Yearly changes in antibacterial activities of cefozopran against various clinical isolates between 1996 and 2000--I. Gram-positive bacteria].

    Science.gov (United States)

    Suzuki, Yumiko; Nishinari, Chisato; Endo, Harumi; Tamura, Chieko; Jinbo, Keiko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

    2002-04-01

    The in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates obtained between 1996 and 2000 were yearly evaluated and compared with those of other cephems, oxacephems, carbapenems, and penicillins. Fifteen species, 1,062 strains, of Gram-positive bacteria were isolated from the clinical materials annually collected from January to December, and consisted of methicillin-susceptible Staphylococcus aureus (MSSA; n = 127), methicillin-resistant Staphylococcus aureus (MRSA; n = 123), Staphylococcus epidermidis (n = 104), Staphylococcus haemolyticus (n = 58), Streptococcus pyogenes (n = 100), Streptococcus agalactiae (n = 50), Streptococcus pneumoniae (n = 125), Enterococcus faecalis (n = 150), Enterococcus faecium (n = 50), Enterococcus avium (n = 50), and Peptostreptococcus spp. (P. anaerobius, P. asaccharolyticus, P. magnus, P. micros, P. prevotii; n = 125). CZOP possessed stable antibacterial activities against all strains tested throughout 5 years. The MIC90 of CZOP against MRSA and S. haemolyticus tended to decrease while against S. pneumoniae and Peptostreptococcus spp., tended to increase year by year. However, the MIC90 just changed a little and were consistent with the results from the studies performed until the new drug application approval. Increases in the MIC90 against S. pneumoniae were also observed with cefpirome (CPR), cefepime (CFPM), flomoxef (FMOX), sulbactam/cefoperazone (SBT/CPZ), and imipenem (IPM). Increases in the MIC90 against Peptostreptococcus spp. were also observed with ceftazidime (CAZ), CPR, CFPM, FMOX, SBT/CPZ, and IPM. The decreases in the sensitivities were not always considered to depend upon generation of resistant bacteria because the annual MIC range of each antibacterial agent was almost generally wide every year and the annual sensitivity of each strain to the agents extremely varied. In conclusion, the annual antibacterial activities of CZOP against the Gram

  2. Isolated Optic Disc Tuberculosis

    Science.gov (United States)

    Mansour, Ahmad M.; Tabbara, Khalid F.; Tabbarah, Zuhair

    2015-01-01

    We present a healthy male subject who developed progressive visual loss in the left eye initially diagnosed as optic neuritis. Upon suspicion of infectious etiology, testing was positive for tuberculosis. There were no signs or symptoms of active systemic tuberculosis infection. The patient responded swiftly to antimycobacterial therapy with return of vision and resolution of disc swelling. Positive purified protein derivative skin test, negative chest radiograph, negative systemic workup, negative workup for other causes of unilateral optic neuritis and quick response to mycobacterial therapy reaffirm the entity of isolated optic disc tuberculosis similar to isolated choroidal tuberculosis without systemic manifestation. PMID:26483675

  3. Isolated Optic Disc Tuberculosis

    Directory of Open Access Journals (Sweden)

    Ahmad M. Mansour

    2015-09-01

    Full Text Available We present a healthy male subject who developed progressive visual loss in the left eye initially diagnosed as optic neuritis. Upon suspicion of infectious etiology, testing was positive for tuberculosis. There were no signs or symptoms of active systemic tuberculosis infection. The patient responded swiftly to antimycobacterial therapy with return of vision and resolution of disc swelling. Positive purified protein derivative skin test, negative chest radiograph, negative systemic workup, negative workup for other causes of unilateral optic neuritis and quick response to mycobacterial therapy reaffirm the entity of isolated optic disc tuberculosis similar to isolated choroidal tuberculosis without systemic manifestation.

  4. Primary isolated hepatic tuberculosis

    International Nuclear Information System (INIS)

    Sheikh, A.S.F.; Qureshi, I.H.; Saba, K.; Bukhari, M.H.

    2013-01-01

    Isolated hepatic tuberculosis without pulmonary or bowel involvement is a diagnostic challenge and can cause considerable morbidity. A young lady from Lahore presented with fever, pain in right hypochondria, nausea and weight loss. CT scan of abdomen showed multiple small hypodense non-enhancing lesions and a heterogeneous texture of liver. Biopsy confirmed the diagnosis of hepatic tuberculosis. It was concluded a case of isolated hepatic tuberculosis without evidence of other primary sites involvement. It is important to consider tuberculosis in the differential diagnosis when suspecting lymphoproliferative or metastatic diseases in a patient with vague symptoms and abnormal hepatic texture on CT. (author)

  5. A note on isolate domination

    Directory of Open Access Journals (Sweden)

    Ismail Sahul Hamid

    2016-04-01

    Full Text Available A set $S$ of vertices of a graph $G$ such that $\\left\\langle S\\right\\rangle$ has an isolated vertex is called an \\emph{isolate set} of $G$. The minimum and maximum cardinality of a maximal isolate set are called the \\emph{isolate number} $i_0(G$ and the \\emph{upper isolate number} $I_0(G$ respectively. An isolate set that is also a dominating set (an irredundant set is an $\\emph{isolate dominating set} \\ (\\emph{an isolate irredundant set}$. The \\emph{isolate domination number} $\\gamma_0(G$ and the \\emph{upper isolate domination number} $\\Gamma_0(G$ are respectively the minimum and maximum cardinality of a minimal isolate dominating set while the \\emph{isolate irredundance number} $ir_0(G$ and the \\emph{upper isolate irredundance number} $IR_0(G$ are the minimum and maximum cardinality of a maximal isolate irredundant set of $G$. The notion of isolate domination was introduced in \\cite{sb} and the remaining were introduced in \\cite{isrn}. This paper further extends a study of these parameters.   

  6. Emotion regulation during isolation

    Czech Academy of Sciences Publication Activity Database

    Poláčková Šolcová, Iva; Šolcová, Iva

    2012-01-01

    Roč. 47, Suppl. 1 (2012) ISSN 0020-7594. [International Congress of Psychology /30./. 22.07.2012-27.07.2012, Cape Town] R&D Projects: GA ČR(CZ) GAP407/11/2226 Institutional support: RVO:68081740 Keywords : emotion regulation * isolation * Mars500 Subject RIV: AN - Psychology

  7. Beyond insulation and isolation

    DEFF Research Database (Denmark)

    Højlund, Marie Koldkjær

    2016-01-01

    are insulation and isolation strategies to reduce measurable and perceptual noise levels. However, these strategies do not actively support the need to feel like an integral part of the shared hospital environment, which is a key element in creating healing environments, according to the paradigm of Evidence-Based...

  8. Fault isolation techniques

    Science.gov (United States)

    Dumas, A.

    1981-01-01

    Three major areas that are considered in the development of an overall maintenance scheme of computer equipment are described. The areas of concern related to fault isolation techniques are: the programmer (or user), company and its policies, and the manufacturer of the equipment.

  9. Proteoglycan isolation and analysis

    DEFF Research Database (Denmark)

    Woods, A; Couchman, J R

    2001-01-01

    Proteoglycans can be difficult molecules to isolate and analyze due to large mass, charge, and tendency to aggregate or form macromolecular complexes. This unit describes detailed methods for purification of matrix, cell surface, and cytoskeleton-linked proteoglycans. Methods for analysis...

  10. Isolated limb perfusion.

    Science.gov (United States)

    Gillespie, Rosalyn; Chantier, Nariane

    1994-12-08

    Growing concern over the rising incidence of malignant melanoma has brought about a need for information on this disorder and the treatment available. Isolated limb perfusion is a relatively new technique used in only a few hospitals. An increased knowledge base will lead to a better understanding of the nursing care required and to a more in-depth care plan.

  11. Broadband Faraday isolator.

    Science.gov (United States)

    Berent, Michał; Rangelov, Andon A; Vitanov, Nikolay V

    2013-01-01

    Driving on an analogy with the technique of composite pulses in quantum physics, we theoretically propose a broadband Faraday rotator and thus a broadband optical isolator, which is composed of sequences of ordinary Faraday rotators and achromatic quarter-wave plates rotated at the predetermined angles.

  12. Activities of Tedizolid and Linezolid Determined by the Reference Broth Microdilution Method against 3,032 Gram-Positive Bacterial Isolates Collected in Asia-Pacific, Eastern Europe, and Latin American Countries in 2014.

    Science.gov (United States)

    Pfaller, Michael A; Flamm, Robert K; Jones, Ronald N; Farrell, David J; Mendes, Rodrigo E

    2016-09-01

    Tedizolid and linezolid in vitro activities against 3,032 Gram-positive pathogens collected in Asia-Pacific, Eastern European, and Latin American medical centers during 2014 were assessed. The isolates were tested for susceptibility by the current reference broth microdilution methods. Due to concern over the effect of MIC endpoint criteria on the results of testing the oxazolidinones tedizolid and linezolid, MIC endpoint values were read by two methods: (i) reading the MIC at the first well where the trailing began without regard for pinpoint trailing, according to CLSI M07-A10 and M100-S26 document instructions for reading linezolid (i.e., 80% inhibition of growth; these reads were designated tedizolid 80 and linezolid 80), and (ii) at 100% inhibition of growth (designated tedizolid 100 and linezolid 100). All Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus anginosus group, and Enterococcus faecalis isolates were inhibited at tedizolid 80 and 100 MIC values of 0.25 and 0.5, 0.25 and 0.25, 0.25 and 0.5, 0.12 and 0.25, and 0.5 and 1 μg/ml, respectively. Generally, MIC50 and MIC90 results for tedizolid 80 and linezolid 80 were one doubling dilution lower than those read at 100% inhibition. Tedizolid was 4- to 8-fold more potent than linezolid against all the isolates tested regardless of the MIC endpoint criterion used. Despite the differences in potency, >99.9% of isolates tested in this survey were susceptible to both linezolid and tedizolid using CLSI and EUCAST interpretive criteria. In conclusion, tedizolid demonstrated greater in vitro potency than linezolid against Gram-positive pathogens isolated from patients in medical centers across the Asia-Pacific region, Eastern Europe, and Latin America. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  13. High voltage isolation transformer

    Science.gov (United States)

    Clatterbuck, C. H.; Ruitberg, A. P. (Inventor)

    1985-01-01

    A high voltage isolation transformer is provided with primary and secondary coils separated by discrete electrostatic shields from the surfaces of insulating spools on which the coils are wound. The electrostatic shields are formed by coatings of a compound with a low electrical conductivity which completely encase the coils and adhere to the surfaces of the insulating spools adjacent to the coils. Coatings of the compound also line axial bores of the spools, thereby forming electrostatic shields separating the spools from legs of a ferromagnetic core extending through the bores. The transformer is able to isolate a high constant potential applied to one of its coils, without the occurrence of sparking or corona, by coupling the coatings, lining the axial bores to the ferromagnetic core and by coupling one terminal of each coil to the respective coating encasing the coil.

  14. Psychopathology of social isolation

    OpenAIRE

    Baek, Sang-Bin

    2014-01-01

    The most important defining factor of being human is the use of symbolic language. Language or communication problem occurs during the growth, the child will have a higher risk of social isolation and then the survival will be threatened constantly. Today, adolescents and youths are familiar with computer and smart-phone devices, and communication with others by these devices is easy than face-to-face communication. As adolescents and youths live in the comfortable and familiar cyber-world ra...

  15. Mechanical beam isolator

    International Nuclear Information System (INIS)

    Post, R.F.; Vann, C.S.

    1996-10-01

    Back-reflections from a target, lenses, etc. can gain energy passing backwards through a laser just like the main beam gains energy passing forwards. Unless something blocks these back-reflections early in their path, they can seriously damage the laser. A Mechanical Beam Isolator is a device that blocks back-reflections early, relatively inexpensively, and without introducing aberrations to the laser beam

  16. Neglected isolated scaphoid dislocation

    Directory of Open Access Journals (Sweden)

    Jong-Ryoon Baek

    2016-01-01

    Full Text Available The authors present a case of isolated scaphoid dislocation in a 40-year-old male that was undiagnosed for 2 months. The patient was treated by open reduction, Kirschner wire fixation, interosseous ligament repair using a suture anchor and Blatt's dorsal capsulodesis. At 6 years followup, his radiographs of wrist showed a normal carpal alignment with a scapholunate gap of 3 mm and no evidence of avascular necrosis (AVN of the scaphoid.

  17. Miniature Optical Isolator, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — To address NASA's need for compact optical isolators, Physical Optics Corporation (POC) proposes to continue the development of a new Miniature Optical Isolator...

  18. Isolation and Cultivation of Anaerobes

    DEFF Research Database (Denmark)

    Aragao Börner, Rosa

    2016-01-01

    Anaerobic microorganisms play important roles in different biotechnological processes. Their complex metabolism and special cultivation requirements have led to less isolated representatives in comparison to their aerobic counterparts.In view of that, the isolation and cultivation of anaerobic...

  19. Padronização da técnica de multiplex PCR para a detecção de Staphylococcus aureus, Streptococcus agalactiae e Escherichia coli em amostras de leite bovino, obtidas de tanques de expansão

    OpenAIRE

    Troncarelli, Marcella Zampoli [UNESP

    2011-01-01

    Com o objetivo de normatizar a cadeia produtiva visando à melhoria da qualidade do leite no Brasil, o MAPA delineou a IN n° 51, em vigor desde julho de 2005. Em linha com estes parâmetros, objetivou-se, no presente estudo, padronizar o método de mPCR para a detecção de S. aureus, S. agalactiae e E. coli em amostras de leite bovino obtidas de tanques de expansão, e avaliar esta técnica como possível recurso diagnóstico a ser empregado em programas de controle de qualidade do leite oferecido pa...

  20. [Antibacterial activity of cefpodoxime against clinical isolates in 2000 and 2001].

    Science.gov (United States)

    Abe, Tomomi; Fukuoka, Takashi; Sato, Yuki; Ito, Kazuyoshi; Sei, Masami

    2002-12-01

    As the post-marketing surveillance of cefpodoxime proxetil (Banan), MICs of cefpodoxime (CPDX, an active form of Banan) against 1090 clinical isolates of 22 species from 15 medical institutions all over Japan from June 2000 to March 2001 were measured using the broth microdilution method approved by the Japanese Society of Chemotherapy and compared with those of oral cephem antibacterials, cefaclor, cefdinir, cefditoren, and cefcapene. In this study, remarkable change in the activity of CPDX was observed in Streptococcus pneumoniae and Haemophilus influenzae compared with the susceptibility in the studies before Banan was launched. This cause is considered to be the increase in the incidence of the following resistant strains: penicillin-intermediate S. pneumoniae (47.3%), penicillin-resistant S. pneumoniae (PRSP, 15.1%), and beta-lactamase-negative ampicillin-resistant (BLNAR) H. influenzae (24.0%), which were scarcely isolated in 1989 when Banan was launched. Other tested drugs also exhibited low activity against these resistant strains. However, CPDX showed comparatively good activity with MIC90 of 2 micrograms/mL against PRSP. Against methicillin-susceptible Staphylococcus spp., Streptococcus pyogenes, Streptococcus agalactiae, and Moraxella catarrhalis, CPDX also showed comparatively good activity with MIC90 of marketing. Against quinolones-resistant Neisseria gonorrhoeae, CPDX showed excellent activity with MIC90 of 0.5 microgram/mL. Against members of the family Enterobacteriaceae except for Citrobacter freundii, Enterobacter spp., Proteus vulgaris, and Morganella morganii, CPDX showed good activity. However, in Escherichia coli, Klebsiella spp. Proteus spp., and Providencia spp., there are some high-resistant strains to all tested drugs including CPDX. Against Peptostreptococcus spp., MIC90 of CPDX was 8 micrograms/mL and its MIC range was widely distributed from 0.03 to 32 micrograms/mL, which were similar to those in the studies before its marketing. In

  1. Diabetes in population isolates

    DEFF Research Database (Denmark)

    Grarup, Niels; Moltke, Ida; Albrechtsen, Anders

    2015-01-01

    Type 2 diabetes (T2D) is an increasing health problem worldwide with particularly high occurrence in specific subpopulations and ancestry groups. The high prevalence of T2D is caused both by changes in lifestyle and genetic predisposition. A large number of studies have sought to identify...... on glucose-stimulated plasma glucose, serum insulin levels, and T2D. The variant defines a specific subtype of non-autoimmune diabetes characterized by decreased post-prandial glucose uptake and muscular insulin resistance. These and other recent findings in population isolates illustrate the value...

  2. Journey from isolation

    Directory of Open Access Journals (Sweden)

    Wim A. Dreyer

    2011-04-01

    Full Text Available Since the Ottawa Consultation in 1982, the relationship between the Nederduitsch Hervormde Kerk van Afrika (NHKA and the World Alliance of Reformed Churches (WARC was nonexistent. In the NHKA it became progressively clear that it would be impossible to travel the road of faith alone. This article examined the factors which contributed to the growing isolation of the NHKA, especially nationalism, a particularistic ecclesiology and the rejection of Apartheid by international ecumenical bodies. It also reflected on efforts of the NHKA to return to the international ecumenical movement.

  3. Waste isolation: a bibliography

    International Nuclear Information System (INIS)

    McLaren, L.H.

    1982-12-01

    This bibliography contains information on waste isolation included in the Department of Energy's Energy Data Base from Janurary 1981 through November 1982. The abstracts are grouped by subject category as shown in the table of contents. Entries in the subject index also facilitate access by subject, e.g., Radioactive Waste Disposal/Salt Deposits. Within each category the arrangement is by report number for reports, followed by nonreports in reverse chronological order. These citations are to research reports, journal articles, books, patents, theses, and conference papers from worldwide sources. Five indexes, each preceded by a brief description, are provided: Corporate Author, Personal Author, Subject, Contract Number, and Report Number

  4. Phenotypic and genotypic antimicrobial susceptibility pattern of Streptococcus spp. isolated from cases of clinical mastitis in dairy cattle in Poland.

    Science.gov (United States)

    Kaczorek, E; Małaczewska, J; Wójcik, R; Rękawek, W; Siwicki, A K

    2017-08-01

    Mastitis of dairy cattle is one of the most frequently diagnosed diseases worldwide. The main etiological agents of mastitis are bacteria of the genus Streptococcus spp., in which several antibiotic resistance mechanisms have been identified. However, detailed studies addressing this problem have not been conducted in northeastern Poland. Therefore, the aim of our study was to analyze, on phenotypic and genotypic levels, the antibiotic resistance pattern of Streptococcus spp. isolated from clinical cases of mastitis from dairy cattle in this region of Poland. The research was conducted using 135 strains of Streptococcus (Streptococcus uberis, n = 53; Streptococcus dysgalactiae, n = 41; Streptococcus agalactiae, n = 27; other streptococci, n = 14). The investigation of the antimicrobial susceptibility to 8 active substances applied in therapy in the analyzed region, as well as a selected bacteriocin (nisin), was performed using the minimum inhibitory concentration method. The presence of selected resistance genes (n = 14) was determined via PCR. We also investigated the correlation between the presence of resistance genes and the antimicrobial susceptibility of the examined strains in vitro. The highest observed resistance of Streptococcus spp. was toward gentamicin, kanamycin, and tetracycline, whereas the highest susceptibility occurred toward penicillin, enrofloxacin, and marbofloxacin. Additionally, the tested bacteriocin showed high efficacy. The presence of 13 analyzed resistance genes was observed in the examined strains [gene mef(A) was not detected]. In most strains, at least one resistance gene, mainly responsible for resistance to tetracyclines [tet(M), tet(K), tet(L)], was observed. However, a relationship between the presence of a given resistance gene and antimicrobial susceptibility on the phenotypic level was not always observed. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  5. Yeast Isolation for Bioethanol Production

    Directory of Open Access Journals (Sweden)

    EKA RURIANI

    2012-09-01

    Full Text Available We have isolated 12 yeast isolates from five different rotten fruits by using a yeast glucose chloramphenicol agar (YGCA medium supplemented with tetracycline. From pre-screening assay, four isolates exhibited higher substrate (glucose-xylose consumption efficiency in the reaction tube fermentation compared to Saccharomyces cerevisiae dan Saccharomyces ellipsoids as the reference strains. Based on the fermentation process in gooseneck flasks, we observed that two isolates (K and SB showed high fermentation efficiency both in sole glucose and mixed glucose-xylose substrate. Moreover, isolates K and SB produced relatively identical level of ethanol concentration compared to the reference strains. Isolates H and MP could only produce high levels of ethanol in glucose fermentation, while only half of that amount of ethanol was detected in glucose-xylose fermentation. Isolate K and SB were identified as Pichia kudriavzeevii (100% based on large sub unit (LSU ribosomal DNA D1/D2 region.

  6. Isolated sulfite oxidase deficiency.

    Science.gov (United States)

    Rupar, C A; Gillett, J; Gordon, B A; Ramsay, D A; Johnson, J L; Garrett, R M; Rajagopalan, K V; Jung, J H; Bacheyie, G S; Sellers, A R

    1996-12-01

    Isolated sulfite oxidase (SO) deficiency is an autosomal recessively inherited inborn error of sulfur metabolism. In this report of a ninth patient the clinical history, laboratory results, neuropathological findings and a mutation in the sulfite oxidase gene are described. The data from this patient and previously published patients with isolated sulfite oxidase deficiency and molybdenum cofactor deficiency are summarized to characterize this rare disorder. The patient presented neonatally with intractable seizures and did not progress developmentally beyond the neonatal stage. Dislocated lenses were apparent at 2 months. There was increased urine excretion of sulfite and S-sulfocysteine and a decreased concentration of plasma cystine. A lactic acidemia was present for 6 months. Liver sulfite oxidase activity was not detectable but xanthine dehydrogenase activity was normal. The boy died of respiratory failure at 32 months. Neuropathological findings of cortical necrosis and extensive cavitating leukoencephalopathy were reminiscent of those seen in severe perinatal asphyxia suggesting an etiology of energy deficiency. A point mutation that resulted in a truncated protein missing the molybdenum-binding site has been identified.

  7. Schizosaccharomyces isolation method

    Directory of Open Access Journals (Sweden)

    Benito Santiago

    2014-01-01

    Full Text Available This study discusses the optimization of a selective and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms to select individuals from this genus for industrial purposes, especially in light of the recent recommendation of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main physiological characteristics of these species, such as their high resistances to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the genus resistance to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid. Malic acid was used as a differential factor due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey and honeycombs.

  8. Isolated Cardiac Hydatid Cyst

    International Nuclear Information System (INIS)

    Shakil, U.; Rehman, A. U.; Shahid, R.

    2015-01-01

    Hydatid cyst disease is common in our part of the world. Cardiac hydatid cyst is its rare manifestation. We report this case of 48-year male having isolated cardiac hydatid cyst, incidentally found on computed tomography. This patient presented in medical OPD of Combined Military Hospital, Lahore with one month history of mild retrosternal discomfort. His general physical and systemic examinations as well as ECG were unremarkable. Chest X-ray showed an enlarged cardiac shadow with mildly irregular left heart border. Contrast enhanced CT scan of the chest showed a large well defined multiloculated non-enhancing cystic lesion with multiple daughter cysts involving wall of left ventricle and overlying pericardium. Serology for echinococcus confirmed the diagnosis of hydatid cyst. Patient was offered the surgical treatment but he opted for medical treatment only. Albendezol was prescribed. His follow-up echocardiography after one month showed no significant decrease in size of the cyst. (author)

  9. The Neuroendocrinology of Social Isolation

    Science.gov (United States)

    Cacioppo, John T.; Cacioppo, Stephanie; Capitanio, John P.; Cole, Steven W.

    2016-01-01

    Social isolation has been recognized as a major risk factor for morbidity and mortality in humans for more than a quarter of a century. Although the focus of research has been on objective social roles and health behavior, the brain is the key organ for forming, monitoring, maintaining, repairing, and replacing salutary connections with others. Accordingly, population-based longitudinal research indicates that perceived social isolation (loneliness) is a risk factor for morbidity and mortality independent of objective social isolation and health behavior. Human and animal investigations of neuroendocrine stress mechanisms that may be involved suggest that (a) chronic social isolation increases the activation of the hypothalamic pituitary adrenocortical axis, and (b) these effects are more dependent on the disruption of a social bond between a significant pair than objective isolation per se. The relational factors and neuroendocrine, neurobiological, and genetic mechanisms that may contribute to the association between perceived isolation and mortality are reviewed. PMID:25148851

  10. The neuroendocrinology of social isolation.

    Science.gov (United States)

    Cacioppo, John T; Cacioppo, Stephanie; Capitanio, John P; Cole, Steven W

    2015-01-03

    Social isolation has been recognized as a major risk factor for morbidity and mortality in humans for more than a quarter of a century. Although the focus of research has been on objective social roles and health behavior, the brain is the key organ for forming, monitoring, maintaining, repairing, and replacing salutary connections with others. Accordingly, population-based longitudinal research indicates that perceived social isolation (loneliness) is a risk factor for morbidity and mortality independent of objective social isolation and health behavior. Human and animal investigations of neuroendocrine stress mechanisms that may be involved suggest that (a) chronic social isolation increases the activation of the hypothalamic pituitary adrenocortical axis, and (b) these effects are more dependent on the disruption of a social bond between a significant pair than objective isolation per se. The relational factors and neuroendocrine, neurobiological, and genetic mechanisms that may contribute to the association between perceived isolation and mortality are reviewed.

  11. Miniature Optical Isolator, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — To address NASA's need for miniature optical isolators in atom interferometry applications, Physical Optics Corporation (POC) proposes to develop a miniature optical...

  12. Inhibitory effect of Lactobacillus rhamnosus on pathogenic bacteria isolated from women with bacterial vaginosis

    Directory of Open Access Journals (Sweden)

    Gita Eslami

    2014-06-01

    Full Text Available Background: Considering the high prevalence of bacterial vaginosis and its association with urinary tract infection in women and treatment of gynecologic problems occur when a high recurrence of bacterial vaginosis is often treated with antibiotics. The purpose of this study is to investigate the inhibitory effect of Lactobacillus rhamnosus on pathogenic bacteria isolated from women with bacterial vaginosis, respectively.Materials and Methods: 96 samples from women with bacterial vaginosis discharge referred to health centers dependent Shahid Beheshti University in 91-92 were taken by a gynecologist with a dacron swab and put in sterile tubes containing TSB broth and Thioglycollate broth and were immediately sent to the lab location in cold chain for the next stages of investigation. From Thioglycollate and TSB medium was cultured on blood agar and EMB and Palkam and Differential diagnosis environments, and then incubated for 24 h at 37°C. Strains of Lactobacillus rhamnosus were cultured in MRSA environment and were transfered to the lab. After purification of pathogenic bacteria, MIC methods and antibiogram, Lactobacillus rhamnosus inhibitory effect on pathogenic bacteria is checked. Statistical analysis was done by SPSS software v.16.Results: The results of this study show the inhibitory effect of Lactobacillus rhamnosus on some pathogenic bacteria that cause bacterial vaginosis, including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Entrococcus, Listeria monocytogenes and E.Coli. Microscopic examination of stained smears of the large number of Lactobacillus and pathogenic bacteria showed reduced. The prevalence of abnormal vaginal discharge, history of drug use means of preventing pregnancy and douching, respectively, 61%, 55%, 42% and 13% respectively. Significant difference was observed between the use and non-use of IUD in women with bacterial vaginosis infection

  13. Canadian geologic isolation program

    International Nuclear Information System (INIS)

    Dyne, P.J.

    1976-01-01

    The Canadian geologic isolation program is directed at examining the potential of (1) salt deposits and (2) hard rock as repositories for radioactive wastes. It was felt essential from the inception that alternative host rocks be evaluated over a fairly large geographical area. The studies on salt deposits to date are based on existing geological information and have identified the areas that show some potential and merit further study. The factors considered include depth, thickness and purity of the deposit, overlying aquifers, and the potential for gas and oil exploration as well as potash recovery. The studies on hard rock are restricted to plutonic igneous rocks in the Ontario part of the Canadian Shield. Because geological information on their nature and extent is sparse, the study is limited to bodies that are well exposed and for which information is available.for which information is available. Field studies in the next two seasons are aimed at mapping the fault and joint patterns and defining the geologic controls on their development. In 1977 and 1978, two or three of the more favorable sites will be mapped in greater detail, and an exploratory drilling program will be established to determine the extent of fracturing at depth and the hydrology of these fractures. Conceptual designs of mined repositories in hard rock are also being made with the hope of identifying, at an early stage in this program, special problems in hard-rock repositories that may require development and study

  14. Isolated Amoebic Abscess of Spleen

    Directory of Open Access Journals (Sweden)

    Kaushik M

    2013-04-01

    Full Text Available Amoebic liver abscess is the most common extraintestinal manifestation of amoebiasis. Extrahepatic amoebic abscesses have occasionally been described in the lung, brain, and skin and presumably result from hematogenous spread. Isolated amoebic abscess of spleen has been reported scarcely in literature. We report here a case of isolated amoebic abscess of spleen.

  15. METHICILLIN RESISTANCE IN STAPHYLOCOCCAL ISOLATES ...

    African Journals Online (AJOL)

    The study assessed the importance of Staphylococcus aureus as a urinary pathogen and the incidence of multidrug resistant (MDR), methicillin-resistant Staphylococcus aureus (MRSA). A total of 86 staphylococcal isolates made up of 50 clinical isolates from urine samples submitted to the Medical Microbiology Laboratory ...

  16. Polyketides isolated from Penicillium herquei

    International Nuclear Information System (INIS)

    Marinho, Andrey Moacir do Rosario; Marinho, Patricia Santana Barbosa; Rodrigues Filho, Edson; Ferreira, Izabel Cristina Piloto

    2011-01-01

    In this work we are reporting the isolation of polyketides citreoserine (1), emodin (2), janthinone (3), dihydrocitrinone (4) and citrinin H-1 (5). The compounds were isolated by chromatographic procedures and identified by spectral methods of NMR 1D and 2D and MS. The compounds 1, 2 and 3 were tested against promastigotes of Leishmania brasiliensis. (author)

  17. Review of nuclear waste isolation

    International Nuclear Information System (INIS)

    Richard, B.H.

    1978-06-01

    On Jun 22 and 23, 1978, Rockwell Hanford Operations assembled a committee of their personnel, subcontractors, and representatives of other waste isolation programs for a review of nuclear waste isolation. Appendix A lists the participants and their affiliations; Appendix B indicates the agenda. The purpose of the review was to gather experts in the areas pertaining to isolation of nuclear waste to discuss three basic issues that must be addressed in isolation studies. These were: the paths of transport to the biosphere; the barriers needed for containment; and the isolation time necessary for each radioactive isotope. In that these issues are media dependent, the basalt medium was emphasized. Conclusions of the review are described

  18. Patients experience of source isolation

    DEFF Research Database (Denmark)

    Johansen, Kamilla; Pedersen, Didde; Kragbak, Nina

    2014-01-01

    , Nursing education in Århus, Hedeager 2, 8200 Aarhus N, Denmark. Background: Medical treatment and care of patients with infections may include source isolation of the patient, to avoid spreading of the infection. However, isolation is a potential physiological and psychological stress factor...... of the patients perspectives of being isolated to identify areas of potential interest for developing new caring strategies to minimize the negative side effects of isolation. Methods: Literature was systematically searched in CINAHL, Nursing Reference Center, Social Care Online, SveMed+, The Cochrane Library...... of Care: The patients felt abandoned and forgotten by the nurses, because of fewer visits and time limited communication. This led to emotions such as frustrations, insecurity and neglect. While isolated the patients felt it difficult to achieve contact and have an optimal relation with the nurses...

  19. Comparative Genomic Analysis of the ICESa2603 Family ICEs and Spread of erm(B- and tet(O-Carrying Transferable 89K-Subtype ICEs in Swine and Bovine Isolates in China

    Directory of Open Access Journals (Sweden)

    Jinhu eHuang

    2016-02-01

    Full Text Available Integrative and conjugative elements (ICEs of the ICESa2603 family have been isolated from several species of Streptococcus spp.; however, the comparative genomic and evolutionary analyses of these particular ICEs are currently only at their initial stages. By investigating 13 ICEs of the ICESa2603 family and two ICESa2603 family-like ICEs derived from diverse hosts and locations, we have determined that ICEs comprised a backbone of 30 identical syntenic core genes and accessory genes that were restricted to the intergenic sites or the 3'-end of the non-conserved domain of core genes to maintain its function. ICESa2603 family integrase IntICESa2603 specifically recognized a 15-bp att sequence (TTATTTAAGAGTAAC at the 3'-end of rplL, which was highly conserved in genus Streptococcus. Phylogenetic analyses suggest that extensive recombination/insertion and the occurrence of a hybrid/mosaic in the ICESa2603 family were responsible for the significant increase in ICE diversity, thereby broadening its host range. Approximately 42.5% and 38.1% of the tested Streptococcus suis and Streptococcus agalactiae clinical isolates respectively contained ICESa2603 family Type Ⅳ secretion system (T4SS genes, and 80.5% and 62.5% of which also respectively carried intICESa2603, indicating that ICESa2603 family is widely distributed across these bacteria. Sequencing and conjugation transfer of a novel sequence type ST303 clinical S. suis isolate HB1011 demonstrated that the 89K-subtype ICESsuHB1011 retained its transferrable function, thereby conferring tetracycline and macrolide resistance.

  20. Dynamic isolation technologies in negative pressure isolation wards

    CERN Document Server

    Xu, Zhonglin

    2017-01-01

    This book presents novel design principles and technologies for dynamic isolation based on experimental studies. These approaches have now become the local standard in Beijing and are currently being promoted for use nationwide. Further, the book provides details of measures and guidelines for the design process. Departing from the traditional understanding that isolation wards should be designed with high negative pressure, airtight doors and fresh air, it establishes the basis for designing biological clean rooms, including isolation wards, using a simple and convenient scientific approach. This book is intended for designers, engineers, researchers, hospital management staff and graduate students in heating ventilation air conditioning (HVAC), air cleaning technologies and related areas.

  1. Isolated Proofs of Knowledge and Isolated Zero Knowledge

    DEFF Research Database (Denmark)

    Damgård, Ivan Bjerre; Nielsen, Jesper Buus; Wichs, Daniel

    2008-01-01

    . This is because the prover may just be forwarding messages between the environment and the verifier while the environment performs all the necessary computation. In this paper we consider an ℓ-isolated prover, which is restricted to exchanging at most ℓ bits of information with its environment. We introduce a new...... notion called ℓ-isolated proofs of knowledge (ℓ-IPoK). These protocols securely ensure that an ℓ-isolated prover knows the witness. To prevent the above-mentioned attack, an ℓ-IPoK protocol has to have communication complexity greater than ℓ. We show that for any relation in NP and any value ℓ...

  2. Tigecycline Susceptibility Trends Among Pathogens Isolated from Complicated Skin and Soft-Tissue Infections in North and Latin America: 2012–2016

    Science.gov (United States)

    Hoban, Daryl; Renteria, Martha; Leister-Tebbe, Heidi; Sahm, Dan

    2017-01-01

    Abstract Background The Tigecycline Evaluation Surveillance Trial (TEST) monitors the activity of tigecycline and other antimicrobials against clinically-relevant pathogens collected globally. This study reports the activity of tigecycline (TGC) against Gram-positive and Gram-negative isolates collected in North and Latin America from patients with complicated skin and soft-tissue infections (CSSTI). Methods Hospital sites from North America (NA) and Latin America (LA) collected non-duplicate clinical Gram-positive and -negative isolates from various complicated skin and skin structure infection sources during 2012–2016. Organism identification and antibiotic susceptibility (S) testing was performed by the local laboratories. Susceptibility testing was determined using the broth microdilution method according to CLSI guidelines and categorical interpretation of results was done using CLSI or FDA (tigecycline) breakpoint criteria where appropriate. Cefoxitin disk testing was performed for all S. aureus to determine methicillin susceptibility (i.e., MRSA and MSSA). Results The table provides %S and MIC90 data for TGC against CSSTI isolates Region, n, %S, MIC 90 (µg/ml) North America Latin America Organism n %S MIC 90 n %S MIC 90 S. aureus 2270 100 0.12 310 100 0.25 Enterobacter spp. 925 96.4 1 159 92.5 2 P. aeruginosa 758 na* > 8 165 na > 8 E. coli 716 99.9 0.25 241 100 0.25 Enterococcus spp. 691 99.3 0.12 135 100 0.12 S. agalactiae 503 100 0.12 58 100 0.06 K. pneumoniae 471 94.9 2 152 91.5 2 S. marcescens 347 96.8 2 67 97.0 2 A. baumannii 310 na* 2 97 na 1 K. oxytoca 204 99.0 0.5 15 100 1 *na = not applicable or no breakpoints available for this species. Conclusion Based on %S and MIC90 data TGC exhibited potent activity against isolates of all organism groups from complicated skin and soft-tissue infections, regardless of the geographic region. However, given the potential many of these organisms have for developing resistance, continued and careful

  3. Antimicrobial susceptibility trends among gram-positive and -negative clinical isolates collected between 2005 and 2012 in Mexico: results from the Tigecycline Evaluation and Surveillance Trial.

    Science.gov (United States)

    Morfin-Otero, Rayo; Noriega, Eduardo Rodriguez; Dowzicky, Michael J

    2015-12-15

    The Tigecycline Evaluation and Surveillance Trial (T.E.S.T) is a global antimicrobial surveillance study of both gram-positive and gram-negative organisms. This report presents data on antimicrobial susceptibility among organisms collected in Mexico between 2005 and 2012 as part of T.E.S.T., and compares rates between 2005-2007 and 2008-2012. Each center in Mexico submitted at least 200 isolates per collection year; including 65 gram-positive isolates and 135 gram-negative isolates. Minimum inhibitory concentrations (MICs) were determined using Clinical Laboratory Standards Institute (CLSI) broth microdilution methodology and antimicrobial susceptibility was established using the 2013 CLSI-approved breakpoints. For tigecycline US Food and Drug Administration (FDA) breakpoints were applied. Isolates of E. coli and K. pneumoniae with a MIC for ceftriaxone of >1 mg/L were screened for ESBL production using the phenotypic confirmatory disk test according to CLSI guidelines. The rates of some key resistant phenotypes changed during this study: vancomycin resistance among Enterococcus faecium decreased from 28.6 % in 2005-2007 to 19.1 % in 2008-2012, while β-lactamase production among Haemophilus influenzae decreased from 37.6 to 18.9 %. Conversely, methicillin-resistant Staphylococcus aureus increased from 38.1 to 47.9 %, meropenem-resistant Acinetobacter spp. increased from 17.7 to 33.0 % and multidrug-resistant Acinetobacter spp. increased from 25.6 to 49.7 %. The prevalence of other resistant pathogens was stable over the study period, including extended-spectrum β-lactamase-positive Escherichia coli (39.0 %) and Klebsiella pneumoniae (25.0 %). The activity of tigecycline was maintained across the study years with MIC90s of ≤2 mg/L against Enterococcus spp., S. aureus, Streptococcus agalactiae, Streptococcus pneumoniae, Enterobacter spp., E. coli, K. pneumoniae, Klebsiella oxytoca, Serratia marcescens, H. influenzae, and Acinetobacter spp. All gram

  4. Waste Isolation Pilot Plant Overview

    Energy Technology Data Exchange (ETDEWEB)

    Weaver, Douglas James [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-03-27

    The mission of Waste Isolation Pilot Plant (WIPP) is to demonstrate the safe, environmentally sound, cost effective, permanent disposal of Transuranic (TRU) waste left from production of nuclear weapons.

  5. Seismic isolation in New Zealand

    International Nuclear Information System (INIS)

    Skinner, R.I.; Robinson, W.H.; McVerry, G.H.

    1989-01-01

    Bridges, buildings, and industrial equipment can be given increased protection from earthquake damage by limiting the earthquake attack through seismic isolation. A broad summary of the seismic responses of base-isolated structures is of considerable assistance for their preliminary design. Seismic isolation as already used in New Zealand consists of a flexible base or support combined with some form of energy-dissipating device, usually involving the hysteretic working of steel or lead. This paper presents examples of the New Zealand experience, where seismic isolation has been used for 42 bridges, 3 buildings, a tall chimney, and high-voltage capacitor banks. Additional seismic response factors, which may be important for nuclear power plants, are also discussed briefly

  6. WIPP: Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    1984-01-01

    The following aspects of the Waste Isolation Pilot Plant are discussed briefly: history and site selection; salt as a disposal medium; transporting waste materials; early key events; impacts on New Mexico; project organization; and site certification profile

  7. Isolation of rat adrenocortical mitochondria

    Energy Technology Data Exchange (ETDEWEB)

    Solinas, Paola [Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Department of Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Fujioka, Hisashi [Electron Microscopy Facility, Department of Pharmacology, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Tandler, Bernard [Department of Biological Sciences, School of Dental Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Hoppel, Charles L., E-mail: charles.hoppel@case.edu [Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Department of Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States)

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer A method for isolation of adrenocortical mitochondria from the adrenal gland of rats is described. Black-Right-Pointing-Pointer The purified isolated mitochondria show excellent morphological integrity. Black-Right-Pointing-Pointer The properties of oxidative phosphorylation are excellent. Black-Right-Pointing-Pointer The method increases the opportunity of direct analysis of adrenal mitochondria from small animals. -- Abstract: This report describes a relatively simple and reliable method for isolating adrenocortical mitochondria from rats in good, reasonably pure yield. These organelles, which heretofore have been unobtainable in isolated form from small laboratory animals, are now readily accessible. A high degree of mitochondrial purity is shown by the electron micrographs, as well as the structural integrity of each mitochondrion. That these organelles have retained their functional integrity is shown by their high respiratory control ratios. In general, the biochemical performance of these adrenal cortical mitochondria closely mirrors that of typical hepatic or cardiac mitochondria.

  8. Isolation Effect in Immediate and Delayed Recall

    Science.gov (United States)

    Bellezza, Francis S.; Cheney, Terry L.

    1973-01-01

    If the hypothesis of selective rehearsal is used to account for the isolation effect, then the recall of isolated items will depend both on the serial position of the isolated item and on whether recall is immediate or delayed. (Author)

  9. Isolated Systolic Hypertension: A Health Concern?

    Science.gov (United States)

    Isolated systolic hypertension: A health concern? Is having a high top number (systolic) blood pressure, but a normal bottom number (diastolic) ... mm Hg, you have a common type of high blood pressure called isolated systolic hypertension. Isolated systolic hypertension can ...

  10. Waste isolation safety assessment program

    International Nuclear Information System (INIS)

    Brandstetter, A.; Harwell, M.A.

    1979-05-01

    Associated with commercial nuclear power production in the United States is the generation of potentially hazardous radioactive wastes. The Department of Energy (DOE), through the National Waste Terminal Storage (NWTS) Program, is seeking to develop nuclear waste isolation systems in geologic formations that will preclude contact with the biosphere of waste radionuclides in concentrations which are sufficient to cause deleterious impact on humans or their environments. Comprehensive analyses of specific isolation systems are needed to assess the expectations of meeting that objective. The Waste Isolation Safety Assessment Program (WISAP) has been established at the Pacific Northwest Laboratory (operated by Battelle Memorial Institute) for developing the capability of making those analyses. Among the analyses required for isolation system evaluation is the detailed assessment of the post-closure performance of nuclear waste repositories in geologic formations. This assessment is essential, since it is concerned with aspects of the nuclear power program which previously have not been addressed. Specifically, the nature of the isolation systems (e.g., involving breach scenarios and transport through the geosphere), and the time-scales necessary for isolation, dictate the development, demonstration and application of novel assessment capabilities. The assessment methodology needs to be thorough, flexible, objective, and scientifically defensible. Further, the data utilized must be accurate, documented, reproducible, and based on sound scientific principles

  11. Geomorphology of Minnesota - Isolated Landform Structures

    Data.gov (United States)

    Minnesota Department of Natural Resources — Geomorphology of Minnesota - Isolated Landform Structures are essentially cartographic arcs representing isolated glacial features that were mapped in conjunction...

  12. Status and phosphorus solubilization potential of bacteria and arbuscular mycorrhizal fungi isolated from various locations of Khyber Pakhtunkhwa province

    International Nuclear Information System (INIS)

    Wahid, F.; Sharif, M.; Khan, M.

    2016-01-01

    The soils of Pakistan are alkaline calcareous in nature and its high pH makes phosphorus (P) unavailable for plants uptake. Chemical sources of P fertilizers are a costly and detrimental practice. Therefore, investigations were conducted to determine the native status of phosphate solubilizing bacteria (PSB) and arbuscular mycorrhizal fungi (AMF) in three different zones of Khyber Pakhtunkhwa province of Pakistan. In order to select the efficient PSB strains for solubility enhancement of P from rock phosphate (RP), rhizosphere soil samples were collected from irrigated and rainfed fields of maize, sorghum, pastures and vegetables. Population density of PSB ranged from 1.7*107 to 2.7*108 CFU g-1 rhizosphere soil. The bacterial strains Coccus, Streptoccocus and Bacillus sp. were identified on the basis of their microscopic, phenotypic and morphological characters. Most of the AM fungal spores identified were belonging to Glomus mosseae and Glomus intradices. A range of 02-35 spores per 20 g air dried soil were recorded. The PSB strains such as Coccus DIM7, Streptococcus PIM6 and Bacillus sp. PIS7 solubilized more P from RP than any other strain in both of the liquid and solid medium. Results show that areas under investigations are rich in P solubilizing micro flora providing a rich source for inoculum production. Moreover, the PSB strains have the capability to solubilize P from RP that can be used as biofertilizers for optimum crop production. (author)

  13. Sensitivity of seismically isolated structures

    International Nuclear Information System (INIS)

    Politopoulos, I.; Hoan, Khac Pham

    2009-01-01

    In this paper we study the sensitivity of seismically isolated structures to a small variability of the earthquake excitation and of some structural properties with respect to the probability of failure and floor spectra. In particular, the influence of the nonlinear behaviour of the isolated superstructure on the vulnerability and on the floor spectra is investigated by means of a series of Monte Carlo simulations of simple two degrees-of-freedom systems. Several types of passive and active isolation systems are examined and three different idealized nonlinear constitutive laws are considered for the superstructure. It is found that, in general, the probability of failure does not depend on the specific cyclic behaviour of the assumed constitutive law and general trends regarding the impact of different isolation devices on vulnerability are established. As for the floor spectra, the influence of moderate nonlinear behaviour of isolated Superstructures, with the exception of the case of a non-dissipative elastic nonlinear law is negligible, contrary to the case of conventional Structures. (authors)

  14. Sensitivity of seismically isolated structures

    Energy Technology Data Exchange (ETDEWEB)

    Politopoulos, I. [CEA Saclay, DEN DANS DM2S, 91 - Gif sur Yvette (France); Hoan, Khac Pham

    2009-07-15

    In this paper we study the sensitivity of seismically isolated structures to a small variability of the earthquake excitation and of some structural properties with respect to the probability of failure and floor spectra. In particular, the influence of the nonlinear behaviour of the isolated superstructure on the vulnerability and on the floor spectra is investigated by means of a series of Monte Carlo simulations of simple two degrees-of-freedom systems. Several types of passive and active isolation systems are examined and three different idealized nonlinear constitutive laws are considered for the superstructure. It is found that, in general, the probability of failure does not depend on the specific cyclic behaviour of the assumed constitutive law and general trends regarding the impact of different isolation devices on vulnerability are established. As for the floor spectra, the influence of moderate nonlinear behaviour of isolated Superstructures, with the exception of the case of a non-dissipative elastic nonlinear law is negligible, contrary to the case of conventional Structures. (authors)

  15. Isolated Echinococcosis of cervical region

    Directory of Open Access Journals (Sweden)

    Pratima Khare

    2014-01-01

    Full Text Available Echinococcosis, commonly called as hydatid disease, is a parasitic infestation caused by the larva of the genus Echinococcus in human. Isolated occurrence of Echinococcosis without any evidence of visceral disease is very rare. A thorough search of the literature revealed only 11 cases of isolated cervical Echinococcosis. We report here a very rare case of isolated hydatid cyst in a 45-year-old female patient, who presented with swelling in right cervical region about 5 cm below the angle of mandible with no evidence of the disease elsewhere in the body. The case was diagnosed on fine needle aspiration cytology. The diagnosis was further supported by histopathology. We propose that the treating physician should also consider the differential diagnosis of Echinococcosis in the presence of an asymptomatic soft tissue mass, especially when the patient lives in an endemic area.

  16. Sympathicotomy for isolated facial blushing

    DEFF Research Database (Denmark)

    Licht, Peter Bjørn; Pilegaard, Hans K; Ladegaard, Lars

    2012-01-01

    Background. Facial blushing is one of the most peculiar of human expressions. The pathophysiology is unclear, and the prevalence is unknown. Thoracoscopic sympathectomy may cure the symptom and is increasingly used in patients with isolated facial blushing. The evidence base for the optimal level...... of targeting the sympathetic chain is limited to retrospective case studies. We present a randomized clinical trial. Methods. 100 patients were randomized (web-based, single-blinded) to rib-oriented (R2 or R2-R3) sympathicotomy for isolated facial blushing at two university hospitals during a 6-year period...... between R2 and R2-R3 sympathicotomy for isolated facial blushing. Both were effective, and QOL increased significantly. Despite very frequent side effects, the vast majority of patients were satisfied. Surprisingly, many patients experienced mild recurrent symptoms within the first year; this should...

  17. VIBRATION ISOLATION SYSTEM PROBABILITY ANALYSIS

    Directory of Open Access Journals (Sweden)

    Smirnov Vladimir Alexandrovich

    2012-10-01

    Full Text Available The article deals with the probability analysis for a vibration isolation system of high-precision equipment, which is extremely sensitive to low-frequency oscillations even of submicron amplitude. The external sources of low-frequency vibrations may include the natural city background or internal low-frequency sources inside buildings (pedestrian activity, HVAC. Taking Gauss distribution into account, the author estimates the probability of the relative displacement of the isolated mass being still lower than the vibration criteria. This problem is being solved in the three dimensional space, evolved by the system parameters, including damping and natural frequency. According to this probability distribution, the chance of exceeding the vibration criteria for a vibration isolation system is evaluated. Optimal system parameters - damping and natural frequency - are being developed, thus the possibility of exceeding vibration criteria VC-E and VC-D is assumed to be less than 0.04.

  18. Isolation, characterization and identification of actinomycetes from ...

    African Journals Online (AJOL)

    A total of 62 isolates of actinomycetes were isolated from 7 soil samples collected from Agriculture Research Center Semongok, Sarawak. All 62 isolates exhibited a range of colony colours (dark grey, grey, dark brown, brownish, whitish and yellowish white). All the isolates were later purified and subjected to a few ...

  19. Isolated petrous apex ectopic craniopharyngioma

    Directory of Open Access Journals (Sweden)

    Julius July

    2015-12-01

    Full Text Available Primary ectopic craniopharyngioma is a rare entity. Isolated petrous apex bone location has not been reported previously. This study reports a case of 26-year-old male with right abducent nerve palsy. CT and MRI imaging reveal right petrous apex cystic lesion. No sellar or suprasellar region involvement was found. Endoscopic endonasal transphenoid approach has been successfully performed. Histopathology examination confirms the diagnosis of adamantinomatous craniopharyngioma. So far, it’s probably the first case report of primary ectopic craniopharyngioma isolated in the petrous apex. This case report supports the premise that primary ectopic craniopharyngioma is a multifactorial process that starts with an error from migrated embryological cells.

  20. Isolation contactor state control system

    Energy Technology Data Exchange (ETDEWEB)

    Bissontz, Jay E.

    2017-05-16

    A controller area network (CAN) installed on a hybrid electric vehicle provides one node with control of high voltage power distribution system isolation contactors and the capacity to energize a secondary electro-mechanical relay device. The output of the secondary relay provides a redundant and persistent backup signal to the output of the node. The secondary relay is relatively immune to CAN message traffic interruptions and, as a result, the high voltage isolation contactor(s) are less likely to transition open in the event that the intelligent output driver should fail.

  1. Seismic isolation of nuclear power plants using sliding isolation bearings

    Science.gov (United States)

    Kumar, Manish

    Nuclear power plants (NPP) are designed for earthquake shaking with very long return periods. Seismic isolation is a viable strategy to protect NPPs from extreme earthquake shaking because it filters a significant fraction of earthquake input energy. This study addresses the seismic isolation of NPPs using sliding bearings, with a focus on the single concave Friction Pendulum(TM) (FP) bearing. Friction at the sliding surface of an FP bearing changes continuously during an earthquake as a function of sliding velocity, axial pressure and temperature at the sliding surface. The temperature at the sliding surface, in turn, is a function of the histories of coefficient of friction, sliding velocity and axial pressure, and the travel path of the slider. A simple model to describe the complex interdependence of the coefficient of friction, axial pressure, sliding velocity and temperature at the sliding surface is proposed, and then verified and validated. Seismic hazard for a seismically isolated nuclear power plant is defined in the United States using a uniform hazard response spectrum (UHRS) at mean annual frequencies of exceedance (MAFE) of 10-4 and 10 -5. A key design parameter is the clearance to the hard stop (CHS), which is influenced substantially by the definition of the seismic hazard. Four alternate representations of seismic hazard are studied, which incorporate different variabilities and uncertainties. Response-history analyses performed on single FP-bearing isolation systems using ground motions consistent with the four representations at the two shaking levels indicate that the CHS is influenced primarily by whether the observed difference between the two horizontal components of ground motions in a given set is accounted for. The UHRS at the MAFE of 10-4 is increased by a design factor (≥ 1) for conventional (fixed base) nuclear structure to achieve a target annual frequency of unacceptable performance. Risk oriented calculations are performed for

  2. Active Fault Isolation in MIMO Systems

    DEFF Research Database (Denmark)

    Niemann, Hans Henrik; Poulsen, Niels Kjølstad

    2014-01-01

    isolation is based directly on the input/output s ignals applied for the fault detection. It is guaranteed that the fault group includes the fault that had occurred in the system. The second step is individual fault isolation in the fault group . Both types of isolation are obtained by applying dedicated......Active fault isolation of parametric faults in closed-loop MIMO system s are considered in this paper. The fault isolation consists of two steps. T he first step is group- wise fault isolation. Here, a group of faults is isolated from other pos sible faults in the system. The group-wise fault...

  3. Comparison of seismic isolation concepts for FBR

    International Nuclear Information System (INIS)

    Shiojiri, H.; Mazda, T.; Kasai, H.; Kanda, J.N.; Kubo, T.; Madokoro, M.; Shimomura, T.; Nojima, O.

    1989-01-01

    This paper seeks to verify the reliability and effectiveness of seismic isolation for FBR. Some results of the preliminary study of the program are described. Seismic isolation concepts and corresponding seismic isolation devices were selected. Three kinds of seismically-isolated FBR plant concepts were developed by applying promising seismic isolation concepts to the non-isolated FBR plant, and by developing plant component layout plans and building structural designs. Each plant was subjected to seismic response analysis and reduction in the amount of material of components and buildings were estimated for each seismic isolation concepts. Research and development items were evaluated

  4. Physical isolation with virtual support

    DEFF Research Database (Denmark)

    Wearne, Susan M.; Teunissen, Pim W.; Dornan, Tim

    2015-01-01

    Purpose: Changing the current geographical maldistribution of the medical workforce is important for global health. Research regarding programs that train doctors for work with disadvantaged, rural populations is needed. This paper explores one approach of remote supervision of registrars in isol...

  5. Emotional isolation in BBC Forum

    International Nuclear Information System (INIS)

    Sienkiewicz, J; Chmiel, A

    2014-01-01

    We analyze emotionally annotated massive data from BBC Forum and examine properties of the isolation phenomenon of negative and positive users. Our results show the existence of a percolation threshold dependent on the average emotional value in the network of negatively charged nodes

  6. Sterols isolated from Tubifex tubifex.

    Science.gov (United States)

    Perez Gutierrez, Rosa Martha; Reyes, Ivan Cordova

    2006-11-01

    Four 3beta-hydroxysterols isolated from chloroform extract from Tubifex tubifex were shown to have the structures stigmast-23-en-3-ol, stigmast-7,14-dien-3-ol, 22-dehydrocholesterol, and 24 methylenecholesterol based on spectroscopic methods. Stigmasterol and beta-sitosterol were also investigated on the basis of mass spectral analyses and compared with known compounds.

  7. High-Voltage Isolation Transformer

    Science.gov (United States)

    Clatterbuck, C. H.; Ruitberg, A. P.

    1985-01-01

    Arcing and field-included surface erosion reduced by electrostatic shields around windings and ferromagnetic core of 80-kilovolt isolation transformer. Fabricated from high-resistivity polyurethane-based material brushed on critical surfaces, shields maintained at approximately half potential difference of windings.

  8. Isolated tricuspid valve infective endocarditis

    African Journals Online (AJOL)

    1990-07-07

    Jul 7, 1990 ... thromycin and cefamandole was isolated from multiple blood. Department of .... through the tricuspid orifice into the right atrium. ..... ('ma' 50) indicating adequate platelet function.) In the ... reponed here failed to prevent spontaneous haemorrhage ... this preparation is in shon supply and is very expensive.

  9. Isolating Lysosomes from Rat Liver.

    Science.gov (United States)

    Pryor, Paul R

    2016-04-01

    This protocol describes the generation of a fraction enriched in lysosomes from rat liver. The lysosomes are rapidly isolated using density-gradient centrifugation with gradient media that retain the osmolarity of the lysosomes such that they are functional and can be used in in vitro assays. © 2016 Cold Spring Harbor Laboratory Press.

  10. Optical isolation by Faraday rotator

    International Nuclear Information System (INIS)

    Kasai, Takeshi; Matsushima, Isao; Nemoto, Fusashi; Yano, Masaaki

    1984-01-01

    Three Faraday rotators designed as optical isolators in a high power glass laser system are described. The spatial fluctuation of applied magnetic field is less than 1% throughout the Faraday glass rod. The Faraday rotators transmit more than 80% of the forward-going laser light and reject more than 96% of the backward-going light. (author)

  11. SYMBIODINIUM ISOLATES FROM STONY CORAL: ISOLATION, GROWTH CHARACTERISTICS AND EFFECTS OF UV IRRADIATION

    Science.gov (United States)

    Symbiodinium spp. Isolates from Stony Coral: Isolation, Growth Characteristics and Effects of UV Irradiation (Abstract). J. Phycol. 37(3):42-43.Symbiodinium species were isolated from Montipora capitata, Acropora palmata and two field samples of Porites porites. Cultures ...

  12. Establishing the isolated Standard Model

    International Nuclear Information System (INIS)

    Wells, James D.; Zhang, Zhengkang; Zhao, Yue

    2017-02-01

    The goal of this article is to initiate a discussion on what it takes to claim ''there is no new physics at the weak scale,'' namely that the Standard Model (SM) is ''isolated.'' The lack of discovery of beyond the SM (BSM) physics suggests that this may be the case. But to truly establish this statement requires proving all ''connected'' BSM theories are false, which presents a significant challenge. We propose a general approach to quantitatively assess the current status and future prospects of establishing the isolated SM (ISM), which we give a reasonable definition of. We consider broad elements of BSM theories, and show many examples where current experimental results are not sufficient to verify the ISM. In some cases, there is a clear roadmap for the future experimental program, which we outline, while in other cases, further efforts - both theoretical and experimental - are needed in order to robustly claim the establishment of the ISM in the absence of new physics discoveries.

  13. Isolated urachal malakoplakia mimicking malignancy

    Directory of Open Access Journals (Sweden)

    Saisriharsha Pakalapati

    2017-01-01

    Full Text Available Malakoplakia is an unusual inflammatory disease with uncertain pathogenesis affecting any organ in the body, but predominantly genitourinary tract, with specific predilection to the bladder. We report a rare case of isolated malakoplakia of the urachus in a 29-year-old male patient who presented with lower urinary tract symptoms without any hematuria. Investigations revealed sterile pyuria with no bacterial growth in urine. Radiological investigations revealed a mass in the urachal region. The patient underwent cystoscopy with biopsy followed by pelvic lymph node dissection and partial cystectomy with excision of the urachal mass. Histopathological examination of the mass revealed malakoplakia. Postoperative course was uneventful. To the best of our knowledge, this is the first ever case report of isolated urachal malakoplakia without any concomitant malignancy or bladder involvement reported in our country and one of the very few reported worldwide.

  14. Isolated vasculitis of the CNS

    International Nuclear Information System (INIS)

    Block, F.; Reith, W.

    2000-01-01

    Vasculitis is a rare cause for disease of the CNS. The isolated vasculitis of the CNS is restricted to the CNS whereas other forms of vasculitis affect various organs including the CNS. Headache, encephalopathy, focal deficits and epileptic seizures are the major symptoms suggestive for vasculitis. One major criterion of the isolated vasculitis of the CNS is the lack of evidence for other vasculitis forms or for pathology of other organs. Angiography displays multifocal segmental stenosis of intracranial vessels. MRI demonstrates multiple lesions which in part show enhancement after gadolinium. A definite diagnosis can only be made on the grounds of biopsy from leptomeninges and parenchyma. Therapy consists of corticosteroids and cyclophosphamid. (orig.) [de

  15. Isolated adrenal paracoccidioidomycosis: Case report

    International Nuclear Information System (INIS)

    Uribe Castro, Jorge Ricardo; Quintana, Humberto; Puentes, Alix Sofia and others

    2011-01-01

    Even though paracoccidioidomycosis has a relatively high prevalence in Latin America in a systemic form, isolated cases, especially compromising the adrenal glands, are uncommon, with only two reported cases. In this article, we report the case of a 55 year-old male with clinical manifestations of adrenal insufficiency. The only imaging finding was the presence of bilateral adrenal masses. The biopsy showed Paracoccidioides brasiliensis infection.

  16. Transformerless dc-Isolated Converter

    Science.gov (United States)

    Rippel, Wally E.

    1987-01-01

    Efficient voltage converter employs capacitive instead of transformer coupling to provide dc isolation. Offers buck/boost operation, minimal filtering, and low parts count, with possible application in photovoltaic power inverters, power supplies and battery charges. In photovoltaic inverter circuit with transformerless converter, Q2, Q3, Q4, and Q5 form line-commutated inverter. Switching losses and stresses nil because switching performed when current is zero.

  17. Preparation and isolation of isobenzofuran

    Directory of Open Access Journals (Sweden)

    Morten K. Peters

    2017-12-01

    Full Text Available The synthesis, isolation and characterization of isobenzofuran are described in this publication. Isobenzofuran is of general interest in synthetic and physical organic chemistry because it is one of the most reactive dienes known. A number of synthetic pathways have been published which all suffer from disadvantages such as low yields and difficult purification. We present a synthetic pathway to prepare isobenzofuran in laboratory scale with high yields, from affordable, commercially available starting materials.

  18. Evolution of Isolated Neutron Stars

    OpenAIRE

    Popov, S. B.

    2001-01-01

    In this paper we briefly review our recent results on evolution and properties of isolated neutron stars (INSs) in the Galaxy. As the first step we discuss stochastic period evolution of INSs. We briefly discuss how an INS's spin period evolves under influence of interaction with turbulized interstellar medium. To investigate statistical properties of the INS population we calculate a {\\it census} of INSs in our Galaxy. Then we show that for exponential field decay the range of minimum value ...

  19. Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

    Science.gov (United States)

    Pringle, M; Landén, A; Franklin, A

    2006-02-01

    There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

  20. Input-output rearrangement of isolated converters

    DEFF Research Database (Denmark)

    Madsen, Mickey Pierre; Kovacevic, Milovan; Mønster, Jakob Døllner

    2015-01-01

    This paper presents a new way of rearranging the input and output of isolated converters. The new arrangement posses several advantages, as increased voltage range, higher power handling capabilities, reduced voltage stress and improved efficiency, for applications where galvanic isolation...

  1. Isolated malignant melanoma metastasis to the pancreas

    DEFF Research Database (Denmark)

    Larsen, Anne K; Krag, Christen; Geertsen, Poul

    2013-01-01

    SUMMARY: Malignant melanomas rarely develop isolated pancreatic metastases. We describe a unique patient who is still alive 22 years following an isolated pancreatic melanoma metastasis, and we review the sparse literature in the field....

  2. Isolated Horizon, Killing Horizon and Event Horizon

    OpenAIRE

    Date, G.

    2001-01-01

    We consider space-times which in addition to admitting an isolated horizon also admit Killing horizons with or without an event horizon. We show that an isolated horizon is a Killing horizon provided either (1) it admits a stationary neighbourhood or (2) it admits a neighbourhood with two independent, commuting Killing vectors. A Killing horizon is always an isolated horizon. For the case when an event horizon is definable, all conceivable relative locations of isolated horizon and event hori...

  3. Behavioural reproductive isolation and speciation in Drosophila

    Indian Academy of Sciences (India)

    In the genus Drosophila, the phenomenon of behavioural reproductive isolation, which is an important type of premating (prezygotic) reproductive isolating mechanisms, has been extensively studied and interesting data have been documented. In many cases incomplete sexual isolation has been observed and the pattern ...

  4. Isolation and morphological characterization of antibiotic producing ...

    African Journals Online (AJOL)

    Purpose: To isolate and characterize antibiotic producing actinomycetes from soil samples in Belgaum, Karnataka, India. Methods: Crowded plate technique was used for the isolation of actinomycetes in media such as soybean – casein digest medium and actinomycetes isolation agar. The morphological and cultural ...

  5. Isolation and characterization of copper tolerant bacterial isolates

    International Nuclear Information System (INIS)

    Farooq, S.; Shoeb, E.; Badar, U.; Akhtar, J.

    2008-01-01

    Twelve bacterial strains were isolated from metal contaminated sites close to the chemical factory, Purification and characterization of these strains was done. Maximum tolerable concentration (MTC) of all the isolated strains was determined against heavy metals cadmium chloride (CdCl/sub 2/), copper sulphate (CuSO/sub 4/), and nickel chloride (NiCI/sub 2/) and antibiotics kanamycin (Km), streptomycin (Sm), and chloramphenicol (Cm). Most promising strain was found to be GESSF012 which showed MTC of 4.5 mM and 1.6 mM against CdCI/sub 2/ in enriched and minimal media respectively; whereas MTC of 750 micro g/ml was against Sm. GESSF012 demonstrated the occurrence of multiple stress tolerance as this strain showed considerable tolerance against other heavy metals including CuSO/sub 4/, (3.0 mM in enriched media and 1.8 mM in minimal media) and NiCl/sub 2/, (2.0 mM in enriched media and 1.8 mM in minimal media) as well as other antibiotics Cm and Km (150 and 125 micro g/ml respectively). Plasmids were detected in most of the strains including GESSF012. (author)

  6. Fungi isolated in school buildings

    Directory of Open Access Journals (Sweden)

    Elżbieta Ejdys

    2013-12-01

    Full Text Available The aim of the study was to determine the species composition of fungi occurring on wall surfaces and in the air in school buildings. Fungi isolated from the air using the sedimentation method and from the walls using the surface swab technique constituted the study material. Types of finish materials on wall surfaces were identified and used in the analysis. Samples were collected in selected areas in two schools: classrooms, corridors, men's toilets and women's toilets, cloakrooms, sports changing rooms and shower. Examinations were conducted in May 2005 after the heating season was over. Fungi were incubated on Czapek-Dox medium at three parallel temperatures: 25, 37 and 40°C, for at least three weeks. A total of 379 isolates of fungi belonging to 32 genera of moulds, yeasts and yeast-like fungi were obtained from 321 samples in the school environment. The following genera were isolated most frequently: Aspergillus, Penicillium and Cladosporium. Of the 72 determined species, Cladosporium herbarum, Aspergillus fumigatus and Penicillium chrysogenum occurred most frequently in the school buildings. Wall surfaces were characterised by an increased prevalence of mycobiota in comparison with the air in the buildings, with a slightly greater species diversity. A certain species specificity for rough and smooth wall surfaces was demonstrated. Fungi of the genera Cladosporium and Emericella with large spores adhered better to smooth surfaces while those of the genus Aspergillus with smaller conidia adhered better to rough surfaces. The application of three incubation temperatures helped provide a fuller picture of the mycobiota in the school environment.

  7. Isolated penile torsion in newborns.

    Science.gov (United States)

    Eroglu, Egemen; Gundogdu, Gokhan

    2015-01-01

    We reported on the incidence of isolated penile torsion among our healthy children and our approach to this anomaly. Between 2011 and 2014, newborn babies with penile torsion were classified according to the angle of torsion. Surgical correction (penile degloving and reattachment for moderate cases and dorsal dartos flap technique in case of resistance) after 6 months was advised to the babies with rotations more than 45°. Among 1000 newborn babies, 200 isolated penile torsions were found, and among these, 43 had torsions more than 45°, and 4 of these had angles greater than 90°. The mean angle of the rotations was found 30.45° (median: 20°). In total, 8 children with 60° torsions were previously circumcised. Surgery was performed on 19 patients, with a mean patient age of 12 ± 2 months. Of these 19, 13 babies were corrected with degloving and reattachment. This technique was not enough on the remaining 6 patients; therefore, derotational dorsal dartos flap was added to correct the torsion. After a mean of 15.6 ± 9.8 months, residual penile rotation, less than 15°, was found only in 2 children. The incidence of isolated penile torsion is 20% in newborns. However, rotation more than 45° angles are seen in 4.3% of male babies. Correction is not necessary in mild degrees, and penile degloving with reattachment is enough in most cases. If the initial correction is insufficient, dorsal dartos flap rotation is easy and effective. Prior circumcision neither disturbs the operative procedure nor affects the outcomes.

  8. Basalt waste isolation project overview

    International Nuclear Information System (INIS)

    Dahlem, D.H.

    1987-01-01

    The proposed candidate site for a high-level nuclear waste repository is located beneath the Hanford Nuclear Reservation in southeastern Washington State. At this point, the Hanford Reservation has been selected as one of three preferred candidates in the draft environmental assessment. Project activities have concentrated on understanding the site location with respect to the 10CFR60, 40CFR191, and 10CFR960, identifying critical parameters for design of water package and repository seals, and identifying parameters for repository design. This paper describes the program to evaluate the site and to identify the natural processes that would effect isolation

  9. [Neuroanatomy of Isolated Body Lateropulsion].

    Science.gov (United States)

    Nakazato, Yoshihiko; Tamura, Naotoshi; Ikeda, Kei; Tanaka, Ai; Yamamoto, Toshimasa

    2016-03-01

    Axial body lateropulsion, a phenomenon where the body is pulled toward the side of the lesion, with tendency of falling down, is the well-known transient feature of lateral medullary syndrome. In some cases, axial body lateropulsion occurs without vestibular and cerebellar symptoms (isolated body lateropulsion:[iBL]). Patients with iBL have a lesion located in the spinocerebellar tract, descending lateral vestibulospinal tract, vestibulo-thalamic pathway, dentatorubrothalamic pathway, or thalamocortical fascicle. This review deals with the anatomic basis and clinical significance of iBL.

  10. Thermodynamic laws in isolated systems.

    Science.gov (United States)

    Hilbert, Stefan; Hänggi, Peter; Dunkel, Jörn

    2014-12-01

    The recent experimental realization of exotic matter states in isolated quantum systems and the ensuing controversy about the existence of negative absolute temperatures demand a careful analysis of the conceptual foundations underlying microcanonical thermostatistics. Here we provide a detailed comparison of the most commonly considered microcanonical entropy definitions, focusing specifically on whether they satisfy or violate the zeroth, first, and second laws of thermodynamics. Our analysis shows that, for a broad class of systems that includes all standard classical Hamiltonian systems, only the Gibbs volume entropy fulfills all three laws simultaneously. To avoid ambiguities, the discussion is restricted to exact results and analytically tractable examples.

  11. Variable stiffness and damping MR isolator

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, X Z; Wang, X Y; Li, W H; Kostidis, K [University of Wollongong, School of Mechanical, Materials and Mechatronic Engineering, NSW 2522 (Australia)], E-mail: weihuali@uow.edu.au

    2009-02-01

    This paper presents the development of a magnetorheological (MR) fluid-based variable stiffness and damping isolator for vibration suppressions. The MR fluid isolator used a sole MR control unit to achieve the variable stiffness and damping in stepless and relative large scope. A mathematical model of the isolator was derived, and a prototype of the MR fluid isolator was fabricated and its dynamic behavior was measured in vibration under various applied magnetic fields. The parameters of the model under various magnetic fields were identified and the dynamic performances of isolator were evaluated.

  12. In vitro activity of Ceftaroline against bacterial pathogens isolated from patients with skin and soft tissue and respiratory tract infections in African and Middle Eastern countries: AWARE global surveillance program 2012-2014.

    Science.gov (United States)

    Karlowsky, James A; Biedenbach, Douglas J; Bouchillon, Samuel K; Hackel, Meredith; Iaconis, Joseph P; Sahm, Daniel F

    2016-10-01

    The objective of this report was to document antimicrobial susceptibility testing surveillance data for ceftaroline and comparative agents from the AWARE (Assessing Worldwide Antimicrobial Resistance Evaluation) global surveillance program for bacterial pathogens causing skin and soft tissue and respiratory tract infections in African and Middle Eastern countries from 2012 through 2014. Pathogen identities were confirmed by MALDI-TOF and antimicrobial susceptibility testing performed by CLSI broth microdilution methodology in a central laboratory. All methicillin-susceptible Staphylococcus aureus (MSSA) (n= 923; MIC90, 0.25 μg/mL) and 91.8% of methicillin-resistant S. aureus (MRSA) (n= 1161; MIC90, 1 μg/mL) tested were susceptible to ceftaroline. The maximum ceftaroline MIC observed for isolates of MRSA was 2 μg/mL. All Streptococcus pyogenes (n= 174; MIC90, 0.008 μg/mL), Streptococcus agalactiae (n= 44; MIC90, 0.015 μg/mL), Streptococcus pneumoniae (n= 351; MIC90, 0.25 μg/mL), and Haemophilus influenzae (n= 84; MIC90, ≤0.015 μg/mL) were susceptible to ceftaroline. Rates of susceptibility to ceftaroline among ESBL-negative Escherichia coli (n= 338), Klebsiella pneumoniae (n= 241), and Klebsiella oxytoca (n= 97) were 89.1% (MIC90, 1 μg/mL), 94.2% (MIC90, 0.5 μg/mL), and 99.0% (MIC90, 0.5 μg/mL), respectively. Copyright © 2016. Published by Elsevier Inc.

  13. Establishing the isolated Standard Model

    Energy Technology Data Exchange (ETDEWEB)

    Wells, James D.; Zhang, Zhengkang [Michigan Univ., Ann Arbor, MI (United States). Michigan Center for Theoretical Physics; Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Zhao, Yue [Michigan Univ., Ann Arbor, MI (United States). Michigan Center for Theoretical Physics

    2017-02-15

    The goal of this article is to initiate a discussion on what it takes to claim ''there is no new physics at the weak scale,'' namely that the Standard Model (SM) is ''isolated.'' The lack of discovery of beyond the SM (BSM) physics suggests that this may be the case. But to truly establish this statement requires proving all ''connected'' BSM theories are false, which presents a significant challenge. We propose a general approach to quantitatively assess the current status and future prospects of establishing the isolated SM (ISM), which we give a reasonable definition of. We consider broad elements of BSM theories, and show many examples where current experimental results are not sufficient to verify the ISM. In some cases, there is a clear roadmap for the future experimental program, which we outline, while in other cases, further efforts - both theoretical and experimental - are needed in order to robustly claim the establishment of the ISM in the absence of new physics discoveries.

  14. Isolation of technogenic magnetic particles

    Energy Technology Data Exchange (ETDEWEB)

    Catinon, Mickaël, E-mail: mickael.catinon@gmail.com [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Ayrault, Sophie, E-mail: sophie.ayrault@lsce.ispl.fr [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Boudouma, Omar, E-mail: boudouma@ccr.jussieu.fr [Service du MEB, UFR928, Université Pierre et Marie Curie, 75252 Paris VI (France); Bordier, Louise, E-mail: Louise.Bordier@lsce.ipsl.fr [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Agnello, Gregory, E-mail: contact@evinrude.fr [Evinrude, Espace St Germain, 38200 Vienne (France); Reynaud, Stéphane, E-mail: stephane.reynaud@ujf-grenoble.fr [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Tissut, Michel, E-mail: michel.tissut@ujf-grenoble.fr [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France)

    2014-03-01

    Technogenic magnetic particles (TMPs) emitted by various industrial sources, such as smelting plants, end up after atmospheric transfer on the soil surface. In the present study, we characterised the origin and composition of such particles emitted by a large iron smelting plant and deposited on particular substrates, namely tombstones, which act as a very interesting and appropriate matrix when compared to soil, tree bark, lichens or attic dust. The isolation and subsequent description of TMPs require a critical step of separation between different components of the sample and the magnetic particles; here, we described an efficient protocol that fulfils such a requirement: it resorts to water suspension, sonication, repeated magnetic extraction, sedimentation, sieving and organic matter destruction at 550 °C in some instances. The isolated TMPs displayed a noticeable crystalline shape with variable compositions: a) pure iron oxides, b) iron + Cr, Ni or Zn, and c) a complex structure containing Ca, Si, Mg, and Mn. Using Scanning Electron Microscope Energy Dispersive X-ray (SEM–EDX), we obtained profiles of various and distinct magnetic particles, which allowed us to identify the source of the TMPs. - Highlights: • The developed method offers a low-cost approach of large-scale dry deposition. • Tombstones are excellent supports for sampling these atmospheric deposits. • Smelted elements crystallise after cooling, giving typical technogenic magnetic particles (TMPs). • Coupling microscopic and bulk analyses allows identifying TMP origin. • Magnetic TMPs issued from steel industry were separated by a new technique.

  15. Power inverter with optical isolation

    Science.gov (United States)

    Duncan, Paul G.; Schroeder, John Alan

    2005-12-06

    An optically isolated power electronic power conversion circuit that includes an input electrical power source, a heat pipe, a power electronic switch or plurality of interconnected power electronic switches, a mechanism for connecting the switch to the input power source, a mechanism for connecting comprising an interconnecting cable and/or bus bar or plurality of interconnecting cables and/or input bus bars, an optically isolated drive circuit connected to the switch, a heat sink assembly upon which the power electronic switch or switches is mounted, an output load, a mechanism for connecting the switch to the output load, the mechanism for connecting including an interconnecting cable and/or bus bar or plurality of interconnecting cables and/or output bus bars, at least one a fiber optic temperature sensor mounted on the heat sink assembly, at least one fiber optic current sensor mounted on the load interconnection cable and/or output bus bar, at least one fiber optic voltage sensor mounted on the load interconnection cable and/or output bus bar, at least one fiber optic current sensor mounted on the input power interconnection cable and/or input bus bar, and at least one fiber optic voltage sensor mounted on the input power interconnection cable and/or input bus bar.

  16. Assessing Social Isolation: Pilot Testing Different Methods.

    Science.gov (United States)

    Taylor, Harry Owen; Herbers, Stephanie; Talisman, Samuel; Morrow-Howell, Nancy

    2016-04-01

    Social isolation is a significant public health problem among many older adults; however, most of the empirical knowledge about isolation derives from community-based samples. There has been less attention given to isolation in senior housing communities. The objectives of this pilot study were to test two methods to identify socially isolated residents in low-income senior housing and compare findings about the extent of isolation from these two methods. The first method, self-report by residents, included 47 out of 135 residents who completed in-person interviews. To determine self-report isolation, residents completed the Lubben Social Network Scale 6 (LSNS-6). The second method involved a staff member who reported the extent of isolation on all 135 residents via an online survey. Results indicated that 26% of residents who were interviewed were deemed socially isolated by the LSNS-6. Staff members rated 12% of residents as having some or a lot of isolation. In comparing the two methods, staff members rated 2% of interviewed residents as having a lot of isolation. The combination of self-report and staff report could be more informative than just self-report alone, particularly when participation rates are low. However, researchers should be aware of the potential discrepancy between these two methods.

  17. In vitro activity of tigecycline against isolates collected from complicated skin and skin structure infections and intra-abdominal infections in Africa and Middle East countries: TEST 2007-2012.

    Science.gov (United States)

    Renteria, M I; Biedenbach, D J; Bouchillon, S K; Hoban, D J; Raghubir, N; Sajben, P; Mokaddas, E

    2014-05-01

    Complicated skin and skin structure infections (cSSSIs) and intra-abdominal infections (IAIs) are problematic due to decreasing therapeutic options available against multidrug-resistant pathogens common among these types of infections. A total of 2245 isolates from African and the Middle Eastern (AfME) countries were collected to determine in vitro activity for tigecycline and comparators during 2007-2012 as part of the Tigecycline Evaluation Surveillance Trial program. Tigecycline was launched in the AfME in 2007 and remains active against a wide range of targeted pathogens worldwide. Isolates were recovered from cSSSI (1990) and IAI (255) from 38 sites in 11 AfME countries. Staphylococcus aureus was the most common species from cSSSI (27.9%), and the methicillin-resistant S. aureus rate was 25%. Enterococcus spp. (7.1%) and Streptococcus agalactiae (2.9%) were other common Gram-positive pathogens represented. Enterobacter spp. (14.5%), Pseudomonas aeruginosa (13.9%), Escherichia coli (11.4%), Klebsiella spp. (10.9%), and Acinetobacter spp. (7.2 %) were the most common Gram-negative species collected. Tigecycline MIC(90) values were 0.25 μg/mL against S. aureus. E. coli and Enterobacter spp. had tigecycline MIC(90) values of 1 and 2 μg/mL, respectively. E. coli was the most frequently collected species from IAI (28.3%), followed by Klebsiella spp. (20.8%), Enterococcus spp. (11.8%), and Stenotrophomonas maltophilia (6.3%). Isolates collected from IAI had the following tigecycline MIC(90) values: E. coli (1 μg/mL), Klebsiella spp. and other Enterobacteriaceae (2 μg/mL), Enterococcus spp. (0.25 μg/mL), and S. maltophilia (1 μg/mL). Tigecycline in vitro activity was observed against a broad spectrum of bacterial species, including strains resistant to other antimicrobial classes. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Oligotrophic bacteria isolated from clinical materials.

    OpenAIRE

    Tada, Y; Ihmori, M; Yamaguchi, J

    1995-01-01

    Oligotrophic bacteria (oligotrophs) are microorganisms that grow in extremely nutritionally deficient conditions in which the concentrations of organic substances are low. Many oligotrophic bacteria were isolated from clinical materials including urine, sputum, swabbings of the throat, vaginal discharges, and others. Seventy-seven strains of oligotrophic bacteria from 871 samples of clinical material were isolated. A relatively higher frequency of isolation of oligotrophic bacteria was shown ...

  19. Isolated trochlear nerve palsy with midbrain hemorrhage

    Directory of Open Access Journals (Sweden)

    Raghavendra S

    2010-01-01

    Full Text Available Midbrain hemorrhage causing isolated fourth nerve palsy is extremely rare. Idiopathic, traumatic and congenital abnormalities are the most common causes of fourth nerve palsy. We report acute isolated fourth nerve palsy in an 18-year-old lady due to a midbrain hemorrhage probably due to a midbrain cavernoma. The case highlights the need for neuroimaging in selected cases of isolated trochlear nerve palsy.

  20. Recent advances in micro-vibration isolation

    Science.gov (United States)

    Liu, Chunchuan; Jing, Xingjian; Daley, Steve; Li, Fengming

    2015-05-01

    Micro-vibration caused by disturbance sources onboard spacecraft can severely degrade the working environment of sensitive payloads. Some notable vibration control methods have been developed particularly for the suppression or isolation of micro-vibration over recent decades. Usually, passive isolation techniques are deployed in aerospace engineering. Active isolators, however, are often proposed to deal with the low frequency vibration that is common in spacecraft. Active/passive hybrid isolation has also been effectively used in some spacecraft structures for a number of years. In semi-active isolation systems, the inherent structural performance can be adjusted to deal with variation in the aerospace environment. This latter approach is potentially one of the most practical isolation techniques for micro-vibration isolation tasks. Some emerging advanced vibration isolation methods that exploit the benefits of nonlinearity have also been reported in the literature. This represents an interesting and highly promising approach for solving some challenging problems in the area. This paper serves as a state-of-the-art review of the vibration isolation theory and/or methods which were developed, mainly over the last decade, specifically for or potentially could be used for, micro-vibration control.

  1. Fault Isolation for Shipboard Decision Support

    DEFF Research Database (Denmark)

    Lajic, Zoran; Blanke, Mogens; Nielsen, Ulrik Dam

    2010-01-01

    Fault detection and fault isolation for in-service decision support systems for marine surface vehicles will be presented in this paper. The stochastic wave elevation and the associated ship responses are modeled in the frequency domain. The paper takes as an example fault isolation of a containe......Fault detection and fault isolation for in-service decision support systems for marine surface vehicles will be presented in this paper. The stochastic wave elevation and the associated ship responses are modeled in the frequency domain. The paper takes as an example fault isolation...... to the quality of decisions given to navigators....

  2. Genetics Home Reference: isolated growth hormone deficiency

    Science.gov (United States)

    ... can be inherited? More about Inheriting Genetic Conditions Diagnosis & Management Resources Genetic Testing (4 links) Genetic Testing Registry: Ateleiotic dwarfism Genetic Testing Registry: Autosomal dominant isolated somatotropin deficiency ...

  3. Loneliness and Social Isolation among Eldely People

    Directory of Open Access Journals (Sweden)

    Engin Püllüm

    2017-11-01

    Full Text Available Social isolation is defined as the process of people losing their contacts with other social resources or their willingness to participate. Social isolation is a grave and widespread problem among seniors in society causing many harmful health conditions. Social isolation may be prevented by defining risk factors in seniors and leading them to suitable resources. The elderly should be encouraged to share their experiences and continue to produce for as long as possible, so that they are prevented from feeling isolated from society and the continuity of their economic productivity is ensured.

  4. Mycoplasma contamination of Chlamydia pneumoniae isolates

    DEFF Research Database (Denmark)

    Huniche, BS; Jensen, Lise Torp; Birkelund, Svend

    1998-01-01

    media, immunoblotting, polymerase chain reaction and pulsed field gel electrophoresis. Two of the 6 ATCC isolates [ATCC VR1355 (TWAR strain 2043) and ATCC VR1356 (TWAR strain 2023)] were infected with Mycoplasma hominis and 1 isolate [ATCC VR2282 (TWAR strain TW183)] was contaminated with both...... Mycoplasma hominis and Mycoplasma orale, whereas 3 of the ATCC isolates [ATCC VR1310, ATCC VR1360 (TWAR strain CM-1) and ATCC 53592 (TWAR strain AR39)] were not contaminated. The Finnish C. pneumoniae isolates Kajaani 6 and Parola were found to be contaminated with M. hominis and M. orale, respectively...

  5. Permanent isolation surface barrier: Functional performance

    International Nuclear Information System (INIS)

    Wing, N.R.

    1993-10-01

    This document presents the functional performance parameters for permanent isolation surface barriers. Permanent isolation surface barriers have been proposed for use at the Hanford Site (and elsewhere) to isolate and dispose of certain types of waste in place. Much of the waste that would be disposed of using in-place isolation techniques is located in subsurface structures, such as solid waste burial grounds, tanks, vaults, and cribs. Unless protected in some way, the wastes could be transported to the accessible environment via transport pathways, such as water infiltration, biointrusion, wind and water erosion, human interference, and/or gaseous release

  6. Isolation and characterization of methanogenic bacteria from ...

    African Journals Online (AJOL)

    Isolation and characterization of methanogenic bacteria from brewery wastewater in Kenya. Sylvia Injete Murunga, Duncan Onyango Mbuge, Ayub Njoroge Gitau, Urbanus Ndungwa Mutwiwa, Ingrid Namae Wekesa ...

  7. Isolated systems with wind power. Main report

    DEFF Research Database (Denmark)

    Lundsager, P.; Bindner, Henrik W.; Clausen, Niels-Erik

    2001-01-01

    The overall objective of this research project is to study the development of methods and guidelines rather than "universal solutions" for the use of wind energy in isolated communities. The main specific objective of the project is to develop and present amore unified and generally applicable...... approach for assessing the technical and economical feasibility of isolated power supply systems with wind energy. As a part of the project the following tasks were carried out: Review of literature, fieldmeasurements in Egypt, development of an inventory of small isolated systems, overview of end...... for Isolated Systems with Wind Power, applicable for international organisations such as donoragencies and development banks....

  8. Update on clinically isolated syndrome.

    Science.gov (United States)

    Thouvenot, Éric

    2015-04-01

    Optic neuritis, myelitis and brainstem syndrome accompanied by a symptomatic MRI T2 or FLAIR hyperintensity and T1 hypointensity are highly suggestive of multiple sclerosis (MS) in young adults. They are called "clinically isolated syndrome" (CIS) and correspond to the typical first multiple sclerosis (MS) episode, especially when associated with other asymptomatic demyelinating lesions, without clinical, radiological and immunological sign of differential diagnosis. After a CIS, the delay of apparition of a relapse, which corresponds to the conversion to clinically definite MS (CDMS), varies from several months to more than 10 years (10-15% of cases, generally called benign RRMS). This delay is generally associated with the number and location of demyelinating lesions of the brain and spinal cord and the results of CSF analysis. Several studies comparing different MRI criteria for dissemination in space and dissemination in time of demyelinating lesions, two hallmarks of MS, provided enough substantial data to update diagnostic criteria for MS after a CIS. In the last revision of the McDonald's criteria in 2010, diagnostic criteria were simplified and now the diagnosis can be made by a single initial scan that proves the presence of active asymptomatic lesions (with gadolinium enhancement) and of unenhanced lesions. However, time to conversion remains highly unpredictable for a given patient and CIS can remain isolated, especially for idiopathic unilateral optic neuritis or myelitis. Univariate analyses of clinical, radiological, biological or electrophysiological characteristics of CIS patients in small series identified numerous risk factors of rapid conversion to MS. However, large series of CIS patients analyzing several characteristics of CIS patients and the influence of disease modifying therapies brought important information about the risk of CDMS or RRMS over up to 20 years of follow-up. They confirmed the importance of the initial MRI pattern of

  9. [Prenatal management of isolated IUGR].

    Science.gov (United States)

    Senat, M-V; Tsatsaris, V

    2013-12-01

    To evaluate the performance of different antenatal tools for the monitoring of fetuses with isolated intrauterine growth restriction (IUGR). To define the prenatal management of IUGR and indications for delivery before and after 32 weeks of gestation. PubMed, Embase and the Cochrane databases were searched using the keywords "IUGR", "fetal growth restriction", "cardiotocography", "amniotic fluid", "ultrasound assessment", "biophysical profile", "Doppler ultrasonography", "randomized trial", "meta-analysis". These terms were also combined together. Fetal monitoring of isolated IUGR should be based on the combined use of fetal heart rate (FHR) and ultrasound Doppler. The use of computerized FHR, with short-term variability (STV) measurement allows longitudinal monitoring and provides objective values upon which to decide very premature delivery (LE3). The use of umbilical Doppler is associated with a decrease in perinatal morbidity, especially in IUGR (LE1). It should be the first-line mean for the monitoring of SGA and IUGR fetuses (LE1). The additional use of cerebral Doppler is associated with a better predictive value for a poor perinatal outcome than the umbilical Doppler alone (LE3). Therefore, cerebral Doppler should be used in fetuses with IUGR, whether the umbilical Doppler is normal or not. As morbidity and mortality is increased in IUGR with pathological ductus venosus, the use of this Doppler should be considered in the monitoring of IUGR at before 32 weeks (professional consensus). Routine hospitalization is not mandatory for the monitoring of fetuses with IUGR/SGA. However, tertiary referral is advisable in cases of severe IUGR at between 26 to 32 weeks (professional consensus). The decision for delivery cannot be standardized and should be based on the combined analysis of gestational age, fetal heart rate analysis and Doppler study (professional consensus). Monitoring of fetuses with IUGR and decision for delivery should be based on the combined

  10. [Post-marketing surveillance of antibacterial activities of cefozopran against various clinical isolates--I. Gram-positive bacteria].

    Science.gov (United States)

    Igari, Jun; Oguri, Toyoko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

    2003-10-01

    As a post-marketing surveillance, the in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates were yearly evaluated and compared with those of other cephems, oxacephems, penicillins, and carbapenems. Changes in the bacterial susceptibility for CZOP were also evaluated with the resistance ratio calculated with breakpoint MIC. Sixteen species (2,363 strains) of Gram-positive bacteria were isolated from the clinical materials annually collected from 1996 to 2001, and consisted of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible Staphylococcus epidermidis (MSSE), methicillin-resistant Staphylococcus epidermidis (MRSE), Staphylococcus haemolyticus, Staphylococcus saprophyticus, Enterococcus faecalis, Enterococcus faecium, Enterococcus avium, Streptococcus pyogenes, Streptococcus agalactiae, penicillin-susceptible Streptococcus pneumoniae (PSSP), penicillin-intermediate resistant S. pneumoniae (PISP), penicillin-resistant S. pneumoniae (PRSP), Streptococcus milleri group and Peptostreptococcus spp. The antibacterial activity of CZOP either against MSSA or MSSE was preferable (MIC90: 2 or 0.5 micrograms/mL) and comparable to those of other cephems. CZOP was also effective on MRSE (MIC90: 16 micrograms/mL) but not on MRSA. CZOP and other cephems had low antibacterial activity against S. haemolyticus (MIC90: 64 micrograms/mL). The antibacterial activity of CZOP against S. saprophyticus was comparable to or higher than those of other cephems, but the MIC90 of CZOP in 2001 was higher than those in 1996-2000 (32 vs 1-2 micrograms/mL). The antibacterial activity of CZOP against E. faecalis was comparable to that of cefpirome (CPR; MIC90: 16 micrograms/mL) and higher than those of other cephems. No antibacterial activity of CZOP against E. faecium and E. avium was observed, like other drugs. The antibacterial activity of CZOP against S. pyogenes

  11. Single Cell Isolation and Analysis

    Directory of Open Access Journals (Sweden)

    Ping Hu

    2016-10-01

    Full Text Available Increasing evidence shows that the heterogeneity of individual cells within a genetically identical population can be critical to their peculiar function and fate. Conventional cell based assays mainly analysis the average responses from a population cells, while the difference within individual cells may often be masked. The cell size, RNA transcripts and protein expression level are quite different within individual cells and these variations are key point to answer the problems in cancer, neurobiology, stem cell biology, immunology and developmental biology. To better understand the cell-to-cell variations, the single cell analysis can provide much more detailed information which may be helpful for therapeutic decisions in an increasingly personalized medicine. In this review, we will focus on the recent development in single cell analysis, including methods used in single cell isolation, analysis and some application examples. The review provides the historical background to single cell analysis, discusses limitations, and current and future possibilities in this exciting field of research.

  12. Isolated polypeptide having arabinofuranosidase activity

    Science.gov (United States)

    Foreman, Pamela; Van Solingen, Pieter; Goedegebuur, Frits; Ward, Michael

    2010-02-23

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry. TABLE-US-00001 cip1 cDNA sequence (SEQ ID NO: 1) GACTAGTTCA TAATACAGTA GTTGAGTTCA TAGCAACTTC 50 ACTCTCTAGC TGAACAAATT ATCTGCGCAA ACATGGTTCG CCGGACTGCT 100 CTGCTGGCCC TTGGGGCTCT CTCAACGCTC TCTATGGCCC AAATCTCAGA 150 CGACTTCGAG TCGGGCTGGG ATCAGACTAA ATGGCCCATT TCGGCACCAG 200 ACTGTAACCA GGGCGGCACC GTCAGCCTCG ACACCACAGT AGCCCACAGC 250 GGCAGCAACT CCATGAAGGT CGTTGGTGGC CCCAATGGCT ACTGTGGACA 300 CATCTTCTTC GGCACTACCC AGGTGCCAAC TGGGGATGTA TATGTCAGAG 350 CTTGGATTCG GCTTCAGACT GCTCTCGGCA GCAACCACGT CACATTCATC 400 ATCATGCCAG ACACCGCTCA GGGAGGGAAG CACCTCCGAA TTGGTGGCCA 450 AAGCCAAGTT CTCGACTACA ACCGCGAGTC CGACGATGCC ACTCTTCCGG 500 ACCTGTCTCC CAACGGCATT GCCTCCACCG TCACTCTGCC TACCGGCGCG 550 TTCCAGTGCT TCGAGTACCA CCTGGGCACT GACGGAACCA TCGAGACGTG 600 GCTCAACGGC AGCCTCATCC CGGGCATGAC CGTGGGCCCT GGCGTCGACA 650 ATCCAAACGA CGCTGGCTGG ACGAGGGCCA GCTATATTCC GGAGATCACC 700 GGTGTCAACT TTGGCTGGGA GGCCTACAGC GGAGACGTCA ACACCGTCTG 750 GTTCGACGAC ATCTCGATTG CGTCGACCCG CGTGGGATGC GGCCCCGGCA 800 GCCCCGGCGG TCCTGGAAGC TCGACGACTG GGCGTAGCAG CACCTCGGGC 850 CCGACGAGCA CTTCGAGGCC AAGCACCACC ATTCCGCCAC CGACTTCCAG 900 GACAACGACC GCCACGGGTC CGACTCAGAC ACACTATGGC CAGTGCGGAG 1000 GGATTGGTTA CAGCGGGCCT ACGGTCTGCG CGAGCGGCAC GACCTGCCAG 1050 GTCCTGAACC CATACTACTC CCAGTGCTTA TAAGGGGATG AGCATGGAGT 1100 GAAGTGAAGT GAAGTGGAGA GAGTTGAAGT GGCATTGCGC TCGGCTGGGT 1150 AGATAAAAGT CAGCAGCTAT GAATACTCTA TGTGATGCTC ATTGGCGTGT 1200 ACGTTTTAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1250 AAAAAAAAAA AAAAAAAAAG GGGGCGGCCG C 1271

  13. Stability of isolated Barchan dunes

    Science.gov (United States)

    Fourrière, Antoine; Charru, François

    2010-11-01

    When sand grains are entrained by an air flow over a non-erodible ground, or with limited sediment supply from the bed, they form isolated dunes showing a remarkable crescentic shape with horns pointing downstream. These dunes, known as Barchan dunes, are commonly observed in deserts, with height of a few meters and velocity of a few meters per year (Bagnold 1941). These dunes also exist under water, at a much smaller, centimetric size (Franklin & Charru 2010). Their striking stability properties are not well understood yet. Two phenomena are likely to be involved in this stability: (i) relaxation effects of the sand flux which increases from the dune foot up to the crest, related to grain inertia or deposition, and (ii) a small transverse sand flux due to slope effects and the divergence of the streamlines of the fluid flow. We reproduced aqueous Barchan dunes in a channel, and studied their geometrical and dynamic properties (in particular their shape, velocity, minimum size, and rate of erosion). Using coloured glass beads (see the figure), we were then able to measure the particle flux over the whole dune surface. We will discuss the stability of these dunes in the light of our measurements.

  14. Cooling system upon reactor isolation

    International Nuclear Information System (INIS)

    Yamamoto, Kohei; Oda, Shingo; Miura, Satoshi

    1992-01-01

    A water level indicator for detecting the upper limit value for a range of using a suppression pool and a thermometer for detecting the temperature of water at the cooling water inlet of an auxiliary device are disposed. When a detection signal is intaken and the water level in the suppression pool reach the upper limit value for the range of use, a secondary flow rate control value is opened and a primary flow rate control valve is closed. When the temperature of the water at the cooling water inlet of the auxiliary device reaches the upper limit value, the primary and the secondary flow rate control valves are opened. During a stand-by state, the first flow rate control valve is set open and the secondary flow rate control valve is set closed respectively. After reactor isolation, if a reactor water low level signal is received, an RCIC pump is actuated and cooling water is sent automatically under pressure from a condensate storage tank to the reactor and the auxiliary device requiring coolants by way of the primary flow rate control valve. Rated flow rate is ensured in the reactor and cooling water of an appropriate temperature can be supplied to the auxiliary device. (N.H.)

  15. Cooling device upon reactor isolation

    International Nuclear Information System (INIS)

    Otsu, Tatsuya

    1995-01-01

    A vacuum breaking valve is disposed to a sucking pipeline of vacuum pumps. A sucking port of the breaking valve is connected with an exhaustion side of a relief valve of a liquid nitrogen-filled tank by way of communication pipes. When a cooling device is operated upon reactor isolation and the vacuum pumps are operated, a three directional electromagnetic valve is operated, and nitrogen discharged out of the exhaustion port of the relief valve of the liquid nitrogen-filled tank is sent to a nitrogen releasing port on the suction side of the vacuum breaking valve by way of the communication pipes and released to atmosphere. When the pressure in the vacuum tank is excessively lowered in this state and the vacuum breaking valve is opened, nitrogen flows from the nitrogen discharge port into the vacuum tank through the breaking valve, and are sent to a pressure suppression chamber by the vacuum pumps. Since a great amount of nitrogen is sent to the pressure suppression chamber, and the inflow of the air is reduced, increase of oxygen concentration in the pressure suppression chamber can be suppressed. (I.N.)

  16. Distinguishing Isolated Photons from Jets

    CERN Document Server

    Pieri, Marco; Branson, James G

    2006-01-01

    We have developed isolation and shower-shape variables that are optimized for reducing the jet background for a high transverse-energy photon signal. To help understand the relative importance of each variable, we have computed the background rejection power as a function of signal efficiency for these variables as well as for some simple combinations of variables. We find that a combination of tracker plus ECAL information can give very significant background rejection power. By adding information from the HCAL, the rejection power can be improved. In addition we find that a very significant reduction in background can be achieved by adding photon shower shape information from the ECAL to the Neural Net inputs. About 1 in 200 jets contains a high E_T electromagnetic shower that is reconstructed by the CMS software and is therefore a potential background for photon detection. An additional jet rejection factor of 100 with 80% photon efficiency (or 400 with 50% efficiency) can be achieved by using the isolatio...

  17. Review of the ISOL Method

    CERN Document Server

    Lindroos, M

    2004-01-01

    The ISOL technique was invented in Copenhagen over 50 years ago and eventually migrated to CERN where a suitable proton drive beam was available at the Syncho-Cyclotron. The quick spread of the technique from CERN to many other laboratories has resulted in a large user community, which has assured the continued development of the method, physics in the front-line of fundamental research and the application of the method to many applied sciences. The technique is today established as one of the main techniques for on-line isotope production of high intensity and high quality beams. The thick targets used allows the production of unmatched high intensity radioactive beams. The fact that the ions are produced at rest makes it ideally suitable for low energy experiments and for post acceleration using well established accelerator techniques. The many different versions of the technique will be discussed and the many facilities spread all over the world will be reviewed. The major developments at the existing faci...

  18. Isolation colling device for reactor

    International Nuclear Information System (INIS)

    Ikehara, Morihiko; Arai, Shigeki.

    1982-01-01

    Purpose: To prevent undesired operation of an emergency core cooling system due to excess lowering of water level in a reactor. Constitution: In an emergency facility adapted to drive a turbine, upon reactor isolation, with the excess steams of the reactor to operate a pump and thereby inject cooling water to the reactor, a water level detector is provided and connected to a pump exhaust valve control circuit, a turbine inlet valve control circuit and a by-pass valve control circuit. Valve ON-OFF is automatically controlled depending on the water level to thereby render the level constant. A by-pass pipe is branched from a pump exhaust pipe and connected to a condensate storage tank. When the water level rises due to water injection, the injecting water is returned to circulate by way of the by-pass pipe to the condensate storage tank under the ON-OFF for each of the valves while the turbine being kept to drive. Then, if the water level is lowered, water injection is started again by the ON-OFF for each of the valves. (Ikeda, J.)

  19. Urine Exosome Isolation and Characterization.

    Science.gov (United States)

    Street, Jonathan M; Koritzinsky, Erik H; Glispie, Deonna M; Yuen, Peter S T

    2017-01-01

    Exosomes are nanometer-scale, membrane-enclosed vesicles that can potentially be used to detect nephrotoxicity, and reveal the subsequent response of the kidney. Epithelial cells of every nephron segment can contribute to the urinary exosome population, which is rich in potential biomarkers, including membrane proteins such as transporters and receptors, transcription factors, and microRNAs. These exosomal biomarkers may be up- or downregulated upon nephrotoxicant exposure. Exosome isolation is an area of ongoing research. Although faster and simpler methods have been developed, ultracentrifugation remains a mainstay for purification. A single ultracentrifugation step provides an enriched preparation suitable for biomarker discovery, and a second ultracentrifugation on a sucrose/D 2 O cushion provides the purest exosome preparation currently available and may be preferred for bioactivity assays. The concentration of exosomes can be determined using Nanosight Nanoparticle Tracking Analysis and their contents studied with a variety of approaches including western blots for proteins and RT-qPCR for microRNAs.

  20. Blackbody Radiation from Isolated Neptunes

    Science.gov (United States)

    Ginzburg, Sivan; Sari, Re'em; Loeb, Abraham

    2016-05-01

    Recent analyses of the orbits of some Kuiper belt objects hypothesize the presence of an undiscovered Neptune-size planet at a very large separation from the Sun. The energy budget of Neptunes on such distant orbits is dominated by the internal heat released by their cooling rather than solar irradiation (making them effectively “isolated”). The blackbody radiation that these planets emit as they cool may provide the means for their detection. Here, we use an analytical toy model to study the cooling and radiation of isolated Neptunes. This model can translate a detection (or a null detection) to a constraint on the size and composition of the hypothesized “Planet Nine.” Specifically, the thick gas atmosphere of Neptune-like planets serves as an insulating blanket that slows down their cooling. Therefore, a measurement of the blackbody temperature, {T}{{eff}}˜ 50 {{K}}, at which a Neptune emits, can be used to estimate the mass of its atmosphere, {M}{{atm}}. Explicitly, we find the relation {T}{{eff}}\\propto {M}{{atm}}1/12. Despite this weak relation, a measurement of the flux at the Wien tail can constrain the atmospheric mass, at least to within a factor of a few, and provide useful limits to possible formation scenarios of these planets. Finally, we constrain the size and composition of Planet Nine by combining our model with the null results of recent all-sky surveys.

  1. Effect of isolation techniques on the characteristics of pigeon pea (Cajanus cajan) protein isolates

    OpenAIRE

    Adenekan, Monilola K.; Fadimu, Gbemisola J.; Odunmbaku, Lukumon A.; Oke, Emmanuel K.

    2017-01-01

    Abstract In this study, the effect of different isolation techniques on the isolated proteins from pigeon pea was investigated. Water, methanol, ammonium sulfate, and acetone were used for the precipitation of proteins from pigeon pea. Proximate composition, and antinutritional and functional properties of the pigeon pea flour and the isolated proteins were measured. Data generated were statistically analyzed. The proximate composition of the water‐extracted protein isolate was moisture 8.30%...

  2. Antimicrobial activity of pure platelet-rich plasma against microorganisms isolated from oral cavity.

    Science.gov (United States)

    Drago, Lorenzo; Bortolin, Monica; Vassena, Christian; Taschieri, Silvio; Del Fabbro, Massimo

    2013-02-25

    Autologous platelet concentrates (PCs) have been extensively used in a variety of medical fields to promote soft and hard tissue regeneration. The significance behind their use lies in the abundance of growth factors in platelets α-granules that promotes wound healing. In addition, antibacterial properties of PCs against various bacteria have been recently pointed out. In this study, the antimicrobial effect of pure platelet-rich plasma (P-PRP) was evaluated against oral cavity microorganisms such as Enterococcus faecalis, Candida albicans, Streptococcus agalactiae, Streptococcus oralis and Pseudomonas aeruginosa. Blood samples were obtained from 17 patients who underwent oral surgery procedures involving the use of P-PRP. The antibacterial activity of P-PRP, evaluated as the minimum inhibitory concentration (MIC), was determined through the microdilution twofold serial method. P-PRP inhibited the growth of Enterococcus faecalis, Candida albicans, Streptococcus agalactiae and Streptococcus oralis, but not of Pseudomonas aeruginosa strains. P-PRP is a potentially useful substance in the fight against postoperative infections. This might represent a valuable property in adjunct to the enhancement of tissue regeneration.

  3. Assessment of effectiveness of geologic isolation systems

    International Nuclear Information System (INIS)

    Soldat, J.K.; Napier, B.A.; Strenge, D.L.; Schreckhise, R.G.; Zimmerman, M.G.

    1981-01-01

    The program for Assessment of Effectiveness of Geologic Isolation Systems (AEGIS) is managed through PNL's Water and Land Resources Department and is funded through the Battelle Office of Nuclear Waste Isolation (ONWI). The Ecological Sciences Department was involved in two subtasks under AEGIS: Dose Methodology Development and Reference Site Initial Analysis (RSIA) for a Salt Dome

  4. Isolation and characterization of extreme halophilic archaea

    Energy Technology Data Exchange (ETDEWEB)

    Franze, Madlen; Cherkouk, Andrea [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). HZDR Young Investigator Group

    2017-06-01

    Extreme halophilic archaea from the family Halobactereacea represent a dominant part of the microbial community present in saline soils as well as rock salts. By using a culture-dependent approach different Haloarchaea could be isolated and were phylogenetic analysed. Interestingly, isolates closely related to different Halobacterium spp. were found in both environments.

  5. ASPECTS OF THE ISOLATION AND CHARACTERIZATON OF ...

    African Journals Online (AJOL)

    A local isolate of Alternaria alternata isolated from soil under the decaying cassava peels heap was screened for the production of a-amylase and hydrolysis of starch. The maximal dextrinizing amylolytic activity of the partially purified enzyme was obtained at temperature of 600C and pH 7.0. Apart from glucose, sucrose and ...

  6. Virus isolation: Specimen type and probable transmission

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. Virus isolation: Specimen type and probable transmission. Over 500 CHIK virus isolations were made. 4 from male Ae. Aegypti (?TOT). 6 from CSF (neurological involvement). 1 from a 4-day old child (transplacental transmission.

  7. Isolation and characterization of extreme halophilic archaea

    International Nuclear Information System (INIS)

    Franze, Madlen; Cherkouk, Andrea

    2017-01-01

    Extreme halophilic archaea from the family Halobactereacea represent a dominant part of the microbial community present in saline soils as well as rock salts. By using a culture-dependent approach different Haloarchaea could be isolated and were phylogenetic analysed. Interestingly, isolates closely related to different Halobacterium spp. were found in both environments.

  8. Isolation and characterization of cellulolytic Bacillus licheniformis ...

    African Journals Online (AJOL)

    Eight cellulose degrading bacteria were isolated from compost and were identified as Bacillus licheniformis by 16S rRNA sequencing. Among the eight isolates, Bacillus licheniformis B4, B7 and B8 showed the highest cellulase activity. B. licheniformis B4 and B8 showed the maximum cellulase activity during the stationary ...

  9. Isolated gallbladder rupture following blunt abdominal injury

    African Journals Online (AJOL)

    2015-05-26

    May 26, 2015 ... Kaohsiung Medical University, Kaohsiung, Taiwan. Abstract. Isolated traumatic gallbladder rupture subsequent to blunt abdominal injury is rare. Most literatures on the subjects consist of case reports. We reported a rare case of isolated gallbladder rupture and discussed the possible predisposing factors to ...

  10. Experiments on seismic isolation in Italy

    International Nuclear Information System (INIS)

    Bonacina, G.; Bettinali, F.; Martelli, A.; Olivieri, M.

    1992-01-01

    Static and dynamic tests have been performed in Italy on high damping steel-laminated elastomer bearings in various scales, rubber specimens and structures isolated by means of such bearings, in the framework of studies in progress to support seismic isolation development. Tests on rubber specimens and bearings have already provided important data (vertical and horizontal stiffness, damping, creep, temperature, aging and scale effects, etc.), necessary for the development and validation of numerical models, comparison with the test results of isolated structure mockups and actual buildings, and improvement of design guidelines. Dynamic experiments of structures concerned both full-scale and scaled isolated structure mock-ups and actual isolated buildings (one of those forming the SIP Administration Center at Ancona, an isolated house at Squillace, Calabria). Both snap-back tests and forced excitation experiments were performed, to rather large displacements. The latter were both sinusoidal and (on a 1/4 scale mock-up) seismic, with one- and multidirectional simultaneous excitations. Test results have already demonstrated the adequacy of seismic isolation and have provided data useful for the comparison with single bearing test results and validation of numerical models for the analysis of isolated structures. This paper reports the main features and results of tests performed or in progress. Further tests planned have been mentioned in the Status Report. Numerical analysis of measured data and guidelines development have been discussed in separate technical papers. (author)

  11. Antagonist potential of Trichoderma indigenous isolates for ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... the severity of insects and pathogens attacks especially .... After 30-days of incubation at 26°C, the isolation of Trichoderma ... Percentage inhibition (I) of colony growth of P. palmivora was ..... In fact, our success in isolating.

  12. K-Basin isolation barrier seal

    International Nuclear Information System (INIS)

    Ruff, E.S.

    1994-10-01

    This report documents various aspects of the design, analysis, procurement, and fabrication of the hydraulic seal on the isolation barriers to be installed in the 100-K Area spent nuclear fuel basin. The isolation barrier is used to keep water in the basin in the event of an earthquake

  13. Noncommunicating Isolated Enteric Duplication Cyst in the ...

    African Journals Online (AJOL)

    Noncommunicating isolated enteric duplications in the abdomen are an extremely rare variant of enteric duplications with their own blood supply. We report a case of a noncommunicating isolated ileal duplication in a 10-month-old boy. He was admitted because of severe abdominal distension and developed irritability ...

  14. Experimental Study of Vibration Isolation Characteristics of a Geometric Anti-Spring Isolator

    Directory of Open Access Journals (Sweden)

    Lixun Yan

    2017-07-01

    Full Text Available In order to realize low-frequency vibration isolation, a novel geometric anti-spring isolator consisting of several cantilever blade springs are developed in this paper. The optimal design parameters of the geometric anti-spring isolator for different nonlinear geometric parameters are theoretically obtained. The transmissibility characteristic of the geometric anti-spring isolator is investigated through mathematical simulation. A geometric anti-spring isolator with a nonlinear geometric parameter of 0.92 is designed and its vibration isolation performance and nonlinearity characteristic is experimentally studied. The experiment results show that the designed isolator has good low-frequency vibration isolation performance, of which the initial isolation frequency is less than 3.6 Hz when the load weight is 21 kg. The jump phenomena of the response of the isolator under linear frequency sweep excitation are observed, and this result demonstrates that the geometric anti-spring isolator has a complex nonlinearity characteristics with the increment of excitation amplitude. This research work provides a theoretical and experimental basis for the application of the nonlinear geometric anti-spring low-frequency passive vibration isolation technology in engineering practice.

  15. Study on three dimensional seismic isolation system

    International Nuclear Information System (INIS)

    Morishita, Masaki; Kitamura, Seiji

    2003-01-01

    Japan Nuclear Cycle Development Institute (JNC) and Japan Atomic Power Company (JAPC) launched joint research programs on structural design and three-dimensional seismic isolation technologies, as part of the supporting R and D activities for the feasibility studies on commercialized fast breeder reactor cycle systems. A research project by JAPC under the auspices of the Ministry of Economy, Trade, and Industry (METI) with technical support by JNC is included in this joint study. This report contains the results of the research on the three-dimensional seismic isolation technologies, and the results of this year's study are summarized in the following five aspects. (1) Study on Earthquake Condition for Developing 3-dimensional Base Isolation System. The case study S2 is one of the maximum ground motions, of which the records were investigated up to this time. But a few observed near the fault exceed the case study S2 in the long period domain, depending on the fault length and conditions. Generally it is appropriate that the response spectra ratio (vertical/horizontal) is 0.6. (2) Performance Requirement for 3-dimensional Base Isolation System and Devices. Although the integrity map of main equipment/piping dominate the design criteria for the 3-dimensional base isolation system, the combined integrity map is the same as those of FY 2000, which are under fv=1Hz and over hv=20%. (3) Developing Targets and Schedule for 3-dimensional Isolation Technology. The target items for 3-dimensional base isolation system were rearranged into a table, and developing items to be examined concerning the device were also adjusted. A development plan until FY 2009 was made from the viewpoint of realization and establishment of a design guideline on 3-dimensional base isolation system. (4) Study on 3-dimensional Entire Building Base Isolation System. Three ideas among six ideas that had been proposed in FY2001, i.e., '3-dimensional base isolation system incorporating hydraulic

  16. Isolation and characterization of phenol degrading yeast.

    Science.gov (United States)

    Patel, Riddhi; Rajkumar, Shalini

    2009-04-01

    A phenol degrading yeast isolate was identified and characterized from the soil sample collected from a landfill site, in Ahmedabad, India, by plating the soil dilutions on Sabouraud's Dextrose Agar. The microscopic studies and biochemical tests indicated the isolate to be Saccharomyces cerevisiae. The phenol degrading potential of the isolate was measured by inoculation of pure culture in the mineral medium containing various phenol concentrations ranging from 100 to 800 mg l(-1 )and monitoring phenol disappearance rate at regular intervals of time. Growth of the isolate in mineral medium with various phenol concentrations was monitored by measuring the turbidity (OD(600) nm). The results showed that the isolated yeast was tolerant to phenol up to 800 mg(-1). The phenol degradation ranged from 8.57 to 100% for the concentration of phenol from 800 mg l(-1 )to 200 mg l(-1), respectively. ((c) 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

  17. Isolation of Pasteurella multocida from broiler chickens

    Directory of Open Access Journals (Sweden)

    Sri Poernomo

    1996-06-01

    Full Text Available Pasteurella multocida, the etiological agent of fowl cholera, was isolated from five, 32 days oldbroilerchickens in the late of 1992. The chickens were from a farm located in Bogor area, raised in cages and each flock consisted of 1,550 broilers . Therewere 230 birds, aging from 28-31 days old, died with clinical signs of lameness and difficulty in breathing. Serological test of the isolate revealed serotype Aof Carter classification . To prove its virulences, the isolate was then inoculated into 3 mice subcutaneously. The mice died less then 24 hours postinoculation and P. multocida can be reisolated . The sensitivity test to antibiotics and sulfa preparations showed that the isolate was sensitive to ampicillin, doxycyclin, erythromycin, gentamycin, sulfamethoxazol-trimethoprim and baytril, but resistance to tetracyclin, kanamycin and oxytetracyclin. This is the first report of P. multocida isolation in broiler chickens in Indonesia, and it is intended to add information on bacterial diseases in poultry in Indonesia.

  18. Effect of isolation techniques on the characteristics of pigeon pea (Cajanus cajan) protein isolates.

    Science.gov (United States)

    Adenekan, Monilola K; Fadimu, Gbemisola J; Odunmbaku, Lukumon A; Oke, Emmanuel K

    2018-01-01

    In this study, the effect of different isolation techniques on the isolated proteins from pigeon pea was investigated. Water, methanol, ammonium sulfate, and acetone were used for the precipitation of proteins from pigeon pea. Proximate composition, and antinutritional and functional properties of the pigeon pea flour and the isolated proteins were measured. Data generated were statistically analyzed. The proximate composition of the water-extracted protein isolate was moisture 8.30%, protein 91.83%, fat 0.25%, ash 0.05%, and crude fiber 0.05%. The methanol-extracted protein isolate composition was moisture 7.87%, protein 91.83%, fat 0.17%, and ash 0.13%, while crude fiber and carbohydrates were not detected. The composition of the ammonium sulfate-extracted protein isolate was moisture 7.73%, protein 91.73%, fat 0.36, ash 0.13%, and crude fiber 0.67%. The acetone-extracted protein isolate composition was moisture 8.03%, protein 91.50%, ash 0.67%, and fat 0.30%, but crude fiber and carbohydrates were not detected. The isolate precipitated with ammonium sulfate displayed the highest foaming capacity (37.63%) and foaming stability (55.75%). Isolates precipitated with methanol and acetone had the highest water absorption capacity (160%). Pigeon pea protein isolates extracted with methanol and ammonium sulfate had the highest oil absorption capacity of 145%. Protein isolates recovered through acetone and methanol had the highest emulsifying capacity of 2.23% and emulsifying stability of 91.47%, respectively. The proximate composition of the recovered protein isolates were of high purity. This shows the efficiency of the extraction techniques. The isolates had desirable solubility index. All the isolation techniques brought significant impact on the characteristics of the isolated pigeon pea protein.

  19. Isolement et identification des levures.

    Directory of Open Access Journals (Sweden)

    S. DOMERCQ

    1967-06-01

    Full Text Available La fermentation du moût de raisin est assurée par les levures qui se trouvent sur la pellicule du raisin, sur les rafles, et aussi, par celles qui envahissent le matériel de vendange et de vinification dès que la récolte est commencée. L'apparition deLa fermentation du moût de raisin est assurée par les levures qui se trouvent sur la pellicule du raisin, sur les rafles, et aussi, par celles qui envahissent le matériel de vendange et de vinification dès que la récolte est commencée. L'apparition des signes de la fermentation est spontanée. La fermentation est rapide dans la majorité des cas, si elle devient trop violente et que la température s'élève dangeureusement, il faut la « canaliser », la calmer. Des accidents peuvent se produire; des arrêts de fermentation laissant des sucres non fermentés pouvant être attaqués par d'autres microorganismes, des développements de levures formant des substances nuisibles à la qualité; par la suite, il s'agit d'obtenir la stabilité du vin, en particulier, éviter les refermentations des vins doux. L'étude de ces problèmes nécessite une étude préliminaire, celle des levures responsables de la fermentation alcoolique du jus de raisin, qui ne peut être faite que sur des cultures pures, isolées au préalable à l'aide de prélèvements soigneusement réalisés.s signes de la fermentation est spontanée. La fermentation est rapide dans la majorité des cas, si elle devient trop violente et que la température s'élève dangeureusement, il faut la « canaliser », la calmer. Des accidents peuvent se produire; des arrêts de fermentation laissant des sucres non fermentés pouvant être attaqués par d'autres microorganismes, des développements de levures formant des substances nuisibles à la qualité; par la suite, il s'agit d'obtenir la stabilité du vin, en particulier, éviter les refermentations des vins doux. L'étude de ces problèmes nécessite une étude pr

  20. Are Isolated Indigenous Populations Headed toward Extinction?

    Directory of Open Access Journals (Sweden)

    Robert S Walker

    Full Text Available At least 50 indigenous groups spread across lowland South America remain isolated and have only intermittent and mostly hostile interactions with the outside world. Except in emergency situations, the current policy of governments in Brazil, Colombia, and Peru towards isolated tribes is a "leave them alone" strategy, in which isolated groups are left uncontacted. However, these no-contact policies are based on the assumption that isolated populations are healthy and capable of persisting in the face of mounting external threats, and that they can maintain population viability in the long-term. Here, we test this assumption by tracking the sizes and movements of cleared horticultural areas made by 8 isolated groups over the last 10-14 years. We used deforestation data derived from remote sensing Landsat satellite sensors to identify clearings, and those were then validated and assessed with high-resolution imagery. We found only a single example of a relatively large and growing population (c. 50 cleared ha and 400 people, whereas all of the other 7 groups exhibited much smaller villages and gardens with no sizable growth through time. These results indicated that the smaller groups are critically endangered, and it prompts an urgent re-thinking of policies toward isolated populations, including plans for well-organized contacts that may help save lives and rescue isolated indigenous populations from imminent extinction.

  1. Are Isolated Indigenous Populations Headed toward Extinction?

    Science.gov (United States)

    Walker, Robert S; Kesler, Dylan C; Hill, Kim R

    2016-01-01

    At least 50 indigenous groups spread across lowland South America remain isolated and have only intermittent and mostly hostile interactions with the outside world. Except in emergency situations, the current policy of governments in Brazil, Colombia, and Peru towards isolated tribes is a "leave them alone" strategy, in which isolated groups are left uncontacted. However, these no-contact policies are based on the assumption that isolated populations are healthy and capable of persisting in the face of mounting external threats, and that they can maintain population viability in the long-term. Here, we test this assumption by tracking the sizes and movements of cleared horticultural areas made by 8 isolated groups over the last 10-14 years. We used deforestation data derived from remote sensing Landsat satellite sensors to identify clearings, and those were then validated and assessed with high-resolution imagery. We found only a single example of a relatively large and growing population (c. 50 cleared ha and 400 people), whereas all of the other 7 groups exhibited much smaller villages and gardens with no sizable growth through time. These results indicated that the smaller groups are critically endangered, and it prompts an urgent re-thinking of policies toward isolated populations, including plans for well-organized contacts that may help save lives and rescue isolated indigenous populations from imminent extinction.

  2. Isolated microalbuminuria indicates a poor medical prognosis.

    Science.gov (United States)

    Scheven, Lieneke; Van der Velde, Marije; Lambers Heerspink, Hiddo J; De Jong, Paul E; Gansevoort, Ron T

    2013-07-01

    Microalbuminuria is often regarded as a sign of end-organ damage due to diabetes and/or hypertension, and as such to be associated with an increased risk for cardiovascular events. It has been questioned whether isolated microalbuminuria, that is microalbuminuria in the absence of a cardiovascular disease (CVD) history, hypertension and diabetes has clinical relevance. Included were 8356 subjects who participated in the first four screening rounds of the PREVEND study, a prospective, community-based, observational cohort study. Isolated microalbuminuria was defined as microalbuminuria (30-300 mg/24 h), in the absence of a CVD history, hypertension (blood pressuredefinition of isolated microalbuminuria, in which 2250 person-years of follow-up were available. In subjects with isolated microalbuminuria, the incidence rates of cardiovascular events and mortality, hypertension and diabetes were 15.3, 28.9 and 8.9 per 1000 person-year follow-up, respectively. Subjects with isolated microalbuminuria had an increased risk for cardiovascular events and mortality [crude HR 2.23 (1.63-3.07); Phypertension [OR 1.95 (1.47-2.59); Phypertension and/or diabetes. This increased risk remained significant after adjustment for age and gender. The relative risk held by isolated microalbuminuria was similar to the relative risk held by microalbuminuria in subjects that did have a CVD history, hypertension and/or diabetes. Isolated microalbuminuria indicates a poor prognosis and warrants medical attention.

  3. Different Babesia canis isolates, different diseases.

    Science.gov (United States)

    Schetters, T P; Moubri, K; Précigout, E; Kleuskens, J; Scholtes, N C; Gorenflot, A

    1997-11-01

    Using surface immunofluorescence isolate-specific antigens were detected on the membrane of erythrocytes infected with Babesia parasites. In addition, the strains reacted differently with Plasmagel in that the European isolate (B.c. canis) could be purified on Plasmagel effectively, whereas infected erythrocytes of the South-African isolate (B.c. rossi) could not. Experimental infection of dogs with Babesia canis isolates from geographically different areas revealed different pathology. The European isolate obtained from France exhibited transient parasitaemia, usually below 1%, associated with low PCV values and congestion of internal organs. Clinical disease was correlated with an effect on the coagulation system, and not with peripheral parasitaemia. Infection of dogs with South-African-derived isolate induced high parasitaemia usually much higher than 1%, which required chemotherapeutic treatment. In these animals clinical disease was correlated with peripheral parasitaemia and not with parameters of the coagulation system. The results show that the etiology of disease caused by these isolates of B.c. canis and B.c. rossi is different. This might have implications for the development of vaccines against these infections.

  4. Preventing social isolation in older people.

    Science.gov (United States)

    Cotterell, Natalie; Buffel, Tine; Phillipson, Christopher

    2018-07-01

    The extent of social isolation amongst older people has emerged as a major concern for health and social policy. Although the social and health outcomes of social isolation are well documented, evidence regarding the prevention of isolation in later life remains scarce. This article addresses this by presenting the findings from a literature review focusing on the identification, assessment, prevention, and intervention strategies relevant to social isolation in older age. The paper first addresses the issues of identification and assessment, using an ecological framework to identify the risk factors for social isolation at four levels: individual, relationship, community, and societal. It then reviews different types of interventions to reduce or prevent social isolation in later life, including one-to-one, group, service provision, technology-based, neighbourhood, and structural interventions. The paper discusses both the opportunities and the constraints associated with these different approaches. The discussion highlights future directions for research, emphasising the need for a cultural change from 'cure' to 'prevention' of social isolation across the life-course, and the importance of acknowledging greater diversity within the ageing population. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. EMPLACEMENT DRIFT ISOLATION DOOR CONTROL SYSTEM

    International Nuclear Information System (INIS)

    N.T. Raczka

    1998-01-01

    The purpose of this analysis is to review and refine key design concepts related to the control system presently under consideration for remotely operating the emplacement drift isolation doors at the potential subsurface nuclear waste repository at Yucca Mountain. This analysis will discuss the key design concepts of the control system that may be utilized for remotely monitoring, opening, and closing the emplacement drift isolation doors. The scope and primary objectives of this analysis are to: (1) Discuss the purpose and function of the isolation doors (Presented in Section 7.1). (2) Review the construction of the isolation door and other physical characteristics of the doors that the control system will interface with (Presented in Section 7.2). (3) Discuss monitoring and controlling the operation of the isolation doors with a digital control system (either a Programmable Logic Controller (PLC) system or a Distributed Control System (DCS)) (Presented in Section 7.3). (4) Discuss how all isolation doors can be monitored and controlled from a subsurface central control center (Presented in Section 7.4). This analysis will focus on the development of input/output (I/O) counts including the types of I/O, redundancy and fault tolerance considerations, and processor requirements for the isolation door control system. Attention will be placed on operability, maintainability, and reliability issues for the system operating in the subsurface environment with exposure to high temperatures and radiation

  6. Cryopreservation of Mycobacterium bovis isolates

    Directory of Open Access Journals (Sweden)

    Cássia Yumi Ikuta

    2016-11-01

    Full Text Available Research, development of new biotechnological methods, diagnostic tests, confirmation of results, and reinvestigations are possible because of the availability of well-preserved living organisms maintained without any changes. Cryopreservation is a simpler, more reliable and long-term stable method for culture maintenance. Storage temperature and composition of the suspending vehicle are factors that affect the viability of mycobacterial strains. Three vehicles and three storage temperatures were evaluated to define a suitable cryoprotective medium for the preservation of Mycobacterium bovis strains. Colonies of sixteen M. bovis isolates were used to prepare the suspensions, which were then added to three vehicles: sterile 0.85% saline solution (SS, Middlebrook 7H9 broth (7H9, and Middlebrook 7H9 broth with sodium pyruvate (7H9p replacing glycerol. Aliquots of these suspensions were frozen by three different methods, directly in the -20°C freezer, directly in the -80°C freezer, and at -196°C by immersion in liquid nitrogen (LN. The frozen aliquots were thawed at room temperature after 45, 90 and 120 days. Mycobacterial viability was assessed by counting the living cells on plates of Stonebrink medium before and after the freezing procedure. Storage at -20°C exhibited a lower recovery of M. bovis compared to storage at -80°C (Dunn’s test, p=0.0018 and LN (Dunn’s test, p=0.0352. There was no statistically significant difference between storage at -80°C and in LN (Dunn’s test, p=0.1403, yet -80°C showed better results than LN. All three suspending vehicles showed no statistically significant difference in terms of viability (Friedman’s test, p=0.7765. Given the low loss proportion of 5% during storage at -20°C and the high cost equipment required for storage at -80°C and LN, we recommend storage at -20°C or -80°C, when this is available, for preservation of M. bovis field strains.

  7. Genetic differences between avian and human isolates of Candida dubliniensis.

    LENUS (Irish Health Repository)

    McManus, Brenda A

    2009-09-01

    When Candida dubliniensis isolates obtained from seabird excrement and from humans in Ireland were compared by using multilocus sequence typing, 13 of 14 avian isolates were genetically distinct from human isolates. The remaining avian isolate was indistinguishable from a human isolate, suggesting that transmission may occur between humans and birds.

  8. Effect of Trichoderma isolates on tomato seedling growth response ...

    African Journals Online (AJOL)

    Trichoderma species are commonly used as biological control agents against phytopathogenic fungi and some isolates are able to improve plant growth. In this study, the effects of three Trichoderma isolates including Trichoderma harzianum isolate T969, T. harzianum isolate T447 and Trichoderma sp. isolate T in tomato ...

  9. [Isolation of C. pyloridis from children].

    Science.gov (United States)

    Spirina, T S; Bokser, G V; Safonova, T B; Taranenko, L A; Levitskaia, S V

    1989-01-01

    The authors analyze the findings of gastroduodenal biopsies in 25 children aged 6 to 15 years suffering from mucosal inflammations and functional abnormalities of the upper portions of the digestive tract. Chocolate agar with various bases, one of which has been suggested by the authors, has been used for the isolation of C. pyloridis. Five cultures have been isolated. Microscopic examinations and biochemical identification of the isolates have lead the authors to a conclusion that campylobacter infections of the upper digestive tract may be diagnosed with certainty only after a comprehensive bacteriologic examination.

  10. Fault Detection and Isolation for Spacecraft

    DEFF Research Database (Denmark)

    Jensen, Hans-Christian Becker; Wisniewski, Rafal

    2002-01-01

    This article realizes nonlinear Fault Detection and Isolation for actuators, given there is no measurement of the states in the actuators. The Fault Detection and Isolation of the actuators is instead based on angular velocity measurement of the spacecraft and knowledge about the dynamics...... of the satellite. The algorithms presented in this paper are based on a geometric approach to achieve nonlinear Fault Detection and Isolation. The proposed algorithms are tested in a simulation study and the pros and cons of the algorithms are discussed....

  11. Improved containment isolation for CANDU plants

    Energy Technology Data Exchange (ETDEWEB)

    Stretch, A H [Atomic Energy of Canada Ltd., Mississauga, ON (Canada)

    1996-12-31

    The publication of Regulatory Policy Statement R- 7 in February 1991 by the Atomic Energy Control Board imposes new requirement for the isolation of fluid piping penetrating the containment boundary. The Appendix of R-7 describes the detailed requirements for metal extensions of the containment envelope, including the code classification qf the pressure retaining portions and isolation requirements for fluid piping and tubing.The application of these new requirements to the existing CANDU 6 design resulted in a number of design changes, including the addition of manual and automatic isolation valves, changes in code classification, and leakage monitoring considerations. (author). 3 refs., 3 figs.

  12. Isolators Including Main Spring Linear Guide Systems

    Science.gov (United States)

    Goold, Ryan (Inventor); Buchele, Paul (Inventor); Hindle, Timothy (Inventor); Ruebsamen, Dale Thomas (Inventor)

    2017-01-01

    Embodiments of isolators, such as three parameter isolators, including a main spring linear guide system are provided. In one embodiment, the isolator includes first and second opposing end portions, a main spring mechanically coupled between the first and second end portions, and a linear guide system extending from the first end portion, across the main spring, and toward the second end portion. The linear guide system expands and contracts in conjunction with deflection of the main spring along the working axis, while restricting displacement and rotation of the main spring along first and second axes orthogonal to the working axis.

  13. STREPTOMYCETE Producing Antibiotics Isolated From Egyptian Soil

    International Nuclear Information System (INIS)

    SALAMA, S.M; MAHMOUD, S.M; ELKABBANY, H.M.

    2010-01-01

    An investigation was concerned the distribution of Streptomyces species in some types of Egyptian soils. Thirty nine actinomycetes isolates were obtained from 7 types of Egyptian soils; 8 isolates exhibited antimicrobial activities against all tested organisms and three of them were identified as Streptomyces griseoluteus (YM23), Streptomyces aurantiogriseus (S15) and Streptomyces nogalator (H12). RAPD-PCR showed correlation between 8 isolates in 3 clades. The active metabolite was extracted with ethyl acetate and concentrated in vacuum and the crude fraction was purified using thin layer and column chromatography

  14. Earthquake response observation of isolated buildings

    International Nuclear Information System (INIS)

    Harada, O.; Kawai, N.; Ishii, T.; Sawada, Y.; Shiojiri, H.; Mazda, T.

    1989-01-01

    Base isolation system is expected to be a technology for a rational design of FBR plant. In order to apply this system to important structures, accumulation of verification data is necessary. From this point of view, the vibration test and the earthquake response observation of the actual isolated building using laminated rubber bearings and elasto-plastic steel dampers were conducted for the purpose of investigating its dynamic behavior and of proving the reliability of the base isolation system. Since September in 1986, more than thirty earthquakes have been observed. This paper presents the results of the earthquake response observation

  15. Isolated jejunal perforation following blunt abdominal trauma

    Directory of Open Access Journals (Sweden)

    Ahmet Pergel

    2012-09-01

    Full Text Available Isolated perforation of the jejunum, following blunt abdominaltrauma, is extremely rare. These injuries aredifficult to diagnose because initial clinical signs are frequentlynonspecific and a delay in treatment increasesmortality and morbidity of the patients. Conventional radiogramsare often inadequate for diagnosing this subsetof trauma. For an accurate and timely diagnosis, thepossibility of bowel perforation and the need for repeatedexaminations should be kept in mind. Herein, we presenta 28-year-old man with isolated jejunal perforation followingblunt abdominal trauma.Key words: Blunt abdominal trauma, isolated jejunal perforation,early diagnosis

  16. The heterogeneity of socially isolated older adults: a social isolation typology.

    Science.gov (United States)

    Machielse, Anja

    2015-01-01

    Recent statistics show a growing number of older adults who are living alone and are socially isolated. It is against this background that, in recent years, many interventions have been developed to address social isolation among the elderly. Evaluative studies show that most interventions are hardly effective, though. An important reason for this is the heterogeneity of the socially isolated. This article offers insight into this heterogeneity by presenting a typology with different profiles of socially isolated older adults and the intervention implications of this typology. The typology is derived from an extensive qualitative study on socially isolated elderly individuals in the Netherlands. The typology imposes some degree of order to a diversity of circumstances, ambitions, and possibilities of the socially isolated elderly, thereby deepening the understanding of the heterogeneity of this population. The definition of social isolation used in this study starts from a societal angle of incidence, namely the current policy context of Western European welfare states, in which governments emphasize the importance of independence and self-reliance of their citizens. Developed from that perspective, the typology provides a theoretical basis for applying interventions aimed at increasing self-reliance of social isolated elderly. This perspective on social isolation also has consequences for the way in which the effectiveness of interventions to alleviate social isolation is assessed.

  17. Optimization of yeast (Saccharomyces cerevisiae) RNA isolation ...

    African Journals Online (AJOL)

    Yomi

    2012-01-16

    Jan 16, 2012 ... isolation method for real-time quantitative PCR and microarray ... disease genes to experimental evolution and systems biology (Landry et al., .... scanning, and preliminary analyses with GeneChip Operating. Software 1.4 ...

  18. Antigenic typing Polish isolates of canine parvovirus

    Energy Technology Data Exchange (ETDEWEB)

    Mizak, B. [National Veterinary Research Institute, Pulawy (Poland); Plucienniczak, A. [Polish Academy ofd Sciences. Microbiology and Virology Center, Lodz (Poland)

    1995-12-31

    Polish strains of canine parvovirus isolated between 1982 and 1993 were examined to determine the extent to which the virus has evolved antigenically and genetically over eleven years. Two CPV isolates obtained in Warsaw in 1982 and Pulawy in 1993, were examined using monoclonal antibody typing, restriction analysis and sequencing VP-2 protein gene. Five other isolates from Warsaw and Pulawy were tested with the panel of monoclonal antibodies specific to CPV-2, CPV-2a and common for canine parvovirus, feline panleukopenia virus and milk enteritis virus. Results of the studies demonstrated that all isolates tested represented CPV-2a antigenic type. Rapid antigenic strain replacement recorded by Parrish and Senda in the U.S.A and Japan was not confirmed in Poland. (author). 30 refs, 2 tabs.

  19. ISOLATION AND IDENTIFICATION OF FUNGI RESPONSIBLE FOR ...

    African Journals Online (AJOL)

    User

    technique. The diseased plant leaves were taken to the laboratory for culture, isolation, and identification ... The tree is native to Asia particularly eastern India, Burma, and the Andaman ..... in Southeastern Nigeria and Biological. Control with ...

  20. Isolation and molecular characterisation of Mycobacterium bovis ...

    African Journals Online (AJOL)

    : Three hundred and six milk samples collected from 102 infected cows in different Tunisian regions were analysed. M. bovis isolates were further characterized by spoligotyping and variable number tandem repeat typing. Results: A total of ...

  1. A case of familial isolated hemihyperplasia

    Directory of Open Access Journals (Sweden)

    Bacino Carlos A

    2004-02-01

    Full Text Available Abstract Background Hemihyperplasia (hemihypertrophy is defined as asymmetric body overgrowth of one or more body parts. Hemihyperplasia can be isolated or be part of well-defined syndromes such as in the case of Beckwith-Wiedemann syndrome (BWS. Isolated hemihyperplasia is usually sporadic, but a number of familial occurrences have been described. Case presentation We describe a Tunisian family in which three maternal cousins and their maternal grandfather present with isolated hemihyperplasia. Conclusions The etiology of isolated hemihyperplasia is unknown although in BWS, genomic imprinting has been shown to play a role in the asymmetric overgrowth. Given the similarity between these two conditions, it is possible that both may share a common pathogenesis. We also discuss the possible genetic mechanisms leading to the production of hemihyperplasia in this family.

  2. Isolation and characterization of biogenic calcium carbonate ...

    Indian Academy of Sciences (India)

    Biogenic calcium carbonate/phosphate were isolated and characterized from oral bacteria (CPOB). The crystalline nature ... XRD analysis revealed the cubic phase of ... subjected to identify upto genus level according to Bergey's. Manual of ...

  3. Isolate resistance of Blastocystis hominis to metronidazole.

    Science.gov (United States)

    Haresh, K; Suresh, K; Khairul Anus, A; Saminathan, S

    1999-04-01

    Isolates of Blastocystis hominis from infected immigrant workers from Indonesia, Bangladesh and infected individuals from Singapore and Malaysia were assessed for growth pattern and degree of resistance to different concentrations of metronidazole. Viability of the cells was assessed using eosin-brillian cresyl blue which stained viable cells green and nonviable cells red. The Bangladeshi and Singaporean isolates were nonviable even at the lowest concentration of 0.01 mg/ml, whereas 40% of the initial inoculum of parasites from the Indonesian isolate at day one were still viable in cultures with 1.0 mg/ml metronidazole. The study shows that isolates of B. hominis of different geographical origin have different levels of resistance to metronidazole. The search for more effective drugs to eliminate th parasite appears inevitable, especially since surviving parasites from metronidazole cultures show greater ability to multiply in subcultures than controls.

  4. Occurrence, isolation and DNA identification of Streptococcus ...

    African Journals Online (AJOL)

    ajl yemi

    2011-11-28

    Nov 28, 2011 ... Streptococcus thermophilus involved in Algerian ... among reference, and wild strains of S. thermophilus and for their differentiation from Enterococcus spp. ..... Isolation and characterization of Lactobacillus delbrueckii ssp.

  5. Connecting Vietnam's isolated communities to improve healthcare ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2018-01-29

    Jan 29, 2018 ... Connecting Vietnam's isolated communities to improve healthcare ... of pregnancy and new motherhood improved their interactions with health workers. ... Return to main page: Overcoming eHealth challenges with social and ...

  6. AEROBIC BACTERIAL ISOLATES FROM INFECTED WOUNDS

    African Journals Online (AJOL)

    boaz

    most frequently reported as the cause of delay wound healing (6-9, 3). ... All isolates were resistant to Ampicillin, Amoxicillin- clavulanate and .... Ulcer bed infection. Report of a case of enlarging venous leg ulcer colonized by ... Ann. Burns Fire.

  7. Isolated torsion of fallopian tube. Radiological findings

    International Nuclear Information System (INIS)

    Tomas Fanjul, L.; Aldea Martinez, J.; Fernandez Matia, G.; Rodrigo Verguizas, J.; Fernandez Alvarez, G.; Galindo Vicente, M.C.

    1993-01-01

    Isolated tubal torsion is a very uncommon disorder that is rarely diagnosed preoperatively. We present a case and review the literature, which only provides ultrasonographic findings in 5 cases reported to date. 11 refs

  8. Genetics Home Reference: isolated sulfite oxidase deficiency

    Science.gov (United States)

    ... and Management Resources (1 link) GeneReview: Isolated Sulfite Oxidase Deficiency General Information from MedlinePlus (5 links) Diagnostic Tests Drug Therapy Genetic Counseling Palliative Care Surgery and ...

  9. Antigenic typing Polish isolates of canine parvovirus

    International Nuclear Information System (INIS)

    Mizak, B.; Plucienniczak, A.

    1995-01-01

    Polish strains of canine parvovirus isolated between 1982 and 1993 were examined to determine the extent to which the virus has evolved antigenically and genetically over eleven years. Two CPV isolates obtained in Warsaw in 1982 and Pulawy in 1993, were examined using monoclonal antibody typing, restriction analysis and sequencing VP-2 protein gene. Five other isolates from Warsaw and Pulawy were tested with the panel of monoclonal antibodies specific to CPV-2, CPV-2a and common for canine parvovirus, feline panleukopenia virus and milk enteritis virus. Results of the studies demonstrated that all isolates tested represented CPV-2a antigenic type. Rapid antigenic strain replacement recorded by Parrish and Senda in the U.S.A and Japan was not confirmed in Poland. (author). 30 refs, 2 tabs

  10. Isolation of entomopathogenic nematodes and control of ...

    African Journals Online (AJOL)

    Jaime Ruiz

    when a new NE is isolated from field samples, laboratory assessments of ..... Tanks to Amanda Hodson, University of California at. Davis, for carrying out ... Manual of techniques in insect pathology. Academic Press, New York. Koppenhöfer A ...

  11. Isolation and characterization of a bacterial celluloseproducing ...

    African Journals Online (AJOL)

    year ripened persimmon vinegar, which belonged to the family of Acetobacteracea based on its morphological and physiological characteristics. The phylogenetic position of the isolated strain was most closely related to Gluconacetobacter ...

  12. Triterpene alcohol isolation from oil shale.

    Science.gov (United States)

    Albrecht, P; Ourisson, G

    1969-03-14

    Isoarborinol, an intact pentacyclic unsaturated alcohol, was isolated from the Messel oil shale (about 50 x 106 years old). Complex organic substances, even those very sensitive to oxidation, reduction, or acidic conditions, can thus survive without alteration for long periods.

  13. Cerveau isolé and pretrigeminal rats.

    Science.gov (United States)

    Zernicki, B; Gandolfo, G; Glin, L; Gottesmann, C

    1984-01-01

    Cortical and hippocampal EEG activity was analysed in 14 cerveau isole and 8 pretrigerninal rats. In the acute stage, waking EEG patterns were absent in the cerveau isole, whereas sleep EEG patterns were absent in the pretrigeminal preparations. However, already on the second day the EEG waking-sleep cycle recovered in the majority of rats. Paradoxically, stimuli directed to the caudal part of preparations evoked stronger cortical and hippocampal EEG arousal than olfactory and visual stimuli. The behavior of the caudal part was observed in 25 preparations. Although in abortive form, the rats did show some locomotor and grooming behavior, and could be fed orally. The peripheral events of paradoxical sleep appeared only on the fourth or fifth day of survival of the cerveau isole rats. It is concluded that the activity of the isolated cerebrum of the rat is similar to that of cat preparations, but that functions of the caudal neuraxis are superior in rats.

  14. Isolated neck-lifting procedure: isolated stork lift.

    Science.gov (United States)

    Barbarino, Sheila C; Wu, Allan Y; Morrow, David M

    2013-04-01

    Many patients desire cosmetic improvement of neck laxity when consulting with a plastic surgeon about their face. Neck laxity and loss of the cervicomental angle can be due to multiple components of aging such as skin quality/elasticity, loss of platysma muscle tone, and submental fat accumulation. Traditionally, the procedure of choice for patients with an aging lower face and neck is a cervicofacial rhytidectomy. However, occasionally, a patient wishes to have no other facial surgery than an improvement of their excessive skin of the anterior, lateral, and/or posterior neck. In other instances, a patient may present with having had a face/neck-lifting procedure that left objectionable vertical/diagonal lines at the lateral neck. In both these instances, a surgeon should consider an isolated stork lift (ISL) procedure. An ISL procedure avoids and/or corrects problematic vertical/diagonal lateral neck folds by "walking" the excess skin flaps around the posterior inferior occipital hairline bilaterally, bringing the flaps together at the lateral and posterior neck, which sometimes involves a midline posterior dart excision of the dog ear. A patient presenting with excessive skin of the neck (anterior, lateral, and/or posterior) and/or residual vertical/diagonal skin folds is an excellent candidate for the ISL. The ISL procedure was performed on 273 patients over a 2-year period at The Morrow Institute. Patients were included if they had excessive skin of the anterior, lateral, and/or posterior neck and/or diagonal/vertical lateral bands and did not desire a full face-lifting procedure. Patients were excluded from this study if they would not accept having longer hair in order to cover the scar along the posterior inferior occipital hairline or a midline T-flap skin closure scar at the base of the posterior midline neck. Under a combination of local anesthesia and IV sedation, a postauricular face-lift incision was made that was extended in a circumoccipital fashion

  15. Using Euler buckling springs for vibration isolation

    CERN Document Server

    Winterflood, J; Blair, D G

    2002-01-01

    Difficulties in obtaining ideal vertical vibration isolation with mechanical springs are identified as being due to the mass of the elastic element which is in turn due to its energy storage requirement. A new technique to minimize this energy is presented - being an Euler column undergoing elastic buckling. The design of a high performance vertical vibration isolation stage based on this technique is presented together with its measured performance.

  16. Using Euler buckling springs for vibration isolation

    International Nuclear Information System (INIS)

    Winterflood, J; Barber, T; Blair, D G

    2002-01-01

    Difficulties in obtaining ideal vertical vibration isolation with mechanical springs are identified as being due to the mass of the elastic element which is in turn due to its energy storage requirement. A new technique to minimize this energy is presented - being an Euler column undergoing elastic buckling. The design of a high performance vertical vibration isolation stage based on this technique is presented together with its measured performance

  17. Base isolation strategies for structures and components

    International Nuclear Information System (INIS)

    Varma, Veto; Reddy, G.R.; Vaze, K.K.; Kushwaha, H.S.

    2003-08-01

    In the present report the effect of laminated rubber bearing (LRB) system on the dynamic response of the structure was studied. A LRB system was designed and tested in the laboratory for its dynamic characteristics. Finite element analysis was also performed and based on this analysis, isolator for PHWR nuclear power plant was designed. Analysis of the building was performed with and without isolator. Comparison of responses was made in terms of frequencies, accelerations and displacements and floor response spectra. (author)

  18. Isolated rib fractures in geriatric patients

    OpenAIRE

    Elmistekawy, Elsayed M.; Hammad, Abd Almohsen M.

    2007-01-01

    Introduction: The goal of this study was to investigate the short-term outcomes in patients older than 60 years with isolated rib fractures and admitted to emergency hospital. Materials and Methods: This study included patients who were 60 years old or more and sustained blunt chest injury and had isolated rib fractures. The following data were obtained from the medical records: age, gender, number of fracture ribs, side of fracture ribs, mechanism and nature of injury, preexisting medical...

  19. Fault Detection and Isolation using Eigenstructure Assignment

    DEFF Research Database (Denmark)

    Jørgensen, R. B.; Patton, R.; Chen, J.

    1994-01-01

    The purpose of this article is to investigate the robustness to model uncertainties of observer based fault detection and isolation. The approach is designed with a straight forward dynamic nad the observer.......The purpose of this article is to investigate the robustness to model uncertainties of observer based fault detection and isolation. The approach is designed with a straight forward dynamic nad the observer....

  20. Isolation of avian influenza virus in Texas.

    Science.gov (United States)

    Glass, S E; Naqi, S A; Grumbles, L C

    1981-01-01

    An avian influenza virus with surface antigens similar to those of fowl plague virus (Hav 1 Nav 2) was isolated in 1979 from 2 commercial turkey flocks in Central Texas. Two flocks in contact with these infected flocks developed clinical signs, gross lesions, and seroconversion but yielded no virus. This was the first recorded incidence of clinical avian influenza in Texas turkeys and only the second time that an agent with these surface antigens was isolated from turkeys in U.S.

  1. Criteria: waste tank isolation and stabilization

    International Nuclear Information System (INIS)

    Metz, W.P.; Ogren, W.E.

    1976-09-01

    The crystallized Hanford high-level wastes stored in single-shell underground tanks consist of sludges and salt cakes covered with supernatural liquor. Purpose of stabilization and isolation is to reduce the releases and losses as a result of a loss of tank integrity. The tanks will be modified so that no inadvertent liquid additions can be made. Criteria for the isolation and stabilization are given and discussed briefly

  2. New approach for isolation of VNTR markers.

    OpenAIRE

    Nakamura, Y; Carlson, M; Krapcho, K; Kanamori, M; White, R

    1988-01-01

    Elsewhere we have reported an efficient method for isolating VNTR (Variable Number of Tandem Repeats) markers. Several of the VNTR markers isolated in those experiments were sequenced, and a DNA sequence of 9 bp (GNNGTGGG) emerged as an apparent consensus sequence for VNTR markers. To confirm this result and to develop more VNTR markers, we synthesized nine different 18-base-long oligonucleotides whose sequences each included GNNGTGGG. When 102 cosmid clones selected by these oligonucleotides...

  3. Probiotic features of two oral Lactobacillus isolates

    OpenAIRE

    Zavisic, Gordana; Petricevic, Sasa; Radulovic, Zeljka; Begovic, Jelena; Golic, Natasa; Topisirovic, Ljubisa; Strahinic, Ivana

    2012-01-01

    In this study, we checked lactobacilli strains of human origin for their potential as probiotic. Samples were collected from oral mucosa of 16 healthy individuals, out of which twenty isolates were obtained and two of them were selected and identified as Lactobacillus plantarum (G1) and L. casei (G3). Both isolates exhibited antagonistic action towards pathogenic microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella abony, and Clostridium sporogene...

  4. Criteria: waste tank isolation and stabilization

    Energy Technology Data Exchange (ETDEWEB)

    Metz, W.P.; Ogren, W.E.

    1976-09-01

    The crystallized Hanford high-level wastes stored in single-shell underground tanks consist of sludges and salt cakes covered with supernatural liquor. Purpose of stabilization and isolation is to reduce the releases and losses as a result of a loss of tank integrity. The tanks will be modified so that no inadvertent liquid additions can be made. Criteria for the isolation and stabilization are given and discussed briefly. (DLC)

  5. The supply solutions for isolated rural consumers

    International Nuclear Information System (INIS)

    Hazi, Gheorghe; Solomon, Petre; Hazi, Aneta

    2004-01-01

    This paper establishes the supply optimal solutions for isolated rural consumers. A complex technical-economical calculation method is developed for selection of the best solutions. This analysis is based on the minimization of the net present value, NPV, criterion. Using the results of this calculation, one can select easily the supply solution for a given active power and for a given distance separating the power source and the isolated consumer. (authors)

  6. Protein import into isolated pea root leucoplasts

    OpenAIRE

    Chu, Chiung-Chih; Li, Hsou-min

    2015-01-01

    Leucoplasts are important organelles for the synthesis and storage of starch, lipids and proteins. However, molecular mechanism of protein import into leucoplasts and how it differs from that of import into chloroplasts remain unknown. We used pea seedlings for both chloroplast and leucoplast isolations to compare within the same species. We further optimized the isolation and import conditions to improve import efficiency and to permit a quantitative comparison between the two plastid types....

  7. Genotoxicological Evaluation of NUTRALYS Pea Protein Isolate

    OpenAIRE

    Aouatif, Chentouf; Looten, Ph.; Parvathi, M. V. S.; Raja Ganesh, S.; Paranthaman, V.

    2013-01-01

    NUTRALYS Pea Protein Isolate, a protein supplement, is a high-quality source of protein which is primarily emulsifying functional protein. We evaluated the genotoxic potential of NUTRALYS isolated from dry yellow pea, using three established genotoxicity tests (AMES test in vitro chromosomal aberration test, and in vivo micronucleus test) employing OECD guidelines under GLP conditions. In the bacterial reverse mutation test, NUTRALYS did not show positive responses in strains detecting point ...

  8. Corynebacterium species isolated from patients with mastitis.

    Science.gov (United States)

    Paviour, Sue; Musaad, Sahar; Roberts, Sally; Taylor, Graeme; Taylor, Susan; Shore, Keith; Lang, Selwyn; Holland, David

    2002-12-01

    Corynebacteria were isolated from breast tissue, pus, or deep wound swabs of 24 women; the most common species isolated was the newly described Corynebacterium kroppenstedtii, followed by Corynebacterium amycolatum and Corynebacterium tuberculostearicum. Gram-positive bacilli were seen in samples sent for culture or in histological specimens for 12 women, and 9 of the 12 women from whom adequate histological specimens were obtained had conditions that met the criteria for granulomatous lobular mastitis, a chronic inflammatory disease of unknown etiology.

  9. Geographically isolated wetlands: Rethinking a misnomer

    Science.gov (United States)

    Mushet, David M.; Calhoun, Aram J.K.; Alexander, Laurie C.; Cohen, Matthew J.; DeKeyser, Edward S.; Fowler, Laurie G.; Lane, Charles R.; Lang, Megan W.; Rains, Mark C.; Walls, Susan

    2015-01-01

    We explore the category “geographically isolated wetlands” (GIWs; i.e., wetlands completely surrounded by uplands at the local scale) as used in the wetland sciences. As currently used, the GIW category (1) hampers scientific efforts by obscuring important hydrological and ecological differences among multiple wetland functional types, (2) aggregates wetlands in a manner not reflective of regulatory and management information needs, (3) implies wetlands so described are in some way “isolated,” an often incorrect implication, (4) is inconsistent with more broadly used and accepted concepts of “geographic isolation,” and (5) has injected unnecessary confusion into scientific investigations and discussions. Instead, we suggest other wetland classification systems offer more informative alternatives. For example, hydrogeomorphic (HGM) classes based on well-established scientific definitions account for wetland functional diversity thereby facilitating explorations into questions of connectivity without an a priori designation of “isolation.” Additionally, an HGM-type approach could be used in combination with terms reflective of current regulatory or policymaking needs. For those rare cases in which the condition of being surrounded by uplands is the relevant distinguishing characteristic, use of terminology that does not unnecessarily imply isolation (e.g., “upland embedded wetlands”) would help alleviate much confusion caused by the “geographically isolated wetlands” misnomer.

  10. Isolated vertigo and possibility of brain ischemia.

    Science.gov (United States)

    Mosarrezai, Arash; Toghae, Mansoureh; Majed, Masoud; Aloosh, Mehdi

    2012-08-01

    In cases of isolated vertigo, physicians are unable to definitely distinguish between central or peripheral vertigo by history and physical examination. Some central causes of isolated vertigo such as cerebellar stroke can be life-threatening and require intervention. Brain infarction can be detected shortly after the onset of clinical symptoms by using diffusion-weighted MRI (DWI). We have conducted this study to perform DWI on isolated vertigo patients with a higher probability of brain infarction. We enrolled 55 consecutive patients with isolated vertigo who had at least one cardiovascular risk factor. A questionnaire that consisted of cardiovascular risk factors was completed and DWI performed for each patient. We analyzed the association of cardiovascular risk factors with infarction as identified by DWI. Using DWI, 5 (9.1%) patients had an acute ischemic stroke. Among cardiovascular risk factors, analysis showed a significant relationship between diabetes mellitus (DM) and infarction. Isolated vertigo may occur due to the occlusion of a small artery in the area of brain circulated by the posterior inferior cerebellar artery. According to our results, DWI may be used in diabetic patients with isolated vertigo to locate a probable infarction.

  11. Isolation of cellulose microfibrils - An enzymatic approach

    Directory of Open Access Journals (Sweden)

    Sain, M.

    2006-11-01

    Full Text Available Isolation methods and applications of cellulose microfibrils are expanding rapidly due to environmental benefits and specific strength properties, especially in bio-composite science. In this research, we have success-fully developed and explored a novel bio-pretreatment for wood fibre that can substantially improve the microfibril yield, in comparison to current techniques used to isolate cellulose microfibrils. Microfibrils currently are isolated in the laboratory through a combination of high shear refining and cryocrushing. A high energy requirement of these procedures is hampering momentum in the direction of microfibril isolation on a sufficiently large scale to suit potential applications. Any attempt to loosen up the microfibrils by either complete or partial destruction of the hydrogen bonds before the mechanical process would be a step forward in the quest for economical isolation of cellulose microfibrils. Bleached kraft pulp was treated with OS1, a fungus isolated from Dutch Elm trees infected with Dutch elm disease, under different treatment conditions. The percentage yield of cellulose microfibrils, based on their diameter, showed a significant shift towards a lower diameter range after the high shear refining, compared to the yield of cellulose microfibrils from untreated fibres. The overall yield of cellulose microfibrils from the treated fibres did not show any sizeable decrease.

  12. Design experience on seismically isolated buildings

    International Nuclear Information System (INIS)

    Giuliani, G.C.

    1991-01-01

    This paper describes the practical problems associated with the structural design of seismically isolated buildings now under construction in Ancona, Italy. These structures are the first seismically isolated buildings in Italy. The Ancona region is in zone 2 of the Italian Seismic Code. It has a design acceleration of 0.07 g which corresponds to a ground surface acceleration of 0.25 g. The last significant earthquake was recorded on June 14, 1972, having a single shock-type wave with a peak acceleration of 0.53 g. Taking into account the aforesaid earthquake, the structural design of these new buildings was performed according to an acceleration spectrum which was different from the zone 2 seismic code and which provided protection for stronger ground motions. To minimize the cost of the structure, the buildings used ribbed plate decks, thus reducing the amount of material and the mass of the structures to be isolated. The design requirements, dynamic analysis performed, structural design, and practical engineering employed are reported in this paper. A comparison between the costs of a conventionally designed and a base-isolated structure is also reported. It shows a net savings of 7% for the base-isolated structure. The tests undertaken for certifying the mechanical properties of the isolators for both static and dynamic loads are also described, as is the full-scale dynamic test which is scheduled for next year (1990) for one of the completed buildings. (orig.)

  13. Design experience on seismically isolated buildings

    International Nuclear Information System (INIS)

    Giuliani, G.C.

    1989-01-01

    This paper describes the practical problems associated with the structural design of a group of seismically isolated buildings now under construction in Ancona, Italy. These structures are the first seismically isolated buildings in Italy. Taking into account previous earthquakes, the structural design of these new buildings was performed according to an acceleration spectrum which was different from its Zone 2 seismic code and which provided protection for stronger ground motions. To minimize the cost of the structure, the buildings used ribbed plate decks, thus reducing the amount of material and the mass of the structures to be isolated. The design requirements, dynamic analysis performed, structural design, and practical engineering employed are reported in this paper. A comparison between the costs of a conventionally designed and a base-isolated structure is also reported. The tests undertaken for certifying the mechanical properties of the isolators for both static and dynamic loads are also described, as is the full-scale dynamic test which is scheduled for next year (1990) for one of the completed buildings. Lessons learned in this design effort are potentially applicable to seismic base isolation for nuclear power plants

  14. The waste isolation safety assessment programme

    International Nuclear Information System (INIS)

    Brandstetter, A.; Harwell, M.A.

    1980-01-01

    Associated with commercial nuclear power production in the USA is the generation of potentially hazardous radioactive wastes. The Department of Energy (DOE), through the National Waste Terminal Storage (NWTS) Programme, is seeking to develop nuclear waste isolation systems in geologic formations that will preclude contact with the biosphere of waste radionuclides in concentrations which are sufficient to cause deleterious impact on humans or their environments. Comprehensive analyses of specific isolation systems are needed to assess the expectations of meeting that objective. The Waste Isolation Safety Assessment Programme (WISAP) has been established at the Pacific Northwest Laboratory (operated by Battelle Memorial Institute) for developing the capability of making those analyses. Among the analyses required for isolation system evaluation is the detailed assessment of the post-closure performance of nuclear waste repositories in geologic formations. This assessment is essential, since it is concerned with aspects of the nuclear power programme which previously have not been addressed. Specifically, the nature of the isolation systems (e.g. involving breach scenarios and transport through the geosphere), and the time-scales necessary for isolation, dictate the development, demonstration and application of novel assessment capabilities. The assessment methodology needs to be thorough, flexible, objective, and scientifically defensible. Further, the data utilized must be accurate, documented, reproducible, and based on sound scientific principles. (author)

  15. Xerophilic mycopopulations isolated from rapeseeds (Brassica napus

    Directory of Open Access Journals (Sweden)

    Škrinjar Marija M.

    2013-01-01

    Full Text Available This paper presents the results of an investigation related to mycological populations of rapeseed samples produced in the Institute of Field and Vegetable Crops in Novi Sad (location: Rimski Šančevi, Novi Sad, with a special emphasis on the potentially toxigenic mycopopulations. Mycological investigations were performed on the samples that were treated with 4% solution of Na-hypochlorite, and on the ones that were not submitted to this treatment. Isolation and determination of total mould count was carried out using Dichloran Glycerol Agar (DG18. The identification of isolated moulds was done according to modern keys for fungal determination. From 20 untreated tested samples, 17 were contaminated with moulds (10.0 to 4.7x102 cfu/g. When the samples were treated with 4% solution of Na-hypochlorite, moulds were isolated only form 4 samples, and the total mould count ranged from 10.0 to 60.0 cfu/g. In the isolated mycopopulations, xerophilic moulds dominated, especially those from the genera Aspergillus, Eurotium and Penicillium. In the isolated mycopopulations, high degree of isolated species belonged to toxigenic species from the genera Alternaria, Aspergillus, Fusarium, Eurotium and Penicillium. [Projekat Ministarstva nauke Republike Srbije, br. III46009 i br.TR31025

  16. Dissemination of clonal Salmonella enterica serovar Typhimurium isolates causing salmonellosis in Mauritius

    DEFF Research Database (Denmark)

    Issack, M. I.; Migura, Lourdes Garcia; Ramsamy, Veemala D.

    2013-01-01

    from foodborne illness outbreaks and sporadic gastroenteritis cases, four blood isolates, one postmortem colon isolate, 14 food isolates, and five poultry isolates. All isolates were pansusceptible to the 16 antibiotics tested, except for two isolates that were resistant to sulfamethoxazole...

  17. Isolation and Characterization of Pollulan Produced by a Local Isolate of Aureobasidium Pullulans

    OpenAIRE

    Kassim, Mohammad Bashir Ismail [محمد بشيراسماعيل قاسم

    2000-01-01

    A local isolate of Aureobasidium pullulans was obtained from fig leaves surface. The isolate was identified as A. pullulans on the basis of appearance of polymorphism in the colonies, melanin pigment and pullulan production in shake flask cultures. Pullulan production by the local isolate of A pullulans was two thirds of that produced by ATCC 42023. The studies of physico — chemical properties, chemical analysis, sugar composition periodate oxidation and smith degradation showed that the pull...

  18. Bacterial isolates from the bryozoan Membranipora membranacea: influence of culture media on isolation and antimicrobial activity.

    Science.gov (United States)

    Heindl, Herwig; Thiel, Vera; Wiese, Jutta; Imhoff, Johannes F

    2012-03-01

    From specimens of the bryozoan Membranipora membranacea collected in the Baltic Sea, bacteria were isolated on four different media, which significantly increased the diversity of the isolated groups. All isolates were classified according to 16S rRNA gene sequence analysis and tested for antimicrobial properties using a panel of five indicator strains and six different media. Each medium featured a unique set of isolated phylotypes, and a phylogenetically diverse collection of isolates was obtained. A total of 96 isolates were assigned to 49 phylotypes and 29 genera. Only one-third of the members of these genera had been isolated previously from comparable sources. The isolates were affiliated with Alpha- and Gammaproteobacteria, Bacilli, and Actinobacteria. A comparable large portion of up to 22 isolates, i.e., 15 phylotypes, probably represent new species. Likewise, 47 isolates (approximately 50%) displayed antibiotic activities, mostly against grampositive indicator strains. Of the active strains, 63.8 % had antibiotic traits only on one or two of the growth media, whereas only 12.7 % inhibited growth on five or all six media. The application of six different media for antimicrobial testing resulted in twice the number of positive hits as obtained with only a single medium. The use of different media for the isolation of bacteria as well as the variation of media considered suitable for the production of antibiotic substances significantly enhanced both the number of isolates obtained and the proportion of antibiotic active cultures. Thus the approach described herein offers an improved strategy in the search for new antibiotic compounds.

  19. SUSCEPTIBILITY OF RESPIRATORY ISOLATES OF STREPTOCOCCUS PNEUMONIAE ISOLATED FROM CHILDREN HOSPITALIZED IN THE CLINICAL CENTER NIS.

    Science.gov (United States)

    Dinić, Marina M; Mladenović Antić, Snezana; Kocić, Branislava; Stanković Dordević, Dobrila; Vrbić, Miodrag; Bogdanović, Milena

    2016-01-01

    Streptococcus pneumoniae is one of the most common causes of respiratory infections. The aim was to study the susceptibility to antimicrobial agents of respiratory isolates ofStreptococcus pneumoniae obtained from hospitalized children. A total of 190 respiratory pneumococcal isolates obtained from children aged from 0 to 14 years were isolated and identified by using standard microbiological methods. Susceptibility to oxacillin, erythromycin, clindamycin, tetracycline, cotrimoxazole, ofloxacin and rifampicin was tested by disc diffusion method. Minimal inhibitory concentrations for amoxicillin and ceftriaxone were determined by means of E test. The macrolide-resistant phenotype was detected by double disc diffusion test. All tested isolates were susceptible to amoxicillin and ceftriaxone. The minimal amoxicillin concentration inhibiting the growth of 50% of isolates and of 90% of isolates was 0.50 microg/ml and 1.0 microg/ml, respectively and the minimal ceftriaxone concentration inhibiting the growth of 50% of isolates and of 90% of isolates was 0.25 microg/ml and 0.50 microg/ml, respectively. Susceptibility to erythromycin and clindamycin was observed in 21.6% and 29.47% of isolates, respectively. The resistence to macrolides-M phenotype was detected in 10.07% of isolates and constitutive macrolide-lincosamide-streptogramin phenotype (constitutive MLS phenotype) was found in 89.93% of isolates. All tested isolates were susceptible to ofloxacin and rifampicin. Amoxicillin could be the therapy of choice in pediatric practice. The macrolides should not be recommended for the empirical therapy of pneumococcal respiratory tract infection in our local area.

  20. Single spore isolation and morphological characterization of local Malaysian isolates of rice blast fungus Magnoporthe grisea

    Science.gov (United States)

    Mishra, Ankitta; Ratnam, Wickneswari; Bhuiyan, Md Atiqur Rahman; Ponaya, Ariane; Jena, Khisord K.

    2015-09-01

    Rice blast is a destructive disease, caused by the fungal pathogen Magnaporthe grisea. It causes considerable damage to rice and leads to crop loss in rice growing regions worldwide. Although fungicides can be used to control rice blast, they generate additional cost in rice production and contamination of environment and food. Therefore, the use of resistant varieties is thought to be one of the most economically and environmentally efficient ways of crop protection from the disease. Six new local Malaysian isolates of M. grisea were isolated using single spore isolation method. Five isolates were from infected leaf samples collected from Kompleks Latihan MADA, Kedah and one was from Kelantan. These isolates were identified using morphological characteristics and microscopic studies and later confirmed by ITSequences. These isolates were induced to sporulate and used for greenhouse screening on two differential rice varieties: Mahsuri (susceptible) and Pongsu Seribu 2 (resistant). Among the 6 isolates, isolate number 3 was found to be the most virulent showing high sporulation while isolate number 4 was very slow growing, and the least virulent.