Sample records for streptoccocus agalactiae isolates

  1. Nile Tilapia Infectivity by Genomically Diverse Streptoccocus agalactiae Isolates from Multiple Hosts

    Streptococcus agalactiae, Lancefield group B Streptococcus (GBS), is recognized for causing cattle mastitis, human neonatal meningitis, and fish meningo-encephalitis. We investigated the genomic diversity of GBS isolates from different phylogenetic hosts and geographical regions using serological t...

  2. Genomic Diversity of Streptoccocus agalactiae Isolates from Multiple Hosts and Their Infectivity in Nile Tilapia

    Streptococcus agalactiae, the Lancefield group B Streptococcus (GBS), has a broad host range and can be pathogenic to numerous animals, including fish. GBS is most recognized for causing cattle mastitis and human neonatal meningitis, it also causes fatal meningo-encephalitis in fish. We investigat...

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

    Vera Eickel; Barbara Kahl; Beatrice Reinisch; Angelika Dübbers; Peter Küster; Claudia Brandt; Barbara Spellerberg


    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 characte...

  4. Molecular typing of Streptococcus agalactiae isolates from fish

    The genetic variability among Streptococcus agalactiae isolates recovered from fish was characterized using single-stranded conformation polymorphisms (SSCP) analysis of the intergenic spacer region (ISR), and amplified fragment length polymorphism (AFLP) fingerprinting. A total of 49 S. agalactiae ...

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

    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.

  6. Human Streptococcus agalactiae isolate in Nile tilapia (Oreochromis niloticus)

    Streptococcus agalactiae, the Lancefield group B Streptococcus (GBS), long recognized as a mammalian pathogen, is an emerging pathogen to fish. We show that a GBS serotype Ia, multilocus sequence type ST-7 isolate from a human neonatal meningitis clinical case causes disease signs and mortality in N...

  7. Human Streptococcus agalactiae Isolate in Nile Tilapia (Oreochromis niloticus)

    Evans, Joyce J.; Phillip H. Klesius; Pasnik, David J.; Bohnsack, John F.


    Streptococcus agalactiae, the Lancefield group B streptococcus (GBS) long recognized as a mammalian pathogen, is an emerging concern with regard to fish. We show that a GBS serotype Ia multilocus sequence type ST-7 isolate from a clinical case of human neonatal meningitis caused disease and death in Nile tilapia (Oreochromis niloticus).

  8. Antibiotic Susceptibilities and Serotyping of Clinical Streptococcus Agalactiae Isolates

    Altay Atalay


    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.

  9. Isolation and identification of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR) from affected sheep to Contagious agalactia of Khuzestan province, Iran

    Pooladgar, A.R.; Looni, R.,; Ghaemmaghami, Sh.; Pourbakhsh, A.; Ashtari, A.; Ali Shirudi, A.


    Mycoplasma agalactiae (M. agalactiae) is one of the main causes of contagious agalactia, an infectious syndrome of sheep and goats in Khuzestan province –southwest of Iran that is characterized by mastitis and subsequent failure of milk production, arthritis, abortion and keratoconjunctivitis. This study was carried out to isolation and identification of M. agalactiae with culture and polymerase chain reaction (PCR) method from sheep in Khuzestan province, Iran. A total of 91 samples were col...

  10. DNA microarray-based typing of Streptococcus agalactiae isolates.

    Nitschke, Heike; Slickers, Peter; Müller, Elke; Ehricht, Ralf; Monecke, Stefan


    Streptococcus agalactiae frequently colonizes the urogenital tract, and it is a major cause of bacterial septicemia, meningitis, and pneumonia in newborns. For typing purposes, a microarray targeting group B streptococcus (GBS) virulence-associated markers and resistance genes was designed and validated with reference strains, as well as clinical and veterinary isolates. Selected isolates were also subjected to multilocus sequence typing. It was observed that putative typing markers, such as alleles of the alpha-like protein or capsule types, vary independently of each other, and they also vary independently from the affiliation to their multilocus sequence typing (MLST)-defined sequence types. Thus, it is not possible to assign isolates to sequence types based on the identification of a single distinct marker, such as a capsule type or alp allele. This suggests the occurrence of frequent genomic recombination. For array-based typing, a set of 11 markers (bac, alp, pil1 locus, pepS8, fbsB, capsule locus, hylB, abiG-I/-II plus Q8DZ34, pil2 locus, nss plus srr plus rogB2, and rgfC/A/D/B) was defined that provides a framework for splitting the tested 448 S. agalactiae isolates into 76 strains that clustered mainly according to MLST-defined clonal complexes. There was evidence for region- and host-specific differences in the population structure of S. agalactiae, as well as an overrepresentation of strains related to sequence type 17 among the invasive isolates. The arrays and typing scheme described here proved to be a convenient tool for genotyping large numbers of clinical/veterinary isolates and thus might help obtain insight into the epidemiology of S. agalactiae.

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

    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.


    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, isolate

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

    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...

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

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


    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.

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

    Liu, Guangjin; Zhang, Wei; Lu, Chengping


    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.

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

    Angela Mariana Lusiastuti


    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.

  16. Complete Genome Sequence of Streptococcus agalactiae Strain S25 Isolated from Peritoneal Liquid of Nile Tilapia

    Mainardi, Rafaella Menegheti; Lima Júnior, Edson Antônio; Ribeiro Júnior, Jose Carlos; Beloti, Vanerli; Carmo, Anderson Oliveira; Kalapothakis, Evanguedes; Gonçalves, Daniela Dib; Padua, Santiago Benites


    Streptococcus agalactiae (Lancefield group B; GBS) is one of the major pathogens in fish production, especially in Nile tilapia (Oreochromis niloticus). The genomic characteristics of GBS isolated from fish must be more explored. Thus, we present here the genome of GBS S25, isolated from Nile tilapia from Brazil. PMID:27491974

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

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


    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.

  18. Complete Genome Sequence of Streptococcus agalactiae Strain S25 Isolated from Peritoneal Liquid of Nile Tilapia.

    Mainardi, Rafaella Menegheti; Lima Júnior, Edson Antônio; Ribeiro Júnior, Jose Carlos; Beloti, Vanerli; Carmo, Anderson Oliveira; Kalapothakis, Evanguedes; Gonçalves, Daniela Dib; Padua, Santiago Benites; Pereira, Ulisses Pádua


    Streptococcus agalactiae (Lancefield group B; GBS) is one of the major pathogens in fish production, especially in Nile tilapia (Oreochromis niloticus). The genomic characteristics of GBS isolated from fish must be more explored. Thus, we present here the genome of GBS S25, isolated from Nile tilapia from Brazil.

  19. Serology based immunological crossreactivity among various isolates of streptococcus agalactiae from mastitic buffaloes

    R. Hussain


    Full Text Available To measure the degree of immunological cross-reactivity among different isolates of Streptococcus agalactiae from mastitic cases of buffaloes and cows in four districts (Faisalabad, Jhang, Toba Tek Singh and Sargodha, two tests were used. The isolates of Streptococcus agalactiae obtained from mastitic animals from these districts were purified on Edward’s medium. Hyper immune sera were raised by injecting 1.2 x 109 cfu/ml I/V of the isolates separately into rabbits. The sera thus raised were tested for in vitro immunological cross-reactivity through counter immuno-electrophoresis and bacterial agglutination test. Eight Streptococcus agalactiae isolates were selected and named A, B, C, D, E, F, G and H. In the bacterial agglutination test, all variants were similar antigenically and showed positive reactions, but a few reactions were negative, so the cross-reactivity among the variants was (93.75 % with respect to the positive reactions. The percentage of negative reactions was (6.25 %. The cross-reactivity among the Str. agalactiae isolates was 98.43% by modified counter immuno-electrophoresis assay.

  20. Differential pathogenicity of five Streptococcus agalactiae isolates of diverse geographic origin in Nile tilapia (Oreochromis niloticus L.)

    Streptococcus agalactiae is an emerging pathogen of fish and has caused significant morbidity amd mortality worldwide. The work in this study assessed whether pathogenic differences exist among isolates from different geographic locations. Nile tilapia (Oreochromis niloticus L.) were administered an...

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

    Ana Beatriz de Almeida Corrêa


    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.

  2. Molecular cloning and bioinformatic analysis of the Streptococcus agalactiae neuA gene isolated from tilapia.

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


    Cytidine monophosphate (CMP) N-acetylneuraminic acid (NeuNAc) synthetase, which is encoded by the neuA gene, can catalyze the activation of sialic acid with CMP, and plays an important role in Streptococcus agalactiae infection pathogenesis. To study the structure and function of the S. agalactiae neuA gene, we isolated it from diseased tilapia, amplified it using polymerase chain reaction (PCR) with specific primers, and cloned it into a pMD19-T vector. The recombinant plasmid was confirmed by PCR and restriction enzyme digestion, and identified by sequencing. Molecular characterization analyses of the neuA nucleotide amino acid sequence were performed using bioinformatic tools and an online server. The results showed that the neuA nucleotide sequence contained a complete coding region, which comprised 1242 bp, encoding 413 amino acids (aa). The aa sequence was highly conserved and contained a Glyco_tranf_GTA_type superfamily and an SGNH_hydrolase superfamily conserved domain, which are related to sialic acid activation catalysis. The NeuA protein possessed many important sites related to post-translational modification, including 28 potential phosphorylation sites and 2 potential N-glycosylation sites, had no signal peptides or transmembrane regions, and was predicted to reside in the cytoplasm. Moreover, the protein had some B-cell epitopes, which suggests its potential in development of a vaccine against S. agalactiae infection. The codon usage frequency of neuA differed greatly in Escherichia coli and Homo sapiens genes, and neuA may be more efficiently expressed in eukaryotes (yeast). S. agalactiae neuA from tilapia maintains high structural homology and sequence identity with CMP-NeuNAc synthetases from other bacteria.

  3. Unexpected genetic diversity of Mycoplasma agalactiae caprine isolates from an endemic geographically restricted area of Spain

    De la Fe Christian


    Full Text Available Abstract Background The genetic diversity of Mycoplasma agalactiae (MA isolates collected in Spain from goats in an area with contagious agalactia (CA was assessed using a set of validated and new molecular typing methods. Validated methods included pulsed field gel electrophoresis (PFGE, variable number of tandem repeats (VNTR typing, and Southern blot hybridization using a set of MA DNA probes, including those for typing the vpma genes repertoire. New approaches were based on PCR and targeted genomic regions that diverged between strains as defined by in silico genomic comparisons of sequenced MA genomes. Results Overall, the data showed that all typing tools yielded consistent results, with the VNTR analyses being the most rapid method to differentiate the MA isolates with a discriminatory ability comparable to that of PFGE and of a set of new PCR assays. All molecular typing approaches indicated that the Spanish isolates from the endemic area in Murcia were very diverse, with different clonal isolates probably restricted to separate, but geographically close, local areas. Conclusions The important genetic diversity of MA observed in infected goats from Spain contrasts with the overall homogeneity of the genomic background encountered in MA from sheep with CA in Southern France or Italy, suggesting that assessment of the disease status in endemic areas may require different approaches in sheep and in goats. A number of congruent sub-typing tools are now available for the differentiation of caprine isolates with comparable discriminatory powers.

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

    Yessica Ortega Asencios


    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.

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

    Yang, Feng; Li, Hongsheng; ZHANG, SHIDONG; WANG, XURONG


    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.

  6. Virulence factors, antimicrobial susceptibility and molecular characterization of Streptococcus agalactiae isolated from pregnant women.

    Beigverdi, Reza; Jabalameli, Fereshteh; Mirsalehian, Akbar; Hantoushzadeh, Sedigheh; Boroumandi, Shahram; Taherikalani, Morovat; Emaneini, Mohammad


    Forty-one Streptococcus agalactiae isolates collected from pregnant women at 35-37 weeks of gestation were analysed for their capsular types, antimicrobial resistance determinants, distribution of virulence factors and genetic relatedness using PCR and multiplex PCR. Capsular type III was predominant (65.8%), followed by capsular type II (14.6%), Ib (7.3%), and V(4.9%). All isolates were susceptible to penicillin, vancomycin, linezolid and quinupristin-dalfopristin. Resistance to tetracycline, erythromycin and clindamycin were found in 97.6%, 24.4%, and 14.6% of isolates, respectively. The most common antimicrobial resistance gene was tetM found in 97.6% of the isolates followed by ermTR and ermB found in 12% and 7.3% of isolates, respectively. The most common virulence gene was hly (100%), followed by scpB (97.6%), bca (97.6%), rib (53.65%) and bac (4.9%). The insertion sequence IS1548 was found in 63.4% of isolates. By multi locus variable number of tandem repeat analysis (MLVA) typing, 30 different allelic profiles or MLVA types (MTs) were identified. The most frequent was the MT1 (5/41, 12.2%) and followed by MT2 (4/41, 9.75%). Our data revealed that population structure of these isolates is highly diverse and indicates different MLVA types.

  7. Characterization and antibiotic susceptibility of Streptococcus agalactiae isolates causing urinary tract infections.

    Piccinelli, Giorgio; Biscaro, Valeria; Gargiulo, Franco; Caruso, Arnaldo; De Francesco, Maria Antonia


    Streptococcus agalactiae (GBS) has been implicated in urinary tract infections but the microbiological characteristics and antimicrobial susceptibility of these strains are poorly investigated. In this study, 87 isolates recovered from urine samples of patients who had attended the Spedali Civili of Brescia (Italy) and had single organism GBS cultured were submitted to antimicrobial susceptibility testing, molecular characterization of macrolide and levofloxacin resistance, PCR-based capsular typing and analysis of surface protein genes. By automated broth microdilution method, all isolates were susceptible to penicillin, cefuroxime, cefaclor, and ceftriaxone; 80%, 19.5% and 3.4% of isolates were non-susceptible to tetracycline, erythromycin, and levofloxacin, respectively. Macrolide resistance determinants were iMLS(B) (n=1), cMLS(B) (n=10) and M (n=5), associated with ermTR, ermB and mefA/E. Levofloxacin resistance was linked to mutations in gyrA and parC genes. Predominant capsular types were III, Ia, V, Ib and IX. Type III was associated with tetracycline resistance, while type Ib was associated with levofloxacin resistance. Different capsular type-surface protein gene combinations (serotype V-alp2, 3; serotype III-rib; serotype Ia-epsilon) were detected. A variety of capsular types are involved in significant bacteriuria. The emergence of multidrug resistant GBS may become a significant public health concern and highlights the importance of careful surveillance to prevent the emergence of these virulent GBS.

  8. Frequency of isolation of streptococus agalactiae in pregnant women and their offspring Frecuencia de aislamiento de Streptococcus agalactiae en un grupo de embarazadas y sus productos

    Nancy E. Agudelo O.


    Full Text Available

    A group of 88 pregnant women and 61 of their newborn Infants was studied In order to determine the frequency of isolation of Streptococcus agalactiae; all genital maternal cultures were negative but 3 mothers and 2 Infants had positive pharyngeal secretions; no evidence of streptococcal disease was found in the colonized Infants. The frequency of colonization found in this study agrees with that reported by other authors but the reduced number of cases does not allow In depth epidemiological considerations.

    Se estudiaron 88 madres y 61 de sus recién nacidos con el fin de determinar la frecuencia de aislamiento de Streptococcus agalactiae (estreptococo beta hemolítico del grupo B; no se aisló el germen de ninguno de los cultivos tomados de las secreciones del cuello uterino pero sí a partir de las muestras faringeas, en tres de las madres (3.4% y en dos de sus hijos. El tercer niño no se estudió porque su nacimiento ocurrió por cesárea. Las cifras encontradas en el presente trabajo están de acuerdo con lo que relata la literatura, pero el bajo número de casos estudiado no permite hacer consideraciones epidemiológicas más profundas.

  9. Identification of high immunoreactive proteins from Streptococcus agalactiae isolates recognized by human serum antibodies.

    Brzychczy-Wloch, Monika; Gorska, Sabina; Brzozowska, Ewa; Gamian, Andrzej; Heczko, Piotr B; Bulanda, Malgorzata


    The aim of the studies was to identify immunogenic proteins of Streptococcus agalactiae (group B streptococcus; GBS) isolates. Investigation of the immunoreactivity with human sera allowed us to determine major immunogenic proteins which might be potential candidates for the development of vaccine. For the study, we have selected 60 genetically different, well-characterized GBS clinical isolates. The proteins immunoreactivity with 24 human sera from patients with GBS infections, carriers, and control group without GBS was detected by SDS-PAGE and Western blotting. As a result, some major immunogenic proteins were identified, of which four proteins with molecular masses of about 45 to 50 kDa, which exhibited the highest immunoreactivity features, were analyzed by LC-MS/MS. The proteins were identified by comparative analysis of peptides masses using MASCOT and statistical analysis. The results showed known molecules such as enolase (47.4 kDa), aldehyde dehydrogenase (50.6 kDa), and ones not previously described such as trigger factor (47 kDa) and elongation factor Tu (44 kDa). The preliminary results indicated that some GBS proteins that elicit protective immunity hold promise not only as components in a vaccine as antigens but also as carriers or adjuvants in polysaccharide conjugate vaccines, but more studies are needed.

  10. In vitro susceptibilities of field isolates of Mycoplasma agalactiae to oxytetracycline, tylosin, enrofloxacin, spiramycin and lincomycin-spectinomycin.

    Loria, G R; Sammartino, C; Nicholas, R A J; Ayling, R D


    The minimum inhibitory concentrations (MICs) of tetracycline, enrofloxacin, tylosin, spiramycin and a lincomycin:spectinomycin 1:2 combination, against 24 Sicilian isolates of Mycoplasma agalactiae, the causative organism of contagious agalactia were determined in vitro by a broth dilution method. Enrofloxacin was the most effective antimicrobial in vitro with a range of MIC values from 0.125 to 0.500 microg/ml and an MIC(50) of 0.203 and MIC(90) of 0.365 microg/ml. Using the MIC(50) and MIC(90) values the remaining four antimicrobials are ranked in order of in vitro effectiveness as follows: tylosin (MIC(50)0.292; MIC(90)0.525 microg/ml) was slightly more effective than tetracycline (MIC(50)0.296; MIC(90)0.533 microg/ml), followed by lincomycin:spectinomycin (MIC(50)0.521; MIC(90)0.938 microg/ml) and spiramycin (MIC(50)1.583; MIC(90)2.850 microg/ml). MIC values above 1.000 microg/ml were obtained using tetracycline, tylosin and spiramycin for some M. agalactiae isolates.

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

    Azizollah Ebrahimi


    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.

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

    Ebrahimi, Azizollah; Moatamedi, Azar; Lotfalian, Sharareh; Mirshokraei, Pejhman


    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.

  13. Dominance of clonal complex 10 among the levofloxacin-resistant Streptococcus agalactiae isolated from bacteremic patients in a Korean hospital.

    Ryu, Hyejin; Park, Yeon-Joon; Kim, Yong-Kyun; Chang, Jiyoung; Yu, Jin Kyung


    Streptococcus agalactiae has emerged as an important cause of invasive infection in adults. Forty-nine S. agalactiae isolates (41 from adults and 8 from neonates) were collected during a 4-year period (2010-2013) and analyzed by multilocus sequence typing (MLST). Antibiotic susceptibility to erythromycin, clindamycin and levofloxacin was determined and the determinants of resistance (ermA, ermB, ermC, mefA, lnuB) were detected by PCR and mutation in gyrA, gyrB, parC and parE gene was investigated by sequence analysis. They were resolved into 14 sequence types (STs) and belonged to five clonal complexes (CCs). The distribution of CC was significantly different according to the age group; CC1 (18/41) and CC10 (13/41) was the most common among the adult isolates but CC19 (5/8) was predominant among the neonatal isolates. The resistance rate to erythromycin, clindamycin was 18.4% and 24.5%, respectively. Among the 13 strains resistant to erythromycin and/or clindamycin, two isolates harbored ermA and 10 isolates harbored ermB. The levofloxacin resistance rate was very high (32.7%) and was significantly higher in CC10 (71.4%). All the levofloxacin-resistant isolates had identical gyrA substitution (Ser81Leu) but parC substitution was different according to the CCs. The additional mutation in parE (His221Tyr) was found only in CC19. Continuous monitoring of the fluoroquinolone resistance and genotypic distribution among S. agalactiae is needed.

  14. Short communication: Lipolytic activity on milk fat by Staphylococcus aureus and Streptococcus agalactiae strains commonly isolated in Swedish dairy herds.

    Vidanarachchi, Janak K; Li, Shengjie; Lundh, Åse Sternesjö; Johansson, Monika


    The objective of this study was to determine the lipolytic activity on milk fat of 2 bovine mastitis pathogens, that is, Staphylococcus aureus and Streptococcus agalactiae. The lipolytic activity was determined by 2 different techniques, that is, thin-layer chromatography and an extraction-titration method, in an experimental model using the most commonly occurring field strains of the 2 mastitic bacteria isolated from Swedish dairy farms. The microorganisms were inoculated into bacteria-free control milk and incubated at 37°C to reflect physiological temperatures in the mammary gland. Levels of free fatty acids (FFA) were analyzed at time of inoculation (t=0) and after 2 and 6h of incubation, showing significant increase in FFA levels. After 2h the FFA content had increased by approximately 40% in milk samples inoculated with Staph. aureus and Strep. agalactiae, and at 6h the pathogens had increased FFA levels by 47% compared with the bacteria-free control milk. Changes in lipid composition compared with the bacteria-free control were investigated at 2 and 6h of incubation. Diacylglycerols, triacylglycerols, and phospholipids increased significantly after 6h incubation with the mastitis bacteria, whereas cholesterol and sterol esters decreased. Our results suggest that during mammary infections with Staph. aureus and Strep. agalactiae, the action of lipases originating from the mastitis pathogens will contribute significantly to milk fat lipolysis and thus to raw milk deterioration.

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

    Gilbert Gwendolyn L


    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.

  16. Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China.

    Li, Liping; Wang, Rui; Liang, Wanwen; Gan, Xi; Huang, Ting; Huang, Yan; Li, Jian; Shi, Yunliang; Chen, Ming; Luo, Honglin


    Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae (GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the first time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in five provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding 36.9% (62/168), 37.5% (63/168) and 19.6% (33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia (95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010, but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during 2007-2011, III only occurred in 2012. 14 different PFGE types, including 4 new types (N, O, P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9% (141/168) of the total GBS strains. Ia corresponded to 11 PFGE types (A-H, N-P), in which type D predominated (51%). Ib represented 3 genotypes (I, J and Q) and III harbored only 2 genotypes (N and F). Type N and F synchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years. Although Ia still predominated, new rare serotype III already occurred in China.

  17. A comparative investigation of Streptococcus agalactiae isolates from fish and cattle

    Streptococcus agalactiae is the causative bacterium of streptococcosis and causes severe economic losses in wild and cultured fish and cattle, worldwide. In fish, infection can result in septicemia with hemorrhages on the body surface and in the external and internal organs. Streptococcus agalacti...

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

    The complete genome of a virulent Streptococcus agalactiae strain 138P is 1838701 bp in size, containing 1831 genes. The genome has 1593 coding sequences, 152 pseudo genes, 16 rRNAs, 69 tRNAs, and 1 non-coding RNA. The annotation of the genome is added by the NCBI Prokaryotic Genome Annotation Pipel...

  19. [Vaginal colonization of the Streptococcus agalactiae in pregnant woman in Tunisia: risk factors and susceptibility of isolates to antibiotics].

    Ferjani, A; Ben Abdallah, H; Ben Saida, N; Gozzi, C; Boukadida, J


    Streptococcus agalactiae or Group B Streptococcus (GBS) is one of the main bacterial causes of serious infections in newborns. We have evaluated prospectively GBS vaginal colonization in pregnant women and we have tried to determine the risk factors of the colonization by GBS and the particularities of the different isolated strains. We have screened 300 pregnant women with vaginal and anal sample in a same swab. Thirty nine (13%) pregnant women are colonized by SGB, 0% in the first trimester, 10.2% in the second trimester and 17% in the third trimester. Different factors are associated significantly with GBS colonization: past history of infection in newborns, genital infection during pregnancy and parity The highest rates of resistance are found in tetracycline (97.4%), erythromycin (51.3%) and lincomycin (46.2%). All the strains were susceptible to amoxicilin and pristinamycin.

  20. In vitro susceptibilities of caprine Mycoplasma agalactiae field isolates to six antimicrobial agents using the E test methodology.

    Filioussis, George; Petridou, Evanthia; Giadinis, Nektarios D; Kritas, Spyridon K


    The minimum inhibitory concentrations (MICs) of enrofloxacin, ciprofloxacin, spectinomycin, tetracycline, spiramycin and erythromycin against 30 caprine Greek isolates of Mycoplasma agalactiae were determined using E test methodology. The E test strips were placed on Eaton's agar medium without antimicrobials and phenol red. MICs were then read by determining where the growth inhibition zone intersected with the MIC scale on the strip. An MIC value of 8 µg/mL was considered as a guide to mycoplasma resistance. All isolates were sensitive to fluoroquinolones (MIC50, 0.19 g/mL; MIC90, 0.38 µg/mL; highest MIC, 0.5 µg/mL), spectinomycin (MIC50, 0.5 µg/mL; MIC90, 1 µg/mL; highest MIC, 1 µg/mL), and spiramycin (MIC50, 1 µg/mL; MIC90, 1.5 µg/mL; highest MIC, 2 µg/mL). Two strains exhibited resistance to tetracycline (MIC 32 µg/mL) but these were not found to carry any of the tet(M), tet(O), and tet(S) resistance genes. Finally all isolates expressed resistance to erythromycin (MIC50, 128 µg/mL; MIC90, >256 µg/mL).

  1. Associations between capsular serotype, multilocus sequence type, and macrolide resistance in Streptococcus agalactiae isolates from Japanese infants with invasive infections.

    Morozumi, M; Wajima, T; Kuwata, Y; Chiba, N; Sunaoshi, K; Sugita, K; Sakata, H; Iwata, S; Ubukata, K


    SUMMARY Streptococcus agalactiae (group B streptococcus; GBS) isolates (n = 150) from infants with invasive infections between 2006 and 2011 were analysed for capsular serotype, multilocus sequence type, and antibiotic susceptibility. In cases with late-onset disease (n = 115), primary meningitis was predominant (62.6%), but represented only 39.1% in cases with early-onset disease (n = 23). The most common serotype was III (58.7%), followed by Ia (21.3%) and Ib (12.7%). Sequence types (STs) of serotype III strains included ST17 (50.0%), ST19 (26.1%), ST335 (18.2%), ST27 (4.5%), and ST1 (1.1%). Predominant STs of serotypes Ia and Ib were ST23 (81.3%) and ST10 (84.2%), respectively. No penicillin-resistant strains were detected, but 22·0% of strains had mef(A/E), erm(A), or erm(B) genes, which mediate macrolide resistance. A new ST335, possessing an mef(A/E) gene belonging to clonal complex 19 gradually increased in frequency. Improved prevention of invasive GBS infections in infants requires timely identification, and ultimately vaccine development.

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

    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.

  3. Genomic comparison between pathogenic Streptococcus agalactiae isolated from Nile tilapia in Thailand and fish-derived ST7 strains.

    Kayansamruaj, Pattanapon; Pirarat, Nopadon; Kondo, Hidehiro; Hirono, Ikuo; Rodkhum, Channarong


    Streptococcus agalactiae, or Group B streptococcus (GBS), is a highly virulent pathogen in aquatic animals, causing huge mortalities worldwide. In Thailand, the serotype Ia, β-hemolytic GBS, belonging to sequence type (ST) 7 of clonal complex (CC) 7, was found to be the major cause of streptococcosis outbreaks in fish farms. In this study, we performed an in silico genomic comparison, aiming to investigate the phylogenetic relationship between the pathogenic fish strains of Thai ST7 and other ST7 from different hosts and geographical origins. In general, the genomes of Thai ST7 strains are closely related to other fish ST7s, as the core genome is shared by 92-95% of any individual fish ST7 genome. Among the fish ST7 genomes, we observed only small dissimilarities, based on the analysis of clustered regularly interspaced short palindromic repeats (CRISPRs), surface protein markers, insertions sequence (IS) elements and putative virulence genes. The phylogenetic tree based on single nucleotide polymorphisms (SNPs) of the core genome sequences clearly categorized the ST7 strains according to their geographical and host origins, with the human ST7 being genetically distant from other fish ST7 strains. A pan-genome analysis of ST7 strains detected a 48-kb gene island specifically in the Thai ST7 isolates. The orientations and predicted amino acid sequences of the genes in the island closely matched those of Tn5252, a streptococcal conjugative transposon, in GBS 2603V/R serotype V, Streptococcus pneumoniae and Streptococcus suis. Thus, it was presumed that Thai ST7 acquired this Tn5252 homologue from related streptococci. The close phylogenetic relationship between the fish ST7 strains suggests that these strains were derived from a common ancestor and have diverged in different geographical regions and in different hosts.

  4. Characterization of Tn5801.Sag, a variant of Staphylococcus aureus Tn916 family transposon Tn5801 that is widespread in clinical isolates of Streptococcus agalactiae.

    Mingoia, Marina; Morici, Eleonora; Tili, Emily; Giovanetti, Eleonora; Montanari, Maria Pia; Varaldo, Pietro E


    Tn5801, originally detected in Staphylococcus aureus Mu50, is a Tn916 family element in which a unique int gene (int5801) replaces the int and xis genes in Tn916 (int916 and xis916). Among 62 tet(M)-positive tetracycline-resistant Streptococcus agalactiae isolates, 43 harbored Tn916, whereas 19 harbored a Tn5801-like element (Tn5801.Sag, ∼20.6 kb). Tn5801.Sag was characterized (PCR mapping, partial sequencing, and chromosomal integration) and compared to other Tn5801-like elements. Similar to Tn5801 from S. aureus Mu50, tested in parallel, Tn5801.Sag was unable to undergo circularization and conjugal transfer.

  5. Draft Genome Sequences of Streptococcus agalactiae Serotype Ia and III Isolates from Tilapia Farms in Thailand.

    Areechon, Nontawith; Kannika, Korntip; Hirono, Ikuo; Kondo, Hidehiro; Unajak, Sasimanas


    Streptococcus agalactiaeserotypes Ia and III were isolated from infected tilapia in cage and pond culture farms in Thailand during 2012 to 2014, in which pathogenicity analysis demonstrated that serotype III showed higher virulence than serotype Ia. Here, we report the draft genome sequencing of piscineS. agalactiaeserotypes Ia and III.


    Group B Streptococcus agalactiae (GBS) is a cause of infectious disease in multiple poikilothermic and homothermic animal species. Epidemiological and zoonotic considerations necessitate an undertaking of a comparison of S. agalactiae isolates from different phylogenetic hosts and geographical regi...

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

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

  8. In vitro antimicrobial activity of Combretum molle (Combretaceae) against Staphylococcus aureus and Streptococcus agalactiae isolated from crossbred dairy cows with clinical mastitis.

    Regassa, Fekadu; Araya, Mengistu


    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.

  9. Serotype distribution and antimicrobial susceptibilities of Streptococcus agalactiae isolated from infected cultured tilapia (Oreochromis niloticus) in Thailand: Nine-year perspective.

    Dangwetngam, Machalin; Suanyuk, Naraid; Kong, Fanrong; Phromkunthong, Wutiporn


    Streptococcus agalactiae (group B Streptococcus, GBS) infection remains a major problem associated with high mortality of cultured tilapia worldwide. The present study reports the serotype distribution and antimicrobial susceptibilities of GBS isolated from infected tilapia cultured in Thailand. One hundred and forty-four GBS isolates were identified by biochemical, serological and molecular analyses. Of these 144 GBS isolates, 126 were serotype Ia and 18 were serotype III. Antimicrobial susceptibilities of the 144 GBS isolates were determined by the disc diffusion method. Most GBS isolates were susceptible to lincomycin, norfloxacin, oxytetracycline, ampicillin, erythromycin and chloramphenicol, but resistant to oxolinic acid, gentamicin, sulfamethoxazole and trimethoprim. However, 17 isolates displayed an oxytetracycline-resistant phenotype and harboured the tet(M) gene. The broth microdilution method was used to determine the minimal inhibitory concentrations (MICs) of 17 oxytetracycline-resistant GBS isolates, and then minimal bactericidal concentrations (MBCs) of these isolates were evaluated. Oxytetracyline-resistant isolates were found to be susceptible to ampicillin, lincomycin, norfloxacin, erythromycin and chloramphenicol, with the MIC and MBC ranging from ≤ 0.125 to 0.5 μg ml- 1 and ≤ 0.125 to 2 μg ml- 1, respectively. Moreover, all 17 oxytetracycline-resistant isolates demonstrated resistance to trimethoprim, oxolinic acid, gentamicin, sulfamethoxazole and oxytetracycline, with the MIC and MBC ranging from 16 to ≥ 128 μg ml- 1 and ≥ 128 μg ml- 1, respectively. These findings are useful information for antibiotic usage in fish aquaculture.

  10. Molecular epidemiology and distribution of serotypes, genotypes, and antibiotic resistance genes of Streptococcus agalactiae clinical isolates from Guelma, Algeria and Marseille, France.

    Bergal, A; Loucif, L; Benouareth, D E; Bentorki, A A; Abat, C; Rolain, J-M


    This study describes, for the first time, the genetic and phenotypic diversity among 93 Streptococcus agalactiae (group B Streptococcus, GBS) isolates collected from Guelma, Algeria and Marseille, France. All strains were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The molecular support of antibiotic resistance and serotyping were investigated by polymerase chain reaction (PCR). The phylogenetic lineage of each GBS isolate was determined by multilocus sequence typing (MLST) and grouped into clonal complexes (CCs) using eBURST. The isolates represented 37 sequence types (STs), 16 of which were novel, grouped into five CCs, and belonging to seven serotypes. Serotype V was the most prevalent serotype in our collection (44.1%). GBS isolates of each serotype were distributed among multiple CCs, including cps III/CC19, cps V/CC1, cps Ia/CC23, cps II/CC10, and cps III/CC17. All isolates presented susceptibility to penicillin, whereas resistance to erythromycin was detected in 40% and tetracycline in 82.2% of isolates. Of the 37 erythromycin-resistant isolates, 75.7% showed the macrolide-lincosamide-streptogramin B (MLSB)-resistant phenotype and 24.3% exhibited the macrolide (M)-resistant phenotype. Constitutive MLSB resistance (46%) mediated by the ermB gene was significantly associated with the Guelma isolates, whereas the M resistance phenotype (24.3%) mediated by the mefA/E gene dominated among the Marseille isolates and belonged to ST-23. Tetracycline resistance was predominantly due to tetM, which was detected alone (95.1%) or associated with tetO (3.7%). These results provide epidemiological data in these regions that establish a basis for monitoring increased resistance to erythromycin and also provide insight into correlations among clones, serotypes, and resistance genes.

  11. 中国南方地区罗非鱼无乳链球菌的分子流行病学研究%Molecular epidemiology of Streptococcus agalactiae isolated from tilapia in Southern China

    郭玉娟; 张德锋; 樊海平; 陈学年; 李彤彤; 李爱华


    从广东省以及海南省等地区养殖的患病罗非鱼体内分离、收集到多株致病菌,经生化分析和分子生物学鉴定,均为无乳链球菌.对这些菌株分别进行了耐药谱测定、分子分型试验以及分子血清型分析.药敏试验结果表明,2007-2010年分离到的无乳链球菌耐药谱基本相似;多位点可变数目串联重复序列分析( MLVA)试验中,选择5个高变异指数的可变数目重复位点(VNTR)进行分子分型,结果表明,所有鱼源无乳链球菌菌株为同一MLVA型,而作为对照的牛源无乳链球菌则明显不同;为了对这些菌株进一步分型,分别进行了分子血清型和表面蛋白抗原基因的检测,结果表明,鱼源无乳链球菌的分子血清型均为Ⅰa型,表面蛋白抗原均为alpha-C蛋白.这进一步说明了不同年份和不同地区的鱼源无乳链球菌在基因水平上为同一分子类型,具有相同的起源或传染源.同时也说明,我国南方地区罗非鱼无乳链球菌在这几年中未发生明显的遗传变异.这些结果为罗非鱼无乳链球菌病疫苗研制,疫病监测及药物防治的研究提供理论依据.%Streptococcus agalactiae strains were isolated and collected from moribund tilapia in Guangdong, Hainan,and Fujian Provinces, and these isolates were identified by biochemical and molecular biological characterization. In the current study, we compared the phylogenetic relationships among S. Agalactiae strains. Furthermore, we compared antibiotic sensitivity, molecular classification and molecular serotype of these strains which were isolated from different years and different geographical regions. Antibiotic sensitivity assays showed that among 20 antibiotics tested,there are 10 antibiotics that had no differences,and the other 10 items showed similar sensitivity. Five most diverse loci (Variable number of tandem repeats, VNTR) were used for the construction of MLVA to evaluate the diversity of these strains

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

    Ayling Roger D


    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.

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

    Pridgeon, Julia W; Klesius, Phillip H


    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 (P<0.001) protection to both 3-5g and 15-20g Nile tilapia against challenges with 30 parent isolates of S. 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.

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

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


    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.

  15. Specific detection by PCR of Streptococcus agalactiae in milk.

    Martinez, G.; Harel, J; Gottschalk, M


    The aim of this study was to develop a simple and specific method for direct detection of Streptococcus agalactiae from cow's milk. The method was based on polymerase chain reaction (PCR) using species-specific and universal primers derived from the 16S rRNA gene. The amplification product was verified by restriction endonuclease digest and sequencing. Specific identification was proven on a collection of 147 S. agalactiae isolates of bovine and human origin. In addition, 17 strains belonging...

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

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


    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

  17. Streptococcus agalactiae endogenous endophthalmitis

    Saffra, Norman; Rakhamimov, Aleksandr; Husney, Robert; Ghitan, Monica


    Streptococcus agalactiae (SA) is a Group B Streptococcus, which is a common pathogen implicated in neonatal and geriatric sepsis. Endogenous bacterial endophthalmitis (EBE) is a condition that results from haematogenous seeding of the globe, during transient or persistent bacteremia. We document a case of a non-septic geriatric patient, who developed EBE after a transient bacteraemia with SA.

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

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


    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...... of bulk tank milk samples with milk from other herds. From June to September 1996, bulk tank milk was sampled from 100 Danish dairy herds seven times, with intervals of 2 wk. The samples were examined for the presence of S. agalactiae by four different methods: 1) by the method approved for the program, 2...... 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...

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

    Mousavi, Seyed Masoud


    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.

  20. 海南罗非鱼无乳链球菌分离鉴定及其特性研究%Isolation, identification and characterization of Streptococcus agalactiae in tilapia in Hainan

    周清; 沈锦玉; 徐洋; 姚嘉赟; 潘晓艺; 郝贵杰; 尹文林


    从海南省患暴发性疾病的罗非鱼(Tilapia)上分离出1株细菌HNLFYL4,对分离菌株进行了鉴定及致病性和药物敏感性研究.通过形态学观察和生理生化鉴定,结果显示,分离菌株为无乳链球菌(Streptococcus agalactiae).对分离菌株的16S rRNA基因进行PCR扩增和测序,所得序列已登录到GenBank,登录号为HQ645983,与GenBank中收录的链球菌16S rRNA 基因进行比对并构建系统进化树,结果表明,分离菌株的16S rRNA基因序列与无乳链球菌同源性高达100%,进一步确定分离菌株为无乳链球菌.人工感染显示分离菌株对小白鼠和罗非鱼均具有致病性,对小白鼠的LD50为1.0×104 CFU/mL,对体重为500g±20g的罗非鱼的LD50为1.729×109CFU/mL.分离菌株对氯霉素、青霉素G、呋喃妥因等敏感,对丁胺卡那、链霉素、卡那霉素等不敏感.%One pathogenic bacteria strain HNLFYL4 was isolated from diseased tilapia in Hainan. In this study, bacteria isolation and identification, artificial infection tests were used for analysis of the cause of the severe disease. The isolate was identified as Streptococcus agalactiae based on morphology, cultural performance and biochemical characteristics. The 16S rRNA gene sequence of the isolate was amplified and compared with those of other Streptococcus retrieved from GenBank. The 16S rRNA gene sequence of the isolate had the most homology with that of S. Agalactiae in GenBank. The phylogenetic tree of Streptococcus showed that the isolate was clustered together with S. Agalactiae. 16S rRNA gene sequence analysis further proved that the isolate belonged to S. Agalactiae. The virulent tests were carried out by using the pure culture of the bacteria artificially infected tilapia and mouse. The isolate was pathogenic to tilapia and mouse. LD50 was 1. 0 x 10 CFU/mL to mouse and 1. 729 x 10 CFU/mL to tilapia. Drug sensitivity experiment showed that the isolate was highly sensitive to chloramphenicol, penicillin

  1. 1株致病性兔无乳链球菌的分离、鉴定与系统发育分析%Isolation, identification and phylogenetic analysis of a pathogenic Streptococcus agalactiae in rabbit

    任思宇; 耿毅; 汪开毓; 罔赵英; 刘星星; 吴春艳


    从患病家兔肝、肺、脑和心血中分离到1株链状革兰阳性球菌P110323,以1×106 CFU/mL菌液灌胃感染健康家兔后,表现出站立不稳、四肢划动、惊厥和呼吸障碍等症状,感染96 h后,死亡率为60%.解剖见肺淤血、出血;肝、脾和肾肿大,坏死;大脑肿胀,脑膜充血,脑回肿胀扁平,脑沟变浅.分离菌在BHI平板上形成湿润、边缘整齐的灰白色菌落;在5%的脱纤维羊血平板上形成透明的β溶血环;V-P试验、水解精氨酸与马脲酸等为阳性;水解七叶苷和利用甘露醇、山梨醇和乳糖产酸为阴性.16S rDNA系统发育分析显示分离菌与Streptococcus difficilis和Streptococcus agalactiae聚在一簇.gyrB基因系统发育分析表明分离菌属于无乳链球菌(S.agalactiae),结合细菌形态特点和生理生化特性鉴定该病原菌为无乳链球菌(S.agalactiae),多重PCR方法确定该菌荚膜多糖类型为Ia型.%A pathogenic bacterium (P110323) was isolated from the liver,lung,cerebrum and heart blood of the diseased rabbit.Artificial infection of rabbits with P110323 proved that it was the pathogen of the disease,disease signs and a mortality rate of 60% were noted after 96 h postinoculation with 106 CFU/mL bacterium.The clinical symptoms of infected rabbits included incoordination,lateral prostration,paddling and convulsions with acute respiratory distress syndrome.Necropsy revealed degeneration and necrotic changes in many organs,especially,serious lesions occurred in the liver and kidney.Hemorrhage and swelling of the lung and spleen were considerable;The surfece of gyri was swollen and flattened with the shallow cerebullar sulci,hyperemia of meninges was obvious.The isolated bacterium was a Gram-positive cocci which possessed β-hemolytic activity on agar media with 5% sterile sheep's blood and had moistening and gray colonies in BHI.Arginine,hippuric acid and V-P test were positive; esculin,sorbitol,lactose and mannitol were

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

    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...

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

    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...

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

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


    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.

  5. Human Streptococcus agalactiae strains in aquatic mammals and fish

    Delannoy Christian MJ


    Full Text Available Abstract 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

  6. 齐口裂腹鱼无乳链球菌的分离鉴定及其感染的病理损伤%Isolation, identification and pathogenicity of Streptococcus agalactiae from Schizothorax prenanti

    余泽辉; 张佳; 耿毅; 邓梦玲; 汪开毓; 黄小丽; 陈德芳; 王均


    In 2012-2013, a serious infectious disease characterized by exophthalmia, hemorrhage, and neurological symptoms emerged in Schizothorax prenanti farms in Sichuan Province. A Gram-positive, chain-forming coccus was found in liver and kidney smears. Two strains of Gram-positive, chain-forming cocci were isolated from the diseased fish. The colonies had a smooth-surface and were white, circular, regular and pin-head sized after incubation at 28℃for 48 h on brain heart infusion (BHI). Artificial infection proved that the isolates were the pathogen of the disease. It was primarily identified to be Streptococcus agalactiae according to morphological, physiological and biochemical characteristics. Furthermore, a 16S rRNA gene sequence analysis by BLAST in GenBank indicated that the two isolates showed high levels of similarity to S. agalactiae (more than 96.5%). A phylogenetic tree was constructed comparing the 16S rDNA sequences of both isolates (GenBank accession number KF773744 and KF761304) to other related bacterial species in the GenBank database. In the phylogenetic tree the two isolates and other S. agalactiae strains constituted a branch. In addition, the two isolates were positive in a specific PCR detection of S. agalactiae based on the cfb gene. Based on the phenotypic characteristics, phylogenetic analysis of the 16S rRNA gene, and specific PCR detection, both isolates were identified as S. agalactiae. The sensitivity results showed that both strains were sensitive to doxycycline, amoxicillin, cephalothin V, ofloxacin and levofloxacin, but showed some difference in sensitivity to neomycin and amikacin. Histopathologically, the S. agalactiae infection could cause serious pathological changes in multi-organs and tissues, especially in liver, kidney and brain. The main pathologic lesions were degeneration, necrosis and infiltration of the inflammation cells. Electron microscopical examination found that bacteria could invade hepatocytes, renal cells and

  7. Isolation and Identification of Streptococcus agalactiae from Dairy Cow Mastitis in Qinghai Province%青海地区奶牛无乳链球菌的分离与鉴定



    Streptococcus agalactiae is one of pathogenic bacterium that inducing mastitis of dairy cow. In order to master the popularity and laid the reference for selection of medicine to treat mastitis, 258 samples were collected from 16 dairy cow farms in Qinghai province, and 137 Streptococcus strains were isolated,including Streptococcus agalactiae 42 strains after subculture, gram straining, plasma-coagulase test, CAMP test, biochemical tests and animal pathogenicity tests. The results indicated that Streptococcus agalactiae was the primary pathogenic bacteria of mastitis. All of Streptococcus agalactiaes were hypersensitive to cephradine, and partially resistant to amikacin ofloxacin, ampicillin and sulfanilamide group after K-B medicine sensitivity test.%无乳链球菌是导致奶牛乳房炎的常见病原菌之一,为了掌握青海地区无乳链球菌的流行情况及常用药物的敏感性,从而为本地区更好的预防与治疗奶牛乳房炎提供保障.本试验在青海地区16个奶牛场共采集258份乳房炎乳样,经革兰氏染色与绵羊鲜血琼脂培养基培养纯化,获得疑似链球菌137株;随后经玻片法血浆凝固酶试验、CAMP试验、生化试验与动物致病性试验对无乳链球菌进行鉴定,结果显示,137株链球菌中含有无乳链球菌42株,分离率为30.7%;经K-B纸片扩散法对无乳链球菌耐药性检测结果显示,该地区无乳链球菌主要对头孢类高度敏感,对氧氟沙星、磺胺类、氨苄青霉素与阿米卡星中度敏感.

  8. 内蒙古无乳链球菌耐药性及四环素耐药基因检测%Drug resistance and tetracycline resistance gene identification of Streptococcus agalactiae isolates from bovine in Inner Mongolia

    杜琳; 郝永清


    为了解内蒙古地区隐性乳房炎无乳链球菌分离株耐药性及耐药基因,采集内蒙古不同地区的规模化养殖场隐性乳房炎乳样,采用 Kirby-Bauer(K-B)纸片扩散法测试分离株对16种抗菌药物的敏感性,PCR 方法检测其耐药基因.结果显示:无乳链球菌对大部分抗生素较敏感.对其有较强抑菌作用的药物为:青霉素 G、头孢噻肟、头孢唑啉、阿莫西林、红霉素、环丙沙星、恩诺沙星、氧氟沙星、林可霉素、呋喃妥因、万古霉素.其敏感性达到90%~100%.该菌株对四环素有较强的耐药性,耐药率达77.27%.PCR 扩增四环素耐药基因 tetM 、tetO 、tetK 、tetL ,结果显示22株无乳链球菌均含有 tetM 基因,其基因的检出率为100%.其中7株同时含有 tetK 基因,tetO 、tetL 基因未检出.%In order to understand drug resistance and tetracycline resistance gene of Streptococcus agalactiae epidemic strains,the prevalent strains of S .agalactiae from the main bovine culture area of Inner Mongolia Autonomous Region were detected through Kirby-B auer(K-B)method and PCR.Drug susceptibility testing indicated that the S .agalactiae isolated strains were sensitive to most antibiotics. The S .agalactiae isolates were susceptible to penicillin,cefotaxime,cefazolin,amoxicillin,erythromy-cin,ciprofloxacin,enrofloxacin,ofloxacin,clindamycin,nitrofurantoin,and vancomycin with susceptibili-ties rates of 90% to 100%.In contrast,the S .agalactiae isolates were resistant to tetracycline with re-sistance rates of 77.27%.Four resistance genes including tetM ,tet O ,tetK ,and tetL were characterized by PCR.All strains carried the resistance gene tetM and 7 strains also carried the resistance gene tetK . The resistance genes tetO and tetL were not detected.The results indicated that the resistance gene were related to the resistant type of antimicrobial agents and was important for further research and clinical therapy resistance mechanisms.

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

    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


    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.

  10. 罗非鱼源γ溶血性无乳链球菌的分离鉴定%Isolation and identification of γ-hemolytic streptococcus agalactiae from tilapia

    黎娅; 罗福广; 左跃; 易弋; 伍时华; 杨军


    On September 5, 2012, a disease outbreak was occurred at a tilapia farm in Liuzhou , Guangxi.Aγ-hemolyt-ic gram-positive streptococcus was isolated from brain of infected tilapia .The isolate was identified as rare Streptococcus agalactiae by biochemical assays, PCR identify and 16S rDNA analysis.The strain was γ-hemolytic and negative in CAMP test which showed it was different from normal Streptococcus agalactia strains.The isolate exhibited high virulence to tilapia by experimental infection .Drug sensitivity experiment showed that it was resistant to norfloxacin , polymyxin B , co-trimoxazole and sensitive to 17 types of antibiotics such as penicillin G , et al.%2012年9月5日广西柳州市某罗非鱼养殖场出现暴发性疾病,从患病罗非鱼的脑组织中分离到1株γ溶血性革兰氏阳性链球菌。通过生理生化试验、基于cfb基因的PCR鉴定及16S rDNA序列分析,将该菌株鉴定为无乳链球菌。该菌株为γ溶血性, CAMP试验阴性,与常见无乳链球菌有较大差别。人工感染试验结果显示分离菌株对罗非鱼具有很强的致病性。药物敏感试验结果显示分离菌株对诺氟沙星、多黏菌素B、复方新诺明耐药,对青霉素G等17种药物均敏感。

  11. ISOLASI BAKTERIOFAGA ANTI Streptococcus agalactiae DARI IKAN NILA (Oreochromis niloticus

    Angela Mariana Lusiastuti


    -hemolytic. Streptococcus iniae, a b-hemolytic bacterium is the main causative agent of septicemia, meningoencephalitis, and fish mortality. Moreover, the other causative agent is S. agalactiae. Bacteriophages which are viruses that live on bacteria can be selected and used as therapy for Streptococcosis. The aim of this research is to isolate bacteriophage of S. agalactiae as therapeutic agent candidate giving protecting effect for Streptococcosis. Phages was isolated from Brain Heart Infusion Agar (BHIA were obtained from cultures of S. iniae and S. agalactiae. Strains S. iniae and S. agalactiae were isolated from Streptococcosis fish. The predominant types of colonies were subcultured and subjected to biochemical and physiological tests such as Gram staining, catalase test, hemolytic activity in blood agar and API 20 Strep System. Phages were isolated by using a double agar layer method. A zona lysis with plaques was removed from the plate, centrifuged and the supernatant was filtered through a 0.45 µm membrane filter and stored at 4oC. From 15 isolates of S. iniae and S. agalactiae, only one isolate PSaT-18 showed lysis zone. The lysis zone developed because the phages give protection to S. agalactiae. The lysis zone is not clear because the phage concentration is too low and the dilution in filtration process is too high. Later, these phages were used for invitro and invivo test.

  12. Streptococcus iniae and Streptococcus agalactiae

    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...

  13. Streptococcus agalactiae Native Valve Endocarditis: Uncommon Presentation of Multiple Myeloma.

    Pinho Oliveira, Ana; Delgado, Anne; Martins, Cláudia; Gama, Pedro


    Adults with chronic immunosuppressive conditions are at an increased risk for Streptococcus agalactiae endocarditis, which is typically characterized by acute onset, presence of large vegetations, rapid valvular destruction and frequent complications. We report a rare case of a 74 years old man presenting with fever, renal infarction, ischemic stroke and uveitis. Infective endocarditis was diagnosed and Streptococcus agalactiae was isolated in blood cultures. A multiple myeloma Ig G-K was also diagnosed. The infective endocarditis was successfully treated with a course of benzylpenicillin and gentamicin. The authors highlight the severity of vascular embolic disease present in this case and the diagnostic challenge. They also intend to remind about the association between Streptococcus agalactiae endocarditis and chronic diseases, despite its low reported prevalence.

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

    Lyhs, Ulrike; Kulkas, Laura; Katholm, Jorgen;


    Streptococcus agalactiae is an emerging pathogen of nonpregnant human adults worldwide and a reemerging pathogen of dairy cattle in parts of Europe. To learn more about interspecies transmission of this bacterium, we compared contemporaneously collected isolates from humans and cattle in Finland...

  15. Heterogeneity of Hemolysin Expression during Neonatal Streptococcus agalactiae Sepsis▿

    Sigge, Anja; Schmid, Manuel; Mauerer, Stefanie; Spellerberg, Barbara


    The β-hemolysin of Streptococcus agalactiae is a major virulence factor; consequently, nonhemolytic strains rarely cause infections. We report on a case of neonatal sepsis caused by a strain displaying heterogeneous hemolysin expression. It was detected by the simultaneous isolation of hemolytic and nonhemolytic colonies from cultures of the infant's blood.

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

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


    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

  17. Agalactiae in uncomplicated urinary tract infections

    Katarzyna Jermakow


    Full Text Available Background . Urinary tract infections (UTIs represent 10-20% of all community-acquired infections. Escherichia coli remains the most commonly isolated microorganism. Because of the dominance of these bacteria, routine laboratory tests are not recommended in making the first UTI diagnosis. However, careful monitoring of all etiological agents of ncomplicated UTIs should be an important strategy for empirical therapy. Objectives . A determination of the incidence of UTI bacterial etiological agents in outpatients. Material and methods. The results of urine cultures were analyzed. The samples were obtained in the period from 01.2011 to 03.2013 from outpatients of all age groups. Results . The rods of Escherichia coli were the most frequently identified microorganisms responsible for uncomplicated UTIs. They were isolated from 52% of infected patients. Every fourth case of UTI indicated the participation of Enterococcus spp. And Streptococcus agalactiae . The next 21% of infections were caused by Gram-negative bacteria other than E. coli . The role of coagulase-negative staphylococci was insignificant and equaled less than 2%. Conclusions . Despite the dominant role of E. coli in urinary tract infections, there has been a decrease in the frequency of its isolation. However, the natural resistance of Enterococcus spp. to antibiotics increases the number of urinary tract infections caused by these bacteria. Streptococcus agalactiae is a UTI cause with similar frequency in women of reproductive age and postmenopausal women. Monitoring of the incidence of uncomplicated infections facilitates empirical treatment and prevents UTI recurrence.

  18. Streptococcus agalactiae Serotype Distribution and Antimicrobial Susceptibility in Pregnant Women in Gabon, Central Africa.

    Belard, Sabine; Toepfner, Nicole; Capan-Melser, Mesküre; Mombo-Ngoma, Ghyslain; Zoleko-Manego, Rella; Groger, Mirjam; Matsiegui, Pierre-Blaise; Agnandji, Selidji T; Adegnika, Ayôla A; González, Raquel; Kremsner, Peter G; Menendez, Clara; Ramharter, Michael; Berner, Reinhard


    Neonatal invasive disease due to Streptococcus agalactiae is life threatening and preventive strategies suitable for resource limited settings are urgently needed. Protective coverage of vaccine candidates based on capsular epitopes will relate to local epidemiology of S. agalactiae serotypes and successful management of critical infections depends on timely therapy with effective antibiotics. This is the first report on serotype distribution and antimicrobial susceptibility of S. agalactiae in pregnant women from a Central African region. Serotypes V, III, and Ib accounted for 88/109 (81%) serotypes and all isolates were susceptible to penicillin and clindamycin while 13% showed intermediate susceptibility to erythromycin.

  19. Identification of a novel insertion sequence element in Streptococcus agalactiae.

    Spellerberg, B; Martin, S; Franken, C; Berner, R; Lütticken, R


    Gain and loss of bacterial pathogenicity is often associated with mobile genetic elements. A novel insertion sequence (IS) element designated ISSa4 was identified in Streptococcus agalactiae (group B streptococci). The 963bp IS element is flanked by 25bp perfect inverted repeats and led to the duplication of a 9bp target sequence at the insertion site. ISSa4 contains one open reading frame coding for a putative transposase of 287 aa and exhibits closest similarities to insertion elements of the IS982 family which has previously not been identified in streptococci. Analysis of different S. agalactiae strains showed that the copy number of ISSa4 in S. agalactiae varies significantly between strains. The S. agalactiae strain with the highest copy number of ISSa4 was nonhemolytic and harbored one copy inserted in cylB, which encodes the membrane-spanning domain of the putative hemolysin transporter (Spellerberg et al., 1999. Identification of genetic determinants for the hemolytic activity of Streptococcus agalactiae by ISS1 transposition. J. Bacteriol. 181, 3212-3219). Determination of the distribution of ISSa4 in different S. agalactiae strains revealed that ISSa4 could be detected only in strains isolated after 1996, which might indicate a recent acquisition of this novel insertion element by S. agalactiae.

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

    Boulyana, M.


    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 int...

  1. 2007-2012年中国罗非鱼无乳链球菌流行菌株血清型分析%Serotype of Streptococcus agalactiae isolated from Nile tilapia Oreochromis niloticus in China from 2007 to 2012

    李莉萍; 甘西; 陈明; 王瑞; 黄婷; 梁万文; 雷爱莹; 李健; 黄维义; 唐佳有; 施金谷


    采用PCR血清型鉴定方法,对2007-2012年从广西、广东、海南、福建和云南等地养殖的罗非鱼Oreochromis niloticus中分离获得的168株无乳链球菌Streptococcus agalactiae流行菌株的血清型进行分析。结果表明:有159株为Ⅰa血清型,6株为Ⅰb血清型,3株为Ⅲ血清型。各区域流行菌株血清型也存在差异:从广西自治区7个地区50个养殖场的罗非鱼中分离的61株无乳链球菌中,有54株为Ⅰa型,4株为Ⅰb型,3株为Ⅲ型;从广东省10个地区59个养殖场的罗非鱼中分离的64株无乳链球菌中,有63株为Ⅰa型,1株为Ⅰb型;从海南省5个地区26个养殖场的罗非鱼中分离的33株无乳链球菌中,有32株为Ⅰa型,1株为Ⅰb型;从福建省漳州6个养殖场的罗非鱼中分离的6株和从云南省文山4个养殖场的罗非鱼中分离的4株均为Ⅰa型。研究表明,2007-2012年中国罗非鱼链球菌病流行菌株血清型存在Ⅰa、Ⅰb和Ⅲ3种血清型,Ⅰa型为主要血清型,研究结果可为准确分析中国罗非鱼链球菌病流行规律和特点提供数据,为该病防治及疫苗候选菌株筛选提供理论基础。%PCR typing assay was used to evaluate serotype of 168 Streptococcus agalactiae prevalent strains isolated from Nile tilapia Oreochromis niloticus in Guangxi,Guangdong,Hainan,Fujian and Yunnan provinces in China from 2007 to 2012. There were 159 strains ofⅠa,6 strains ofⅠb, and 3 strains of Ⅲ, with differences in serotypes of isolates from different regions. In the 61 strains isolated from 50 farms in 7 regions in Guangxi,54 strains wereⅠa, 4 strains Ⅰb, and 3 strains Ⅲ. There were 64 strains isolated from 59 farms in 10 regions in Guangdong, with 63 strains of Ⅰa,1 strain of Ⅰb. Thirty-three strains were isolated from 26 farms in 5 regions in Hannan, including 32 strains of Ⅰa, and 1 strains ofⅠb. Four strains ofⅠa were isolated from 6 farms in Zhangzhou in Fujian and Wenshan in Yunnan

  2. Streptococcus agalactiae infection in tilapia (Oreochromis niloticus

    Suanyuk, N.


    Full Text Available Streptococcus agalactiae was isolated from cultured tilapia in Surat Thani province. Isolates were Gram-positive cocci, catalase negative, alpha-haemolytic and serogroup B. Streptococcal-infected fish showed various swimming abnormalities such as swimming on their side, erratic surface or bottom swimming including serpentine movement, exophthalmia and opacity. Internally, splenomegaly, ascites as well as pale liver discoloration were observed. Fish experimentally infected by peritoneal injection using 101 -108 CFU/ fish showed 20-90% mortality within 10 days and the LD50 was 3.60x101 - 1.72x107 CFU. Haematocrit, haemoglobin, plasma protein and blood cell values of infected and moribund fish were significantly decreased. Histopathological findings included the occurrence of inflammation, cells necrosis, infiltration of lymphocytes and the formation of granulomas in the infected organs.

  3. Clinical distribution and drug resistance of 179 isolates of Streptococcus agalactia%179株无乳链球菌的标本分布及耐药性分析

    林宇岚; 黄尔; 欧启水; 杨滨; 陈守涛; 荀振; 张泽佳; 闫翔宇; 郭少英; 留玉芬; 高丽钦; 甘龙杰


    Objective To investigated the clinical distributions and antimicrobial susceptibility of Streptococcus agalactia strains isolated from the patients .Methods The identification and susceptibility of the strains were mainly measured by automatic VITEK‐Ⅱ system ,the K‐B disc diffusion tests were used for the resistance test of erythromycin ,meropenem ,and D‐test .Results The iso‐lates were mainly from urine (63 .1% ) ,genital tract(7 .8% ) and wound secretion(6 .7% ) .They were obtained from patients in dif‐ferent situations ,including 110 patients who were older than 50 years old (61 .5% ) ,113 female patients (63 .1% ) ,12 gravidas (6 .7% ) ,3 vertical transmitted newborns(1 .7% ) ,and 82 patients with cancer ,undergoing chemo radiotherapy ,with diabetes ,tuber‐culosis or after operations(45 .8% ) .The resistant rates of the isolated Streptococcus agalactia to erythromycin and clindamycin were 42 .9% -93 .3% and 41 .9% -80 .0% respectively .The positive rate of D‐test was 4 .1% .The strains were highly resistant to tet‐racycline(>80% ) ,while the resistance to penicillin was below 10% except in 2008 .All isolates were susceptible to vancomycin and meropenem .Only one strain was resistant to Quinupristin‐dalfopristin .Conclusion Streptococcus agalactia infection in adults most‐ly cause genitourinary tract ,skin and soft tissue infections .There were more females than males with Streptococcus agalactia infec‐tion .Penicillin andβ‐lactams are still the first choice for the treatment .Erythromycin ,clindamycin and tetracycline should be used with caution under the guidance of laboratory susceptibility test results .%目的:分析临床分离无乳链球菌的标本分布和对常用抗菌药物的耐药性。方法收集不同来源标本,在VIT EKⅡ微生物分析仪上采用GPI卡和GP67卡进行菌种鉴定和药敏实验,红霉素、美罗培南的药敏实验及D‐试验采用K‐B纸片法检测。

  4. Drug resistance analysis on the drug resistance of Streptococcus agalactiae isolated from female urogenital tract%女性泌尿生殖道无乳链球菌的耐药性分析

    高晶; 刘晓艳


    目的:对女性泌尿生殖道分离的无乳链球菌对常用抗菌药物的耐药情况做回顾性分析,了解常用抗菌药物的耐药趋势变化,指导临床合理使用抗菌药物。方法对2009年1月至2012年12月送检的女性泌尿生殖道标本进行常规培养,使用VITEK32全自动细菌鉴定分析仪对分离出的细菌进行鉴定和药敏试验,同时对红霉素耐药、克林霉素敏感或中介的无乳链球菌菌株做D-抑菌圈试验。结果妇科患者检出569株无乳链球菌,产科患者检出192株无乳链球菌,二者年龄差异有统计学意义(P<0.01)。761株无乳链球菌中无万古霉素耐药菌株,青霉素和利奈唑胺耐药率均<3%;呋喃妥因的耐药率2009年至2011年均<0.5%,2012年为7.2%;四环素耐药率>90%;红霉素、克林霉素及左氧氟沙星的耐药率较高,且呈逐年上升趋势;D-抑菌圈试验阳性率为21.7%。结论临床须根据体外药敏试验结果合理使用抗菌药物,同时须加强耐药性监测。%Objective To analyze retrospectively the drug resistance of Streptococcus agalactiae isolated from female urogenital tract,to understand the drug resistance tendency change of commonly-used antibiotics,and to guide the rational use of antibiotics in clinical application.Methods The routine culture was performed for female urogenital tract specimens collected from January 2009 to December 2012,and the identification and drug sensitivity test were performed for Streptococcus agalactiae by VITEK32 automatic bacterium identification analyzer.The D-inhibition zone trial was tested for isolates which were drug-resistant to erythromycin and sensitive or intermediary to clindamycin. Results There was a statistical significance in age distribution among 569 specimens from Department of Gynecology and 192 specimens from Department of Obstetrics (P<0.01).Drug resistance rate was 0 to vancomycin and <3% to penicillin

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

    Esti Handayani Hardi


    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.

  6. Molecular epidemiology of Streptococcus agalactiae recovered from significant bacteriuria.

    Schuab, Rôde B B; Arêas, Glauber P; Souza, Viviane C; Barros, Rosana R


    Streptococcus agalactiae has been implicated in urinary tract infections, but the molecular epidemiology of such infections is poorly characterized. In this study, 194 isolates recovered from significant bacteriuria of non-pregnant individuals were submitted to antimicrobial susceptibility testing, molecular characterization of macrolide resistance, PCR-based capsular typing and analysis of genetic diversity by pulsed-field gel electrophoresis (PFGE). By disk diffusion, all isolates were susceptible to ceftriaxone, levofloxacin, penicillin G and vancomycin; 87.6% and 9.3% of isolates were non-susceptible to tetracycline and clindamycin, respectively. The minimum inhibitory concentration (MIC) confirmed that 11.3% of isolates were resistant to erythromycin. Macrolide resistance determinants were iMLSB (n = 9), cMLSB (n = 9) and M (n = 4), associated with ermA, ermB and mefA/E. Predominant capsular types were V, Ia, II and III. No significant association was observed between any capsular type and the occurrence of pyuria. However, type III was associated with erythromycin resistance, while type II was associated with erythromycin-susceptible isolates. Distinct PFGE profiles were observed among different types, but identical profiles were found among erythromycin-susceptible and -resistant isolates of the same type. A variety of capsular and PFGE types are involved in significant bacteriuria. Although capsular types found here are prevalent in different infections, the frequency of each type seems to be unique. Erythromycin resistance is due to polyclonal origin instead of the expansion of few clones of S. agalactiae.

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

    M. Boulyana


    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.

  8. Complete Genome Sequence of Streptococcus agalactiae CNCTC 10/84, a Hypervirulent Sequence Type 26 Strain

    Hooven, Thomas A.; Randis, Tara M.; Daugherty, Sean C.; Narechania, Apurva; Planet, Paul J.; Tettelin, Hervé; Ratner, Adam J.


    Streptococcus agalactiae (group B Streptococcus [GBS]) is a human pathogen with a propensity to cause neonatal infections. We report the complete genome sequence of GBS strain CNCTC 10/84, a hypervirulent clinical isolate frequently used to study GBS pathogenesis. Comparative analysis of this sequence may shed light on novel pathogenic mechanisms.

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

    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


    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.

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

    Lutfiana Safitri


    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.

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

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


    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.

  12. Optimal Culture Conditions and Medium of Streptococcus agalactiae Isolated from Tilapia%罗非鱼源无乳链球菌培养基及培养条件的优化

    陈贺; 吴灶和; 王蓓; 黄郁葱; 鲁义善; 简纪常


    The culture and the characteristics of growth of Streptococcus agalactiae (ZJGY08) were studied as the model strain. The effects of different factors, including nutritional factors, initial pH value, dissolved oxygen and temperature, on its growth in the fluid culture medium named Brian Heart Infusion (BHI) were explored to optimize its growth parameters. The results indicated that the initial pH 7 - 8 and the temperatures 28 - 37 degrees and the shake speed 180 - 200 r/min were optimal for the Streptococcus agalactiae. The maximal bacteria's concentration could be got by supplying 20 mg/mL yeast powder and 2 mg/mL glucose in the BHI.%研究无乳链球菌(Streptococcus agalactiae)ZQGY08的培养与生长特性,探讨脑心浸液(Brian Heart Infusion,BHI)培养基的营养因子、初始pH值、溶氧、温度等因素对无乳链球菌生长的影响,并采用正交实验法对无乳链球菌培养基营养因子添加量进行优化.结果表明,无乳链球菌生长的最适初始pH为7~8,培养温度为28 ~ 37℃,振荡转速为180~ 200 r/min.在BHI培养基中添加20 mg/mL的酵母粉和2 mg/mL的葡萄糖即可在对数期获得最大细菌量.

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

    Laith, AA; 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


    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, hyperemic and

  14. Transcriptomic and genomic evidence for Streptococcus agalactiae adaptation to the bovine environment


    Background Streptococcus agalactiae is a major cause of bovine mastitis, which is the dominant health disorder affecting milk production within the dairy industry and is responsible for substantial financial losses to the industry worldwide. However, there is considerable evidence for host adaptation (ecotypes) within S. agalactiae, with both bovine and human sourced isolates showing a high degree of distinctiveness, suggesting differing ability to cause mastitis. Here, we (i) generate RNAseq data from three S. agalactiae isolates (two putative bovine adapted and one human) and (ii) compare publicly available whole genome shotgun sequence data from an additional 202 isolates, obtained from six host species, to elucidate possible genetic factors/adaptations likely important for S. agalactiae growth and survival in the bovine mammary gland. Results Tests for differential expression showed distinct expression profiles for the three isolates when grown in bovine milk. A key finding for the two putatively bovine adapted isolates was the up regulation of a lactose metabolism operon (Lac.2) that was strongly correlated with the bovine environment (all 36 bovine sourced isolates on GenBank possessed the operon, in contrast to only 8/151 human sourced isolates). Multi locus sequence typing of all genome sequences and phylogenetic analysis using conserved operon genes from 44 S. agalactiae isolates and 16 additional Streptococcus species provided strong evidence for acquisition of the operon via multiple lateral gene transfer events, with all Streptococcus species known to be major causes of mastitis, identified as possible donors. Furthermore, lactose fermentation tests were only positive for isolates possessing Lac.2. Combined, these findings suggest that lactose metabolism is likely an important adaptation to the bovine environment. Additional up regulation in the bovine adapted isolates included genes involved in copper homeostasis, metabolism of purine, pyrimidine

  15. Expression and Immunogenicity Analysis of Hemolysin Gene in Streptococcus agalactiae Isolated from Oreochromis niloticus%罗非鱼源无乳链球菌溶血素基因体外表达及其免疫原性

    王蓓; 李桂欢; 王培; 汤菊芬; 鲁义善; 吴灶和; 简纪常


    Streptococcus is a genus of spherical gram-positive bacteria that can infect a wide range of hosts. Streptococcal diseases of cultured fish caused by Streptococcus agalactiae have been frequently reported in South China, which incurred catastrophic economic losses. Vaccine is one of effective ways to control and eradicate infectious diseases, and so far no effective subunit vaccine for S. agalactiae diseases has been developed. To test whether Hemolysin (Hly) is a potential candidate for vaccine development of Oreochromis niloticus, we cloned hly gene based on bioinformatics analysis and constructed and expressed it. The open reading frame of hly gene contains 1335 bp that encodes 444 amino acid residues. Alignment analysis indicated that the Hly protein was highly homologous to Hly proteins from other Streptococcus(99%). SDS-PAGE confirmed the expression of Hly (51.7 kD) in Escherichia coli BL21(DE3). Then, the recombinant Hly protein was purified by affinity chromatography, which was used for vaccination in Tilapia, O. niloticus. Immunogenicity was confirmed by subsequent western blotting. Enzyme-linked immunosorbent assay (ELISA) analysis demonstrated that Hly produced an observable antibody response in all vaccinated fish, and the maximum antibody titers in the sera reached 1:4096. The RPS value for the Sip vaccine was 70. Tilapia vaccinated with Hly was highly resistant to the infection of the contaminant S. agalactiae. These results indicate that Hly is an effective vaccine candidate against S. agalactiae for tilapia, O. niloticus.%为研究罗非鱼源无乳链球菌溶血素(Hemolysin, Hly)对鱼体的免疫保护作用,根据已获得的无乳链球菌ZQ0910全基因组序列设计引物扩增hly基因,定向克隆于原核表达载体pET-28a中,构建原核重组质粒pET-28a-hly,经IPTG诱导表达后,制成亚单位疫苗免疫吉富罗非鱼,并分析疫苗的免疫保护力。结果显示,hly基因产物大小1335 bp,编码444个氨

  16. Molecular detection of Streptococcus agalactiae in bovine raw milk samples obtained directly from bulk tanks.

    Elias, Acácia Orieth; Cortez, Adriana; Brandão, Paulo Eduardo; da Silva, Rodrigo Costa; Langoni, Helio


    Mastitis is the most common infectious disease affecting dairy cattle; in addition, it remains the most economically important disease of dairy industries around the world. Streptococcus agalactiae, a contagious pathogen associated with subclinical mastitis, is highly infectious. This bacterium can cause an increase in bulk tank bacterial counts (BTBC) and bulk tank somatic cell counts (BTSCC). The microbiological identification of S. agalactiae in samples from bulk tanks is an auxiliary method to control contagious mastitis. Thus, there are some limitations for time-consuming cultures or identification methods and additional concerns about the conservation and transport of samples. Bulk tank samples from 247 dairy farms were cultured and compared through polymerase chain reaction (PCR), directed to 16S rRNA genes of S. agalactiae, followed by BTBC and S. agalactiae isolation. The mean value of BTBC was 1.08×10(6) CFU mL(-1) and the bacterium was identified through the microbiological method in 98 (39.7%; CI(95%)=33.8-45.9%) and through PCR in 110 (44.5%; CI(95%)=38.5-50.8%) samples. Results indicated sensitivity of 0.8571±0.0353 (CI(95%)=0.7719-0.9196) and specificity of 0.8255±0.0311 (CI(95%)=0.7549-0.8827). The lack of significant difference between microbiological and molecular results (κ=0.6686±0.0477 and CI(95%)=0.5752-0.7620) indicated substantial agreement between the methods. This suggests that PCR can be used for bulk tank samples to detect contagious mastitis caused by S. agalactiae.

  17. Could β-hemolytic, group B Enterococcus faecalis be mistaken for Streptococcus agalactiae?

    Savini, Vincenzo; Gherardi, Giovanni; Marrollo, Roberta; Franco, Alessia; Pimentel De Araujo, Fernanda; Dottarelli, Samuele; Fazii, Paolo; Battisti, Antonio; Carretto, Edoardo


    A β-hemolytic Enterococcus faecalis strain agglutinating Lancefield group A, B, C, D, F, and G antisera was observed from a rectovaginal swab, in the context of antenatal screening for Streptococcus agalactiae (group B Streptococcus [GBS]). This is the first multi-Lancefield antisera-agglutinating isolate of this species, and it raised particular concern, as it may mimic GBS, leading to false reporting and useless receipt of intrapartum antibiotics.

  18. SEQUENCE ANALYSIS OF Streptococcus agalactiae, A PATHOGEN CAUSING STREPTOCOCCOSIS IN TILAPIA (Oreochromis niloticus

    Angela Mariana Lusiastuti


    Full Text Available Pathogen identification based on biochemical properties can barely differentiate Streptococcus iniae and S. agalactiae. Beside that, this technique is also limited by the length of time required to complete the assays. Therefore, rapid diagnosis is necessary to initiate prompt therapeutic and prophylactic measures in order to limit any potential economic losses caused by such pathogens. The aim of the present study was to identify Streptococcosis species using amplification of S. agalactiae DNA sequence with species-specific primer Sdi 61 AGGAAACCTGCCATTTGCG and Sdi 252 CAATCTATTTCTAGATCGTGG and perform phylogenetic analysis based on DNA nucleotide sequence data. The sequencing of PCR products was performed at BPPT Puspiptek Serpong by using the respective PCR primers, Big Dye Terminator Chemistry and AmpliTaq-FS DNA polymerase. The sequencing reactions were run on the ABI Prism version 3103 – Avant Genetic Analyzer (USA and the result was read by Sequence Navigator program (Applied Biosystem. Alignment multiple analysis was done based on the data from Genebank with BLASTN ( on the nucleotide level. Neighbor-joining phylogenetic trees were generated with Genetyx programme version 7 with UPGMA and MEGA software version 4.0. The result revealed that the isolates from brain, eye, and kidney of diseased Tilapia were infected by S. agalactiae and it has 99% similarity with Genebank. It has close relationship with S. agalactiae at genebank with UPGMA method. These isolates showed high variation in the first sequence which is similar to S. iniae. The information of S. agalactiae genomes suggests that gene acquisition, duplication, and reassortment have played an important role in genetic diversity and evolution of S. agalactiae. Screening of breeder fish stocks with the developed PCR methodology, followed by elimination of infected stocks, would provide an efficient strategy to control fish

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

    Manuel Díaz Álvarez


    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.

  20. 宝石鲈无乳链球菌的分离鉴定及耐药性分析%Isolation, Identification and Drug Susceptibility Analysis of Streptococcus agalactiae in Scortum barcoo

    黄木珍; 黎炯; 潘忠超; 石和荣


    2013年高温季节在广东广州2个养殖场患病宝石鲈(Scortum barcoo)病灶处共分离到2株细菌Py1和Py2.宝石鲈攻毒试验结果显示2株分离菌均具有较强毒力(半致死量分别为7.42×106和1.72×106 CFU/mL),且都对罗非鱼具有高度感染性(半致死量分别为3.39×106和8.66×105 CFU/mL).经ATB生化鉴定及cfb基因序列分析,确定分离株为无乳链球菌(Streptococcus agalactiae),并对其进行了相关药敏试验检测,以期为该病的预防提供参考.

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

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


    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 < 0.01), and the antibody reached their maximum levels 14-21 days after the immunization but decreased significantly after 28 days of vaccination. YM001 bacteria were isolated from the brain, liver, kidney, and spleen tissues of fish after oral immunization and the bacteria existed for the longest time in the spleen (up to 15 days). Taken together, this study obtained a safe, stable, and highly

  2. Mycoplasma agalactiae, an Etiological Agent of Contagious Agalactia in Small Ruminants: A Review

    Amit Kumar


    Full Text Available Mycoplasma agalactiae is one of the causal agents of classical contagious agalactia (CA, a serious, economically important but neglected enzootic disease of small ruminants. It occurs in many parts of the world and most notably in the Mediterranean Basin. Following the infection common complications are septicaemia, mastitis, arthritis, pleurisy, pneumonia, and keratoconjunctivitis. Primary or tentative diagnosis of the organism is based upon clinical signs. Various serological tests, namely, growth precipitation, immunofluorescence, complement fixation test, haemagglutination inhibition, agglutination, immunodiffusion, enzyme immunoassays, immunoelectrophoresis, blotting techniques, and others, are available. Molecular tools seem to be much more sensitive, specific, and faster and help to differentiate various strains. The real-time PCR, multiplex PCR, quantitative PCR, PCR-RFLP, MLST, and gene probes, complementary to segments of chromosomal DNA or 16S ribosomal RNA (rRNA, have strengthened the diagnosis of M. agalactiae. Both live attenuated and adjuvant (alum precipitated or saponified inactivated vaccines are available with greater use of inactivated ones due to lack of side effects. The present review discusses the etiology, epidemiology, pathogenesis, and clinical signs of contagious agalactia in small ruminants along with trends and advances in its diagnosis, treatment, vaccination, prevention, and control strategies that will help in countering this disease.

  3. Antimicrobial Susceptibility of Streptococcus agalactiae Isolated from Dairy Cows with Clinical Mastitis in Tongliao City of Inner Mongolia%内蒙古通辽市某奶牛场乳房炎无乳链球菌的分离、鉴定及药敏试验



    旨在调查引起内蒙古通辽地区奶牛乳房炎发生的优势病原菌。采用常规方法对采集自该地区某奶牛场的3份临床型乳房炎患牛奶样进行优势致病菌的分离鉴定,采用纸片扩散法(K-B法)评价分离菌对兽医临床常用抗菌药物的敏感性。结果表明,从3份乳样中共分离到14株菌,经生化试验鉴定为无乳链球菌;药敏试验结果表明,分离菌对链霉素及磺胺嘧啶表现出较强的耐药性,而卡那霉素、庆大霉素及麦迪霉素对分离菌则表现出较好的抑菌效果。%The aim of the present study was to determine the dominant species of bacteria associated with bovine mastitis in Tongliao City of Inner Mongolia. Three quarter milk samples were collected from dairy cows with clinical mastitis and were used to isolate pathogenic bacteria by conventional method. The antimicrobial susceptibility test of the isolates was conducted using Kirby-Bauer disk diffusion method. The results revealed that a total of 14 strains were isolated from the 3 samples and they were subsequently phenotypically identified as Streptococcus agalactiae. While reduced susceptibility to streptomycin and sulfadiazine were observed, kanamycin, gentamicin and midecamycin are found as susceptible drugs for the isolated Streptococcus strains.

  4. Molecular characterization of Streptococcus agalactiae from vaginal colonization and neonatal infections: a 4-year multicenter study in Greece.

    Liakopoulos, Apostolos; Mavroidi, Angeliki; Vourli, Sofia; Panopoulou, Maria; Zachariadou, Levantia; Chatzipanagiotou, Stylianos; Spiliopoulou, Iris; Zerva, Loukia; Petinaki, Efthymia


    A multicenter collection comprising of 171 Streptococcus agalactiae isolates from pregnant women recovered between 2007 and 2010 and 46 from unmatched neonates with invasive infections was subjected to antimicrobial susceptibility testing and genetic characterization. High rates of erythromycin resistance (20.47%) were observed only in isolates from pregnant women. ST1 was dominant in the vaginal colonization, whereas the hypervirulent ST-17 clone was detected in 67.39% of neonatal infections.

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

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


    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.

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

    Mereghetti Laurent


    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.

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

    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.

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

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


    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.

  9. The liposoluble proteome of Mycoplasma agalactiae: an insight into the minimal protein complement of a bacterial membrane

    Cacciotto Carla


    Full Text Available Abstract Background Mycoplasmas are the simplest bacteria capable of autonomous replication. Their evolution proceeded from gram-positive bacteria, with the loss of many biosynthetic pathways and of the cell wall. In this work, the liposoluble protein complement of Mycoplasma agalactiae, a minimal bacterial pathogen causing mastitis, polyarthritis, keratoconjunctivitis, and abortion in small ruminants, was subjected to systematic characterization in order to gain insights into its membrane proteome composition. Results The selective enrichment for M. agalactiae PG2T liposoluble proteins was accomplished by means of Triton X-114 fractionation. Liposoluble proteins were subjected to 2-D PAGE-MS, leading to the identification of 40 unique proteins and to the generation of a reference 2D map of the M. agalactiae liposoluble proteome. Liposoluble proteins from the type strain PG2 and two field isolates were then compared by means of 2D DIGE, revealing reproducible differences in protein expression among isolates. An in-depth analysis was then performed by GeLC-MS/MS in order to achieve a higher coverage of the liposoluble proteome. Using this approach, a total of 194 unique proteins were identified, corresponding to 26% of all M. agalactiae PG2T genes. A gene ontology analysis and classification for localization and function was also carried out on all protein identifications. Interestingly, the 11.5% of expressed membrane proteins derived from putative horizontal gene transfer events. Conclusions This study led to the in-depth systematic characterization of the M. agalactiae liposoluble protein component, providing useful insights into its membrane organization.

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

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


    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.

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

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


    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.

  12. Prokaryotic expression and immunogenicity analysis of C5a peptidase (ScpB) of Streptococcus agalactiae isolated from tilapia%罗非鱼无乳链球菌C5a肽酶(ScpB)的原核表达及其免疫原性

    李庆勇; 可小丽; 卢迈新; 朱华平; 高风英; 刘志刚


    Streptococcus agalactiae is a major bacterial pathogen that has caused severe economic losses in many spe-cies of freshwater, marine and estuarine fish worldwide. ScpB is a highly conserved surface protein among all group B streptococcus (GBS) strains and is an attractive surface-exposed antigen for inclusion among vaccine components against GBS. In this study, the scpB gene was amplified from the genomic DNA of an S. agalactiae strain isolated from diseased tilapia farmed in Guangdong province, China. The scpB gene contains a 2 799 bp open reading frame (ORF), encoding 932 amino acids. The molecular mass of the deduced protein was 103 kD. Blast analysis showed that it shares high similarity with scpB sequences of human GBSs registered in GenBank. Prokaryotic expression vector pET32a (+) was used to construct a recombinant expression vector pET32a (+)-scpB, which was transformed into Escherichia coli BL21 (DE3). Colonies containing the recombinant expression vector pET32a (+)-scpB were selected and then induced to express using 0.5 mmol/L IPTG for 8 h at 37℃. The expressed recombinant protein was purified by nickel chelate affinity chromatography and ultrafiltration tube enrichment. SDS-PAGE analysis and western blotting showed a spe-cific protein band of about 121 kD. The recombinant strain could produce large amounts of ScpB protein, mainly in the form of an inclusion body. To analyze the immunogenicity of the recombinant protein, the purified fusion protein and Freund’s adjuvant were mixed according to a certain proportions to produce vaccines. Three immune dosages, 1μg/g(F1), 3μg/g(F3) and 5μg/g(F5) were used. Four weeks after immunization, tilapia were challenged by artificial infection with the GBS ZP-N strain, which was previously isolated and confirmed to be a tilapia pathogen by our labo-ratory. The recorded relative percent survivals (RPS) of the vaccinated groups ranged from 69.99%to 89%, of which group F5 has the highest RPS. The lysozyme

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

    Jafarizadeh Amirreza


    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.

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

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


    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.

  15. [The Bayesian framework of detection of cariesgenic Streptoccocus in dental plaque in children with distal occlusion under orthodontic treatment].

    Chesnokov, V A; Chesnokova, M G; Mironov, A Iu; Turchaninov, D V; Kriga, A S


    The application of Bayes theorem in medical diagnostic includes such important step as derivation for every symptom and diagnosis data values of finite or a posteriori probability of presence of germ, for instance S. sanguis, in patients receiving orthodontic treatment. This value expresses importance of the given symptoms for detection of germ presence. In the implemented studies the Bayes theorem was applied to evaluate probability of presence of particular germ in particular concentration (degree of semination, CO) under concrete symptom group. The rates were used to calculate probability of presence of cariesgenic streptococcus S. mutans and S. sanguis in prospect patient. The rates were calculated for the group with fixed orthodontic apparatuses. The high degree of risk of development of caries in children under orthodontic treatment is conditioned by a whole complex of existing unfavorable factors present in oral cavity. Hence, a powerful negative potential to develop expressed cariesgenic situation in oral cavity is present. The analysis of medical data of patients with distal occlusion was applied using Bayes theorem from the point of view their diagnostic value. The study established 36 symptoms, factors, risk factors and background diseases common in case of distal occlusion in children. The probability to detect the mentioned characteristics and likelihood ratio under different degree of concentration of cariesgenic Streptococcus mutans and Streptococcus sanguis in dental plaque of children was considered. The establishment of character of likelihood ratio for different qualitative content of streptococcus in case of isolation from biotope of dental plaque permitted to determine the informative characteristics. The Bayesian model can be applied in clinics as a computer program to process incoming information about patients with distal occlusion under active orthodontic treatment. The developed diagnostic algorithm in a fast and simple mode of clinical

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

    Franciane Rios Senger


    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.

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

    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...

  18. Clinical analysis of cases of neonatal Streptococcus agalactiae sepsis.

    Zeng, S J; Tang, X S; Zhao, W L; Qiu, H X; Wang, H; Feng, Z C


    With the advent of antibiotic resistance, pathogenic bacteria have become a major threat in cases of neonatal sepsis; however, guidelines for treatment have not yet been standardized. In this study, 15 cases of neonatal Streptococcus agalactiae sepsis from our hospital were retrospectively analyzed. Of these, nine cases showed early-onset and six cases showed late-onset sepsis. Pathogens were characterized by genotyping and antibiotic sensitivity tests on blood cultures. Results demonstrated that in cases with early-onset sepsis, clinical manifestations affected mainly the respiratory tract, while late-onset sepsis was accompanied by intracranial infection. Therefore, we suggest including a cerebrospinal fluid examination when diagnosing neonatal sepsis. Bacterial genotyping indicated the bacteria were mainly type Ib, Ia, and III S. agalactiae. We recommend treatment with penicillin or ampicillin, since bacteria were resistant to clindamycin and tetracycline. In conclusion, our results provide valuable information for the clinical treatment of S. agalactiae sepsis in neonatal infants.

  19. Casereport - Agalactia of mare, agammaglobulinemia and arthritis in foal

    Spasojević Jovan


    Full Text Available Lactation is physiological state of the organism and the final process of the female reproductive cycle. Milk in the first days after birth (colostrum, in addition to the necessary nutrients contains antibodies, with whose ingesting only a newborn individual receives passive immunity that protects it from various infections over time. Mammary gland dysfunction and a lack of breast milking is called agalactia. Due to the occurrence of agalactia in mother, newborn animal is denied of intake of colostrum in its body. Thus prevents ingestion of nutrients and passive immunity, which results in the occurrence of various diseases especially infectious etiology. This paper describes the treatment of agammaglobulinemia in foal after ascertaining the occurrence of primary agalactia in mare. There is described the possibility of substitution, ie. benefits of breast milk substitutes, and the procedure of diagnosis and treatment of carpal arthritis in foal.

  20. Neonatal necrotizing fasciitis of the scrotum caused by Streptococcus agalactiae.

    Kuroda, Junpei; Inoue, Nobuaki; Satoh, Hiroyuki; Fukuzawa, Ryuji; Terakawa, Toshiro; Hasegawa, Yukihiro


    We herein describe the case of a 27-day-old male infant who was brought to the emergency room for intermittent crying, and swelling of the left scrotum. Based on the clinical findings, necrotizing fasciitis was suspected, and surgical intervention was successfully completed within a few hours of admission. Streptococcus agalactiae type Ia was cultured from the drained abscess, and was considered the causative pathogen. To our knowledge, this is the first report of neonatal necrotizing fasciitis caused by S. agalactiae. Prompt diagnosis and immediate surgical debridement are crucial in the initial management of this disease.

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

    Elhaig, Mahmoud Mohey; Selim, Abdelfattah


    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.

  2. Cross-Resistance to Lincosamides, Streptogramins A, and Pleuromutilins Due to the lsa(C) Gene in Streptococcus agalactiae UCN70▿

    Malbruny, Brigitte; Werno, Anja M.; Murdoch, David R.; Leclercq, Roland; Cattoir, Vincent


    Streptococcus agalactiae UCN70, isolated from a vaginal swab obtained in New Zealand, is resistant to lincosamides and streptogramins A (LSA phenotype) and also to tiamulin (a pleuromutilin). By whole-genome sequencing, we identified a 5,224-bp chromosomal extra-element that comprised a 1,479-bp open reading frame coding for an ABC protein (492 amino acids) 45% identical to Lsa(A), a protein related to intrinsic LSA resistance in Enterococcus faecalis. Expression of this novel gene, named lsa(C), in S. agalactiae BM132 after cloning led to an increase in MICs of lincomycin (0.06 to 4 μg/ml), clindamycin (0.03 to 2 μg/ml), dalfopristin (2 to >32 μg/ml), and tiamulin (0.12 to 32 μg/ml), whereas no change in MICs of erythromycin (0.06 μg/ml), azithromycin (0.03 μg/ml), spiramycin (0.25 μg/ml), telithromycin (0.03 μg/ml), and quinupristin (8 μg/ml) was observed. The phenotype was renamed the LSAP phenotype on the basis of cross-resistance to lincosamides, streptogramins A, and pleuromutilins. This gene was also identified in similar genetic environments in 17 other S. agalactiae clinical isolates from New Zealand exhibiting the same LSAP phenotype, whereas it was absent in susceptible S. agalactiae strains. Interestingly, this extra-element was bracketed by a 7-bp duplication of a target site (ATTAGAA), suggesting that this structure was likely a mobile genetic element. In conclusion, we identified a novel gene, lsa(C), responsible for the acquired LSAP resistance phenotype in S. agalactiae. Dissection of the biochemical basis of resistance, as well as demonstration of in vitro mobilization of lsa(C), remains to be performed. PMID:21245447

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

    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


    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.

  4. Characterization of Afb, a novel bifunctional protein in Streptococcus agalactiae

    Sanaz Dehbashi


    Full Text Available Background and Objectives: Streptococcus agalactiae is the leading cause of bacterial sepsis and meningitis in newborns and results in pneumonia and bacteremia in adults. A number of S. agalactiae components are involved in colonization of target cells. Destruction of peptidoglycan and division of covalently linked daughter cells is mediated by autolysins. In this study, autolytic activity and plasma binding ability of AFb novel recombinant protein of S. agalactiae was investigated.Materials and Methods: The gbs1805 gene was cloned and expressed. E. coli strains DH5α and BL21 were used as cloning and expression hosts, respectively. After purification, antigenicity and binding ability to plasma proteins of the recombinant protein was evaluated.Results: AFb, the 18KDa protein was purified successfully. The insoluble mature protein revealed the ability to bind to fibrinogen and fibronectin. This insoluble mature protein revealed that it has the ability to bind to fibrinogen and fibronectin plasma proteins. Furthermore, in silico analysis demonstrated the AFb has an autolytic activity.Conclusions: AFb is a novel protein capable of binding to fibrinogen and fibronectin. This findings lay a ground work for further investigation of the role of the bacteria in adhesion and colonization to the host. Keywords: Streptococcus agalactiae, Fibrinogen, Fibronectin, Autolysin

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

    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.

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

    Haoqin Jiang


    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.

  7. Infection and pathology in Queensland grouper, Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes.

    Delamare-Deboutteville, J; Bowater, R; Condon, K; Reynolds, A; Fisk, A; Aviles, F; Barnes, A C


    Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery-reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re-isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high-dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.

  8. Identification and serotyping of Streptococcus agalactiae by PCR%无乳链球菌及其血清型的PCR方法鉴定

    汤炜; 周雪; 于立权; 吴志军; 朱战波; 崔玉东


    Streptococcus agalactiae is an important pathogen that causes bovine mastitis and life-threatening invasive infections (septicemia,pneumonia,and meningitis) during the neonatal period.To rapidly identify capsular serotypes of S.agalactiae,a total of 22 bovine Streptococcus isolates from different regions of China were typed by biochemical tests and multiplex PCR to amplify the conserved sip and cps genes of the S.agalactiae.The results shown 17 of the 22 isolates were positive for sip gene,whereas,15 of the 17 S.agalactiae were positive identified by biochemical tests.Furthermore,the multiplex PCR also revealed that among the 17 isolates,the cps gene serotypes were 4 of Ia,11 of Ⅱ and 2 of Ⅲ,repectively.This study may provide a reference for identification and serotyping of S.agalactiae.%无乳链球菌(S.agalactiae)是引起奶牛乳房炎和新生儿期侵袭性感染(包括败血症、肺炎和脑膜炎)的重要病原菌.为快速准确地鉴定S.agalactiae及其血清型,本研究对22株从国内不同地区分离的牛源链球菌进行生化鉴定以及针对S.agalactiae保守基因sip进行PCR鉴定,并采用多重PCR技术扩增sip基因阳性菌株的cps基因进行血清分型.结果显示,PCR扩增sip基因阳性17株,其中15株生化试验与PCR结果一致;多重PCR扩增cps基因显示,其中Ia型4株、Ⅱ型11株、Ⅲ型2株.本研究为鉴定S.agalactiae及其血清型提供了参考.

  9. Streptococcus agalactiae clones infecting humans were selected and fixed through the extensive use of tetracycline.

    Da Cunha, Violette; Davies, Mark R; Douarre, Pierre-Emmanuel; Rosinski-Chupin, Isabelle; Margarit, Immaculada; Spinali, Sebastien; Perkins, Tim; Lechat, Pierre; Dmytruk, Nicolas; Sauvage, Elisabeth; Ma, Laurence; Romi, Benedetta; Tichit, Magali; Lopez-Sanchez, Maria-José; Descorps-Declere, Stéphane; Souche, Erika; Buchrieser, Carmen; Trieu-Cuot, Patrick; Moszer, Ivan; Clermont, Dominique; Maione, Domenico; Bouchier, Christiane; McMillan, David J; Parkhill, Julian; Telford, John L; Dougan, Gordan; Walker, Mark J; Holden, Matthew T G; Poyart, Claire; Glaser, Philippe


    Streptococcus agalactiae (Group B Streptococcus, GBS) is a commensal of the digestive and genitourinary tracts of humans that emerged as the leading cause of bacterial neonatal infections in Europe and North America during the 1960s. Due to the lack of epidemiological and genomic data, the reasons for this emergence are unknown. Here we show by comparative genome analysis and phylogenetic reconstruction of 229 isolates that the rise of human GBS infections corresponds to the selection and worldwide dissemination of only a few clones. The parallel expansion of the clones is preceded by the insertion of integrative and conjugative elements conferring tetracycline resistance (TcR). Thus, we propose that the use of tetracycline from 1948 onwards led in humans to the complete replacement of a diverse GBS population by only few TcR clones particularly well adapted to their host, causing the observed emergence of GBS diseases in neonates.

  10. Survival of Mycoplasma agalactiae and Mycoplasma mycoides subspecies capri in heat treated goat colostrum.

    Paterna, Ana; Sánchez, Antonio; Amores, Joaquín; Gómez-Martín, Angel; Corrales, Juan C; Contreras, Antonio; de la Fe, Christian


    The viability of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri (Mmc) was assessed in goat colostrum treated at different temperatures. Samples of colostrum were inoculated with reference strains of M. agalactiae (PG2) and Mmc (PG3) and heated at 56°C or 60°C for 0, 30, 60, 90 or 120 min. Viable colonies of M. agalactiae were recovered after all treatments and there was a significant reduction in the concentration of viable M. agalactiae after 30 min at 56°C and 60°C. No viable colonies of Mmc were observed after 60 min at 60°C.

  11. Detection of Streptococcus agalactiae colonization in pregnant women by using combined swab cultures: cross-sectional prevalence study

    Camila Marconi

    Full Text Available CONTEXT AND OBJECTIVE: Maternal Streptococcus agalactiae colonization and early-onset neonatal sepsis have aroused interest in the worldwide literature. Streptococcal neonatal disease is associated with significant morbidity and mortality in the perinatal period, especially among premature neonates. The aim of this study was to assess the prevalence of maternal streptococcal colonization by using combined swab cultures, compared with swab collection from a single site. DESIGN AND SETTING: Cross-sectional study at Faculdade de Medicina de Botucatu, Universidade Estadual Paulista. METHODS: Samples were obtained from 405 patients at gestational ages of 35 to 37 weeks. Swabs from the perianal (rectal region, vaginal introitus and upper lateral vaginal vault were cultured in Todd-Hewitt selective broth. Colonies suggestive of Streptococcus agalactiae were subjected to the catalase and CAMP (Christie, Atkins, Munch-Petersen tests. To evaluate the positivity of combined swab cultures, Tukey's test was used for comparison of proportions. RESULTS: The prevalence of streptococcal colonization was 25.4%. Among the patients with positive cultures, 28.1% had this at only one collection site, 24.2% simultaneously at two sites and 47.5% at all three sites. Associating the swabs from two collection sites significantly increased streptococcal isolation, compared with a single swab (P < 0.05, except for perianal (rectal collection. Use of combined swabs from three collection sites showed statistically higher isolation rates. CONCLUSION: In combined swab cultures collected from three collection sites, the prevalence of maternal Streptococcus agalactiae colonization was higher than in swabs collected from a single site.

  12. Colonization by Streptococcus agalactiae during pregnancy: maternal and perinatal prognosis

    Patrícia El Beitune; Geraldo Duarte; Cláudia Maria Leite Maffei


    We reviewed colonization by group B Streptococcus beta-haemolyticus of Lancefield (SGB), or Streptococcus agalactiae, in pregnant women, and the consequences of infection for the mother and newborn infant, including factors that influence the risk for anogenital colonization by SGB. We also examined the methods for diagnosis and prophylaxis of SGB to prevent early-onset invasive neonatal bacterial disease. At present, it is justifiable to adopt anal and vaginal SGB culture as part of differen...

  13. Chromosomally and Extrachromosomally Mediated High-Level Gentamicin Resistance in Streptococcus agalactiae.

    Sendi, Parham; Furitsch, Martina; Mauerer, Stefanie; Florindo, Carlos; Kahl, Barbara C; Shabayek, Sarah; Berner, Reinhard; Spellerberg, Barbara


    Streptococcus agalactiae (group B Streptococcus [GBS]) is a leading cause of sepsis in neonates. The rate of invasive GBS disease in nonpregnant adults also continues to climb. Aminoglycosides alone have little or no effect on GBS, but synergistic killing with penicillin has been shown in vitro. High-level gentamicin resistance (HLGR) in GBS isolates, however, leads to the loss of a synergistic effect. We therefore performed a multicenter study to determine the frequency of HLGR GBS isolates and to elucidate the molecular mechanisms leading to gentamicin resistance. From eight centers in four countries, 1,128 invasive and colonizing GBS isolates were pooled and investigated for the presence of HLGR. We identified two strains that displayed HLGR (BSU1203 and BSU452), both of which carried the aacA-aphD gene, typically conferring HLGR. However, only one strain (BSU1203) also carried the previously described chromosomal gentamicin resistance transposon designated Tn3706. For the other strain (BSU452), plasmid purification and subsequent DNA sequencing resulted in the detection of plasmid pIP501 carrying a remnant of a Tn3 family transposon. Its ability to confer HLGR was proven by transfer into an Enterococcus faecalis isolate. Conversely, loss of HLGR was documented after curing both GBS BSU452 and the transformed E. faecalis strain from the plasmid. This is the first report showing plasmid-mediated HLGR in GBS. Thus, in our clinical GBS isolates, HLGR is mediated both chromosomally and extrachromosomally.

  14. Fecal strings Associated with Streptococcus agalactiae Infection in Nile Tilapia, Oreochromis niloticus

    Nile tilapia (Oreochromis niloticus) were experimentally-infected with Streptococcus agalactiae for several infectivity and vaccine studies. Some of the S. agalactiae-infected tilapia produced considerably longer (up to 20 cm in length) fecal waste strings than historically observed from tilapia at...

  15. Complete genome sequence and comparative genomic analysis of an emerging human pathogen, serotype V Streptococcus agalactiae

    Tettelin, H; Masignani, [No Value; Cieslewicz, MJ; Eisen, JA; Peterson, S; Paulsen, IT; Nelson, KE; Margarit, [No Value; Read, TD; Madoff, LC; Beanan, MJ; Brinkac, LM; Daugherty, SC; DeBoy, RT; Durkin, AS; Kolonay, JF; Madupu, R; Lewis, MR; Radune, D; Fedorova, NB; Scanlan, D; Khouri, H; Mulligan, S; Carty, HA; Cline, RT; Van Aken, SE; Gill, J; Scarselli, M; Mora, M; Iacobini, ET; Brettoni, C; Galli, G; Mariani, M; Vegni, F; Maione, D; Rinaudo, D; Rappuoli, R; Telford, JL; Kasper, DL; Grandi, G; Fraser, CM


    The 2,160,267 bp genome sequence of Streptococcus agalactiae, the leading cause of bacterial sepsis, pneumonia, and meningitis in neonates in the U.S. and Europe, is predicted to encode 2,175 genes. Genome comparisons among S. agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and the oth

  16. Seroepidemiological survey of sheep flocks from Northern Japan for Mycoplasma ovipneumoniae and Mycoplasma agalactiae.

    Giangaspero, Massimo; Nicholas, Robin A J; Hlusek, Miroslav; Bonfini, Barbara; Osawa, Takeshi; Orusa, Riccardo; Tatami, Shingo; Takagi, Eishu; Moriya, Hiroaki; Okura, Norimoto; Kato, Kazuo; Kimura, Atsushi; Harasawa, Ryô; Ayling, Roger D


    Sheep flocks from Hokkaido, Iwate and Aomori, three northern prefectures of Japan, were screened for antibodies to Mycoplasma ovipneumoniae and Mycoplasma agalactiae by ELISA. Sixty four animals out of 246 (26%) were seropositive to M. ovipneumoniae, with positive results obtained from all three prefectures. None of the sera tested were serologically positive to M. agalactiae.

  17. Streptococcus agalactiae Serotype Distribution and Antimicrobial Susceptibility in Pregnant Women in Gabon, Central Africa

    Sabine Belard; Nicole Toepfner; Mesküre Capan-Melser; Ghyslain Mombo-Ngoma; Rella Zoleko-Manego; Mirjam Groger; Pierre-Blaise Matsiegui; Agnandji, Selidji T.; Adegnika, Ayôla A; Raquel González; Kremsner, Peter G.; Clara Menendez; Michael Ramharter; Reinhard Berner


    Neonatal invasive disease due to Streptococcus agalactiae is life threatening and preventive strategies suitable for resource limited settings are urgently needed. Protective coverage of vaccine candidates based on capsular epitopes will relate to local epidemiology of S. agalactiae serotypes and successful management of critical infections depends on timely therapy with effective...

  18. Complete genome sequence and comparative genomic analysis of an emerging human pathogen, serotype V Streptococcus agalactiae

    Tettelin, H; Masignani, [No Value; Cieslewicz, MJ; Eisen, JA; Peterson, S; Paulsen, IT; Nelson, KE; Margarit, [No Value; Read, TD; Madoff, LC; Beanan, MJ; Brinkac, LM; Daugherty, SC; DeBoy, RT; Durkin, AS; Kolonay, JF; Madupu, R; Lewis, MR; Radune, D; Fedorova, NB; Scanlan, D; Khouri, H; Mulligan, S; Carty, HA; Cline, RT; Van Aken, SE; Gill, J; Scarselli, M; Mora, M; Iacobini, ET; Brettoni, C; Galli, G; Mariani, M; Vegni, F; Maione, D; Rinaudo, D; Rappuoli, R; Telford, JL; Kasper, DL; Grandi, G; Fraser, CM


    The 2,160,267 bp genome sequence of Streptococcus agalactiae, the leading cause of bacterial sepsis, pneumonia, and meningitis in neonates in the U.S. and Europe, is predicted to encode 2,175 genes. Genome comparisons among S. agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and the

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

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


    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.

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

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


    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.

  1. 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

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


    Background 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. aga...

  2. Viability of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in goat milk samples stored under different conditions.

    Amores, Joaquín; Sánchez, Antonio; Martín, Angel Gómez; Corrales, Juan C; Contreras, Antonio; de la Fe, Christian


    Control programs for contagious agalactia (CA) involve monitoring milk samples to detect this disease. This study was designed to establish the effects of the preservatives generally used in dairy laboratories and storage temperature on the viability of Mycoplasma (M.) agalactiae (Ma) and M. mycoides subsp. capri (Mmc) in goat milk samples. In total, 1440 determinations were conducted for each mycoplasma species in milk samples subjected to different storage temperatures (refrigeration at 4°C or freezing at -20°C), preservation strategies (no preservative, NP; azidiol, AZ; or bronopol, BR) and storage times at each temperature (0, 2, 4, 6, 8, 10 and 24h at 4°C and 48h, 1 week, 2 weeks and 4 weeks at -20°C). Our findings reveal the similar viability of Mmc in milk samples stored at 4°C for 24h under the three preservation conditions examined. In contrast, the isolation of Ma in refrigerated milk samples was compromised by the presence of BR, and in smaller measure by the treatments AZ and NP. Freezing milk samples considerably reduced the viability of both mycoplasmas. Given the different sensitivity of the two mycoplasma species to BR, refrigerated milk samples treated with AZ could be used to detect infections caused by both species through culture-based methods.

  3. 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

    Domingo C. Balderramo


    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

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

    Aline Gerato Dibbern


    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 ...

  5. Streptococcus agalactiae mural infective endocarditis in a structurally normal heart

    Nobuhiro Ariyoshi


    Full Text Available A 38-year-old Caucasian man with uncontrolled diabetes mellitus type 2 was admitted with a 1-week duration of fevers, chills, and a non-productive cough. He had a left ischiorectal abscess 1 month prior to admission. Physical examination revealed caries on a left upper molar and a well-healed scar on the left buttock, but no heart murmur or evidence of micro-emboli. Blood cultures grew Streptococcus agalactiae. A transesophageal echocardiogram revealed a mobile mass in the right ventricle that attached to chordae tendineae without valvular disease or dysfunction. A computed tomography (CT with contrast revealed the mass within the right ventricle, a left lung cavitary lesion, and a splenic infarction. He was initially treated with penicillin G for a week. Subsequently, ceftriaxone was continued for a total of 8 weeks. A follow-up CT showed no evidence of right ventricular mass 8 weeks after discharge. This is the first reported case of S. agalactiae mural infective endocarditis in a structurally normal heart.

  6. Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae.

    Pinto, Tatiana Castro Abreu; Costa, Natália Silva; Corrêa, Ana Beatriz de Almeida; de Oliveira, Ivi Cristina Menezes; de Mattos, Marcos Correa; Rosado, Alexandre Soares; Benchetrit, Leslie Claude


    Streptococcus agalactiae (GBS) is a major source of human perinatal diseases and bovine mastitis. Erythromycin (Ery) and tetracycline (Tet) are usually employed for preventing human and bovine infections although resistance to such agents has become common among GBS strains. Ery and Tet resistance genes are usually carried by conjugative transposons (CTns) belonging to the Tn916 family, but their presence and transferability among GBS strains have not been totally explored. Here we evaluated the presence of Tet resistance genes (tetM and tetO) and CTns among Ery-resistant (Ery-R) and Ery-susceptible (Ery-S) GBS strains isolated from human and bovine sources; and analyzed the ability for transferring resistance determinants between strains from both origins. Tet resistance and int-Tn genes were more common among Ery-R when compared to Ery-S isolates. Conjugative transfer of all resistance genes detected among the GBS strains included in this study (ermA, ermB, mef, tetM and tetO), in frequencies between 1.10(-7) and 9.10(-7), was possible from bovine donor strains to human recipient strain, but not the other way around. This is, to our knowledge, the first report of in vitro conjugation of Ery and Tet resistance genes among GBS strains recovered from different hosts.

  7. Validation of absolute quantitative real-time PCR for the diagnosis of Streptococcus agalactiae in fish.

    Sebastião, Fernanda de A; Lemos, Eliana G M; Pilarski, Fabiana


    Streptococcus agalactiae (GBS) are Gram-positive cocci responsible for substantial losses in tilapia fish farms in Brazil and worldwide. It causes septicemia, meningoencephalitis and mortality of whole shoals that can occur within 72 h. Thus, diagnostic methods are needed that are rapid, specific and sensitive. In this study, a pair of specific primers for GBS was generated based on the cfb gene sequence and initially evaluated by conventional PCR. The protocols for absolute quantitative real-time PCR (qPCR) were then adapted to validate the technique for the identification and quantification of GBS isolated by real-time detection of amplicons using fluorescence measurements. Finally, an infectivity test was conducted in tilapia infected with GBS strains. Total DNA from the host brain was subjected to the same technique, and the strains were re-isolated to validate Koch's postulates. The assay showed 100% specificity for the other bacterial species evaluated and a sensitivity of 367 gene copies per 20 mg of brain tissue within 4 h, making this test a valuable tool for health monitoring programs.

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

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


    -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......Contagious mastitis pathogens continue to pose an economic threat to the dairy industry. An understanding of their frequency and transmission dynamics is central to evaluating the effectiveness of control programmes. The objectives of this study were twofold: (1) to estimate the annual herd...... 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...

  9. [Vertebral osteomyelitis caused by Streptococcus agalactiae in healthy adults. Description of 2 new cases].

    Rivero Marcotegui, M; Hidalgo Ovejero, A; Lecumberri, M Cía; Otermin Maya, I; Pereda García, A


    The importance of Streptococcus agalactiae as a pathogen in nonpregnant adults has been widely recognized in recent years, especially in the elderly or immunocompromised patients. Two cases of vertebral osteomyelitis caused by S. agalactiae in young patients with no known underlying diseases or predisposing factor to infection are reported. A systematic review of the literature (MEDLINE, 1976-May 2008) was performed, 10 cases previously reported in the literature of vertebral osteomyelitis due to S. agalactiae being found in adults under 65 years of age with no predisposing risk factors for infection, most of which have been reported over the last 10 years. We believe that this microbial etiology should be considered in patients of any age and immunological status. Other factors, other than increasing the number of patients with chronic diseases, explaining the increased rate of S. agalactiae infections in adults need to be studied.

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

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


    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

  11. Cystine antagonism of the antibacterial action of lactoperoxidase-thiocyanate-hydrogen peroxide on Streptococcus agalactiae.

    Mickelson, M. N.; Anderson, A J


    Cystine reduction in Streptococcus agalactiae, resulting in sulfhydryl formation, may account for antagonism of the antibacterial effect of lactoperoxidase-thiocyanate-hydrogen peroxide when cystine is present in excess of the amount needed for maximum growth. Accumulation of cystine by S. agalactiae and its reduction to form sulfhydryl compounds were demonstrated. The reduction of cystine appeared to occur by a couple reaction between glutathione reductase and glutathione-disulfide transhydr...

  12. The novel fibrinogen-binding protein FbsB promotes Streptococcus agalactiae invasion into epithelial cells.

    Gutekunst, Heike; Eikmanns, Bernhard J; Reinscheid, Dieter J


    Streptococcus agalactiae is a major cause of bacterial sepsis and meningitis in human newborns. The interaction of S. agalactiae with host proteins and the entry into host cells thereby represent important virulence traits of these bacteria. The present report describes the identification of the fbsB gene, encoding a novel fibrinogen-binding protein that plays a crucial role in the invasion of S. agalactiae into human cells. In Western blots and enzyme-linked immunosorbent assay (ELISA) experiments, the FbsB protein was demonstrated to interact with soluble and immobilized fibrinogen. Binding studies showed the N-terminal 388 residues of FbsB and the Aalpha-subunit of human fibrinogen to recognize each other. By reverse transcription (RT)-PCR, the fbsB gene was shown to be cotranscribed with the gbs0851 gene in S. agalactiae. Deletion of the fbsB gene in the genome of S. agalactiae did not influence the binding of the bacteria to fibrinogen, suggesting that FbsB does not participate in the attachment of S. agalactiae to fibrinogen. In tissue culture experiments, however, the fbsB deletion mutant was severely impaired in its invasion into lung epithelial cells. Bacterial invasion could be reestablished by introducing the fbsB gene on a shuttle plasmid into the fbsB deletion mutant. Furthermore, treatment of lung epithelial cells with FbsB fusion protein blocked S. agalactiae invasion of epithelial cells in a dose-dependent fashion. These results suggest an important role of the FbsB protein in the overall process of host cell entry by S. agalactiae.

  13. Surface Proteins of Streptococcus agalactiae and Related Proteins in Other Bacterial Pathogens

    Lindahl, Gunnar; Stålhammar-Carlemalm, Margaretha; Areschoug, Thomas


    Streptococcus agalactiae (group B Streptococcus) is the major cause of invasive bacterial disease, including meningitis, in the neonatal period. Although prophylactic measures have contributed to a substantial reduction in the number of infections, development of a vaccine remains an important goal. While much work in this field has focused on the S. agalactiae polysaccharide capsule, which is an important virulence factor that elicits protective immunity, surface proteins have received incre...

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

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


    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...

  15. Phenotypes, genotypes, serotypes and molecular epidemiology of erythromycin-resistant Streptococcus agalactiae in Italy.

    De Francesco, M A; Caracciolo, S; Gargiulo, F; Manca, N


    The purpose of this investigation was to analyse Streptococcus agalactiae (group B Streptococcus, GBS) isolates collected in Italy from vaginal and urine samples in respect to their clonality, distribution of virulence factors and antimicrobial resistance determinants. Three hundred and eighty-eight GBS were recovered from clinical samples. They were analysed for antibiotic resistance profiling. Erythromycin-resistant strains were further characterised by multilocus sequence typing (MLST), serotyping and the detection of alp genes of the alpha-like protein (Alp) family. GBS isolates represented 40 different sequence types (STs), grouped in five clonal complexes (CCs) and belonged to seven serotypes. Most serotype V strains (81%) possessed alp2-3; serotype Ia carried mainly epsilon, while the serotype III mainly rib. All isolates were susceptible to penicillin, whereas resistance to erythromycin was detected in 15% of isolates. Most erythromycin-resistant GBS strains were of serotype V (56.8%) and belonged to the CC-1 group (50%). Macrolide resistance phenotypes were the cMLS(B) (46.5%) and the M phenotypes (46.5%) due to the presence of ermB and mefA/E genes, respectively. These results provide data which establish a baseline for monitoring erythromycin resistance in this region and also provide an insight into the correlation among clonal types, serotypes, surface protein and resistance genes. The increased prevalence of strains that displayed the M phenotype strengthens the importance of the epidemiological surveillance of macrolide resistance in GBS, which may also represent an important reservoir of resistance genes for other species.

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

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


    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.

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

    du Merle, Laurence; Rosinski-Chupin, Isabelle; Gominet, Myriam; Bellais, Samuel; Poyart, Claire; Trieu-Cuot, Patrick


    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 indirect

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

    Renata Perugini Biasi-Garbin


    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.

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

    Perugini Biasi-Garbin, Renata; Saori Otaguiri, Eliane; Fernandes da Silva, Mayara; Belotto Morguette, Ana Elisa; Armando Contreras Lancheros, César; Kian, Danielle; Perugini, Márcia Regina Eches; Durán, Nelson; Nakamura, Celso Vataru; Yamauchi, Lucy Megumi; Yamada-Ogatta, Sueli Fumie


    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. PMID:25945115

  20. 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

    Irina Lermontov Borger


    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

  1. Streptococcus agalactiae from pregnant women: antibiotic and heavy-metal resistance mechanisms and molecular typing.

    Rojo-Bezares, B; Azcona-Gutiérrez, J M; Martin, C; Jareño, M S; Torres, C; Sáenz, Y


    We investigated the antibiotic and heavy-metal resistance mechanisms, virulence genes and clonal relationships of macrolide- and/or lincosamide-resistant (M+/-LR) Streptococcus agalactiae (group B Streptococcus, GBS) isolates from pregnant women in La Rioja in Northern Spain, a region with a significant immigrant population. In total 375 GBS isolates were recovered during 2011. About three-quarters of isolates were from European nationals and the remainder distributed among 23 other nationalities. Seventy-five (20%) were classified as M+/-LR strains and 28 (37%) of these were resistant to ⩾3 classes of antibiotics. Capsular serotypes III (29·3%), V (21·3%) and II (12%) were the most frequent. A wide variety of antibiotic resistance genes were detected in M+/-LR strains; notably, 5·3% harboured the lsa(C) gene associated with cross-resistance, and tet(W) was identified in a single strain. We report, for the first time, the detection of cadmium and copper resistance encoded by tcrB + cadA + cadC genes in 20 M+/-LR strains, which raises the possibility of co-selection of antibiotic and heavy-metal resistance disseminated through mobile genetic elements. The M+/-LR strains were highly diverse by DNA macrorestriction profiles (65 patterns) and 16 multilocus sequence types (STs) distributed among six clonal complexes; the most frequent were ST1, ST19, and ST12, and two strains were novel (ST586 and ST601). In conclusion, a wide diversity of genetic lineages of macrolide, lincosamide and heavy-metal- resistant GBS strains was observed in an ethnically diverse maternal population.

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

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


    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...

  3. Non-infectivity of Cattle Streptococcus agalactiae in Nile Tilapia, Oreochromis niloticus and Channel Catfish, Ictalurus punctatus

    Streptococcus agalactiae is classified as a Lancefield’s group B Streptococcus (GBS). It is the causative bacterium of streptococcosis that is responsible for severe economic losses in wild and cultured fish, worldwide. Streptococcus agalactiae also causes bovine mastitis. Only limited comparativ...

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

    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.

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

    Amrullah Amrullah


    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.

  6. Infecção congênita em cabritos por Mycoplasma agalactiae

    N.S. Silva


    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.

  7. Application of the California mastitis test in intramammary Streptococcus agalactiae and Staphylococcus aureus infections of camels (Camelus dromedarius) in Kenya.

    Younan, M; Ali, Z; Bornstein, S; Müller, W


    A study was conducted on 207 lactating camels in six herds in Kenya to evaluate the California mastitis test (CMT) for the detection of intramammary infections (IMIs) caused by Streptococcus agalactiae and Staphylococcus aureus and to investigate the prevalence of both the pathogens in the camel udder. IMI with S. agalactiae was found in 12% of all camels sampled. IMI with S. aureus was present in 11% of all camels sampled. The herd-level prevalence of IMI varied between 0 and 50% for S. agalactiae and between 0 and 13% for S. aureus. Longitudinal observations over 10-12 months confirmed persistent infections for both pathogens. Observations in one herd suggested that camel pox was a contributing factor in spreading and exacerbating S. agalactiae udder infections.The CMT had quarter-level sensitivities of 77 and 68% for S. agalactiae and S. aureus in camels, respectively. The CMT specificities were 91% for both the pathogens.

  8. [Culture media for the detection and the identification of Streptococcus agalactiae].

    de la Rosa, M; Pérez, M; Carazo, C; Pareja, L; Orts, A; Cantudo, P


    Streptococcus agalactiae, a Group B streptococcus, is the main cause of bacterial perinatal infection and is also an important opportunistic pathogen. Detection and identification of S. agalactiae are straight forward with special culture media, where Group B streptococci show a specific, typical pink or red pigment. To quickly and easily detect the pigment, culture media should contain: (i) starch; (ii) an inhibitor of the folate pathway; (iii) animal serum; (iv) a pepsic proteic hydrolysate; and (v) glucose, together with a high-capacity buffer. When selective antibiotics are added to culture media designed in this way, it is possible to detect S. agalactiae directly from clinical samples by observation of its pigment after less than 12 hours of aerobic incubation.

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

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


    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.

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

    Brewer, Linda


    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.

  11. Update on control of Staphylococcus aureus and Streptococcus agalactiae for management of mastitis.

    Keefe, Greg


    The primary method of spread for S agalactiae and S aureus is from cow to cow, so prevention focuses on within and between herd biosecurity to reduce or eliminate the reservoir of infection. S agalactiae is an obligate pathogen of the mammary gland, whereas S aureus is more widespread on other cow body sites and in the environment. Both organisms cause persistent infections, with S agalactiae typically causing higher SCC and bacteria counts in milk. Conventional methods of detection through culture perform well at the cow level. In bulk tanks, augmented procedures should be considered. PCR methods show promise of high sensitivity and specificity, at both the cow and bulk tank level. In developed dairy industries, prevalence of infection has decreased dramatically over the past 30 years for S agalactiae. For S aureus, the herd level of infection remains very high, although with rigorous, consistent application of control measures, within-herd prevalence has decreased. Because the milking time is the primary period for new IMI, it is the focal point of most prevention activities. Premilking and postmilking teat disinfection and proper stimulation and milk-out with adequately functioning equipment are key factors. There is growing evidence that the use of milking gloves is an integral part of contagious mastitis control and the production of high-quality milk. Treatment success is dramatically different between the 2 pathogens. For S agalactiae, eradication can be completed rapidly through a culture and treatment program with minimal culling. For S aureus, treatment success, particularly during lactation, is often disappointing and depends on cow, pathogen, and treatment factors. These factors should be reviewed prior to initiating any treatment to determine the potential for cure. Blanket dry cow therapy and strategic culling are important control procedures for contagious mastitis pathogens. Maintaining a closed herd or, at minimum, adhering to clearly defined

  12. 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

    Domingo C. Balderramo; Ana M. Bertoli; Miguel A. PaganiniI; Abel Zárate; Zlocowski, Juan C.; Alejandro Alvarellos; Francisco Caeiro; Caeiro, Juan P.


    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 tenosin...

  13. 罗非鱼组织内无乳链球菌实时荧光定量 PCR 检测方法建立%Real-time quantitative PCR for detection of Streptococcus agalactiae from tilaPia tissue

    王瑞; 李莉萍; 黄婷; 梁万文; 梁聪; 雷爱莹; 陈明


    为建立一种罗非鱼组织内无乳链球菌(Streptococcus agalactiae)的定量检测方法,以无乳链球菌 cfb 基因为靶标建立了 TaqMan 荧光探针实时定量 PCR 方法,并对该方法的特异性、敏感性及实用性进行了验证。应用建立的方法检测无乳链球菌标准菌株和阳性菌株均为阳性,阴性对照菌株均为阴性;无乳链球菌基因组 DNA 最低检测质量浓度为3.42×10-7 ng·μL -1,每个反应体系检测下限低于10个细胞;人工感染罗非鱼肝、脾、中肠和肾组织样品无乳链球菌检测结果均为阳性,组织样品每微克 DNA 可检测到无乳链球菌细胞数量分别为2.95×103个、2.45×107个、2.34×103个和4.54×104个,对照组为阴性。结果表明建立的罗非鱼组织内无乳链球菌实时荧光定量 PCR 检测方法具有准确性高、特异性和敏感性强等特点,可对罗非鱼组织内无乳链球菌进行快速定量检测,可用于罗非鱼无乳链球菌病的监测与预防。%By designing primers and probe based on the conserved region of cfb gene of Streptococcus agalactiae isolated from tilapia, we developed a real-time fluorescent PCR assay for detection of S. agalactiae from tilapia tissue and verified the specificity,sensitivity and practicability of the assay. The standard and positive strains of S. agalactiae were positive,and all negative controls were negative. The minimum detectable concentration of S. agalactiae genome DNA was 3. 42 × 10 - 7 ng·μL - 1 ,and the detection limit of the assay was less than 10 cells per reaction system. The genome DNA samples of midgut,liver,spleen and kidney which were collected from tilapia artificially infected with S. agalactiae were tested by real-time PCR and were positive. The quantities of S. agalactiae detected in per μg DNA of tissue samples were 2. 95 × 103 cells,2. 45 × 107 cells,2. 34 × 103 cells and 4. 54 × 104 cells,respectively,and the controls were negative

  14. Efficacy of an experimentally inactivated Streptococcus agalactiae vaccine in Nile tilapia (Oreochromis niloticus) reared in Brazil

    Tilapia aquaculture is one of the fastest growing segments of fish production in Brazil. Nile tilapia (Oreochromis niloticus) is largely cultivated in the state of Parana, where Streptococcus agalactiae is the cause of severe disease outbreaks. The objective of this paper was to evaluate an inactiva...

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

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


    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 v

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

    Fléchard, Maud; Gilot, Philippe


    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.

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

    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...

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

    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...

  19. Acidic pH strongly enhances in vitro biofilm formation by a subset of hypervirulent ST-17 Streptococcus agalactiae strains.

    D'Urzo, Nunzia; Martinelli, Manuele; Pezzicoli, Alfredo; De Cesare, Virginia; Pinto, Vittoria; Margarit, Immaculada; Telford, John Laird; Maione, Domenico


    Streptococcus agalactiae, also known as group B Streptococcus (GBS), is a primary colonizer of the anogenital mucosa of up to 40% of healthy women and an important cause of invasive neonatal infections worldwide. Among the 10 known capsular serotypes, GBS type III accounts for 30 to 76% of the cases of neonatal meningitis. In recent years, the ability of GBS to form biofilm attracted attention for its possible role in fitness and virulence. Here, a new in vitro biofilm formation protocol was developed to guarantee more stringent conditions, to better discriminate between strong-, low-, and non-biofilm-forming strains, and to facilitate interpretation of data. This protocol was used to screen the biofilm-forming abilities of 366 GBS clinical isolates from pregnant women and from neonatal infections of different serotypes in relation to medium composition and pH. The results identified a subset of isolates of serotypes III and V that formed strong biofilms under acidic conditions. Importantly, the best biofilm formers belonged to serotype III hypervirulent clone ST-17. Moreover, the abilities of proteinase K to strongly inhibit biofilm formation and to disaggregate mature biofilms suggested that proteins play an essential role in promoting GBS biofilm initiation and contribute to biofilm structural stability.

  20. Multiplex PCR for simultaneous detection of Streptococcus agalactiae, Streptococcus iniae and Lactococcus garvieae: a case of S. agalactiae infection in cultured Nile tilapia (Oreochromis niloticus) and red tilapia (Oreochromis niloticus x Oreochromis mossambicus)

    Chutima Tantikitti; Naraid Suanyuk; Akkarawit Itsaro


    A multiplex PCR (m-PCR) technique was developed for simultaneous detection of the causative agents responsible forstreptococcosis of cultured fish in Thailand i.e., Streptococcus agalactiae, Streptococcus iniae, and Lactococcus garvieae.The study on the sensitivity of the technique indicated that the minimum detected DNA concentration was 9.76, 39.06, and19.53 pg for S. agalactiae, S. iniae and L. garvieae, respectively. Detection of streptococcosis in healthy and diseased Niletilapia (Oreoch...

  1. Analysis of Streptococcus agalactiae pan-genome for prevalence, diversity and functionality of integrative and conjugative or mobilizable elements integrated in the tRNA(Lys CTT) gene.

    Puymège, Aurore; Bertin, Stéphane; Guédon, Gérard; Payot, Sophie


    Streptococcus agalactiae is the first cause of invasive infections in human neonates and is also a major bovine and fish pathogen. High genomic diversity was observed in this species that hosts numerous mobile genetic elements, in particular elements transferable by conjugation. This works aims to evaluate the contribution of these elements to GBS genome diversity. Focusing on genomic islands integrated in the tRNA(Lys) (CTT) gene, a known hotspot of recombination, an extensive in silico search was performed on the sequenced genome of 303 strains of S. agalactiae isolated from different hosts. In all the isolates (except 9), whatever their origin (human, bovine, camel, dog, gray seal, dolphin, fish species or bullfrog), this locus carries highly diverse genomic islands transferable by conjugation such as integrative and conjugative elements (ICEs), integrative and mobilizable elements (IMEs), CIs-mobilizable elements (CIMEs) or composite elements. Transfer of an ICE from an ST67 bovine strain to a phylogenetically distant ST23 human isolate was obtained experimentally indicating that there was no barrier to ICE transfer between strains from different hosts. Interestingly, a novel family of putative IMEs that site-specifically integrate in the nic site of oriT of ICEs belonging to Tn916/ICESt3 superfamily was detected in silico. These elements carry an antibiotic resistance gene (lsa(C)) already described to confer cross-resistance to lincosamides, streptogramins A and pleuromutilins. Further work is needed to evaluate the impact of these IMEs on the transfer of targeted ICEs and the mobility and the dissemination of these IMEs.

  2. Záchyt Streptococcus agalactiae u těhotných žen a novorozenců

    HÁLOVÁ, Eliška


    S. agalactiae are common commensals of the nasopharynx, vagina and gastrointestinal tract. Now digestive tract is considered a very likely source of vaginal colonization. Colonization by the bacteria is in many cases asymptomatic. S. agalactiae causes a variety of infections in neonates and is a leading cause of neonatal morbidity and mortality. Transmission takes place vertically from colonized mother to fetus. A potential source of GBS infections may also be mother's milk. This type of tran...

  3. Detection and Enumeration of Streptococcus agalactiae from Bovine Milk Samples by Real-Time Polymerase Chain Reaction.

    de Carvalho, Nara Ladeira; Gonçalves, Juliano Leonel; Botaro, Bruno Garcia; Silva, Luis Felipe de Prada E; dos Santos, Marcos Veiga


    The aim of this study was to evaluate the use of real-time polymerase chain reaction (qPCR) combined with DNA extraction directly from composite milk and bulk tank samples for detection and enumeration of Streptococcus agalactiae (SAG) causing subclinical mastitis. Dilutions of sterile reconstituted skim milk inoculated with SAG ATCC 13813 were used to establish a standard curve (cfu/mL) for the qPCR assay targeting SAG. The analytical sensitivity and repeatability of the qPCR assay were determined. Bulk tank (BTM; n = 38) and composite milk samples (CM; n = 26) collected from lactating cows with positive isolation of SAG were submitted to the qPCR protocol and SAG plate counting, with results from both methods compared. Amplification of DNA was not possible in two out of 64 samples, indicating that qPCR was able to detect SAG in 96 and 97% of BTM and CM samples, respectively. The inter-assay coefficient of variation was <5%, showing that the technique had adequate repeatability. The qPCR protocol can be a high-throughput and rapid diagnostic assay to accurately detect SAG from BTM and CM samples compared with conventional microbiological culture method. However, the evaluated qPCR protocol is not accurate for enumerating SAG in milk samples, probably due to quantification of DNA of non-viable cells.

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

    Clément eLier


    Full Text Available 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 colonisation and infections in humans since the 1960s, two CRISPR-Cas systems have been described. A type II-A system, characterised 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 colonisation or infection. The CRISPR-Cas locus was analysed 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 colonisation specificities of this lineage.

  5. 牛源性无乳链球菌血清型分布及抗生素耐药性研究%Serotype Distribution of Bovine Streptococcus agalactiae and its Drug Resistance to Antibiotics in China

    李宏胜; 郁杰; 罗金印; 李新圃; 徐继英; 王旭荣; 张礼华


    本研究旨在查明牛源性无乳链球菌血清型分布及对常见抗生素的耐药情况,指导临床合理用药.对从中国部分地区奶牛场采集的临床型乳房炎病牛乳中分离鉴定出78株无乳链球菌地方菌株,制备沉淀反应抗原及6株标准血清型无乳链球菌单因子血清抗体,采用环状沉淀试验,对78株无乳链球菌地方菌株进行了血清学分型鉴定;同时采用K-B纸片法测定了这些菌株对抗生素的耐药情况.结果表明,引起奶牛乳房炎的无乳链球菌血清型主要为X型(60.26%),其次为Ⅲ型(10.26%)、R型(7.69%)、Ⅱ型(7.69%)和Ⅰb型(5.13%),Ⅰ a型尚未发现.无乳链球菌对目前临床上使用的大部分抗生素,如头孢唑啉、头孢噻肟、丁胺卡那霉素、卡那霉素、庆大霉素、四环素、强力霉素、氟苯尼考、多黏菌素B、环丙沙星、氟哌酸和头孢他啶/棒酸均较敏感;但对氨苄青霉素、链霉素、恩诺沙星、阿莫西林/棒酸和复方新诺明,有一定的耐药性,其耐药率达50%~100%.本研究对进一步研制有效的药物及疫苗,指导临床合理用药具有重要的意义.%To identify serotype distribution of bovine Streptococcus agalactiae and its drug resistance to common antibiotics, so that the correct antibiotics was applied during clinical therapy, 78 local strains of Streptococcus agalactiae were isolated from milk samples of dairy cow with clinical mastitis throughout the country. We prepared antigens for precipitation reaction and hyperimmune serum antibodies of 6 standard serotype Streptococcus agalactiae strains, then 78 local strains of Streptococcus agalactiae were identified by ring precipitation test and antibiotic resistance were analyzed. In China, the predominant serotype of Streptococcus agalactiae in bovine mastitis were X-type (60. 26%), Ⅲ-type (10. 26%), R-type (7.69%), Ⅱ -type (7.69%) and I b-type (5. 13%) respectively. However, I a-type was not


    Angela Mariana Lusiastuti


    Full Text Available The successful effect of phages in phage therapy has been proven by an increase of the presence of phages in survivors. Seven previously isolated phages were used to examine the effects of storage temperature and types of media on their survival rate. Phages PSaT-18, PSaT-19, PSaT-20 and PSaT-21 isolated from a Streptococcosis-infected Tilapia (Oreochromis niloticus were used in this study. Phages PSaG-1, PSaG-2 and PSaG-3 were isolated from a diseased Gouramy (Osphronemus gouramy. All of the phages were stored in refrigerator at temperature of 8oC and kept in two types of media, Brain Heart Infusion Broth (BHIB and BHIB enriched with glycerol. To determine the lytic activity of the phages, 20 μL of each phage suspension was poured onto BHIA plate and incubated at 25oC for 24 to 48 hours. The result showed that phages PSaT-18, PSaT-19, PSaT-20 and PSaT-21 have lytic activity against S. agalactiae and S. iniae than phages PSaG-1, PSaG-2, and PSaG-3. Phages PSaT-18, PSaT-19, PSaT-20, and PSaT-21 always survived up to three weeks and phage PSaT-18 in BHIB have the highest potential as a candidate of phage therapy for Streptococcosis.

  7. The MCP-8 gene and its possible association with resistance to Streptococcus agalactiae in tilapia.

    Fu, Gui Hong; Wan, Zi Yi; Xia, Jun Hong; Liu, Feng; Liu, Xiao Jun; Yue, Gen Hua


    Mast cell proteases play an important role in the regulation of the immune response. We identified the cDNA of the mast cell protease 8 (MCP-8) gene and analyzed its genomic structure in tilapia. The ORF of the MCP-8 was 768 bp, encoding 255 amino acids. Quantitative real-time PCR revealed that the MCP-8 gene was expressed predominantly in spleen, moderately in liver, blood, brain, gill, intestine, skin, and weakly expressed in kidney, muscle and eye. After a challenge with Streptococcus agalactiae, the gene was induced significantly (p 0.05). These results suggest that the MCP-8 gene play an important role in the resistance to S. agalactiae in tilapia. The SNP markers in the MCP-8 gene associated with the resistance to the bacterial pathogen may facilitate selection of tilapia resistant to the bacterial disease.

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

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


    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.

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

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


    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 challenged with Ma. High-throughput mRNA sequencing was performed to reveal differentially expressed genes (DEG) at different time-points (3 h, 12 h, and 24 h) post infection (PI). The pathway enrichment analysis of the DEG showed that infection significantly affected pathways associated with immune response...... of the complement system and apoptosis pathways, and expression of genes coding for antimicrobial molecules and peptides. In our study we attempted to interpret the detected transcriptomic changes in a biological context and infer mammary infection resistance candidate genes, interesting for further validation...

  10. Evaluation of chromID strepto B as a screening media for Streptococcus agalactiae

    Morita, Toyohisa; Feng, Dongyun; Kamio, Yoko; Kanno, Isao; Somaya, Teruo; Imai, Kazuhito; Inoue, Misaki; Fujiwara, Mutsunori; Miyauchi, Akihito


    Background Streptococcus agalactiae (Group B Streptococcus, GBS), a leading cause of sepsis and meningitis in infants, can be transmitted vertically from mother to infant during passage through the birth canal. Detection of GBS colonization in perinatal women is a major strategy for the prevention of postpartum neonatal disease. The U.S. Centers for Disease Control and Prevention recommends that all women undergo vaginal-rectal screening for GBS colonization at 35-37 weeks of gestation. Chrom...

  11. Use of a Serotype-Specific DNA Microarray for Identification of Group B Streptococcus (Streptococcus agalactiae)

    Wen, Linyan; Wang, Quan; Li, Yayue; Kong, Fanrong; Gilbert, Gwendolyn L.; Cao, Boyang; Wang, Lei; Feng, Lu


    Group B Streptococcus (GBS; Streptococcus agalactiae) is an important cause of sepsis and meningitis. Nine GBS serotypes, based on capsular polysaccharide (CPS) antigens, have been described. Their distribution varies worldwide and needs to be monitored to understand the epidemiology of GBS disease and inform the development of vaccines. In this study, we sequenced cpsH of GBS serotype II (cpsHII) and compared it with that of the other eight serotypes to identify serotype-specific regions. We...

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

    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.

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

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


    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.

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

    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.


    Angela Mariana Lusiastuti


    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.

  16. Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions.

    Hegde, Shrilakshmi; Gabriel, Cordula; Kragl, Martin; Chopra-Dewasthaly, Rohini


    Appropriate infection models are imperative for the understanding of pathogens like mycoplasmas that are known for their strict host and tissue specificity, and lack of suitable cell and small animal models has hindered pathogenicity studies. This is particularly true for the economically important group of ruminant mycoplasmas whose virulence factors need to be elucidated for designing effective intervention strategies. Mycoplasma agalactiae serves as a useful role model especially because it is phylogenetically very close to M. bovis and causes similar symptoms by as yet unknown mechanisms. Here, we successfully prepared and characterized four different primary sheep cell lines, namely the epithelial and stromal cells from the mammary gland and uterus, respectively. Using immunohistochemistry, we identified vimentin and cytokeratin as specific markers to confirm the typical cell phenotypes of these primary cells. Furthermore, M. agalactiae's consistent adhesion and invasion into these primary cells proves the reliability of these cell models. Mimicking natural infections, mammary epithelial and stromal cells showed higher invasion and adhesion rates compared to the uterine cells as also seen via double immunofluorescence staining. Altogether, we have generated promising in vitro cell models to study host-pathogen interactions of M. agalactiae and related ruminant pathogens in a more authentic manner.

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

    Bruno H.L.S. Alves


    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.

  18. Comparison of culture and PCR to detect Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in ear swabs taken from goats.

    Amores, Joaquín; Corrales, Juan C; Martín, Angel Gómez; Sánchez, Antonio; Contreras, Antonio; de la Fe, Christian


    This study was designed to evaluate the validity of PCR for the direct detection of Mycoplasma (M.) agalactiae and Mycoplasma mycoides subsp. capri (Mmc), as the two species most frequently causing contagious agalactia (CA) in goats. The PCR method was compared with the traditional culture technique to determine which method was most efficient at identifying all auricular carriers present in herds. The samples analyzed were 307 ear swabs taken from goats reared in a CA endemic area. We assessed the validity of each technique to detect each species and agreement between both methods. For each species, the result was taken as true-positive when at least one of the two tests was positive. Of the swabs tested, 246 were scored positive by PCR (235 and 11 for Mmc and M. agalactiae, respectively) and 117 showed a positive culture result (113 for Mmc and 4 for M. agalactiae). 133 of the PCR-positive samples (124 and 9 for Mmc and M. agalactiae, respectively) yielded negative culture results and 4 culture-positive samples tested negative using PCR (2 for each species). Sensitivity and negative predictive values for PCR were 84.62 and 99.32 (for M. agalactiae) and 99.16 and 97.22% (for Mmc) respectively, and for culture were 30.77 and 97.03 (for M. agalactiae) and 47.08 and 36.08% (for Mmc), respectively. PCR proved to be a rapid and sensitive method for the detection of mycoplasmas in the external ear of asymptomatic carriers. Tools such as this are needed to adopt efficient control measures against CA.

  19. 鱼类无乳链球菌病的研究进展%Research Progress on Fish Streptococcus agalactiae Disease

    邓永强; 汪开毓


    无乳链球菌是常见的人兽共患病原菌,常导致新生儿肺炎、脑膜炎、败血症及奶牛乳腺炎。近年来,在水生动物体内也频繁分离到该菌,尤其是自2007年以来,该菌每年都在中国广东、海南、福建、广西等罗非鱼主要养殖区暴发流行,严重威胁着罗非鱼养殖业的健康发展。作者综述了当前国内外对鱼类无乳链球菌病的病原学、流行病学、症状、病理变化、毒力因子(荚膜多糖、CAMP 因子、表面免疫原性蛋白、α蛋白、C5a 肽酶)和防控等方面的研究进展,以期为中国鱼类无乳链球菌病的防控提供参考。%Streptococcus agalactiae is common zoonotic bacteria,which cause neonatal pneumonia, meningitis,septicemia and mastitis in dairy cows.In recent years,it was also be isolated from aquatic animals frequently.Especially since 2007,diseases caused by the bacteria outbreak in Guangdong,Hainan,Fujian,Guangxi and other tilapia aquaculture areas and had a serious threat to the healthy development of tilapia aquaculture in china.In this paper,research progresses on fish Streptococcus agalactiae disease were reviewed.The etiology,epidemiology,symptoms,path-ological changes,virulence factors (capsular polysaccharide,CAMP factor,Sip,protein alpha,C5a peptidase)and prevention and control were included in the review,which would provide refer-ences for the fish Streptococcus agalactiae disease prevention and control.

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

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


    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

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

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


    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

  2. 孕妇生殖道无乳链球菌带菌状况及药物敏感性分析%Analysis of carrying status and drug susceptibility of Streptococcus agalactiae in genital tract of pregnant women

    林爱心; 袁春雷; 汪伟山; 谭南; 杨晓华; 李丽莲


    目的:了解孕妇生殖道无乳链球菌携带及耐药状况,为临床用药提供依据。方法取孕妇阴道分泌物标本进行细菌培养,分离出的无乳链球菌经 VITEK 2全自动细菌鉴定分析仪进行鉴定和药物敏感试验。结果1042份标本中分离出无乳链球菌56例,分离率为5.4%。药物敏感试验结果显示无乳链球菌对青霉素、头孢唑啉、万古霉素、利奈唑烷、呋喃妥因均高度敏感,未发现中介或耐药菌株;左旋氧氟沙星、莫西沙星及克林霉素、红霉素、四环素的敏感率分别为84.0%、84.0%、71.4%、39.3%和8.9%。结论该院孕妇无乳链球菌的携带率不高,青霉素和头孢唑啉可作为治疗的首选药。临床和实验室应重视孕妇无乳链球菌的检测。%Objective To understand the carrying status and drug resistance of streptococcus agalactiae in the genital tract of pregnant women.Methods The vaginal discharge from pregnant women was collected for conducting the bacterial culture.The iso-lated streptococcus agalactiae was performed the identification and the drugs susceptibility test by the VITEK 2 fully automatice bacterial identification analyzer.Results Among 1 042 samples,streptococcus agalactiae was isolated in 6 cases with the separation rate of 5.4%.The drug sensitivity test showed the Streptococcus agalactiae was highly sensitive to penicillin,cefazolin,vancomycin, linezolid and nitrofurantoin.The intermediate strains or drug resistant strains were not found.The sensitive rate to levofloxacin, moxifloxacin and clindamycin,erythromycin,tetracycline were 84.0%,84.0%,71.4%,39.3% and 8.9% respectively.Conclusion The carrying rate of streptococcus agalactiae in pregnant women is not high in our hospital.Penicillin and cefazolin should be as the drug of first choice for therapy and the clincal and laboratory nould payattention to detection of streptococcus agalactiae in pregnant women.

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

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


    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.

  4. Isolation

    Agerholm, Frank Juul


    Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!......Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!...

  5. 孕妇泌尿生殖道无乳链球菌感染的耐药性分析%Drug resistance of Streptococcus agalactiae causing urogenital tract infections in pregnant women

    徐水清; 周俊; 王先利


    目的:分析孕妇泌尿生殖道无乳链球菌的耐药性,为临床感染的预防、合理用药提供参考依据。方法收集医院2009年2月-2013年11月门诊及住院3000例孕产妇的临床资料,培养鉴定阴道分泌物、宫腔内容物、中段尿以及羊水培养中的细菌,并对分离的无乳链球菌进行药敏试验,统计分析数据采用WHONET 5.5软件进行。结果分离出无乳链球菌281株占9.37%,其中阴道分泌物检出率居首位,阳性率为12.61%,羊水检出率最低,阳性率为0.93%;所有无乳链球菌对利奈唑胺、头孢唑林、万古霉素敏感率为100.00%;对头孢曲松、青霉素无耐药株,但有中介株,中介率分别为2.85%、1.78%;对红霉素、克林霉素、左氧氟沙星耐药率分别为60.50%、55.87%、47.69%。结论对孕妇泌尿生殖道进行无乳链球菌药敏试验,不仅能有效地预防该菌感染,还可以为临床提供治疗及指导依据,进一步预防其危害性。%OBJECTIVE To analyze the drug resistance of the Streptococcus agalactiae causing urogenital tract in‐fections in pregnant women so as to provide guidance for prevention of clinical infection and reasonable use of anti‐biotics .METHODS The clinical data were collected from 3 000 pregnant women who were treated in the outpatient department or hospitalized from Feb 2009 to Nov 2013 .The vaginal secretions ,uterine cavity contents ,midstream urine ,and amniotic fluid were cultured for bacteria ,the drug susceptibility testing of the isolated S . agalactiae strains was carried out ,and the statistical analysis of data was performed with the use of WHONET 5 .5 software . RESULTS Totally 281 (9 .37% ) strains of S .agalactiae were isolated ,of which 12 .61% were isolated from the vaginal secretions and 0 .93% were isolated from the amniotic fluid .The drug susceptibility rates of all the S . agalactiae strains to linezolid

  6. A four-year monitoring of community-acquired Streptococcus agalactiae infection in urogenital tract%社区获得性泌尿生殖道无乳链球菌感染4年监测

    李国钢; 周杨霄; 李佳俊; 方美芳; 万汝根


    OBJECTIVE To understand the status of the community-acquired Streptococcus agalactiae infection of urogenital tract and drug resistance for the rational use of antibiotics. METHODS A retrospective analysis and statistics were performed for the S. Agalactiae isolated from the genitourinary tract specimens of outpatient in our hospital from Jan 2007 to Dec 2010. RESULTS The resistance rates of S. Agalactiae to erythromycin and clindamycin showed an increasing trend year by years the resistance rates to the tetracycline and levofloxacin had no significant difference among four years; the resistance rate to penicillin was less than 10. 0% ; The resistance to vancomycin and linezolid had not be found. The positive rate of D-test was 28. 6%. CONCLUSION Use of macrolides and clindamycin should be minimized in the treatment of S. Agalactiae infection. Penicillin antibiotics can not be used in preventive and empirical therapy. Clinicians should have a reasonable choice in antibiotics based on the susceptibility results.%目的 了解社区获得性泌尿生殖道无乳链球菌感染现状及耐药性分析,以指导临床合理用药.方法 对2007年1月-2010年12月送检的泌尿生殖道标本所分离的无乳链球菌,进行回顾性分析.结果 红霉素、克林霉素对无乳链球菌的耐药率呈逐年上升趋势,且增长速度很快;四环素与左氧氟沙星的耐药率4年间无明显差异;青霉素耐药率均<10.0%;万古霉素及利奈唑胺未发现耐药;D试验阳性率为28.6%.结论 在诊治无乳链球菌感染的过程中,应尽量减少大环内酯类药物及克林霉素的使用,青霉素类抗菌药物已不能作为预防和经验治疗用药,临床医师应根据药敏结果合理选择抗菌药物.

  7. On the agalactia post partum in the sow. A clinical study.

    Hermansson, I; Einarsson, S; Larsson, K; Bäckström, L


    The purpose of the present investigation was to study the clinical symptoms of agalactic sows post partum in randomly selected swine herds. The study comprised 71 sows affected with agalactia post partum and 71 healthy sows from the same herds used as controls. Average morbidity of the disease in these 50 herds, consisting of an average of 25 sows, was 12.8%. Sixty-three per cent of the agalactic sows were affected within one day after farrowing. There was no difference in gestation length between healthy sows and agalactic sows. The body temperature was significantly higher for affected than for healthy sows. Mastitis was diagnosed in 35 out of 71 agalactic sows. The criteria for mastitis were swelling and herdening of one or several udder glands. Vaginal discharge was recorded for affected sows (55/57) as well as for healthy sows (51/58) and is therefore not significantly associated with agalactia. The temperament of the agalactic sows was moderately or severely affected in 62 out of 71 agalactic sows. Constipation was recorded for 13/59 agalactic sows and for 3/57 normal sows. The agalactic sows had significantly higher number of piglets per litter at birth while the litter size was higher for healthy sows at weaning. Numerically, more sows were culled among agalactic sows (16/48) than among normal sows (6/42). Moreover, sows affected with agalactia at the previous parturition were more inclined to meet with the disease at the next parturition. Further studies comprising large groups of animals are, however, necessary for evaluation of this question.


    Alessandro Abramo


    Full Text Available Necrotizing fasciitis is a fulminant infection that affects the deep and superficial fascia while initially sparing the overlying skin and underlying muscle. The involvement of the cervicalcompartment is associated with a high morbidity and mortality rate. Immunocompromised patients are prone to this infection. We present a case of a 80 year-old diabetic woman suffering from streptococcal cervical NF (probably secondary to a arm injury with an uncommon involvement of the posterior cervical compartment, highlighting the atypical responsible microorganism (Streptococcus Agalactiae, the role of imaging for early diagnosis and the timely surgical and medical treatment for a successful outcome.

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

    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


    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 als

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

    Masoumeh Malek-Jafarian


    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.

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

    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


    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 als

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

    Mavenyengwa R


    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.

  13. Taxa de infecção e sorotipos de Streptococcus agalactiae em amostras de recém-nascidos infectados na cidade de Campinas (SP, Brasil Infection rate and Streptococcus agalactiae serotypes in samples of infected neonates in the city of Campinas (São Paulo, Brazil

    Kateli Fiolo


    Full Text Available OBJETIVO: Descrever os casos e o perfil microbiológico dos sorotipos de Streptococcus agalactiae provenientes de recém-nascidos em um Centro de Referência da Saúde da Mulher na cidade de Campinas, São Paulo, Brasil. MÉTODOS: Estudo transversal clínico-laboratorial realizado de janeiro de 2007 a dezembro de 2011. As cepas suspeitas, isoladas em amostras de sangue e líquor, foram identificadas a partir da hemólise em ágar sangue, coloração de Gram, provas de catalase, teste de CAMP, hidrólise do hipurato ou por automação microbiológica Vitek 2 BioMerieux®. A seguir, estas cepas foram tipadas por PCR utilizando sucessivamente primers específicos para espécie e para nove sorotipos de S. agalactiae. RESULTADOS: Foram isoladas sete amostras de sangue, uma de líquor e uma de secreção ocular provenientes de nove recém-nascidos com infecções causadas pelo S. agalactiae, sendo sete casos de infecção de início precoce e duas de início tardio. Apenas um destes casos foi positivo para amostras pareadas mãe-filho. Para um total de 13.749 partos no período, os 7 casos correspondem a 0,5 caso de infecção precoce por Streptococcus do Grupo B a cada 1 mil nascidos vivos (ou 0,6 casos por 1 mil, incluindo os 2 de infecção tardia, tendo ocorrido 1, 3, 2, nenhum e 3 casos (um precoce e dois tardios, respectivamente, nos anos de 2007 a 2011. Foi possível realizar o PCR para sete amostras, sendo duas de cada um dos sorotipos Ia e V e o sorotipo III em três amostras, uma delas em um recém-nascido e outras duas em amostra pareada mãe-filho. CONCLUSÕES: Embora com casuística limitada, os sorotipos encontrados coincidem com os mais prevalentes na literatura mundial, mas diferem dos estudos brasileiros, exceto para o sorotipo Ia.PURPOSE: To describe the epidemiological cases and microbiological profile of Streptococcus agalactiae serotypes isolated from infected newborns of a Women's Health Reference Centre of Campinas, São Paulo

  14. Presence of contagious agalactia causing mycoplasmas in Spanish goat artificial insemination centres.

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


    Male goats admitted to artificial insemination centres come from herds that have shown no clinical symptoms of contagious agalactia (CA) for the last 6 mo. However, prior reports suggest that this control measure may not be completely effective. This study was designed to detect the presence of CA-causing mycoplasmas in 9 Spanish centres, comprising 159 goats (147 males and 12 teaser does) of 8 different breeds. A microbiological study was conducted during 8 mo on 448 samples (318 ear swabs, 119 semen samples and 11 milk samples). In 86 samples (84 swabs, 1 semen sample and 1 milk sample), CA-causative mycoplasmas were detected by PCR or culture, and 52 animals (49 goat males and 3 teaser does) tested positive. Most of these positive animals were auricular carriers (n = 50), mainly of Mycoplasma mycoides subsp. capri (Mmc), although some M. agalactiae (Ma) and, interestingly, M. capricolum subsp. capricolum (Mcc) carriers were also identified. At least 1 animal infected by CA-causing mycoplasmas was detected in 8 of the 9 centres (88.8%) although in most (66.7%) no infected animals or only 1 or 2 positive animals were identified. Our results indicate the presence of CA carriers as asymptomatic animals in reproductive programmes. These findings have already prompted efficient measures to detect and avoid the entry of these carriers in Spanish centres. We recommend similar measures for all centres in areas where CA is endemic.

  15. Latent infection of male goats with Mycoplasma agalactiae and Mycoplasma mycoides subspecies capri at an artificial insemination centre.

    de la Fe, Christian; Gómez Martín, Angel; Amores, Joaquín; Corrales, Juan C; Sánchez, Antonio; Poveda, José B; Contreras, Antonio


    Contagious agalactia affects goats and is caused by several species of mycoplasma including Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri (Mmc). Male goats, latently infected with M. agalactiae and Mmc, were identified at a dairy goat breeding artificial insemination centre. In three samplings, conducted over 1 year, ear swabs were assessed for both of the above organisms using culture and PCR techniques. Serological examination for antibodies against these organisms was performed at each time-point and conjunctival, nasal, rectal and preputial swabs were taken from a sub-sample of animals. Mycoplasma mycoides subsp. capri and M. agalactiae were detected in 80 and four ear swabs, respectively and serology confirmed the presence of both agents. A point prevalence of 0.06 goats infected with Mmc at the first sampling point increased to 0.97 at the last sampling, suggesting spread of infection. Both organisms were also detected in preputial and conjunctival swabs suggesting the shedding of these pathogens by other routes. These findings should inform World Organisation for Animal Health (OIE) guidelines on avoiding the introduction of such pathogens into artificial insemination centres and suggest the need to review current recommendations.

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

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


    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.

  17. Combination of selective enrichment and MALDI-TOF MS for rapid detection of Streptococcus agalactiae colonisation of pregnant women.

    Ábrók, Marianna; Arcson, Ágnes; Lázár, Andrea; Urbán, Edit; Deák, Judit


    Sample preparation was optimized for MALDI-TOF MS directly from selective enrichment broth to detect Streptococcus agalactiae. The method was tested on 100 vaginal samples of pregnant women; positive and negative predictive values were 100 and 91%, respectively. If it indicates positivity, colonisation can be reported 18-24h after sample collection.

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

    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.

  19. Antimicrobial resistance in 42 cases of neonate septicemia caused by Streptococcus agalactiae infection%42例新生儿无乳链球菌败血症耐药性分析

    樊卫红; 赵满仓; 刘杰


    Objective To investigate clinical features and drug resistance of neonate septicemia caused by Streptococcus agalac-tiae in the hospital and to provide clinical guidance for treatment.Methods 42 Streptococcus agalactiae isolates in the hospital were identified and their department distribution as well as drug susceptibility was determined by using automatic VITEK-2 system and criteria released by CLSI in 2012.Results The number of cases in preterm neonatal intensive care unit,full-term neonatal intensive care unit,pediatric intensive care unit and extremely preterm neonatal intensive care unit were 3,26,7 and 6,respectively.The drug resistance rates of the isolated Streptococcus agalactiae to ampicillin,erythromycin,clindamycin,quinupristin/dalfopristin,linezolid, penicillin,tetracycline,tigecycline,vancomycin and levofloxacin were 0.0%,69.0%,71.4%,0.0%,0.0%,0.0%,74.4%,0.0%, 0.0% and 38.1%,respectively.Conclusion The antimicrobial resistance rates of Streptococcus agalactiae which caused neonatal septicemia in our hospital are relatively high.Ampicillin and penicillin are recomended in treatment for neonate septicemia caused by Streptococcus agalactiae .%目的:了解该院住院新生儿无乳链球菌败血症的临床特点及无乳链球菌的耐药性,为临床用药提供参考。方法采用 VITEK-2全自动微生物分析鉴定系统和临床实验室标准化协会(CLSI)2012年的判断标准,对该院临床分离的42株无乳链球菌的来源科室分布和药敏试验情况进行统计分析。结果早产新生儿监护病房3例、足月新生儿监护病房26例、儿童重症监护病房7例、极早产新生儿监护病房6例;无乳链球菌对氨苄西林、红霉素、克林霉素、喹努普汀/达福普汀、利奈唑烷、青霉素、四环素、替加环素、万古霉素、左氧氟沙星的耐药率分别为0.0%、69.0%、71.4%、0.0%、0.0%、0.0%、74.4%、0.0%、0.0%和38.1%。结论

  20. Anatomic location of Mycoplasma mycoides subsp. capri and Mycoplasma agalactiae in naturally infected goat male auricular carriers.

    Gómez-Martín, Angel; De la Fe, Christian; Amores, Joaquín; Sánchez, Antonio; Contreras, Antonio; Paterna, Ana; Buendía, Antonio J; Corrales, Juan C


    This study sought to determine whether male goat auricular carriers of mycoplasmas known to cause contagious agalactia could harbour these microorganisms at anatomical sites other than the ears. A microbiological study was conducted in 6 naturally infected bucks that had been diagnosed as chronic auricular asymptomatic carriers of Mycoplasma (M.) mycoides subsp. capri (Mmc) more than one year previously. To detect mycoplasmas, cultures and PCR were performed on 46 samples taken from each goat from the cardio-respiratory, digestive, nervous, lymph and genitourinary systems and several joints. Of a total of 274 samples analyzed, 28 were positive for mycoplasmas (10.1%): Mmc was detected in 17 (6.1%), Mycoplasma (M.) agalactiae in 12 (4.3%) and both microorganisms were identified in one of the samples. In all 6 goats, mixed infection was observed despite none being auricular carriers of M. agalactiae. Mycoplasma spp. were identified at 15 different sites; the most frequent sites being the joints (31.2%, 5 positive samples), lymph nodes (25%, 4 positive samples) and respiratory tract (25%, 4 positive samples). Positive results were also obtained in three brain tissue (18.7%), two cardiac tissue (12.5%) and one ileum, urethra, testicle and bulbourethral gland (6.25%) samples. The histopathological findings may suggest the presence of mild chronic conditions in some of the organs where the bacteria were found. Our findings reveal for the first time the capacity of Mmc and M. agalactiae to colonize several other organ systems in chronically naturally infected auricular carriers, possibly representing an added risk factor for the spread of these microorganisms. In the case of M. agalactiae, colonization seemed to be independent of the animal's auricular carrier state.

  1. Antigenic determinants of alpha-like proteins of Streptococcus agalactiae.

    Maeland, Johan A; Bevanger, Lars; Lyng, Randi Valsoe


    The majority of group B streptococcus (GBS) isolates express one or more of a family of surface-anchored proteins that vary by strain and that form ladder-like patterns on Western blotting due to large repeat units. These proteins, which are important as GBS serotype markers and as inducers of protective antibodies, include the alpha C (Calpha) and R4 proteins and the recently described alpha-like protein 2 (Alp2), encoded by alp2, and Alp3, encoded by alp3. In this study, we examined antigenic determinants possessed by Alp2 and Alp3 by testing of antibodies raised in rabbits, mainly by using enzyme-linked immunosorbent assays (ELISA) and an ELISA absorption test. The results showed that Alp2 and Alp3 shared an antigenic determinant, which may be a unique immunological marker of the Alp variants of GBS proteins. Alp2, in addition, possessed an antigenic determinant which showed specificity for Alp2 and a third determinant which showed serological cross-reactivity with Calpha. Alp3, in addition to the determinant common to Alp2 and Alp3, harbored an antigenic site which also was present in the R4 protein, whereas no Alp3-specific antigenic site was detected. These ELISA-based results were confirmed by Western blotting and a fluorescent-antibody test. The results are consistent with highly complex antigenic structures of the alpha-like proteins in a fashion which is in agreement with the recently described structural mosaicism of the alp2 and alp3 genes. The results are expected to influence GBS serotyping, immunoprotection studies, and GBS vaccine developments.

  2. Development of primer sets for loop-mediated isothermal amplification that enables rapid and specific detection of Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae

    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 ...

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

    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...

  4. Structural basis of lantibiotic recognition by the nisin resistance protein from Streptococcus agalactiae.

    Khosa, Sakshi; Frieg, Benedikt; Mulnaes, Daniel; Kleinschrodt, Diana; Hoeppner, Astrid; Gohlke, Holger; Smits, Sander H J


    Lantibiotics are potent antimicrobial peptides. Nisin is the most prominent member and contains five crucial lanthionine rings. Some clinically relevant bacteria express membrane-associated resistance proteins that proteolytically inactivate nisin. However, substrate recognition and specificity of these proteins is unknown. Here, we report the first three-dimensional structure of a nisin resistance protein from Streptococcus agalactiae (SaNSR) at 2.2 Å resolution. It contains an N-terminal helical bundle, and protease cap and core domains. The latter harbors the highly conserved TASSAEM region, which lies in a hydrophobic tunnel formed by all domains. By integrative modeling, mutagenesis studies, and genetic engineering of nisin variants, a model of the SaNSR/nisin complex is generated, revealing that SaNSR recognizes the last C-terminally located lanthionine ring of nisin. This determines the substrate specificity of SaNSR and ensures the exact coordination of the nisin cleavage site at the TASSAEM region.

  5. Changes in immunoglobulin levels in whey during experimental Streptococcus agalactiae mastitis.

    Mackie, D P; Logan, E F


    Total protein and immunoglobulin levels in the wheys of eight first lactation heifers, four vaccinated and four unvaccinated, were measured during three consecutive experimental intramammary infections with Streptococcus agalactiae. There were no significant differences between infections 1, 2 and 3 in the protein or immunoglobulin content of the uninfected quarters. Peak whey total protein of the infected quarters came earlier with each infection, until by the third they were seen after eight hours. During this acute phase a reversal of the normal milk IgG1/IgG2 ratio in all infected quarters was measured. Increases in whey IgA and IgM in the infected quarters of the vaccinates were also noted. A similar response only occurred following the third infection of the unvaccinated animals. All whey immunoglobulin levels returned to normal by 48 hours after infection, after which only IgG1 levels increased in infected quarters.

  6. Streptococcus agalactiae Endophthalmitis in Boston Keratoprosthesis in a Patient with Steven–Johnson Syndrome

    Al-Otaibi, Humoud M.; Talea, Mohammed; Kirat, Omar; Stone, Donald U.; May, William N.; Kozak, Igor


    A 25-year-old Syrian male with a previous episode of Stevens-Johnson syndrome with bilateral corneal cicatrization previously underwent surgery for Type 1 Boston Keratoprosthesis (K-Pro). Sixteen months after the K-Pro surgery, the patient presented with decreased vision to hand motion and microbial keratitis of the graft around the K-Pro with purulent discharge. Corneal scrapings were nonrevealing. B-scan in 3 days showed increased debris in the vitreous cavity and thickened retinochoroidal layer. Intravitreal tap and injections of vancomycin and ceftazidime were performed. The vitreous culture revealed β-hemolytic Streptococcus agalactiae; fungal cultures were negative. Repeat B-scan 3 days later demonstrated decreased vitreous opacity, and the patient felt more comfortable and was without pain. His visual acuity improved to 20/70, ocular findings have been stable for 9 months, and the patient continues to be monitored. PMID:27994401

  7. L-proline increases survival of tilapias infected by Streptococcus agalactiae in higher water temperature.

    Zhao, Xian-Liang; Han, Yi; Ren, Shi-Tong; Ma, Yan-Mei; Li, Hui; Peng, Xuan-Xian


    Streptococcosis causes massive tilapia kills, which results in heavy economic losses of tilapia farming industry. Out of the Streptococcosis, Streptococcus agalactiae is the major pathogen. The bacterium causes higher mortality of tilapias in higher than lower temperatures. However, effect of temperature on metabolic regulation which is related to the mortality is largely unknown. The present study showed 50% and 70% mortality of tilapias cultured in 25 °C and 30 °C, respectively, in comparison with no death in 20 °C following infection caused by S. agalactiae. Then, GC/MS based metabolomics was used to investigate a global metabolic response of tilapia liver to the two higher water temperatures compared to 20 °C. Thirty-six and forty-five varied abundance of metabolites were identified in livers of tilapias cultured at 25 °C and 30 °C, respectively. More decreasing abundance of amino acids and increasing abundance of carbohydrates were detected in 30 °C than 25 °C groups. On the other hand, out of the pathways enriched, the first five biggest impact pathways belong to amino acid metabolism. Decreasing abundance of l-proline was identified as a crucial biomarker for indexing higher water temperature and a potential modulator to reduce the high death. This was validated by engineering injection or oral addition of l-proline. Exogenous l-proline led to elevated amino acid metabolism, which contributes to the elevated survivals. Our findings provide a potential metabolic modulator for controlling the disease, and shed some light on host metabolic prevention to infectious diseases.

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

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


    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.

  9. 2006-2011年无乳链球菌的临床分布及药敏监测%Clinical distribution and drug sensitivities of Streptococcus agalactiae in hospital

    郭远瑜; 张福林; 倪克明; 楚旭


    目的 调查分析无乳链球菌(GBS)的耐药情况及其临床分布,为临床合理用药提供依据.方法 对临床各科室送检各类标本进行无乳链球菌的培养,经ATB-Expression细菌分析系统进行鉴定和药敏试验;所有资料采用WHONET 5.6软件进行回顾性分析.结果 共分离448株GBS,临床标本分离率最高的是泌尿生殖道分泌物(86.8%),科室检出最多的是妇产科(85.0%);药敏结果显示:耐药率最高的是四环素94.9%,其次是红霉素80.1%和克林霉素62.7%,其他耐药率分别为左旋氧氟沙星36.4%,奎奴普丁/达福普汀20.3%,氯霉素16.3%,头孢噻肟1.1%,青霉素0.2%;呋喃妥因、万古霉素和利奈唑胺耐药率均为0%;克林霉素耐药率表现出下降趋势,左旋氧氟沙星呈现上升趋势.结论 加强无乳链球菌的培养检测,关注无乳链球菌的耐药趋势;提高临床重视并指导临床合理用药和治疗.%Objective To investigate the drug susceptibility and distribution of the Streptococcus agalactiaes and provide the evidence for clinical drug usage. Methods Various types of clinical specimens were cultured for Streptococcus agalactiae examination. 448 clinical isolates of Streptococcus agalactiaes ( GBS) were identified and their drug sensitivities were determined by ATB-Expression system, and all of the data was analyzed by WHONE T5. 6 Software. Results A total of 448 strains were isolated; the isolation rate of urogenital tract secretion was the highest (86. 8% ) , and most of the strains came from the department of Obstetrics and Gynecology (85. 0% ) . The resistance rates of GBS to tetracycline was the the highest (94. 9% ) , followed by erythromycin (80. 1% ) and clindamycin (62. 7% ). The resistance rates to other antibiotics were as follows; levo-floxacin 36. 4% , moxifloxacin 20. 8% , quinupristin/dalfopristin 20. 3% , chloramphenicol 16. 3% , cefotaxime 1.1%, penicillin 0.2%; nitrofurantoin, vancomycin and

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

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


    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.

  11. Successful off-label use of the Cepheid Xpert GBS in a late-onset neonatal meningitis by Streptococcus agalactiae

    Savini, Vincenzo; Marrollo, Roberta; Coclite, Eleonora; Fusilli, Paola; D’Incecco, Carmine; Fazii, Paolo


    We report the case of a late-onset neonatal meningitis by Streptococcus agalactiae (group B Streptococcus - GBS) that was diagnosed with a latex agglutination assay (on cerebrospinal fluid, CSF), as well as by using, for the first time, Xpert GBS (Cepheid, US) on CSF. Due to empirical antibiotics given before sampling, both CSF and blood culture were negative, so the abovementioned diagnostics was crucial. Moreover, the Xpert GBS assay, performed according to an off-label, modified protocol (...

  12. Streptococcus agalactiae in pregnant women in Zimbabwe: Epidemiology and serotype marker characteristics

    Mavenyengwa, Rooyen Tinago


    BackgroundStreptococcus agalactiae (group B streptococcus, GBS) is one of the leading causes of serious bacterial infections such as septicaemia, pneumonia and meningitis in newborns, causing 0.5 to more than 2 cases per 1,000 live births. Children may be infected from their mothers during delivery from GBS colonizing the birth canal. GBS is also an important cause of premature delivery and may cause infections in parturient women. Significant differences in colonization rates have been shown...

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

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


    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

  14. Comprehensive RNA-Seq Profiling to Evaluate the Sheep Mammary Gland Transcriptome in Response to Experimental Mycoplasma agalactiae Infection

    Chopra-Dewasthaly, Rohini; Korb, Melanie; Brunthaler, René; Ertl, Reinhard


    Mycoplasma agalactiae is a worldwide serious pathogen of small ruminants that usually spreads through the mammary route causing acute to subacute mastitis progressing to chronic persistent disease that is hard to eradicate. Knowledge of mechanisms of its pathogenesis and persistence in the mammary gland are still insufficient, especially the host-pathogen interplay that enables it to reside in a chronic subclinical state. This study reports transcriptome profiling of mammary tissue from udders of sheep experimentally infected with M. agalactiae type strain PG2 in comparison with uninfected control animals using Illumina RNA-sequencing (RNA-Seq). Several differentially expressed genes (DEGs) were observed in the infected udders and RT-qPCR analyses of selected DEGs showed their expression profiles to be in agreement with results from RNA-Seq. Gene Ontology (GO) analysis revealed majority of the DEGs to be associated with mycoplasma defense responses that are directly or indirectly involved in host innate and adaptive immune responses. Similar RNA-Seq analyses were also performed with spleen cells of the same sheep to know the specific systemic transcriptome responses. Spleen cells exhibited a comparatively lower number of DEGs suggesting a less prominent host response in this organ. To our knowledge this is the first study that describes host transcriptomics of M. agalactiae infection and the related immune-inflammatory responses. The data provides useful information to further dissect the molecular genetic mechanisms underlying mycoplasma mastitis, which is a prerequisite for designing effective intervention strategies. PMID:28081235

  15. Characterization of two novel gadd45a genes in hybrid tilapia and their responses to the infection of Streptococcus agalactiae.

    Shen, Yubang; Ma, Keyi; Liu, Feng; Yue, Gen Hua


    Diseases are one of the major challenges in tilapia aquaculture. Identification of DNA markers associated with disease resistance may facilitate the acceleration of the selection for disease resistance. Gadd45a (growth arrest and DNA damage 45 A), a stress-inducible gene in humans and mice, has not been studied in fish. We characterized the two prologues of Gadd45a genes in hybrid tilapia. Gadd45a1 and Gadd45a2 shared an identical gene structure and showed an amino acid sequence identity of 73.8%. Their expressions were detected in all 10 tissues examined, with the kidney and gill having high transcriptional expressions. The expression levels of Gadd45a1 were significantly lower than those of Gadd45a2 in all examined tissues. After a challenge with a bacterial pathogen Streptococcus agalactiae, the expressions of the two genes were up-regulated significantly in the spleen, kidney, liver and intestine. These findings suggest that the two Gadd45a genes play an important role in the resistance to S. agalactiae in tilapia. We identified 10 SNPs in the two genes. The SNP markers in the two Gadd45a genes could be used to examine whether they are associated with resistance to S. agalactiae.

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

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


    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.

  17. Nucleotide composition bias and codon usage trends of gene populations in Mycoplasma capricolum subsp. capricolum and M. agalactiae

    Xiao-Xia Ma; Yu-Ping Feng; Jia-Ling Bai; De-Rong Zhang; Xin-Shi Lin; Zhong-Ren Ma


    Because of the low GC content of the gene population, amino acids of the two mycoplasmas tend to be encoded by synonymous codons with an A or T end. Compared with the codon usage of ovine, Mycoplasma capricolum and M. agalactiae tend to select optimal codons, which are rare codons in ovine. Due to codon usage pattern caused by genes with key biological functions, the overall codon usage trends represent a certain evolutionary direction in the life cycle of the two mycoplasmas. The overall codon usage trends of a gene population of M. capricolum subsp. capricolum can be obviously separated from other mycoplasmas, and the overall codon usage trends of M. agalactiae are highly similar to those of M. bovis. These results partly indicate the independent evolution of the two mycoplasmas without the limits of the host cell’s environment. The GC and AT skews estimate nucleotide composition bias at different positions of nucleotide triplets and the protein consideration caused by the nucleotide composition bias at codon positions 1 and 2 largely take part in synonymous codon usage patterns of the two mycoplasmas. The correlation between the codon adaptation index and codon usage variation indicates that the effect of codon usage on gene expression in M. capricolum subsp. capricolum is opposite to that of M. agalactiae, further suggesting independence of the evolutionary process influencing the overall codon usage trends of gene populations of mycoplasmas.


    A. S. Rozhdestvenskaya


    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.

  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

    Mahmmod, Yasser; Toft, Nils; Katholm, Jørgen


    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......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...

  20. Uncaria tomentosa increases growth and immune activity in Oreochromis niloticus challenged with Streptococcus agalactiae.

    Yunis-Aguinaga, Jefferson; Claudiano, Gustavo S; Marcusso, Paulo F; Manrique, Wilson Gómez; de Moraes, Julieta R Engrácia; de Moraes, Flávio R; Fernandes, João B K


    Cat's claw (Uncaria tomentosa) is an Amazon herb using in native cultures in Peru. In mammals, it has been described several effects of this herb. However, this is the first report of its use on the diet of fish. The aim of this study was to determinate the effect of this plant on the growth and immune activity in Oreochromis niloticus. Nile tilapia (81.3 ± 4.5 g) were distributed into 5 groups and supplemented with 0 (non-supplement fish), 75, 150, 300, and 450 mg of U. of diet for a period of 28 days. Fish were inoculated in the swim bladder with inactivated Streptococcus agalactiae and samples were taken at 6, 24, and 48 h post inoculation (HPI). Dose dependent increases were noted in some of the evaluated times of thrombocytes and white blood cells counts (WBC) in blood and exudate, burst respiratory activity, lysozyme activity, melanomacrophage centers count (MMCs), villi length, IgM by immunohistochemistry in splenic tissue, and unexpectedly on growth parameters. However, dietary supplementation of this herb did not affect red blood cells count (RBC), hemoglobin, and there were no observed histological lesions in gills, intestine, spleen, and liver. The current results demonstrate for the first time that U. tomentosa can stimulate fish immunity and improve growth performance in Nile tilapia. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    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.

  2. Biofilm formation by Streptococcus agalactiae: influence of environmental conditions and implicated virulence factors.

    Rosini, Roberto; Margarit, Immaculada


    Streptococcus agalactiae (Group B Streptococcus, GBS) is an important human pathogen that colonizes the urogenital and/or the lower gastro-intestinal tract of up to 40% of healthy women of reproductive age and is a leading cause of sepsis and meningitis in the neonates. GBS can also infect the elderly and immuno-compromised adults, and is responsible for mastitis in bovines. Like other Gram-positive bacteria, GBS can form biofilm-like three-dimensional structures that could enhance its ability to colonize and persist in the host. Biofilm formation by GBS has been investigated in vitro and appears tightly controlled by environmental conditions. Several adhesins have been shown to play a role in the formation of GBS biofilm-like structures, among which are the protein components of pili protruding outside the bacterial surface. Remarkably, antibodies directed against pilus proteins can prevent the formation of biofilms. The implications of biofilm formation in the context of GBS asymptomatic colonization and dissemination to cause invasive disease remain to be investigated in detail.

  3. Structure of Streptococcus agalactiae glyceraldehyde-3-phosphate dehydrogenase holoenzyme reveals a novel surface.

    Ayres, Chapelle A; Schormann, Norbert; Senkovich, Olga; Fry, Alexandra; Banerjee, Surajit; Ulett, Glen C; Chattopadhyay, Debasish


    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a conserved cytosolic enzyme, which plays a key role in glycolysis. GAPDH catalyzes the oxidative phosphorylation of D-glyceraldehyde 3-phosphate using NAD or NADP as a cofactor. In addition, GAPDH localized on the surface of some bacteria is thought to be involved in macromolecular interactions and bacterial pathogenesis. GAPDH on the surface of group B streptococcus (GBS) enhances bacterial virulence and is a potential vaccine candidate. Here, the crystal structure of GBS GAPDH from Streptococcus agalactiae in complex with NAD is reported at 2.46 Å resolution. Although the overall structure of GBS GAPDH is very similar to those of other GAPDHs, the crystal structure reveals a significant difference in the area spanning residues 294-307, which appears to be more acidic. The amino-acid sequence of this region of GBS GAPDH is also distinct compared with other GAPDHs. This region therefore may be of interest as an immunogen for vaccine development.

  4. Biofilm formation by Streptococcus agalactiae: influence of environmental conditions and implicated virulence factors

    Imma eMargarit


    Full Text Available Streptococcus agalactiae (Group B Streptococcus, GBS is an important human pathogen that colonizes the urogenital and/or the lower gastro-intestinal tract of up to 40% of healthy women of reproductive age and is a leading cause of sepsis and meningitis in the neonates. GBS can also infect the elderly and immuno-compromised adults, and is responsible for mastitis in bovines. Like other Gram-positive bacteria, GBS can form biofilm-like three-dimensional structures that could enhance its ability to colonize and persist in the host. Biofilm formation by GBS has been investigated in vitro and appears tightly controlled by environmental conditions. Several adhesins have been shown to play a role in the formation of GBS biofilm-like structures, among which are the protein components of pili protruding outside the bacterial surface. Remarkably, antibodies directed against pilus proteins can prevent the formation of biofilms. The implications of biofilm formation in the context of GBS asymptomatic colonization and dissemination to cause invasive disease remain to be investigated in detail.

  5. Significance, management and prevention of Streptococcus agalactiae infection during the perinatal period.

    Berner, Reinhard


    The highest annual death rate during the first five decades of life occurs in the first year, particularly during the perinatal period between the onset of labor and 72 h after birth. Invasive bacterial disease evoking the severe inflammatory response syndrome is a leading cause of perinatal morbidity and mortality. Group B streptococcus (Streptococcus agalactiae) is the most important pathogen in this period of life, although the concept of intrapartum antimicrobial prophylaxis has impressively reduced the rate of culture-proven invasive infection in neonates. This strategy, however, has considerable limitations since group B streptococcus-related stillbirths or prematurity and late-onset sepsis cannot be prevented. Moreover, the use of intrapartum antimicrobial prophylaxis has significantly increased the use of antibiotics during labor and therefore may select for intrapartum infections caused by other bacteria, including those resistant to antibiotics. Several advances in the development of vaccines and research on virulence factors and pathways involved in the immune response to group B streptococcus have been accomplished within the last years, including complete sequencing of the group B streptococcus genome. Development of effective vaccines and implementation of vaccination strategies will be one of the key challenges in the future for prevention of neonatal group B Streptococcus infections.

  6. Recombination between Streptococcus suis ICESsu32457 and Streptococcus agalactiae ICESa2603 yields a hybrid ICE transferable to Streptococcus pyogenes.

    Marini, Emanuela; Palmieri, Claudio; Magi, Gloria; Facinelli, Bruna


    Integrative conjugative elements (ICEs) are mobile genetic elements that reside in the chromosome but retain the ability to undergo excision and to transfer by conjugation. Genes involved in drug resistance, virulence, or niche adaptation are often found among backbone genes as cargo DNA. We recently characterized in Streptococcus suis an ICE (ICESsu32457) carrying resistance genes [tet(O/W/32/O), tet(40), erm(B), aphA, and aadE] in the 15K unstable genetic element, which is flanked by two ∼1.3kb direct repeats. Remarkably, ∼1.3-kb sequences are conserved in ICESa2603 of Streptococcus agalactiae 2603V/R, which carry heavy metal resistance genes cadC/cadA and mer. In matings between S. suis 32457 (donor) and S. agalactiae 2603V/R (recipient), transconjugants were obtained. PCR experiments, PFGE, and sequence analysis of transconjugants demonstrated a tandem array between ICESsu32457 and ICESa2603. Matings between tandem array-containing S. agalactiae 2603V/R (donor) and Streptococcus pyogenes RF12 (recipient) yielded a single transconjugant containing a hybrid ICE, here named ICESa2603/ICESsu32457. The hybrid formed by recombination of the left ∼1.3-kb sequence of ICESsu32457 and the ∼1.3-kb sequence of ICESa2603. Interestingly, the hybrid ICE was transferable between S. pyogenes strains, thus demonstrating that it behaves as a conventional ICE. These findings suggest that both tandem arrays and hybrid ICEs may contribute to the evolution of antibiotic resistance in streptococci, creating novel mobile elements capable of disseminating new combinations of antibiotic resistance genes.

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

    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.


    AP Jiménez


    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.

  9. Conjugative Transfer and cis-Mobilization of a Genomic Island by an Integrative and Conjugative Element of Streptococcus agalactiae

    Puymège, Aurore; Bertin, Stéphane; Chuzeville, Sarah; Guédon, Gérard; Payot, Sophie


    Putative integrative and conjugative elements (ICEs), i.e., genomic islands which could excise, self-transfer by conjugation, and integrate into the chromosome of the bacterial host strain, were previously identified by in silico analysis in the sequenced genomes of Streptococcus agalactiae (M. Brochet et al., J. Bacteriol. 190:6913–6917, 2008). We investigated here the mobility of the elements integrated into the 3′ end of a tRNALys gene. Three of the four putative ICEs tested were found to ...

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

    Sybille eLandwehr-Kenzel


    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

  11. Streptococcus agalactiae capsule polymer length and attachment is determined by the proteins CpsABCD.

    Toniolo, Chiara; Balducci, Evita; Romano, Maria Rosaria; Proietti, Daniela; Ferlenghi, Ilaria; Grandi, Guido; Berti, Francesco; Ros, Immaculada Margarit Y; Janulczyk, Robert


    The production of capsular polysaccharides (CPS) or secreted exopolysaccharides is ubiquitous in bacteria, and the Wzy pathway constitutes a prototypical mechanism to produce these structures. Despite the differences in polysaccharide composition among species, a group of proteins involved in this pathway is well conserved. Streptococcus agalactiae (group B Streptococcus; GBS) produces a CPS that represents the main virulence factor of the bacterium and is a prime target in current vaccine development. We used this human pathogen to investigate the roles and potential interdependencies of the conserved proteins CpsABCD encoded in the cps operon, by developing knock-out and functional mutant strains. The mutant strains were examined for CPS quantity, size, and attachment to the cell surface as well as CpsD phosphorylation. We observed that CpsB, -C, and -D compose a phosphoregulatory system where the CpsD autokinase phosphorylates its C-terminal tyrosines in a CpsC-dependent manner. These Tyr residues are also the target of the cognate CpsB phosphatase. An interaction between CpsD and CpsC was observed, and the phosphorylation state of CpsD influenced the subsequent action of CpsC. The CpsC extracellular domain appeared necessary for the production of high molecular weight polysaccharides by influencing CpsA-mediated attachment of the CPS to the bacterial cell surface. In conclusion, although having no impact on cps transcription or the synthesis of the basal repeating unit, we suggest that these proteins are fine-tuning the last steps of CPS biosynthesis (i.e. the balance between polymerization and attachment to the cell wall).

  12. Functional analysis of the CpsA protein of Streptococcus agalactiae.

    Hanson, Brett R; Runft, Donna L; Streeter, Cale; Kumar, Abhin; Carion, Thomas W; Neely, Melody N


    Streptococcal pathogens, such as the group B streptococcus (GBS) Streptococcus agalactiae, are an important cause of systemic disease, which is facilitated in part by the presence of a polysaccharide capsule. The CpsA protein is a putative transcriptional regulator of the capsule locus, but its exact contribution to regulation is unknown. To address the role of CpsA in regulation, full-length GBS CpsA and two truncated forms of the protein were purified and analyzed for DNA-binding ability. Assays demonstrated that CpsA is able to bind specifically to two putative promoters within the capsule operon with similar affinity, and full-length protein is required for specificity. Functional characterization of CpsA confirmed that the ΔcpsA strain produced less capsule than did the wild type and demonstrated that the production of full-length CpsA or the DNA-binding region of CpsA resulted in increased capsule levels. In contrast, the production of a truncated form of CpsA lacking the extracellular LytR domain (CpsA-245) in the wild-type background resulted in a dominant-negative decrease in capsule production. GBS expressing CpsA-245, but not the ΔcpsA strain, was attenuated in human whole blood. However, the ΔcpsA strain showed significant attenuation in a zebrafish infection model. Furthermore, chain length was observed to be variable in a CpsA-dependent manner, but could be restored to wild-type levels when grown with lysozyme. Taken together, these results suggest that CpsA is a modular protein influencing multiple regulatory functions that may include not only capsule synthesis but also cell wall associated factors.

  13. Importancia clínica del Streptococcus agalactiae como causante de infección.

    María del Pilar Crespo Ortiz


    Full Text Available El Streptococcus agalactiae o estreptococo ß-hemolítico del grupo b (S. b es causa importante de sepsis neonatal y de infecciones en gestantes y adultos inmunocomprometidos. Se trata de una bacteria encapsulada cuya virulencia se atribuye a una toxina polisacárida y se caracteriza por presentar una alta concentración inhibitoria mínima (CIM a la penicilina. Su papel como patógeno potencial se ha reconocido ampliamente en países industrializados donde en la actualidad se desarrollan estrategias de diagnóstico y prevención dada su alta morbimortalidad; sin embargo, en países en desarrollo no se informa con frecuencia la infección por este germen, básicamente debido a la circulación de serotipos menos virulentos y la falta de una búsqueda y diagnóstico adecuados. La experiencia de la Fundación Valle del Lili, Cali, Colombia, revela en poco tiempo la presencia frecuente de esta bacteria como colonizante y como origen de infecciones severas en neonatos y adultos, con una resistencia moderada a la penicilina y alta a la gentamicina y tetraciclina. Por las implicaciones de esta infección y por lo imperativo de conocer más acerca de ella, se revisan las características de la infección y de los casos, con lo que se pretende alertar sobre la presencia de este germen en Cali a fin de orientar hacia estrategias adecuadas para mejorar su descubrimiento en el laboratorio y hacer un buen diagnóstico y un manejo terapéutico eficaz.

  14. Molecular characterization and expression of CD2 in Nile tilapia (Oreochromis niloticus) in response to Streptococcus agalactiae stimulus.

    Gan, Zhen; Wang, Bei; Tang, Jufen; Lu, Yishan; Jian, JiChang; Wu, Zaohe; Nie, Pin


    The cluster of differentiation 2 (CD2), functioning as a cell adhesion and costimulatory molecule, plays a crucial role in T-cell activation. In this paper, the CD2 gene of Nile tilapia, Oreochromis niloticus (designated as On-CD2) was cloned and its expression pattern under the stimulation of Streptococcus agalactiae was investigated. Sequence analysis showed On-CD2 protein consists of two extracellular Ig-like domains, a transmembrane region, and a long proline-rich cytoplasmic tail, which is a hallmark of CD2, and several important structural characteristics required for T-cell activation were detected in the deduced amino acid sequence of On-CD2. In healthy tilapia, the On-CD2 transcripts were mainly detected in the head kidney, spleen, blood and thymus. Moreover, there was a clear time-dependent expression pattern of On-CD2 after immunized by formalin-inactivated S. agalactiae and the expression reached the highest level at 12 h in the brain and head kidney, 48 h in the spleen, and 72 h in the thymus, respectively. This is the first report on the expression of CD2 induced by bacteria vaccination in teleosts. These findings indicated that On-CD2 may play an important role in the immune response to intracellular bacteria in Nile tilapia.

  15. Contagious agalactia due to Mycoplasma spp. in small dairy ruminants: epidemiology and prospects for diagnosis and control.

    Gómez-Martín, Angel; Amores, Joaquín; Paterna, Ana; De la Fe, Christian


    Contagious agalactia (CA) is a serious disease of small dairy ruminants that has a substantial economic impact on the goat and sheep milk industries. The main aetiological agent of the disease is Mycoplasma agalactiae, although other species, such as Mycoplasma mycoides subsp. capri, Mycoplasma capricolum subsp. capricolum and Mycoplasma putrefaciens, are pathogenic in goats. There are two clinical-epidemiological states of CA in sheep and goats; herds and flocks may exhibit outbreaks of CA or may be chronically infected, the latter with a high incidence of subclinical mastitis and only occasional clinical cases. The complex epidemiology of CA is related to the genetic characteristics and mechanisms of molecular variation of the Mycoplasma spp. involved, along with presence of CA-mycoplasmas in wild ruminant species. In goats, the situation is particularly complex and asymptomatic carriers have been detected in chronically infected herds. The coexistence of other non-pathogenic mycoplasmas in the herd further complicates the diagnosis of CA and the design of efficient strategies to control the disease. Routes of infection, such as the venereal route, may be involved in the establishment of chronic infection in herds. Current challenges include the need for improved diagnostic methods for detection of chronic and subclinical infections and for the design of more efficient vaccines.

  16. Antigen I/II encoded by integrative and conjugative elements of Streptococcus agalactiae and role in biofilm formation.

    Chuzeville, Sarah; Dramsi, Shaynoor; Madec, Jean-Yves; Haenni, Marisa; Payot, Sophie


    Streptococcus agalactiae (i.e. Group B streptococcus, GBS) is a major human and animal pathogen. Genes encoding putative surface proteins and in particular an antigen I/II have been identified on Integrative and Conjugative Elements (ICEs) found in GBS. Antigens I/II are multimodal adhesins promoting colonization of the oral cavity by streptococci such as Streptococcus gordonii and Streptococcus mutans. The prevalence and diversity of antigens I/II in GBS were studied by a bioinformatic analysis. It revealed that antigens I/II, which are acquired by horizontal transfer via ICEs, exhibit diversity and are widespread in GBS, in particular in the serotype Ia/ST23 invasive strains. This study aimed at characterizing the impact on GBS biology of proteins encoded by a previously characterized ICE of S. agalactiae (ICE_515_tRNA(Lys)). The production and surface exposition of the antigen I/II encoded by this ICE was examined using RT-PCR and immunoblotting experiments. Surface proteins of ICE_515_tRNA(Lys) were found to contribute to GBS biofilm formation and to fibrinogen binding. Contribution of antigen I/II encoded by SAL_2056 to biofilm formation was also demonstrated. These results highlight the potential for ICEs to spread microbial adhesins between species.

  17. Streptococcus agalactiae infective endocarditis complicated by large vegetations at aortic valve cusps along with intracoronary extension: An autopsy case report.

    Ro, Ayako


    Streptococcus agalactiae infective endocarditis is a rare condition with high mortality owing to complications of large vegetations and systemic emboli. A 49-year-old man was found dead in his house. He had a history of hepatic cirrhosis and had been diagnosed with type 2 diabetes 2years previously. He had presented with a high fever 10days before his death. An autopsy revealed 50mL of purulent pericardial effusion, and S. agalactiae was detected from the culture of this pericardial effusion. Two slender rope-like vegetations were present at the right aortic valve cusp and noncoronary aortic valve cusp. The vegetation at the right aortic valve cusp extended into the right coronary artery. The right coronary artery was broadly occluded by white rod-like material. The mitral valves were also affected, and the posterior papillary muscle was ruptured. Myocardial infarction was not observed. Systemic microscopic Gram-positive bacterial masses were observed in several organs. The death was attributed to acute myocardial ischemia caused by occlusive intracoronary extension of the vegetation at the proximal right coronary artery.

  18. Conjugative transfer and cis-mobilization of a genomic island by an integrative and conjugative element of Streptococcus agalactiae.

    Puymège, Aurore; Bertin, Stéphane; Chuzeville, Sarah; Guédon, Gérard; Payot, Sophie


    Putative integrative and conjugative elements (ICEs), i.e., genomic islands which could excise, self-transfer by conjugation, and integrate into the chromosome of the bacterial host strain, were previously identified by in silico analysis in the sequenced genomes of Streptococcus agalactiae (M. Brochet et al., J. Bacteriol. 190:6913-6917, 2008). We investigated here the mobility of the elements integrated into the 3' end of a tRNA(Lys) gene. Three of the four putative ICEs tested were found to excise but only one (ICE_515_tRNA(Lys)) was found to transfer by conjugation not only to S. agalactiae strains but also to a Streptococcus pyogenes strain. Transfer was observed even if recipient cell already carries a related resident ICE or a genomic island flanked by attL and attR recombination sites but devoid of conjugation or recombination genes (CIs-Mobilizable Element [CIME]). The incoming ICE preferentially integrates into the 3' end of the tRNA(Lys) gene (i.e., the attR site of the resident element), leading to a CIME-ICE structure. Transfer of the whole composite element CIME-ICE was obtained, showing that the CIME is mobilizable in cis by the ICE. Therefore, genomic islands carrying putative virulence genes but lacking the mobility gene can be mobilized by a related ICE after site-specific accretion.

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

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


    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.

  20. Germicidal activity of a chlorous acid-chlorine dioxide teat dip and a sodium chlorite teat dip during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae.

    Boddie, R L; Nickerson, S C; Adkinson, R W


    Three postmilking teat dips were tested for efficacy against Staphylococcus aureus and Streptococcus agalactiae in two separate studies using experimental challenge procedures that were recommended by the National Mastitis Council. The first study evaluated a barrier teat dip product containing chlorous acid-chlorine dioxide as the germicidal agent, and the second study evaluated a sodium chlorite product with a barrier component as well as a sodium chlorite product without a barrier component. The chlorous acid-chlorine dioxide teat dip reduced new intramammary infections (IMI) caused by Staph. aureus by 91.5% and reduced new IMI caused by Strep. agalactiae by 71.7%. The barrier dip containing sodium chlorite reduced new IMI caused by Staph. aureus and Strep. agalactiae by 41.0 and 0%, respectively. The nonbarrier dip containing sodium chlorite reduced new IMI caused by Staph. aureus by 65.6% and reduced new IMI caused by Strep. agalactiae by 39.1%. Teat skin and teat end conditions were evaluated before and after the second study; no deleterious effects among dipped quarters compared with control quarters were noted for the two sodium chlorite products.

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

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


    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.

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

    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: [Institute of Biotechnology, University of Helsinki, PO Box 65, FIN-00014, Helsinki (Finland)


    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.


    汪开毓; 黄锦炉; 肖丹; 王均; 黄凌远


    Streptococcus agalactiae is an important zoonotic pathogen in human, animal and fish. The capsular poly-saccharides on the cell surface have been recognized as virulence factors, of which the forming processes are regulated by the capsular polysaccharide biosynthesis (cps) genes. The capsular polysaccharide biosynthesis genes exist in capsular polysaccharide operon of S. agalactiae genome, which were predicted to participate in the origination of capsular polysaccharide synthesis, polymerization of oligosaccharides and polysaccharides, transport and anchor the product to the cell surface. Therefore, the cps genes achieve new applications of diagnostic techniques and the construction of attenuated mutant strains. In this paper, the basic properties of cps genes, as well as their transcription regulation, coding proteins and biological functions, regulation mechanism of capsular polysaccharide biosynthesis and serotyping, and application of construction of mutant strains are analyzed and discussed. We looked forward to providing a theoretical reference on the new functions research and innovative applications of S. agalactiae cps genes.%无乳链球菌是一种人畜鱼共患的重要病原菌,菌体表面的荚膜多糖是公认的毒力因子,其合成过程受荚膜多糖合成基因的调控.荚膜多糖合成基因存在于荚膜多糖操纵子中,参与无乳链球菌荚膜多糖的合成启动、寡糖和多糖的聚合以及外输并锚定于菌体表面,在新型诊断技术和减毒突变株的构建方面取得良好的应用.本文首次就cps基因的基本属性、转录调节、编码蛋白及其生物功能、对荚膜多糖合成的调控机理、在血清分型和突变株构建的应用这六个方面进行深入分析和讨论,以期为GBS cps基因的新功能研究和创新应用提供理论参考.

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

    Leonardo Lodolo


    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.

  5. Global transcriptional profiling reveals Streptococcus agalactiae genes controlled by the MtaR transcription factor

    Cvek Urska


    Full Text Available Abstract Background Streptococcus agalactiae (group B Streptococcus; GBS is a significant bacterial pathogen of neonates and an emerging pathogen of adults. Though transcriptional regulators are abundantly encoded on the GBS genome, their role in GBS pathogenesis is poorly understood. The mtaR gene encodes a putative LysR-type transcriptional regulator that is critical for the full virulence of GBS. Previous studies have shown that an mtaR- mutant transports methionine at reduced rates and grows poorly in normal human plasma not supplemented with methionine. The decreased virulence of the mtaR mutant was correlated with a methionine transport defect; however, no MtaR-regulated genes were identified. Results Microarray analysis of wild-type GBS and an mtaR mutant revealed differential expression of 12 genes, including 1 upregulated and 11 downregulated genes in the mtaR mutant. Among the downregulated genes, we identified a cluster of cotranscribed genes encoding a putative methionine transporter (metQ1NP and peptidase (pdsM. The expression of four genes potentially involved in arginine transport (artPQ and arginine biosynthesis (argGH was downregulated and these genes localized to two transcriptional units. The virulence factor cspA, which encodes an extracellular protease, was downregulated. Additionally, the SAN_1255 locus, which putatively encodes a protein displaying similarity to plasminogen activators, was downregulated. Conclusion To our knowledge, this is the first study to describe the global influence of MtaR on GBS gene expression. This study implicates the metQ1NP genes as encoding the MtaR-regulated methionine transporter, which may provide a mechanistic explanation for the methionine-dependent growth defect of the mtaR mutant. In addition to modulating the expression of genes involved in metabolism and amino acid transport, inactivation of mtaR affected the expression of other GBS genes implicated in pathogenesis. These findings

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

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


    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.

  7. Pericarditis purulenta con taponamiento cardíaco por asociación de Streptococcus agalactiae y Salmonella enterica no typhi Purulent pericarditis with pericardial tamponade caused by Streptococcus agalactiae and Salmonellaenterica no typhi

    Lourdes Arruvito


    Full Text Available La pericarditis purulenta (PP es una condición infrecuente, pero con elevada mortalidad. Previo a la era antibiótica, los agentes etiológicos más comúnmente hallados eran Streptococcus pneumoniae y Staphylococcus aureus. Se presenta el caso de un paciente de sexo masculino de 75 años de edad con un cuadro de shock, PP y taponamiento cardíaco, producto de una sepsis por Streptococcus agalactiae y Salmonella entérica no typhi. No se ha hallado ningún caso similar en la literatura. Se destaca el antecedente previo inmediato de la realización de una endoscopia digestiva alta con toma de biopsia de esófago como posible causa de bacteriemia e impacto pericárdico. El curso evolutivo fue malo y el paciente falleció a los 34 días. Esta inusual asociación bacteriana en un huésped debilitado e inmunodeprimido, debería ser incluida dentro de los diagnósticos etiológicos diferenciales de la pericarditis purulenta.Purulent pericarditis (PP is an uncommon condition with high mortality. In the preantibiotic period, Staphyloccocus aureus and Streptococcus pneumoniae were the most common etiologic agents. We describe the case of a 75-year old man with septic shock, PP and cardiac tamponade caused by Streptococcus agalactiae and Salmonella enterica no-typhi. To our knowledge this association of pathogenic organisms has not been previously reported in the literature. The pathogenesis is here reviewed, and in our patient presumably, purulent pericarditis occurred via hematogeneus spread undergoing upper gastrointestinal endoscopy. The patient’s course was complicated and he died on 34 th hospital day. After this case report it is considered that differential etiologic diagnosis of PP should include these agents, especially in immunodepressed patients with predisposing factors.

  8. Overexpression, purification, crystallization and preliminary X-ray diffraction of the nisin resistance protein from Streptococcus agalactiae.

    Khosa, Sakshi; Hoeppner, Astrid; Kleinschrodt, Diana; Smits, Sander H J


    Nisin is a 34-amino-acid antimicrobial peptide produced by Lactococcus lactis belonging to the class of lantibiotics. Nisin displays a high bactericidal activity against various Gram-positive bacteria, including some human-pathogenic strains. However, there are some nisin-non-producing strains that are naturally resistant owing to the presence of the nsr gene within their genome. The encoded protein, NSR, cleaves off the last six amino acids of nisin, thereby reducing its bactericidal efficacy. An expression and purification protocol has been established for the NSR protein from Streptococcus agalactiae COH1. The protein was successfully crystallized using the vapour-diffusion method in hanging and sitting drops, resulting in crystals that diffracted X-rays to 2.8 and 2.2 Å, respectively.

  9. Structure of the Response Regulator NsrR from Streptococcus agalactiae, Which Is Involved in Lantibiotic Resistance.

    Khosa, Sakshi; Hoeppner, Astrid; Gohlke, Holger; Schmitt, Lutz; Smits, Sander H J


    Lantibiotics are antimicrobial peptides produced by Gram-positive bacteria. Interestingly, several clinically relevant and human pathogenic strains are inherently resistant towards lantibiotics. The expression of the genes responsible for lantibiotic resistance is regulated by a specific two-component system consisting of a histidine kinase and a response regulator. Here, we focused on a response regulator involved in lantibiotic resistance, NsrR from Streptococcus agalactiae, and determined the crystal structures of its N-terminal receiver domain and C-terminal DNA-binding effector domain. The C-terminal domain exhibits a fold that classifies NsrR as a member of the OmpR/PhoB subfamily of regulators. Amino acids involved in phosphorylation, dimerization, and DNA-binding were identified and demonstrated to be conserved in lantibiotic resistance regulators. Finally, a model of the full-length NsrR in the active and inactive state provides insights into protein dimerization and DNA-binding.

  10. Controlling contagious agalactia in artificial insemination centers for goats and detection of Mycoplasma mycoides subspecies capri in semen.

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


    Many goat artificial insemination (AI) centers in Spain have adopted new measures to control contagious agalactia (CA). To avoid the introduction of male goats carrying mycoplasma organisms subclinically in their external ear canal (auricular carriers) in these centers, two ear swabs and a blood sample are obtained from all candidate animals for polymerase chain reaction (PCR), culture (swabs) and serologic tests to detect the presence of mycoplasmas. In addition, the semen produced at these centers is routinely cultured and PCR tested also to detect the presence of mycoplasmas. One y after the introduction of this program, we tested 48 ear swabs and 24 blood samples from 24 candidates for admission to these AI Centers. Three of these ear swab samples (3/48, 6.25%) scored positive for the presence of mycoplasmas; Mycoplasma agalactiae (Ma) was detected in two samples and Mycoplasma mycoides subsp. capri (Mmc) in one. All animals were serologically negative for Ma. Also, out of 173 semen samples obtained from 137 admitted animals (2 and 3 samples were obtained in 16 and 10 bucks, respectively), one (1/173, 0.56%) was positive for Mmc. Our findings suggest that ear swab and semen samples are useful tools to control CA at AI Centers. The introduction of this program has also resulted in the first detection of Mmc in semen from a naturally infected goat, confirming the ability of this mycoplasma to colonize the reproductive tract of male goats. These results highlight the need to improve control measures in semen producing centers to minimize the risk of CA transmission.

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

    Rogério Salvador


    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.

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

    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: [University of Alabama at Birmingham, Birmingham, AL 35294 (United States); UNESCO Regional Centre for Biotechnology (RCB), Gurgaon 122 016, Haryana (India)


    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.

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

    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.

  14. Ostéomyélites et ostéoarthrites à streptocoque béta-hémolytique du groupe B ("Streptococcus agalactiae")

    Altrichter, Loan Catherine


    Le "streptococcus agalactiae" est un pathogène retrouvé de façon quasi exclusive chez le nourrisson et la femme enceinte. Les données récentes de la littérature suggèrent une augmentation de sa responsabilité dans les ostéomyélites. De janvier 1999 à juillet 2003, 22 patients hospitalisés à Genève ont présenté une ostéomyélite à "streptococcus agalactiae", prouvée par biopsie osseuse. Tous les patients présentaient un facteur de risque favorisant l'infection, dont 16 (72%) un diabète de type ...

  15. Short communication: comparing real-time PCR and bacteriological cultures for Streptococcus agalactiae and Staphylococcus aureus in bulk-tank milk samples.

    Zanardi, G; Caminiti, A; Delle Donne, G; Moroni, P; Santi, A; Galletti, G; Tamba, M; Bolzoni, G; Bertocchi, L


    For more than 30 yr, a control plan for Streptococcus agalactiae and Staphylococcus aureus has been carried out in more than 1,500 dairy herds of the province of Brescia (northern Italy). From 2010 to 2011, the apparent prevalence of Strep. agalactiae has been relatively stable around 10%, but the apparent prevalence of Staph. aureus has been greater than 40% with an increasing trend. The aim of this paper was to estimate and compare the diagnostic accuracy of 3 assays for the detection of Strep. agalactiae and Staph. aureus in bulk-tank milk samples (BTMS) in field conditions. The assays were a qualitative and a quantitative bacteriological culture (BC) for each pathogen and a homemade multiplex real-time PCR (rt-PCR). Because a gold standard was not available, the sensitivities (Se) and specificities (Sp) were evaluated using a Bayesian latent class approach. In 2012 we collected one BTMS from 165 dairy herds that were found positive for Strep. agalactiae in the previous 2-yr campaigns of eradication plan. In most cases, BTMS collected in these herds were positive for Staph. aureus as well, confirming the wide spread of this pathogen. At the same time we also collected composite milk samples from all the 8,624 lactating cows to evaluate the within-herd prevalence of Strep. agalactiae. Streptococcus agalactiae samples were cultured using a selective medium Tallium Kristalviolette Tossin, whereas for Staph. aureus, we used Baird Parker modified medium with added Rabbit Plasma Fibrinogen ISO-Formulation. In parallel, BTMS were tested using the rt-PCR. Regarding Strep. agalactiae, the posterior median of Se and Sp of the 2 BC was similar [qualitative BC: Se=98%, posterior credible interval (95%PCI): 94-100%, and Sp=99%, 95%PCI: 96-100%; quantitative BC: Se=99%, 95%PCI: 96-100%, and Sp=99%, 95%PCI: 95-100%] and higher than those of the rt-PCR (at 40 cycle threshold, Se=92%, 95%PCI: 85-97%; Sp=94%, 95%PCI: 88-98%). Also in case of Staph. aureus, the posterior medians

  16. Ostéomyélites et ostéoarthrites à Streptocoque béta-hémolytique du groupe B (Streptococcus agalactiae)

    Altrichter, Loan Catherine; Bernard, Louis


    Le «streptococcus agalactiae» est un pathogène retrouvé de façon quasi exclusive chez le nourrisson et la femme enceinte. Les données récentes de la littérature suggèrent une augmentation de sa responsabilité dans les ostéomyélites. De janvier 1999 à juillet 2003, 22 patients hospitalisés à Genève ont présenté une ostéomyélite à «streptococcus agalactiae», prouvée par biopsie osseuse. Tous les patients présentaient un facteur de risque favorisant l’infection, dont 16 (72%) un diabète de type ...

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

    Deguo Wang


    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.

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

    Wang, Deguo; Liu, Yanhong


    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.

  19. An examination of teat drying with disinfectant impregnated cloths on the bacteriological quality of milk and on the transfer of Streptococcus agalactiae before milking.

    McKinnon, C H; Higgs, T M; Bramley, A J


    Total bacterial counts of the milk from individual cows were measured for three groups of ten winter housed cows at three milkings. The teats were either (i) left unwashed or (ii) washed with disinfected water (60 ppm available iodine) and dried with individual paper towels or (iii) washed with plain water and then dried with a single fabric cloth impregnated with a polymeric bisguanide and a quaternary ammonium compound. The mean total bacterial counts/ml for the groups were 5820, 2108 and 1116 respectively. Treatments (ii) and (iii) were also compared for their ability to prevent the inter-teat transfer of bacteria. Before teat washing and drying, one teat of each cow was deliberately contaminated with Streptococcus agalactiae. Significantly fewer teats (5/30) became contaminated with Str. agalactiae when treatment (iii) was used for teat washing and drying compared with treatment (ii) (20/30).

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

    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


    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.

  1. O-Glycosylation of the N-terminal Region of the Serine-rich Adhesin Srr1 of Streptococcus agalactiae Explored by Mass Spectrometry *

    Chaze, Thibault; Guillot, Alain; Valot, Benoît; Langella, Olivier; Chamot-Rooke, Julia; Di Guilmi, Anne-Marie; Trieu-Cuot, Patrick; Dramsi, Shaynoor; Mistou, Michel-Yves


    International audience; Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. Lectin-binding experiments have previously shown that Srr proteins are heavily glycosylated. We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in colli...

  2. A commercial rapid optical immunoassay detects Streptococcus agalactiae from aquatic cultures and clinical specimens

    The BioStar STREPT B Optical ImmunoAssay (OIA) (BioStar® OIA® Strep B Assay Kit; Biostar Incorporation; Louisville, CO, USA) was used to identify 32 known group B streptococcus (GBS) isolates of fish, dolphin, bovine, and human origin. Thirteen non-GBS isolates from fish and other animals were test...

  3. 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.

    Mahmmod, Yasser S; Mweu, Marshal M; Nielsen, Søren S; Katholm, Jørgen; Klaas, Ilka C


    The use of PCR tests as diagnostics for intramammary infections (IMI) based on composite milk samples collected in a non-sterile manner at milk recordings is increasing. Carryover of sample material between cows and non-aseptic PCR sampling may be incriminated for misclassification of IMI 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 milk recordings. Following the farmers' routine pre-milking preparation, 411 of 794 cows were randomly selected for the PSP treatment. These procedures included removing the first streams of milk and 70% alcohol teat disinfection. Composite milk samples were then collected from all cows and tested using PCR. Data on milking order were used to estimate the correlation between consecutively milked cows in each milking unit. Factors associated with the PCR-positivity for S. agalactiae were analyzed using generalized estimating equations assuming a binomially-distributed outcome with a logit link function. Presampling procedures were only significant using cut-off 37. A first-order autoregressive correlation structure provided the best correlation between consecutively milked cows. The correlation was 13%, 11%, 9% at cut-offs <40, 37, and 34, respectively. PSP did not reduce the odds of cows being PCR-positive for S. agalactiae. In conclusion, carryover and non-aseptic sampling affected the PCR results and should therefore be considered when samples from routine milk

  4. Prostaglandin induced parturition in swine with some aspects on prevention of the MMA (metritis, mastitis, agalactia) syndrome.

    Einarsson, S; Gustafsson, B; Larsson, K


    The effect of PGF2alpha to induce parturition in sows was studied in two herds. In one herd with a total number of 150 sows and with a good health status 12 sows were injected intramuscularly with 12.5 mg PGF2alpha during late pregnancy (110--114 days). During the same period 12 sows were treated intramuscularly with saline solution at corresponding gestation stage. The results are given in Tables I and II. The mean age calculated as number of litters was 2.6 in the experimental group and 4.3 in the control group. All sows treated with PGF2alpha farrowed within 30 hours after injection at an average time of 25.4 hours. The controls farrowed at an average 80.2 hours after injection. The mean farrowing time was 4.4 hours for the PG-treated sows while the corresponding figure for the controls was 6.6 hours. The litter size at birth, after 3 weeks and at weaning was somewhat higher in the experimental group. The same applies to the litter weight at the age of 3 weeks. The course of parturition was normal in all sows and the health condition during lactation was good. In another herd -- numbering 18 sows -- all sows which hade farrowed during the last 4 months before the present investigation, had developed the Mastitis-Metritis-Agalactia syndrome (MMA). The symptoms were a prolonged gestation length (116--117 days), fever, mastitis and agalactia. During the present study 3 sows were left untreated while 5 sows were given an intramuscular injection with 12.5 mg PGF2alpha during late pregnancy (112--114 days). Four of the five PG-treated sows farrowed within 32 hours, while the fifth sow farrowed 100 hours after injection. The sow not responding to the PG-treatment and the untreated sows all developed MMA while the four sows responding to treatment remained healthy. The gestation length in treated sows responding to treatment was 113--115 days. In the sows developing MMA the gestation length was 116--117 days. The potential effect for prevention of MMA of shortening the

  5. Molecular Cloning and Expression Analysis of IgD in Nile Tilapia (Oreochromis niloticus) in Response to Streptococcus agalactiae Stimulus.

    Wang, Bei; Wang, Pei; Wu, Zao-He; Lu, Yi-Shan; Wang, Zhong-Liang; Jian, Ji-Chang


    IgD is considered to be a recently-evolved Ig and a puzzling molecule, being previously found in all vertebrate taxa, except for birds. Although IgD likely plays an important role in vertebrate immune responses, the function of IgD in Nile tilapia (Oreochromis niloticus) is virtually unknown. In the present study, a membrane form of IgD (mIgD) heavy chains were cloned from the GIFT strain of Nile tilapia (designated On-mIgD). The On-mIgD heavy chain's cDNA is composed of 3347 bp with a 31 bp of 5'-UTR, 3015 bp open reading frame (ORF) and 301 bp 3'-UTR, encoding a polypeptide of 1004 amino acids (GenBank accession no: KF530821). Phylogenetic analysis revealed that On-mIgD heavy chains showed the highest similarity to Siniperca chuatsi. Quantitative real-time PCR (qRT-PCR) analysis showed that On-mIgD expression occurred predominately in head kidney, thymus, spleen, and kidney. After Streptococcus agalactiae infection, transcripts of On-mIgD increased and reached its peak at 48 h in the head kidney and thymus, and 72 h in the spleen, respectively. Taken together, these results collectively indicated that IgD could possibly have a key role to play in the immune response when bacterial infections in Nile tilapia.

  6. Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.

    Preethi Ragunathan

    Full Text Available Lmb is a 34 kDa laminin binding surface adhesin of Streptococcus agalactiae. The structure of Lmb reported by us recently has shown that it consists of a metal binding crevice, in which a zinc ion is coordinated to three highly conserved histidines. To elucidate the structural and functional significance of the metal ion in Lmb, these histidines have been mutated to alanine and single, double and triple mutants were generated. These mutations resulted in insolubility of the protein and revealed altered secondary and tertiary structures, as evidenced by circular dichroism and fluorescence spectroscopy studies. The mutations also significantly decreased the binding affinity of Lmb to laminin, implicating the role played by the metal binding residues in maintaining the correct conformation of the protein for its binding to laminin. A highly disordered loop, proposed to be crucial for metal acquisition in homologous structures, was deleted in Lmb by mutation (ΔLmb and its crystal structure was solved at 2.6 Å. The ΔLmb structure was identical to the native Lmb structure with a bound zinc ion and exhibited laminin binding activity similar to wild type protein, suggesting that the loop might not have an important role in metal acquisition or adhesion in Lmb. Targeted mutations of histidine residues confirmed the importance of the zinc binding crevice for the structure and function of the Lmb adhesin.

  7. Cloning and Prokaryotic Expression of Streptococcus Agalactiae Sip Gene%无乳链球菌Sip基因的克隆与原核表达

    杨学云; 王蒴; 吴建勇; 王登峰; 李建军


    Streptococcus Agalactiae surface immunogenic protein (Sip) is a highly conservative important target for the study of Streptococcus Agalactiae vaccine. The Sip gene was amplified by PCR and be inserted into vector pET-22b from a recombinant vector pET-22b-Sip. After identified by PCR, restriction enzyme digestion and sequencing methods, the recombinant plasmid was transformed into BL21 (DE3) and induced to express by IPTG. The size and reactionogenicity of recombinant protein was detected. The results showed that the sip gene was 1300bp and was 99.16% in homology with reference sequence of GeneBank. The recombinant protein was found to be about 50KDa as expected and showed good reactionogenicity. This research provided the basis and foundation for the immunization against Streptococcus Agalactiae.%无乳链球菌表面免疫相关蛋白(Surface immunogenic proteins, Sip)具有高度保守性,是无乳链球菌疫苗研究的重要靶标.通过PCR技术扩增Sip基因,将其插入pET-22b载体构建重组质粒pET-22b-Sip,并对重组质粒进行双酶切、PCR鉴定及测序;转入BL21(DE3)中诱导表达,检测重组蛋白的大小及反应原性.结果表明,所扩增Sip基因大小为1 300bp,与GeneBank参考序列同源性为99.16%;蛋白分析表明,目的蛋白大小为50KDa,具有良好的反应原性,为无乳链球菌的免疫预防提供依据和基础.

  8. Artritis séptica de rodilla por Streptococcus agalactiae: presentación de un caso y revisión de la literatura

    Sesma Solis, P. J.; Fernandez Garcia, L.; García de Quevedo Puerta, D.; García López, A.


    Se describe un caso de artritis séptica de rodilla en un paciente diabético de 69 años. En el momento de la presentación se hallaba sometido a curas locales de una úlcera crónica en el mismo miembro y a antibioterapia oral inespecífica por supuesta infección urinaria. En el cultivo del líquido sinovial se identificó un Streptococcus agalactiae (Estreptococo del grupo B). La infección no remitió con antibioterapia intravenosa electiva e irrigación-aspiración articular repetidas, re...

  9. 鱼源无乳链球菌致病机理研究进展%Review on Pathogenesis of Streptococcus agalactiae from Fish

    祝璟琳; 杨弘


    从无乳链球菌的感染与传播途径,引起的病理学变化,致病机制,毒力因子和理化因子的影响等5个方面对鱼源无乳链球菌的致病机理作一综述。%The studies on pathogenesis of Streptococcus agalactiae from fish were summarized as viewed from the following five factors: transmission studies, pathogenic mechanism, Group B Streptococcus (GBS) virulence and environmental factors.

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

    Pinheiro, Sandra Marisa de Oliveira Sequeira


    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...

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

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


    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...... 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....

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

    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.

  13. 广东地区感染养殖罗非鱼的无乳链球菌分子分型研究%Molecular epidemiology of Streptococcus agalactiae in tilapia in Guangdong

    方伟; 梁宇恒; 宁丹; 柯碧霞; 柯昌文; 邓小玲; 彭华林; 孙九峰; 王云新


    以广东省内5个罗非鱼 Oreochromis niloticus 主要养殖区内分离的无乳链球菌 Streptococcus agalactiae 为研究目标,通过毒力基因 PCR 检测、多位点序列分型、脉冲场凝胶电泳技术研究广东地区感染罗非鱼的无乳链球菌分子特征。结果发现170株分离无乳链球菌中毒力基因的携带率介于60.59%~100%之间,毒力基因携带形成10个谱系,dltR-bca-sodA-spb1-cfb-bac (38.24%,65/170),dltr-bca-sodA-spb1-bac (23.53%,40/170)和dltr-bca-sodA-spb1-cfb-bac-scpB (22,35%,38/170)为主要的谱系,dltr-bca-sodA-spb1-bac-scpb (8.24%,14/170)和 dltr-bca-sodA-spb1(4.71%,8/170)次之。多位点序列分型研究证实170株菌在7个位点均为单一的等位基因,为 adhP (10),pheS (1),atr (2),glnA (1),sdhA (3),glcK (2),tkt (2),序列型为 ST7型。146株菌脉冲场凝胶电泳结果表明146株菌分为6个簇(A-D,F,G),相似性介于86%~100%之间。A (18),B (16),C (103),D (7)为主要的遗传簇,占菌株数的98.63%(144/146)。基因簇 A,C 在4个采样区均有发现;基因簇 B 分布在吴川、茂名和阳江;基因簇 D 分布在茂名、珠海和阳江;基因簇 F 和 G 分别来自茂名和珠海。在 A-D 遗传簇中均有发现来自不同地区的菌株呈现100%的相似性,为同一克隆株。结果表明广东省主要罗非鱼养殖区内感染链球菌携带毒力基因比率高,序列型均为高致病的 ST7型别,不同地区间具有相同的 PFGE 型别,不同地区间交叉传播感染的可能高。%Tilapia samples were collected from five main tilapia farms in Guangdong.The characteristics of the isolates were determined by virulence gene detection,Multilocus Sequencing Type (MLST),and Pulsed-Field Gel Electrophoresis (PFGE).The frequency of virulence gene in all isolates varied from 60.59% to 1 00

  14. Development of nested PCR for detection of Streptococcus agalactiae from tilapia%罗非鱼无乳链球菌巢式 PCR 检测方法的建立

    邵辰; 易弋; 黎娅; 黄荷; 伍时华; 杨军


    Based on the cfb gene sequence of Streptococcus agalactiae, two pairs of primers were designed and synthesized and the amplification conditions were optimized to establish the nested PCR method which could identify S.agalactiae with high sensitively.The results showed that S.agalactiae could be directly detected from blood sample at a minimum concentra -tion as 8.7 ×10 4 CFU/ml by this method.Nineteen clinical S.agalactiae -infected tilapia samples collected from Guangxi province were applied to verify this method.The results showed that 260 bp fragment could be amplified in 18 samples and the sequencing analysis revealed that all the amplified fragment were cfb gene of S.agalactiae.The detection accuracy of this method was 94.7%.%根据无乳链球菌(Streptococcus agalactiae)cfb 基因序列,设计、合成2对引物,优化扩增条件,建立了快速高灵敏度鉴别无乳链球菌的巢式 PCR 方法。结果显示:使用该方法对罗非鱼(Oreochromis mossambicus)血液样品进行检测,可检测到活菌浓度为8.7×104 CFU/mL 的无乳链球菌。对采自广西地区受无乳链球菌感染的19份罗非鱼样品进行检测,18份可获得目的片段,扩增到的序列均为无乳链球菌 cfb 基因序列,检测准确度达到94.7%。

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

    S. A. Hussein


    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.

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

    Liliana Serna Cock


    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.

  17. 新生儿无乳链球菌败血症临床分析%Clinical analysis of 11 cases of neonatal streptococcus agalactiae sepsis

    曾淑娟; 丘惠娴


    目的:探讨新生儿无乳链球菌败血症的临床特点及耐药性.方法:对11例新生儿无乳链球菌败血症患儿的临床资料和治疗结果进行回顾性分析.结果:2012年确诊新生儿败血症25例中无乳链球菌感染占44.0%,其中早发败血症7例,晚发败血症4例;临床表现发热6例,黄疸5例,肺炎5例 (早发感染占4例),颅内感染4例 (晚发感染占3例);所有血培养对青霉素、头孢类抗生素和万古霉素均无耐药性,对四环素耐药100.0%,对克林霉素耐药72.7%.结论:无乳链球菌败血症早发感染临床以呼吸道表现为主,晚发感染常合并颅内感染,需注意完善脑脊液检查.治疗上首选青霉素、头孢菌素,克林霉素耐药性高不宜临床选用.%Objective:To investigate the neonatal streptococcus agalactiae sepsis clinical characteristics and resistance in order to guide clinical treatment.Methods:The clinical features and resistance of 11 neonatal streptococcus agalactiae sepsis which admitted in our hospital from January 2012 to December 2012 were retrospectively analyzed.Results:25 cases neonatal sepsis were diagnozed in 2012,streptococcus agalactiae infections accounted for 44%,including 7 cases of early-onset sepsis,4 cases of late-onset sepsis.Clinical manifestations:6 cases got fever,5 cases got jaundice,5 cases got pneumonia (including 4 cases of early-onset infection),4 cases got intracranial infection (including 3 cases late-onset infection).All blood cultures had no resistance to penicillin,cephalosporins and vancomycin,100% resistance to tetracycline,72.7% resistance to clindamycin.Conclusion:The trend of streptococcus agalactiae sepsis is growing in 2012,early-onset infection is often associated with respiratory symptoms,late-onset infection is often associated with intracranial infection,we need to pay attention to improve the cerebrospinal fluid examination.We can choose penicillin,cephalosporin for treatment.Clindamycin is not

  18. Comprehensive identification and profiling of Nile tilapia (Oreochromis niloticus) microRNAs response to Streptococcus agalactiae infection through high-throughput sequencing.

    Wang, Bei; Gan, Zhen; Cai, Shuanghu; Wang, Zhongliang; Yu, Dapeng; Lin, Ziwei; Lu, Yishan; Wu, Zaohe; Jian, Jichang


    MicroRNAs are a kind of small non-coding RNAs that participate in various biological processes. Deregulated microRNA expression is associated with several types of diseases. Tilapia (Oreochromis niloticus) is an important commercial fish species in China. To identify miRNAs and investigate immune-related miRNAs of O. niloticus, we applied high-throughput sequencing technology to identify and analyze miRNAs from tilapia infected with Streptococcus agalactiae at a timescale of 72 h divided into six different time points. The results showed that a total of 3009 tilapia miRNAs were identified, including in 1121 miRNAs which have homologues in the currently available databases and 1878 novel miRNAs. The expression levels of 218 tilapia miRNAs were significantly altered at 6 h-72 h post-bacterial infection (pi), and these miRNAs were therefore classified as differentially expressed tilapia miRNAs. For the 1121 differentially expressed tilapia miRNAs target 41961 genes. GO and KEGG enrichment analysis revealed that some target genes of tilapia miRNAs were grouped mainly into the categories of apoptotic process, signal pathway, and immune response. This is the first report of comprehensive identification of O. niloticus miRNAs being differentially regulated in spleen in normal conditions relating to S. agalactiae infection. This work provides an opportunity for further understanding of the molecular mechanisms of miRNA regulation in O. niloticus host-pathogen interactions.

  19. Comparison of carrot broth- and selective Todd-Hewitt broth-enhanced PCR protocols for real-time detection of Streptococcus agalactiae in prenatal vaginal/anorectal specimens.

    Block, Timothy; Munson, Erik; Culver, Anne; Vaughan, Katharine; Hryciuk, Jeanne E


    The reporting of accurate Streptococcus agalactiae screening results in a short time frame is of tremendous clinical benefit. A total of 203 consecutive primary vaginal/anorectal specimens were cultured in selective Todd-Hewitt broth (LIM broth) and with the StrepB carrot broth kit (carrot broth). One-day broth cultures were subjected to both centrifugation and clarification of a 500-mul aliquot prior to sample lysis (protocol A) and direct lysis of a 50-mul aliquot (protocol B). The lysates were subsequently analyzed by the BD GeneOhm StrepB assay. The results were compared to the carrot broth culture results derived from visualization of pigment on day 1 or from a subculture of carrot broth. Thirty-four carrot broth cultures (16.7%) generated diagnostic pigment following overnight incubation; an additional 26 (12.8%) were positive for S. agalactiae upon subculture. Carrot broth-enhanced PCR by the use of either protocol A or protocol B trended toward a higher rate of positive results (33.0%) than the rate observed by either the LIM broth-enhanced PCR (30.5%) or full carrot broth culture analysis (29.6%). In the context of the result on day 1, both carrot broth- and LIM broth-enhanced PCRs generated more true-positive results (P protocols of carrot broth- or LIM broth-enhanced PCR were >/=95.4%. Whereas protocol A resolved the results for 99.8% of the specimens in the evaluation upon initial testing, a 5.7% initial unresolved rate and a 1.5% final unresolved rate were determined by the use of protocol B. The use of carrot broth within a rapid and highly accurate molecular reflex testing algorithm can limit follow-up testing to cultures without evidence of pigmentation.

  20. 罗非鱼无乳链球菌LAMP快速检测方法的建立%Development of loop-mediated isothermal amplification for detection of Streptococcus agalactiae from Tilapia

    郑磊; 樊海平; 吴斌; 曾占壮; 钟全福; 张新艳


    针对无乳链球菌高度保守的表面免疫相关蛋白基因,采用环介导等温扩增技术( LAMP),设计4条特异性引物,优化反应体系,建立罗非鱼无乳链球菌快速诊断方法。研究结果表明, LAMP方法检测灵敏度达到30 CFU· mL-1,且对2株罗非鱼源无乳链球菌均能扩增出特异性片段,而对14株非无乳链球菌均未扩增出相应片段。建立的LAMP检测方法具有高度的特异性和灵敏性,反应在1 h内完成,为罗非鱼无乳链球菌病的快速诊断提供了一种重要的技术手段。%To develop a technique for rapid detection of Streptococcus agalactiae from Tilapia, the loop-mediated isothermal amplification( LAMP) was introduced to amplify the surface immunogenic protein ( SIP) gene.The detection method was established by using optimum reaction system with four specif-ic primers of the target gene.The results showed that the method had perfect specificity, and the sen-sitivity was 30 CFU · mL-1 , as the DNA of 2 strains of Streptococcus agalactiae were able to be amplified, while the other 14 strains of non -Streptococcus agalactiae were not.The amplification process can be finished in 1 hour.The method provides a better choice for rapid detection of Strepto-coccus agalactiae.

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

    Oscar Fabián Porras Pinto


    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

  2. 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

    Maria Aparecida V. P. Brito


    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.

  3. Increasing of temperature induces pathogenicity of Streptococcus agalactiae and the up-regulation of inflammatory related genes in infected Nile tilapia (Oreochromis niloticus).

    Kayansamruaj, Pattanapon; Pirarat, Nopadon; Hirono, Ikuo; Rodkhum, Channarong


    Temperature strongly affects the health of aquatic poikilotherms. In Nile tilapia (Oreochromis niloticus), elevated water temperatures increase the severity of streptococcosis. Here we investigated the effects of temperature on the vulnerability and inflammatory response of Nile tilapia to Streptococcus agalactiae (Group B streptococci; GBS). At 35 and 28 °C, GBS took 4 and 7h, respectively to reach the log-phase and, when incubated with tilapia whole blood, experienced survival rates of 97% and 2%, respectively. The hemolysis activity of GBS grown at 35 °C was five times higher than that of GBS grown at 28 °C. GBS expressed cylE (β-hemolysin/cytolysin), cfb (CAMP factor) and PI-2b (pili-backbone) much more strongly at 35 °C than at 28 °C. Challenging Nile tilapia reared at 35 and 28 °C with GBS resulted in accumulated mortalities of about 85% and 45%, respectively. At 35 °C, infected tilapia exhibited tremendous inflammatory responses due to a dramatic up-regulation (30-40-fold) of inflammatory-related genes (cyclooxygenase-2, IL-1β and TNF-α) between 6 and 96 h-post infection. These results suggest that the increase of GBS pathogenicity to Nile tilapia induced by elevated temperature is associated with massive inflammatory responses, which may lead to acute mortality.

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

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


    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.

  5. Integrated analysis neurimmiRs of tilapia (Oreochromis niloticus) involved in immune response to Streptococcus agalactiae, a pathogen causing meningoencephalitis in teleosts.

    Wang, Bei; Gan, Zhen; Wang, Zhongliang; Yu, Dapeng; Lin, Ziwei; Lu, Yishan; Wu, Zaohe; Jian, Jichang


    MicroRNAs (miRNAs) are a class of noncoding RNA molecules and play important roles in a wide spectrum of biological processes, including in immune response. Recent years have witnessed considerable amount of research interest in studies on miRNA-mediated modulation gene function during neuroinflammation. Here, we evaluated Streptococcus agalactiae infected tilapia (Oreochromis niloticus) brain for the expression profile of miRNAs, potential functions and their correlation with genes involved in inflammatory pathways. A total of 1981 miRNAs were identified, including in 486 miRNAs which have homologues in the currently available databases and 1945 novel miRNAs. The expression levels of 547 miRNAs were significantly altered at 6 h-48 h post-bacterial infection, and these miRNAs were therefore classified as differentially expressed tilapia miRNAs. Real-time PCR were implemented for 14 miRNAs co-expressed in five samples, and agreement was confirmed between the high-throughput sequencing and real-time PCR data. For the 486 differentially expressed miRNAs target 41,820 genes. GO and KEGG enrichment analysis revealed that some target genes of miRNAs were grouped mainly into the categories of apoptotic, signal pathwayand immune response. This is the first report of comprehensive identification of teleost miRNAs being differentially regulated in brain in normal conditions relating to bacterial infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Evaluation of the brain-derived neurotrophic factor, nerve growth factor and memory in adult rats survivors of the neonatal meningitis by Streptococcus agalactiae.

    Barichello, Tatiana; Lemos, Joelson C; Generoso, Jaqueline S; Carradore, Mirelle M; Moreira, Ana Paula; Collodel, Allan; Zanatta, Jessiele R; Valvassori, Samira S; Quevedo, João


    Streptococcus agalactiae (GBS) is a major cause of severe morbidity and mortality in neonates and young infants, causing sepsis, pneumonia and meningitis. The survivors from this meningitis can suffer serious long-term neurological consequences, such as, seizures, hearing loss, learning and memory impairments. Neurotrophins, such as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) control the neuronal cell death during the brain development and play an important role in neuronal differentiation, survival and growth of neurons. Neonate Wistar rats, received either 10μL of sterile saline as a placebo or an equivalent volume of GBS suspension at a concentration of 1×10(6)cfu/mL. Sixty days after induction of meningitis, the animals underwent behavioral tests, after were killed and the hippocampus and cortex were retired for analyze of the BDNF and NGF levels. In the open-field demonstrated no difference in motor, exploratory activity and habituation memory between the groups. The step-down inhibitory avoidance, when we evaluated the long-term memory at 24h after training session, we found that the meningitis group had a decrease in aversive memory when compared with the long-term memory test of the sham group. BDNF levels decreased in hippocampus and cortex; however the NGF levels decreased only in hippocampus. These findings suggest that the meningitis model could be a good research tool for the study of the biological mechanisms involved in the behavioral alterations secondary to GBS meningitis.

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

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


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

    George K. Gitau


    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.

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

    Gitau, George K; Bundi, Royford M; Vanleeuwen, John; Mulei, Charles M


    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.

  10. Nationwide German Multicenter Study on the Prevalence of Antibiotic Resistance in Streptococcal Blood Isolates from Neutropenic Patients and Comparative In Vitro Activities of Quinupristin-Dalfopristin and Eight Other Antimicrobials

    Reinert, Ralf René; von Eiff, Christof; Kresken, Michael; Brauers, Johannes; Hafner, Dieter; Al-Lahham, Adnan; Schorn, Holger; Lütticken, Rudolf; Peters, Georg


    In a prospective multicenter study (1996 to 1999), 156 episodes of bacteremic streptococcal infections of neutropenic patients were evaluated. Streptococcus oralis (26.3%), S. pneumoniae (26.3%), S. agalactiae (11.5%), S. mitis (9%), and S. pyogenes (5.8%) were the predominant species. Four strains (2.6%) were found to be intermediately resistant to penicillin. One strain (0.6%) was found to be highly resistant to penicillin (MIC, 8 mg/liter). Reduced susceptibility to penicillin was detected among S. oralis (14.6%), S. mitis (7.1%), and S. pneumoniae (4.9%) isolates but was not recorded among S. agalactiae and S. pyogenes. Resistance rates and intermediate resistance rates for other antimicrobials were as follows (all species): amoxicillin, 1.3 and 3.2%; erythromycin, 16 and 2.6%; clindamycin, 5.8 and 0%; ciprofloxacin, 1.9 and 7.7%. Quinupristin-dalfopristin showed good in vitro activity against most streptococcal isolates (MIC at which 50% of the isolates were inhibited [MIC50], 0.5 mg/liter; MIC90, 1 mg/liter, MIC range, 0.25 to 4 mg/liter). PMID:11326015

  11. 新生儿无乳链球菌感染的危险因素及药敏分析%The study of risk factors for neonatal Streptococcus agalactiae infection and sensitivity analysis of antibacterials

    王莹超; 陆文香; 周莉靖


    目的:分析新生儿感染无乳链球菌的危险因素及药物敏感性,为预防和治疗新生儿无乳链球菌感染提供依据。方法收集2013年1~12月1200份临床送检的新生儿血液、胃液、脓液标本,以及相应母体泌尿生殖道标本进行细菌培养与药敏试验,并回顾性分析患儿疾病类型、母体感染状况、分娩方式、孕晚期用药情况、新生儿死亡情况等临床资料。结果共80例新生儿感染无乳链球菌,其中败血症、脐炎、早产、宫内感染、吸入性肺炎患儿分别占8.75%、10.00%、15.00%、22.50%、43.75%;其母体生殖道标本无乳链球菌阳性率达51.25%,且细菌药敏结果与患儿一致;患儿以自然分娩为主,共71例(88.75%),仅9例(11.25%)为剖宫产。80株无乳链球菌对万古霉素、利奈唑胺、青霉素和头孢曲松的敏感率均为100.00%,对红霉素、克林霉素、左氧氟沙星的耐药率较高,分别为77.50%、57.50%、33.75%。结论母体带菌与分娩方式可能是新生儿无乳链球菌感染的重要危险因素,产科医生应重视对围产期孕妇无乳链球菌的常规筛查,实验室应提高对无乳链球菌的检测能力并及时提供药敏检测结果,为临床合理用药提供重要依据。%Objective To analyze the related risk factors of neonatal Streptococcus agalactiae infection and sensitivity of antibac‐terials ,in order to provide for provide evidence for the prevention and treatment of neonatal infection .Methods A total of 1 200 neonatal blood ,gastric juice ,pus specimens ,and maternal reproductive tract specimens were collected from Jan .2013 to Dec .2013 for bacterial culture and drug sensitive test .And clinical data about types of neonatal diseases ,maternal infection status ,mode of de‐livery ,medication in late pregnancy ,situation of neonatal death were retrospectively analyzed .Results A total of

  12. Identificação de Escherichia coli e Streptococcus agalactiae em amostras placentárias de gestantes a termo com e sem suspeita de infecção

    Duarte, Pâmella Oliveira


    A infecção neonatal pode ser responsável por consequências graves ao recém-nascido, caracterizada no nascimento como sepse precoce ou sepse tardia, pela elevada taxa de morbi-mortalidade neonatal. Os agentes infecciosos bacterianos mais frequentes descritos são os estreptococos (Streptococcus agalactiae, Streptococcus constellatus, Streptococcus anginosus e Streptococcus pneumoniae) e a Escherichia coli, cuja incidência ainda é pouco conhecida por não apresentar notificação compulsória e os e...

  13. Bacteriocinogenic Bacteria Isolated from Raw Goat Milk and Goat Cheese Produced in the Center of México.

    Hernández-Saldaña, Oscar F; Valencia-Posadas, Mauricio; de la Fuente-Salcido, Norma M; Bideshi, Dennis K; Barboza-Corona, José E


    Currently, there are few reports on the isolation of microorganisms from goat milk and goat cheese that have antibacterial activity. In particular, there are no reports on the isolation of microorganisms with antibacterial activity from these products in central Mexico. Our objective was to isolate bacteria, from goat products, that synthesized antimicrobial peptides with activity against a variety of clinically significant bacteria. We isolated and identified Lactobacillus rhamnosus, L. plantarum, L. pentosus, L. helveticus and Enterococcus faecium from goat cheese, and Aquabacterium fontiphilum, Methylibium petroleiphilum, Piscinobacter aquaticus and Staphylococcus xylosus from goat milk. These bacteria isolated from goat cheese were able to inhibit Staphylococcus aureus, Bacillus cereus, Escherichia coli, Listeria monocytogenes, L. inoccua, Pseudomona aeruginosa, Shigella flexneri, Serratia marcescens, Enterobacter cloacae and Klebsiella pneumoniae. In addition, bacteria from goat milk showed inhibitory activity against B. cereus, L. lactis, E. coli, S. flexneri, E. cloacae and K. pneumonia; S. aureus, L. innocua, S. agalactiae and S. marcescens. The bacteriocins produced by these isolates were shown to be acid stable (pH 2-6) and thermotolerant (up to 100 °C), but were susceptible to proteinases. When screened by PCR for the presence of nisin, pediocin and enterocin A genes, none was found in isolates recovered from goat milk, and only the enterocin A gene was found in isolates from goat cheese.

  14. Properties of an Arcanobacterium haemolyticum strain isolated from a donkey.

    Sammra, Osama; Balbutskaya, Anna; Nagib, Samy; Alber, Jörg; Lämmler, Christoph; Abdulmawjood, Amir; Timke, Markus; Kostrzewa, Markus; Prenger-Berninghoff, Ellen


    The present study was designed to characterize phenotypically and genotypically an Arcanobacterium haemolyticum strain (A. haemolyticum P646) isolated from a purulent nasal discharge of a donkey. A. haemolyticum P646 showed, compared to sheep blood, an enhanced hemolytic reaction on rabbit blood agar, a synergistic CAMP-like reaction with Streptococcus agalactiae and Rhodococcus equi as indicator strains, a reverse CAMP reaction in the zone of Staphylococcus aureus beta-hemolysin and the typical biochemical properties of this species. The species identity could be confirmed by MALDI-TOF MS analysis, by sequencing the 16S rDNA and glyceraldehyde-3-phosphate dehydrogenase encoding gene gap and by amplification of A. haemolyticum specific parts of 16S-23S rDNA intergenic spacer region and 23S rDNA. A. haemolyticum P646 and the reference strain A. haemolyticum DSM 20595 were further characterized by amplification of the putative virulence genes encoding arcanolysin, phospholipase D, hemolysin A, CAMP factor family protein, a collagen binding protein and two neuraminidases which were present for A. haemolyticum DSM 20595. A. haemolyticum P646 showed a comparable gene spectrum but was negative for the genes encoding collagen binding protein and neuraminidase H. To our knowledge, the present study is the first phenotypic and genotypic characterization of an A. haemolyticum strain isolated from a donkey.

  15. Isolation and characterization of bacteriocinogenic lactic bacteria from M-Tuba and Tepache, two traditional fermented beverages in México.

    de la Fuente-Salcido, Norma M; Castañeda-Ramírez, José Cristobal; García-Almendárez, Blanca E; Bideshi, Dennis K; Salcedo-Hernández, Rubén; Barboza-Corona, José E


    Mexican Tuba (M-Tuba) and Tepache are Mexican fermented beverages prepared mainly with pineapple pulp and coconut palm, respectively. At present, reports on the microbiota and nutritional effects of both beverages are lacking. The purpose of this study was to determine whether M-Tuba and Tepache contain cultivable lactic acid bacteria (LAB) capable of producing bacteriocins. Tepache and M-Tuba contain mesophilic aerobic bacteria, LAB, and yeast. Bacillus subtilis, Listeria monocytogenes, Listeria innocua, Streptococcus agalactiae, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium, and Salmonella spp, were the microorganisms most susceptible to metabolites produced by bacterial isolates. M-Tuba and Tepache contain bacteria that harbor genes coding for nisin and enterocin, but not pediocin. The presence of Lactococcus lactis and E. faecium in M-Tuba and Tepache, was identified by 16S rDNA. These bacteria produced bacteriocins of ∼3.5 kDa and 4.0-4.5 kDa, respectively. Partial purified bacteriocins showed inhibitory effect against Micrococcus luteus, L. monocytogenes, L. innocua, Str. agalactiae, S. aureus, Bacillus cereus, B. subtilis, E. faecalis, and K. pneumoniae. We characterized, for the first time, cultivable microbiota of M-Tuba and Tepache, and specifically, identified candidate lactic bacteria (LAB) present in these beverages that were capable of synthesizing antimicrobial peptides, which collectively could provide food preservative functions.

  16. 罗非鱼无乳链球菌 EsxA 基因克隆与原核表达%Clonging and Protokaryon Expression of EsxA Gene of Streptococcus agalactiae in Tilapia

    马艳平; 梁志凌; 马江耀; 刘振兴; 郝乐; 柯浩


    Ⅶ型分泌系统(T7SS)是近年来发现的分泌系统,分泌两种胞外蛋白,EsxA 基因编码的 ESAT6蛋白和 EsxB 基因编码的 CFP-10蛋白。分泌蛋白具有良好的免疫原性,促进细菌从巨噬细胞吞噬体逃逸、影响巨噬细胞凋亡及裂解细胞等生物学功能,与致病性密切相关。以罗非鱼源无乳链球菌为模板,克隆到294 bp 的 EsxA 基因,将目的序列克隆到 pMD18-T 载体,经测序,目的序列与无乳链球菌 EsxA 基因同源率在99%以上。EsxA 基因克隆到 pET32a 载体中,重组载体在28℃、0.5 mmol/L IPTG 诱导条件下表达量最大,可溶性表达。将纯化的 ESAT6蛋白免疫 Balb/c 鼠,成功制备 ESAT6蛋白鼠多克隆抗体,经 Western blot,ESAT6蛋白具有良好的反应原性。研究结果为进一步进行无乳链球菌 ESAT6蛋白的免疫学功能研究奠定了基础。%New discovered type Ⅶ secretion system (T7SS)secrets two extracellular proteins including Es-xA coded ESAT6 and EsxB coded CFP-10,which is related to pathogenicity closely.ESAT6 and CFP-10 proteins have diverse functions such as promoting bacteria from phagosome of macrophage,impacting mac-rophage apoptosis and lysis.The two secreted proteins in T7SS of Streptococcus agalactiae form the ho-mologous dimmers,collectively known as ESAT6 protein,may be bacterial adhesion protein showing im-portant for host-cell attachment and invasion in S.agalactiae through bioinformatics analysis.But ESAT6 protein whether has the virulence and its pathogenesis is not clear,therefore it is necessary to study on it. We cloned 294 bp sequence from tilapia Streptococcus agalactiae .And the sequence was cloned into pMD18-T vector,the homology is 99% with tilapia Streptococcus agalactiae EsxA gene.EsxA gene was cloned into pET32a vector.The best solouble expression was under 0.5 mmol/L IPTG,28℃.In addition, the purified protein was used to immunize Balb/c mice to prepare mouse anti

  17. Genetic diversity and antimicrobial resistance  in Streptococcus agalactiae strains recovered from female carriers in the Bucharest area

    Codru?a-Romani?a Usein


    Full Text Available For the first time, we used multilocus sequence typing (MLST to understand how Romanian group B streptococcus (GBS strains fit into the global GBS population structure. Colonising isolates recovered from adult human females were tested for antibiotic resistance, were molecularly serotyped based on the capsular polysaccharide synthesis (cps gene cluster and further characterised using a set of molecular markers (surface protein genes, pilus-encoded islands and mobile genetic elements inserted in the scpB-lmb intergenic region. Pulsed-field gel electrophoresis was used to complement the MLST clonal distribution pattern of selected strains. Among the 55 strains assigned to six cps types (Ia, Ib, II-V, 18 sequence types (STs were identified by MLST. Five STs represented new entries to the MLST database. The prevalent STs were ST-1, ST-17, ST-19 and ST-28. Twenty molecular marker profiles were identified. The most common profiles (rib+GBSi1+PI-1, rib+GBSi1+PI-1, PI-2b and alp2/3+PI-1, PI-2a were associated with the cps III/ST-17 and cps V/ST-1 strains. A cluster of fluoroquinolone-resistant strains was detected among the cps V/ST-19 members; these strains shared alp1 and IS1548 and carried PI-1, PI-2a or both. Our results support the usefulness of implementing an integrated genotyping system at the reference laboratory level to obtain the reliable data required to make comparisons between countries.

  18. Experimental infection of goats with an unusual strain of Mycoplasma mycoides subsp. capri isolated in Jordan: comparison of different diagnostic methods

    Massimo Scacchia


    Full Text Available Ten goats were experimentally infected with a Mycoplasma identified by biomolecular methods as Mycoplasma mycoides subsp. capri, strain Irbid which was isolated from goats in an outbreak of contagious agalactia in north Jordan and defined as ‘unusual’, due to its serological characteristics. Two groups of goats infected by the endotracheal route and by aerosol, respectively, were placed in contact with a third group of naive animals. Six weeks after infection, some animals from both the infected and contact groups presented fever and nasal discharge, followed by severe respiratory signs and polyarthritis. Organs were taken from animals that died during the trial or those that were sacrificed at the end of the trial. The results of microbiological isolation and immunohistochemical tests conducted on the organs were compared after a description of the clinical picture and anatomopathological and histopathological signs.

  19. Isolation, identification and screening of antimicrobial thermophilic Streptomyces sp. Al-Dhabi-1 isolated from Tharban hot spring, Saudi Arabia.

    Al-Dhabi, Naif Abdullah; Esmail, Galal Ali; Duraipandiyan, Veeramuthu; Valan Arasu, Mariadhas; Salem-Bekhit, Mounir M


    The strain Streptomyces sp. Al-Dhabi-1 was isolated from soil sediments collected from Tharban hot spring in the southern west of Saudi Arabia using actinomycetes isolation agar and starch casein agar at 55 °C. Identification of the isolate was done according to morphological, physiological and biochemical characteristics and 16S rRNA sequence similarity as well. 16S rRNA sequence and blast analyses confirmed that the isolate belonging to the genus Streptomyces. The sequence was submitted to GenBank with accession number (KF815080). Ethyl acetate extract of Streptomyces sp. Al-Dhabi-1 showed good antimicrobial activities against tested pathogenic microbes. Minimum inhibitory concentration results showed that the best values were observed against S. agalactiae (<0.039 mg/ml) and Klebsiella pneumonia (0.125 mg/ml). Minimum inhibitory concentration of Al-Dhabi-1 against fungi; Cryptococcus neoformans (0.078 mg/ml), C. albicans (0.156 mg/ml), A. niger (0.625 mg/ml), and T. mentagrophytes (0.156 mg/ml). GC-MS analysis was used for the chemical profile of ethyl acetate extract. Benzeneacetic acid (16.02 %) and acetic acid 2-phenylethyl ester (10.35 %) were the major compounds among 31 substances found the ethyl acetate extract. According to the results of antimicrobial activity against pathogenic microbes, it is clear that the actinomycetes from hot springs with extreme environments are promising source for antimicrobial compounds.

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

    Nagalakshmi Narayana-Swamy


    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.

  1. Evidence for the Sialylation of PilA, the PI-2a Pilus-Associated Adhesin of Streptococcus agalactiae Strain NEM316.

    Morello, Eric; Mallet, Adeline; Konto-Ghiorghi, Yoan; Chaze, Thibault; Mistou, Michel-Yves; Oliva, Giulia; Oliveira, Liliana; Di Guilmi, Anne-Marie; Trieu-Cuot, Patrick; Dramsi, Shaynoor


    Streptococcus agalactiae (or Group B Streptococcus, GBS) is a commensal bacterium present in the intestinal and urinary tracts of approximately 30% of humans. We and others previously showed that the PI-2a pilus polymers, made of the backbone pilin PilB, the tip adhesin PilA and the cell wall anchor protein PilC, promote adhesion to host epithelia and biofilm formation. Affinity-purified PI-2a pili from GBS strain NEM316 were recognized by N-acetylneuraminic acid (NeuNAc, also known as sialic acid) specific lectins such as Elderberry Bark Lectin (EBL) suggesting that pili are sialylated. Glycan profiling with twenty different lectins combined with monosaccharide composition by HPLC suggested that affinity-purified PI-2a pili are modified by N-glycosylation and decorated with sialic acid attached to terminal galactose. Analysis of various relevant mutants in the PI-2a pilus operon by flow-cytometry and electron microscopy analyses pointed to PilA as the pilus subunit modified by glycosylation. Double labeling using PilB antibody and EBL lectin, which specifically recognizes N-acetylneuraminic acid attached to galactose in α-2, 6, revealed a characteristic binding of EBL at the tip of the pilus structures, highly reminiscent of PilA localization. Expression of a secreted form of PilA using an inducible promoter showed that this recombinant PilA binds specifically to EBL lectin when produced in the native GBS context. In silico search for potentially glycosylated asparagine residues in PilA sequence pointed to N427 and N597, which appear conserved and exposed in the close homolog RrgA from S. pneumoniae, as likely candidates. Conversion of these two asparagyl residues to glutamyl resulted in a higher instability of PilA. Our results provide the first evidence that the tip PilA adhesin can be glycosylated, and suggest that this modification is critical for PilA stability and may potentially influence interactions with the host.

  2. An analysis of 181 cases with blood stream infection caused by Streptococcus agalactiae in children from 2011 to 2015: a multi-center retrospective study%2011至2015年181例儿童无乳链球菌血流感染多中心研究

    华春珍; 俞蕙; 庄捷秋; 李小露; 许红梅; 罗巧二; 卢洪萍; 俞惠民; 曹云


    目的 了解儿童无乳链球菌血流感染的临床特点及细菌耐药模式,为临床经验诊治提供科学依据.方法 通过实验室信息管理系统查询7家三甲医院(浙江4家、上海2家、重庆1家)2011年1月1日至2015年12月31日血液和脑脊液培养为无乳链球菌的患儿,回顾分析临床特点、耐药模式、治疗及转归.组间比较采用x2检验,t检验或非参数检验.结果 确诊无乳链球菌血流感染181例,男86例,女95例.患儿年龄1d~9岁,中位数13 d,早产儿30例.自然分娩127例(70.2%).早发型(<7 d)患儿自然分娩[81.7% (58/71)比62.3% (66/106)]、胎膜早破[25.4%(18/71)比3.8% (4/106)]、气促呻吟[43.7%(31/71)比15.1% (16/106)的发生率显著高于晚发型(7~89 d),而发热[25.4%(18/71)比85.8% (91/106)]、合并化脓性脑膜炎[29.6% (21/71)比53.8% (57/106)]的发生率显著低于晚发型(P均<0.05).78例(43.1%)患儿合并化脓性脑膜炎.159例(87.8%)患儿血培养前未使用过抗生素.菌株对青霉素、头孢曲松或头孢噻肟的敏感率分别为98.9%、99.0%、99.0%,所有菌株对万古霉素敏感,对克林霉素和红霉素的耐药率分别高达68.0%和34.0%.研究中仅39例(22.0%)采用青霉素类或头孢菌素类单药治疗,80例(45.2%)用含酶抑制剂的复合制剂,61例(34.5%)病原明确后仍应用美罗培南或头孢哌酮-舒巴坦.154例治愈或好转,19例死亡,其中13例合并化脓性脑膜炎,8例自动出院后失访.结论 儿童无乳链球菌血流感染合并化脓性脑膜炎发生率高,病死率较高,青霉素是治疗该菌感染的有效抗生素,临床应根据该菌的耐药特点合理选择抗生素.%Objective To analyze the clinical characteristics of blood stream infection caused by Streptococcus agalactiae in children and the drug-resistance of the isolates.Method All cases with Streptococcus agalactiae growth in blood or cerebrospinal fluid cultures from January 1

  3. Clinical Features and Therapeutic Strategies of Late-onset Streptococcus Agalactiae Meningitis%晚发型无乳链球菌脑膜炎的临床特征及治疗策略

    张胜; 罗序峰; 周涛; 付四毛; 朱建萍


    Streptococcus agalactiae is common pathogenic bacteria for infant suppurative meningitis. In recent years, the prevalence of streptococcus agalactiae meningitis is increasing. The paper made a retrospective analysis of the clinical data of 6 children with streptococcus agalactiae meningitis and investigated the clinical features and therapeutic strategies of the disease. The clinical manifestations of the 6 children included fever,convulsion,decreased responsiveness,tensity or swell in bregma and irregular respiration. Subdural effusion,cerebral hemorrhage,liver function damage and hyponatremia as complication also occurred in some of the children. Antibiotic therapy was determined according to the results of drug sensitive test and cerebrospinal fluid examination. One child was cured and 4 children improved after 4 - week to 6 - week treatment. One child died due to the halt of treatment following septic shock.%无乳链球菌是婴儿期化脓性脑膜炎最为常见的致病菌之一,近年来,无乳链球菌脑膜炎发病率呈上升趋势。本研究回顾性分析6例无乳链球菌脑膜炎患儿临床资料,探讨无乳链球菌脑膜炎的临床特点及治疗策略。患儿临床表现为发热、惊厥、反应低下、前囟紧张或膨隆及呼吸不规则,部分患儿伴有硬膜下积液、脑出血、肝功能损害、低钠血症并发症。经药敏试验和脑脊液检查结果确定抗生素治疗方案。治疗4~6周6例患儿中治愈1例、好转4例、因感染性休克放弃治疗死亡1例。

  4. [Susceptibility of clinically-isolated bacteria strains to respiratory quinolones and evaluation of antimicrobial agent efficacy by Monte Carlo simulation].

    Kosaka, Tadashi; Yamada, Yukiji; Kimura, Takeshi; Kodama, Mai; Fujitomo, Yumiko; Masaki, Nakanishi; Toshiaki, Komori; Keisuke, Shikata; Fujita, Naohisa


    Respiratory quinolones (RQs) are broad-spectrum antimicrobial agents used for the treatment of a wide variety of community-acquired and nosocomial infections. However, bacterial resistance to quinolones has been on the increase. In this study, we investigated the predicted efficacy of RQs for various strains of 9 bacterial species clinically isolated at our university hospital using the Monte Carlo simulation (MCS) method based on pharmacokinetics/pharmacodynamics modeling. In addition, the influence of the patients' renal function on the efficacy of RQs was evaluated. We surveyed antimicrobial susceptibility testing of 9 bacterial species (n = number of strains) [Streptococcus pneumoniae (n = 15), Streptococcus pyogenes (n = 14), Streptococcus agalactiae (n = 19), methicillin-susceptible Staphylococcus aureus (MSSA) (n = 24), Escherichia coli (n = 35), Haemophilus influenzae (n = 17), Klebsiella pneumoniae (n = 14), Pseudomonas aeruginosa (n = 31), and Moraxella catarrhalis (n = 11)] to 4 RQs [garenoxacin (GRNX), levofloxacin (LVFX), sitafloxacin (STFX), and moxifloxacin (MFLX)]. We found that compared with the other RQs, Gram-positive cocci was most resistant to LVFX, and that the minimum inhibitory concentration (MIC₉₀) values for S. pneumoniae, S. pyogenes, S. agalactiae, and MSSA were high (2, 16, > 16, and 8 µg/mL, respectively). In regard to Gram-negative rods, the susceptibility of E. coli to RQs was found to be decreased, with the MIC₉₀ values of GRNX, LVFX, STFX, and MFLX being > 16, 16, 1, and 16 µg/mL, respectively. MCS revealed that the target attainment rate of the area under the unbound concentration-time curve divided by the MIC₉₀ (ƒ · AUC/MIC ratio), against S. pneumoniae was 86.9-100%, but against E. coli was low (52.1-66.2%). The ƒ · AUC/MIC target attainment rate of LVFX against S. pneumoniae, S. pyogenes, and S. agalactiae tended to decrease due to increased creatinine clearance, and that of LVFX and STFX against MSSA also

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

    Celso Pianta


    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.

  6. 无乳链球菌检测及其奶牛乳腺炎防治的研究进展%Research Progress of the Detection Method of Streptococcus agalactiae and Control of Bovine Mastitis

    胡瑞萍; 刘磊; 边艳青; 赵宝华


    本文就近年来国内外在无乳链球菌的鉴定、检测方法、防御等方面的研究进展进行了系统性的综述,为控制该病原微生物的传播、蔓延和由无乳链球菌引起的奶牛乳腺炎的防治提供参考.无乳链球菌(Streptococcus agalactiae)属于B群链球菌,是影响人畜健康的重要病原微生物之一.它可以引起新生儿脑膜炎、肺炎、败血病等,又是奶牛乳腺炎的主要病原体之一,给奶牛业造成了巨大的损失.

  7. SIP亚单位疫苗对无乳链球菌诱导小鼠乳腺炎的保护力评价%Evaluation of Immunization with the SIP Against Streptococcus Agalactiae-induced Mouse Mastitis

    胡长敏; 徐海洋; 陈颖钰; 彭清洁; 郭爱珍


    无乳链球菌是导致奶牛乳腺炎的重要病原菌,本研究评价了SIP(surfaceimmunogenicprotein)亚单位疫苗对小鼠无乳链球菌乳腺炎的免疫保护效果。制备无乳链球菌SIP亚单位疫苗和灭活疫苗,对小鼠进行免疫,并设PBS阴性对照。免疫前后采血测定IgG及IgG亚类的抗体滴度。免疫小鼠分娩后第4天,进行无乳链球菌乳腺攻毒试验。24h后扑杀攻毒鼠,进行乳腺内CFU的测定,制作并观察乳腺组织病理切片。结果显示,SIP亚单位疫苗免疫组IgG及IgG亚类抗体滴度显著高于灭活疫苗组(P〈O.01)。免疫哺乳小鼠乳腺攻毒后,SIP亚单位疫苗免疫组小鼠乳腺CFU显著低于灭活疫苗组及对照组(P〈O.001)o乳腺病理切片显示SIP亚单位疫苗组乳腺组织结构较对照组完整,且中性粒细胞浸润程度最小。本研究表明,无乳链球菌SIP亚单位疫苗有望作为奶牛无乳链球菌乳腺炎的候选亚单位疫苗。%Streptococcus agalactiae (S. agalactiae) is a major cause of bovine mastitis. In the present study we assessed the efficacy of the surface immunogenic protein (SIP) as a mastitis vaccine in a mouse model. Three groups of mice were immunized with either SIP, inactivated S. agalactiae, or PBS as a control. IgG, IgG subtype and the capability of adhesion were evaluated. IgG and IgG subtype titers of mice in the SIP vaccine group were significantly higher than those in the killed vaccine group (P 〈0.01). Immunized lactating mice were challenged with S. agalactiae via the intramammary route. Significantly fewer bacteria were recovered from mice in the SIP group than from those in the killed vaccine group (P 〈0.001). Histopathology indicated that the mammary structure showed greater integrity and a milder inflammatory response in the SIP group as compared with both the inactivated vaccine and PBS control groups. These results suggested that the SIP protein may

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

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


    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.

  9. Haemolytic differential identification of Arcanobacterium haemolyticum isolated from a patient with diabetic foot ulcers

    Kang, Hyesook; Park, Gyunam; Kim, Hyeran


    Introduction: Arcanobacterium haemolyticum (formerly known as Corynebacterium haemolyticum) is the causative agent of sore throat and also causes skin and soft tissue infections in diabetes patients. A. haemolyticum is a Gram-positive, catalase-negative, β-haemolytic bacillus. A. haemolyticum poses a diagnostic challenge in the hospital laboratory because most coryneform bacilli are considered as normal flora or contaminants, and it is therefore difficult to differentiate from β-haemolytic streptococci by colony characteristics. Case presentation: A. haemolyticum was isolated from a diabetic patient with foot ulcers and the isolate was identified by using a VITEK-2 system, CAMP inhibition test, reverse CAMP test and a 23S rRNA gene sequence analysis. The isolated A. haemolyticum inhibited haemolysis of Staphylococcus aureus in the CAMP test and enhanced haemolysis of Streptococcus agalactiae in the reverse CAMP test. The diabetic patient was treated with teicoplanin and imipenem, and the ulcers healed within 2 weeks. Conclusion: The present study suggests that a haemolytic differential method using the CAMP inhibition and reverse CAMP tests can be useful for differentiating A. haemolyticum from β-haemolytic streptococci. PMID:28348747

  10. Construction and Prokaryotic Expression Sip-GAPDH Fusion Gene of Streptococcus agalactiae from GIFT Strain of Nile Tilapia (Oreochromis niloticus)%吉富罗非鱼源无乳链球菌Sip-GAPDH融合基因的构建及其原核表达

    王蓓; 李桂欢; 鲁义善; 汤菊芬; 黄郁葱; 吴灶和; 简纪常


    利用重叠延伸(SOEing)PCR技术,将吉富罗非鱼(Oreochromis niloticus)源无乳链球菌(Streptococcus agalactiae) ZQ0910株表面免疫原性蛋白(Surface immunogenic protein,Sip)基因与甘油醛-3-磷酸脱氢酶(GAPDH)基因通过Linker序列融合,构建成为Sip-GAPDH融合基因。将其定向克隆至原核表达载体pET-32a(+)中,在大肠杆菌BL21(DE3)中进行诱导表达。结果表明,重组质粒pET-32a-Sip-GAPDH在大肠杆菌BL21(DE3)中表达后所获得的融合蛋白Sip-GAPDH分子量为102.01 kD,表达最佳条件为37℃,IPTG浓度0.1 mmol/L,诱导5 h。Western blot鉴定结果表明融合基因得到了成功表达。%The DNA fragment encoding the Sip and GAPDH gene of Streptococcus agalactiae strain ZQ0910 isolated from GIFT Strain of Nile Tilapia(Oreochromis niloticus)were obtained by PCR. The Sip-GAPDH gene was constructed in a Sip-linker-GAPDH format with the standard 15-amino acid linker(Gly4Ser)3 by SOEing PCR technique, and the final full length product was cloned into the pET-32a(+)vector for protein expression in Escherichia coli strain BL21(DE3). The result showed that the expression fusion protein Sip-GAPHD were about 102.01 kD and Western blotting analysis confirmed that the 102.01 kD protein was the fusion protein, because it was specifically recognized by mouse anti-His monoclonal antibody. Inducing the cells at 37℃in 0.1 mmol/L of IPTG for 5 hours was the optimal conditions for expression of the recombinant Sip-GAPDH fusion protein.

  11. Isoler nu

    Andersen, N.E.; Ankerstjerne, P.; Jørgensen, B.

    Omfattende vejledning, der angiver, hvordan energiforbruget i eksisterende huse kan nedbringes ved isolering, tætning og forbedring af varmeanlægget, og hvor meget det koster. Anvisningen indeholder detaljerede eksempler på efterisolering af ydervægge, tage, gulve og vinduer. Henvender sig til...

  12. 奥尼罗非鱼无乳链球菌的鉴定、致病性及药物敏感性研究%Identification, Pathogenicity and Drug Sensitivity of Streptococcus agalactiae from Hybrid Tilapia, Oreochromis niloticus×Oreochromis aureus

    谭晶晶; 陈昌福; 高宇; 刘振兴; 王美珍; 李革雷


    从海南省某水产养殖场患病奥尼罗非鱼(Oreochromis aureus×Oreochromis niloticus)中,分离到1个编号为LFY-08-23的菌株.经过对分离菌株进行生理生化特性鉴定、Biolog系统鉴定以及利用原核生物16S rRNA基因通用引物进行 16S rRNA基因的克隆及序列分析,结果发现LFY-08-23菌株与无乳链球菌(Streptococcus agalactiae)的生理生化特性相似,16S rRNA基因克隆及序列分析结果与S.agalactiae(EF092913,DQ303183,AF459432)自然构成一个分支,相似性高达99.8%.用分离的S.agalactiae LFY-08-23菌株对异育银鲫和斑点叉尾鮰进行人工感染试验,结果表明,当注射菌浓度达到1.0×109cfu/mL时,2种鱼全部死亡.S.agalactiae LFY-08-23菌株对头孢氨噻肟、丙氟哌酸、利福平等13种药物高度敏感,对呋喃妥因、新生霉素、强力霉素等7种药物中度敏感,对卡那霉素、链霉素、磺胺异恶唑等5种药物不敏感.

  13. Escherichia coli Isolated from Urinary Tract Infections of Lebanese Patients between 2000 and 2009: Epidemiology and Profiles of Resistance

    Ziad Daoud


    Full Text Available The purpose of this study was to investigate the bacterial etiology of urinary tract infections in one of the busiest hospitals of Lebanon and to examine the epidemiologic and microbiologic properties of Escherichia coli isolated from urinary tract infections of Lebanese patients over a 10-year period. Methods. This retrospective study analyzed the data generated between 2000 and 2009 (10,013 Gram-positive and Gram-negative bacteria. Bacterial identification was based on standard culture and biochemical characteristics of isolates. Antimicrobial susceptibility was tested by the disk diffusion method, and ESBL production was detected by synergy with third-generation cephalosporins and amoxiclav. Results. E. coli was the most frequent isolate throughout the ten years (60.64% of the total isolates. It was followed by Klebsiella pneumoniae and Proteus sp., Pseudomonas aeruginosa, Enterococcus sp., and Streptococcus agalactiae. E. coli occurred more frequently in women (69.8% than in men (61.4%. The lowest percentage of susceptibility of E. coli was manifested against piperacillin and ampicillin. An increase in the production of ESBL was observed (2.3% in 2000 to 16.8% in 2009. Conclusions. The etiology of urinary tract infections and their susceptibility profiles are important to be evaluated in countries like Lebanon where a severe misuse of antibiotics at all levels is observed.

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

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


    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.

  15. A retrospective analysis of isolates from patients with vaginitis in a private Greek obstetric/gynecological hospital (2003-2006).

    Iavazzo, Christos; Vogiatzi, Christine; Falagas, Matthew E


    Vaginitis is a common cause of complaints in women in various parts of the world. Through our literature search, we identified noteworthy differences in the etiology of vaginitis based on the geographic location. This study was conducted to describe microbial etiology in a population of symptomatic women with vaginitis in Greece. Data for pathogens isolated from cultures of vaginal fluid specimens or identified with microscopy testing of such specimens of women with symptomatic vaginitis who were examined at "Lito" Private Obstetric/Gynecological Hospital, Athens, Greece, during the period 1/2003-12/2006 were analyzed retrospectively. During this period, 1632 women were examined. The mean age of the patients was 28 years, ranging from 18 to 57 years. Three hundred and eighty five from 1632 cultures/tests contributed no pathogen, whereas 1247/1632 cultures/tests were positive. Of the isolates, in which twelve species were identified, 504 (40.4%) were Gardnerella vaginalis, 530 (42.5%) Candida spp, and 101 (8.1%) Trichomonas vaginalis. Less usual isolates were Escherichia coli, Streptococcus agalactiae, Enterococcus spp., Streptococcus viridans, Staphylococcus epidermidis, Peptostreptococcus spp., and Staphylococcus saprophyticus. The authors believe that this study adds to the relevant literature of information regarding the pathogens implicated in patients with symptomatic vaginitis by reporting data from a cohort of patients in Greece. Ninety-one percent of the isolated and identified pathogens from vaginal cultures of symptomatic Greek women included Candida albicans, Gardnerella vaginalis, and Trichomonas vaginalis, with the first two species representing the great majority.

  16. Isolation World


    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.

  17. 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

    José Juvenal Linhares


    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

  18. LAMP检测无乳链球菌方法的建立及应用%Development and Application of Loop-Mediated Isothermal Amplification for Detection of Streptococcus agalactiae

    王永; 赵新; 景海春; 兰青阔; 朱珠; 程奕


    以无乳链球菌纤连蛋白fbs基因为主要研究对象,采取环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对fbs基因的6个区域设计4条特异性引物,利用一种链置换DNA聚合酶(Bst DNA polymerase)在63℃保温1 h,通过荧光显色即可完成对无乳链球菌的检测工作.结果显示,LAMP方法能够特异性检测fbs基因,其检测灵敏度是常规PCR方法的100倍,并与实时荧光定量PCR方法相当.所建立的针对无乳链球菌fbs基因的LAMP检测方法具有高度的特异性及稳定性,结果可靠,非常适合无乳链球菌的快速检测.%To develop a technique for detecting the fbsB gene of Streptococcus agalactiae using a novel DNA amplification method designated Loop-Mediated Isothermal Amplification (LAMP).The detection assay is based on the loop-mediated isothermal amplification (LAMP) reaction.The fbsB gene was amplified by a set of four specially primers that recognize six distinct sequences of the target.The amplification can be obtained in 1 h by incubating all of the reagents in a single tube with Bst DNA polymerase at 63℃.Results showed that the developed LAMP assay demonstrated an exceptionally higher than the conventional PCR.This assay results was found to be 100 times more sensitive than the PCR assay,and consistent with the results of real-time PCR.Our results clearly demonstrate that the LAMP-based assay is a sensitive and reliable means for the detection of Streptococcus agalactiae.

  19. The most common isolated microorganisms and its clinical manifestation in hospitalised women suffering from urogenital diseases in Latvia

    Anatolijs Naum and #269;iks


    Full Text Available Background: The monitoring of genital tract infections, especially sexually transmitted infections, is a vital part of health priority in many countries. One of the most common vaginal infections in women of childbearing age is bacterial vaginosis. Methods: This is a laboratory based retrospective study. We analysed 774 patients' cases (age range 18-35 years. Patients were subcategorized into three groups: patients with inflammation in genital tract, patients without inflammatory processes in genital tract, threatened abortion women. Laboratory tests also were divided depending on the type of examination. Results: There were 49 patients in the first group, 60 women in the second group and 236 pregnant women with threatened abortion in the third group. Gram negative bacteria (Escherichia coli and Enterococcus spp. were the most common isolated bacteria in the first and second group. Coagulase-negative staphylococci were predominant in the third group. Streptococcus agalactiae was isolated in 11.8 % cases. In our study, Trichomonas vaginalis infection was found in 1.2%. Conclusions: The most common isolated bacteria were Coagulase-negative staphylococci and gram negative bacteria. Ureaplasma urealyticum was detected in a decisive majority (38.9% of Mycoplasma species infections. In the first patients group and ndash; women with inflammatory process in genital tract were not found 3 bacteria, opposite to the second and third group. [Int J Reprod Contracept Obstet Gynecol 2015; 4(4.000: 959-962

  20. In vitro activity of Oritavancin against gram-positive pathogens isolated in Canadian hospital laboratories from 2011 to 2015.

    Karlowsky, James A; Walkty, Andrew J; Baxter, Melanie R; Arhin, Francis F; Moeck, Gregory; Adam, Heather J; Zhanel, George G


    Gram-positive bacterial pathogens isolated from patient specimens submitted to 15 Canadian hospital laboratories from 2011 to 2015 were tested in the coordinating laboratory for susceptibility to oritavancin and comparative antimicrobial agents using the Clinical and Laboratory Standards Institute M07-A10 (2015) broth microdilution method. Oritavancin's in vitro activity was equivalent to, or more potent than, vancomycin, daptomycin, linezolid, and tigecycline against methicillin-susceptible Staphylococcus aureus (n=2680; oritavancin MIC90, 0.12μg/mL; 99.9% oritavancin-susceptible), methicillin-resistant S. aureus (n=728; oritavancin MIC90, 0.12μg/mL; 99.7% oritavancin-susceptible), Streptococcus pyogenes (n=218; oritavancin MIC90, 0.25μg/mL; 100% oritavancin-susceptible), Streptococcus agalactiae (n=269; oritavancin MIC90, 0.12μg/mL; 100% oritavancin-susceptible), and vancomycin-susceptible Enterococcus faecalis (n=508; oritavancin MIC90, 0.06μg/mL; 100% oritavancin-susceptible). Oritavancin, dalbavancin, and telavancin demonstrated equivalent in vitro activities (MIC90, μg/mL) against 602 isolates of MSSA (0.06, 0.06, 0.06, respectively) and 144 isolates of MRSA (0.12, 0.06, 0.06, respectively) collected in 2015.

  1. Antimicrobial activity of pure platelet-rich plasma against microorganisms isolated from oral cavity

    Drago, Lorenzo; Bortolin, Monica; Vassena, Christian; Taschieri, Silvio; Del Fabbro, Massimo


    .... 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...

  2. Sensitivities of major causative organisms isolated from patients with acute uncomplicated cystitis against various antibacterial agents: results of subanalysis based on the presence of menopause.

    Matsumoto, Tetsuro; Hamasuna, Ryoichi; Ishikawa, Kiyohito; Takahashi, Satoshi; Yasuda, Mitsuru; Hayami, Hiroshi; Tanaka, Kazushi; Muratani, Tetsuro; Monden, Koichi; Arakawa, Soichi; Yamamoto, Shingo


    We investigated whether the presence of menopause influenced the species and distribution of causative bacteria isolated from patients with acute uncomplicated cystitis (the most common urinary tract infection), and we also investigated the sensitivity of the isolated species to antibacterial agents. Using multivariate analysis, we also investigated risk factors for infection with quinolone-resistant Escherichia coli, because its frequency has increased and it is now a clinical problem in Japan. Six hundred and thirty-four strains were isolated from 489 premenopausal patients (mean age 32.3 ± 10.1 years). Major causative bacteria detected were Escherichia coli (65.0 %), Enterococcus faecalis (12.0 %), Streptococcus agalactiae (5.5 %), and Klebsiella pneumoniae (1.6 %). From 501 postmenopausal patients (mean age 68.7 ± 10.29 years), 657 strains were isolated, and the major causative bacteria detected were E. coli (61.5 %), E. faecalis (13.7 %), K. pneumoniae (5.2 %), and S. agalactiae (4.0 %). The sensitivities to fluoroquinolones (FQs) and cephems of E. coli isolated from premenopausal patients were both ≥90 %, while the sensitivities to FQs of E. coli isolated from postmenopausal patients were about 5 % lower. In regard to infection with quinolone-resistant E. coli (minimal inhibitory concentration of levofloxacin [LVFX] ≥4 μg/mL), significant risk factors were observed in patients with more than two episodes of cystitis within a year (p = 0.0002), patients to whom antibacterial agents were used previously for this episode of cystitis (p = 0.0175), and patients who had a history of FQ administration within 1 month. Although the species and distribution of causative bacteria of acute uncomplicated cystitis were the same regardless of the presence of menopause, the sensitivities to FQs of E. coli detected in postmenopausal patients were significantly lower than those in the premenopausal women. The major risk factors for infection with quinolone-resistant E

  3. 20味中药提取物对无乳链球菌的体外抑菌活性研究%Effects of twenty traditional Chinese medicine extracts against Streptococcus agalactiae in vitro

    彭练慈; 殷中琼; 贾仁勇; 李莉; 代如意; 曲径; 刘明辉; 陈萍


    【目的】筛选对无乳链球菌Streptococcus agalactiae具有良好体外抑菌效果的中药.【方法】通过乙醇回流、超声波方法提取20味中药的抗菌活性部位,采用试管二倍稀释法测定黄连Coptis chinensis等20味中药提取物对无乳链球菌的最低抑菌浓度( Minimal inhibitory concentration ,MIC)和最低杀菌浓度( Minimal bactericidal concentration , MBC),并评价抗菌活性较强的几味中药的体外联合抑菌效果.【结果和结论】乌梅Prunus mume、黄连、黄芩Scutel-laria baicalensis、虎杖Polygonum cuspidatum和地榆Sanguisorba officinalis 5味中药提取物对无乳链球菌的MIC范围为7.80~31.25 mg/mL.蒲公英Taraxacum mongolicum、茵陈Artemisia scoparia、秦皮Fraxinus rhynchophylla、艾叶Arte-misia argyi、黄柏 Phellodendron chinense、鱼腥草 Houttuynia cordata、紫花地丁 Violae yedoensis 和白头翁 Pulsatilla chinensis 8味中药提取物的MIC范围为62.50~125.00 mg/mL.何首乌Po.multiflorum、苦参Sophora flavescens、穿心莲Andrographis paniculata、杜仲Eucommia ulmoides、连翘Forsythia suspensa、金银花Lonicera japonica和夏枯草Prunel-la vulgaris 7味中药提取物的MIC范围为250.00~500.00 mg/mL.联合抑菌试验结果表明,黄芩、虎杖、地榆和黄连的联合抑菌指数(FICI)≤1,虎杖、黄芩和地榆的FICI>2.黄连、黄芩、虎杖和地榆对无乳链球菌均具有很强的体外抗菌活性,地榆、黄连为相加作用,黄连、黄芩和虎杖为协同作用,地榆与黄芩、虎杖为拮抗作用.%Objective] Screening for the traditional Chinese medicines which have good antibacterial effects on Streptococcus agalactiae in vitro.[Method] Using ethanol reflux and ultrasonic method , ex-tracts of 20 different traditional Chinese medicines were obtained .Minimal inhibitory concentration ( MIC) and minimal bactericidal concentration ( MBC) of Coptis chinensis and other 19

  4. [Antibiotic sensitivity of beta-hemolytic streptococci isolated from throat swabs and purulent material].

    Jachna-Sawicka, Katarzyna; Pietrzak, Anna; Bogiel, Tomasz; Gospodarek, Eugenia


    The aim of this study was to evaluate the prevalence and susceptibility of beta-hemolytic streptococci isolated from throat swabs (142--29.9%) and purulent material (333--70.1%) taken from patients treated at University Hospital dr. A. Jurasz in Bydgoszcz Collegium Medicum. L. Rydygier in Bydgoszcz, Nicolaus Copernicus University in Torun in 2005-2009. Of the 475 tested strains, 156 (32.8%) were identified as S. pyogenes. This species accounted for 38.8% of strains isolated from purulent material and 19.0% of swabs from the throat. Among the strains isolated from throat swabs of 62 (43.7%) were identified as Streptococcus group C. Only 5.1% strains were identified as Streptococcus group F. All strains of beta-hemolytic streptococci were susceptible to ampicillin or penicillin, fluoroquinolones, vancomycin and linezolid. Erythromycin-susceptible strains was 83.8%, and 89.1% for clindamycin. A total of 51.3% of erythromycin resistance strains had the cMLS(B) phenotype (63.3% for strains from throat swabs and 46.3% of the purulent materials). Sensitivity to tetracycline was characterized by 51.2% of strains of beta-hemolytic streptococci. The percentage of strains susceptible to this antibiotic among isolates from throat swabs was 63.1%, and purulent material--48.0%. The lowest percentage of strains susceptible to tetracycline (14.1%) were found among S. agalactiae and Streptococcus group G (33.6%) strains. During the study time, saw an increase in the percentage of strains susceptible to tetracycline and erythromycin.

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

    Cosmina Bouari


    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. Antimicrobial susceptibility profiles, serotype distribution and virulence determinants among invasive, non-invasive and colonizing Streptococcus agalactiae (group B streptococcus) from Malaysian patients.

    Eskandarian, N; Ismail, Z; Neela, V; van Belkum, A; Desa, M N M; Amin Nordin, S


    A total of 103 group B streptococci (GBS) including 22 invasive, 21 non-invasive, and 60 colonizing isolates were collected in a Malaysian hospital (June 2010-October 2011). Isolates were characterized by conventional and molecular serotyping and analyzed for scpB, lmb, hylB, cylE, bac, bca and rib gene content. Antimicrobial susceptibility to penicillins, macrolides, lincosamides, quinolones and tetracyclines was determined using disk diffusion and the MICs for penicillin were determined by E-test. Molecular serotyping for all eight serotypes (Ia, Ib, II-VII) was in full accordance with conventional serotyping. Overall, taking CS and MS together, serotype VI was the most common capsular type (22.3 %) followed by VII (21.4 %), III (20.4 %), Ia (17.5 %), V (9.7 %), II (7.7 %) and IV (1 %). Susceptibility to beta-lactam antimicrobials was prevalent (100 %). Resistance rates for erythromycin, clindamycin and tetracycline were 23.3 %, 17.5 % and 71.8 %, respectively. PCR-virulence gene screening showed the presence of cylE, lmb, scpB and hylB in almost all the isolates while rib, bca, and bac genes were found in 29.1 %, 14.6 % and 9.7 % of the isolates. Certain genes were significantly associated with specific serotypes, namely, rib with serotypes Ia, II, III and VI; bca and bac with serotypes II and III. Furthermore, serotype Ia was significantly more common among patients with invasive infections (p < 0.01) and serotype VI isolates were significantly more common among carriers (p < 0.05). In summary, serotype distribution correlates with virulence gene content will be useful in epidemiological studies and design of vaccines.

  7. 奶牛乳房炎病原分离鉴定及其治疗方法研究%Contrast Study on Treatment Methods of Isolation and Identification of Dairy Cattle Mastitis Pathogens

    胡菊梅; 温嘉琪; 郑晓培; 王娟


    In order to formulate prevention and treatment measures for dairy cattle mastitis,50 mastitis samples from 6 different dairy farms in Tianshui area were collected and pathogen isolation,identification and drug sensitive test,106 strains of bacteria belong to 3 categories were isolated.They were showed as follows,Staphylococcus aureus,Staphylococcus epidermidis,Staphylococcus saprophyticus,Staphylococcus haemolyticus,Streptococcus agalactiae,Streptococcus breast,stop Streptococcus agalactiae,Escherichia coli,Salmonella,sand Ray and Proteus bacteria.The detection rate of Staphylococcus aureus was 24.5%,the highest of them.Meanwhile,treatment effects of different measures were compared.Therefore,therapy of combining traditional Chinese and Western medicine is the better measure to cure mastitis,which has high cure rate and quick effect.%为进一步制定奶牛乳房炎的防治措施,对天水地区6个不同奶牛场50份乳房炎奶样,进行病原分离、鉴定和药敏试验,共分离得到3类106株细菌,分别为:金黄色葡萄球菌、表皮葡萄球菌、腐生葡萄球菌、溶血葡萄球菌、无乳链球菌、乳房链球菌、停乳链球菌、大肠杆菌、沙门菌、沙雷菌和变形杆菌。金黄色葡萄球菌检出率最高,为24.5%。同时比较了不同治疗方法对奶牛乳房炎的治疗效果,认为中西医结合治疗乳房炎,其治愈率高,见效快。

  8. Antibacterial Activity of Herbal Preparations against Staphylococcus aureus and Streptococcus agalactiae in Cow Mastitis%中药制剂对奶牛隐性乳房炎金黄色葡萄球菌和无乳链球菌抑菌试验

    高桂生; 高光平; 李正本; 史秋梅; 张艳英; 邵新华; 梁银聚


    [目的]研究中药制剂对奶牛隐性乳房炎金黄色葡萄球菌和无乳链球菌的抑菌效果。[方法]根据中兽医学理论和中药特性,组成复方中药制剂“增乳健牛散”4种不同的方剂,制备水煎液对奶牛乳房炎的主要病原菌进行抑菌试验。[结果]4种不同的方剂水煎液都有抑菌效果;对金黄色葡萄球菌和无乳链球菌,方剂3为极敏,其他3种不同的方剂为高敏;方剂3抑菌作用最强。[结论]该研究为治疗奶牛隐性乳房炎相关中草药方剂的开发提供了理论依据。%[Objective] This study aimed to analyze the antibacterial activity of herbal preparations against Staphylococcus aureus and Streptococcus agalactiae in cow mastitis. [Method] The crude drug solutions of four different prescriptions for Zen-grujianniusan were prepared through reflux extraction. Their antibacterial activity in vitro against Staphylococcus aureus and Streptococcus agalactiae in cow mastitis were investigated. [Result] Al the four different prescriptions exhibited antibacterial activity against S. aureus and S. agalactiae. Among them, prescription Ⅲ was ex-tremely sensitive, and had the best bactericidal effect. The other three prescriptions were highly sensitive. [Conclusion] This study provides a theoretical basis for the development of herbal preparations for the treatment of cow mastitis.

  9. Development and application of loop-mediated isothermal amplification for detection of Streptococcus agalactiae in green terror Aequidens rivulatus%红尾皇冠鱼无乳链球菌病LAMP检测方法的建立与应用

    姚学良; 徐晓丽; 李贺密; 包海岩; 任涵玮; 郑艳坤


    以cfb基因保守序列的6个区域设计4条特异性引物,利用链置换DNA聚合酶( BstDNA polymer:ase)在恒温(65℃)下保温40 min,建立了无乳链球菌 Streptococcus agalactiae 的 LAMP 技术。建立的LAMP方法能够特异性地检测无乳链球菌,检测灵敏度为3·7×101~3·7×102 cfu/mL,反应前加入显色剂钙黄绿素避免二次污染。现场应用中采用FTA卡采集红尾皇冠鱼Aequidens rivulatus病鱼肝脏组织病原菌,操作简便、快捷。研究表明,针对无乳链球菌cfb基因建立的LAMP检测方法具有较高的特异性及稳定性,尤其是具有快速、简便、成本低等优点,可用于无乳链球菌的野外现场快速检测。%The cfb gene of Streptococcus agalactiae was amplified by a set of four specific primers that recognize six distinct sequences of the target in 40 min by incubating all of the reagents in a single tube with BstDNA polymerase at a constant 65 ℃. A loop-mediated isothermal amplification ( LAMP) assay was developed and validated for the specific detection of Streptococcus agalactiae, with detection limits of 3. 7í101-3. 7í102 cfu/mL. The resulting am-plicons were visualized by adding calcein to the reaction tube before the reaction. FTA cards was used for collection and purification of nucleic acids from a liver of green terror Aequidens rivulatus with pathogenic bacteria in field ap-plications. The findings demonstrate that the LAMP-based assay is a sensitive and reliable means for the detection of Streptococcus agalactiae with rapidity, simplicity and low cost, especially suitable for on-site rapid diagnosis.

  10. Isolated sleep paralysis

    Sleep paralysis - isolated; Parasomnia - isolated sleep paralysis ... Episodes of isolated sleep paralysis last from a few seconds to 1 or 2 minutes. During these episodes the person is unable to move ...

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

    Gamil S. G. Zeedan


    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

  12. 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

    Salvador Rogério


    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.

  13. Evaluation of Antimicrobial Activity of Cuminum Cyminum Essential Oil and Extract against Bacterial Strains Isolated from Patients with Symptomatic Urinary Tract Infection

    Yasaman Saee


    Full Text Available Background: Many efforts have been done to find effective agents against resistant pathogens. Cuminum cyminum L. (Cumin is an aromatic plant within the Apiaceae family. It has a variety of purposes and demonstrates antimicrobial and antioxidant properties. This study evaluated the activity of C. cyminum extract and essential oil against bacterial isolates which cause urinary tract infection, including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus agalactiae, group A streptococci, Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus and Staphylococcus saprophyticus isolated from patients with urinary tract infection.Materials and Methods: Extract was prepared by maceration and essential oil was prepared by hydrodistillation from C. cyminum seeds. The study population was 95 patients with urinary tract infection without malignant diseases, diabetes and immunosupression. After identification of organism, susceptibility testing was carried out by disc diffusion method and MIC values by broth microdilution testing.Results: C. cyminum essential oil can have a better effect on the gram-negative bacteria causing urinary tract infection than gram-positive bacteria. In addition, C. cyminum extract have good activity against both gram- positive and gram-negative bacteria. Our findings also showed that essential oil and extract of C. cyminum has better antibacterial activity on uropathogen isolates than amoxicillin and the difference was significant (P value<0.05 but the activity is not superior to other antibiotics.Conclusion: These results suggest that the essential oil and extract of C. cyminum seeds might be considered as interesting sources of antibacterial components against uropathogenic bacteria.

  14. The isolation and identification of Bacillus subtilis from Suaeda salsa and its bacteriostatic activity detection%碱蓬内生枯草芽孢杆菌分离鉴定及抑菌活性检测

    姜华; 修玉萍; 郑音; 任伟重; 侯婧; 张琳; 王有福


    Abstract:Seven endogenous strains were isolated from Liaoning Suaeda salsa using automatic microorganism identification system (VITEK) and molecular biology, including Bacillus stutilis, Bacillus circulans, Bacillus megatherium , Pseudomonas aeruginosa, Staphylococcus lentus, Staphylococcus sciuri and Streptococcus agalactiae. The results showed that Bacillus stutilis could inhibit the growth of 10 species of fungi in this experiment, especially Penicillium oxalicum, on which the inhibition rate reached 86. 4%, and the inhibition rate on Fusarium solani and Fusarium moniliforme were 52. 2% and 41. 4% respectively.%采用全自动微生物鉴定系统(VITEK)和分子生物学鉴定方法,从辽宁碱蓬中分离纯化出7种细菌:枯草芽孢杆菌(Bacillus stutilis)、环状芽孢杆菌(Bacillus circulans)、巨大芽孢杆菌(Bacillus megatherium)、铜绿假单胞菌(Pseudomonas aeruginosa)、缓慢葡萄球菌(Staphylococcus lentus)、松鼠葡萄球菌(Staphylococcus sciuri)和无乳链球菌(Streptococcus agalactiae).其中,枯草芽孢杆菌(Bacillus stutilis)对供试的10种真菌有抑菌作用,尤其对草酸青霉菌丝干物质抑制率可达86.4%,对茄病镰孢菌和串珠镰孢菌的抑制率分别为52.2%和41.4%.

  15. Biological Characteristic and Pathogenicity Research of Streptococcus agalactiae in Tilapia%罗非鱼无乳链球菌生物学特性与致病力研究

    田珂; 贾爱卿; 陈善真; 李守军; 王贵平; 石和荣


    本试验分离纯化1株致使罗非鱼具有链球菌发病症状的病原菌,药敏试验结果表明,该菌对氯霉素等10种药物敏感.PCR扩增该菌的cfb基因,将扩增片段测序后与GenBank上登录的罗非鱼无乳链球菌的cfb基因序列进行比对.同时对该病原菌的生物学特性进行研究,包括培养时间和pH对该菌生长的影响;培养温度对其致病力的影响;探讨了不同攻毒方式(腹腔、胸腔和背部肌肉注射)与鱼发病快慢的关系;并对不同规格(100和10 g左右)的罗非鱼对该菌的易感性进行分析.试验结果表明,该病原菌为无乳链球菌(Streptococcus agalactiae);该菌的最适培养时间为ll.5h,最适培养pH为7.5;腹腔、胸腔和背部肌肉注射后,其发病时间几乎一致,但胸腔注射的试验鱼死亡速度最慢,背部肌肉注射是较安全可行的攻毒方式;33℃培养的无乳链球菌攻毒的罗非鱼发病最快,死亡最集中;100 g的试验鱼比10 g的试验鱼更易感染.

  16. Immune Protective Effect of Streptococcus agalactiae Inactivated Vaccine on Tilapia(Oreochromis niloticus)%无乳链球菌灭活疫苗对罗非鱼的免疫效果

    陈贺; 吴灶和; 王蓓; 鲁义善; 黄郁葱; 简纪常


      制备无乳链球菌(Streptococcus agalactiae)临床分离株ZJ02的灭活疫苗,采用注射和浸泡2种方法,研究疫苗对吉富罗非鱼(Oreochromis niloticus)的免疫保护效果和血清抗体效价.注射组以4个不同浓度的白油佐剂灭活疫苗免疫罗非鱼,浸泡组以相同免疫剂量的灭活疫苗分别与消旋山莨菪碱、脂肪醇聚氧乙烯醚和氯化钠3种佐剂免疫罗非鱼.结果显示,无乳链球菌疫苗对罗非鱼具有较强的免疫原性,与对照组相比,免疫组血清中的抗体效价水平均显著性提高(p<0.05),注射组和浸泡组的抗体效价最高分别达到1∶256和1∶128;注射组中不同剂量的疫苗免疫组免疫保护率较高,高浓度组可达到95%,浸泡组中使用3种佐剂的免疫组免疫保护率分别为59%、64%、55%.表明无乳链球菌灭活疫苗对罗非鱼具有很好的免疫保护性,免疫剂量在5×108~4×109 cfu/mL之间对注射免疫效果有显著性影响,消旋山莨菪碱、脂肪醇聚氧乙烯醚和氯化钠对浸泡免疫效果具有不同的免疫保护率.

  17. 黄河裸裂尻鱼无乳链球菌的分离鉴定与多位点序列分型%Identification of the pathogen Streptococcus agalactiae in Schizopygopsis pylzovi (Kessler) by multilocus sequence typing

    张雨薇; 耿毅; 余泽辉; 汪开毓; 李亚军; 牟维豪; 曾毅仁; 叶梦婷


    2015年10月四川省雅安市某养殖场的黄河裸裂尻鱼(Schizopygopsis pylzovi Kessler)暴发传染病,临床表现为神经症状,眼球、下颌和鳃盖等出血或无明显临床表现而突然死亡,内脏涂片见革兰氏阳性球菌.从病鱼的肝、脾和肾组织中分离到2株病原菌(ZYW151011,ZYW151012),通过人工感染、无乳链球菌种属特异性cfb基因(GBS-specific gene cfb,CAMP factor)的PCR检测和16S rRNA基因序列分析等方法对病原菌进行鉴定,发现2株病原菌均为无乳链球菌(Streptococcus agalactiae).2株病原菌16S rRNA序列(GenBank登录号:KU308396和KU308397)与GenBank中无乳链球菌16S rRNA序列同源性达99.8%.药敏试验结果表明,2株病原菌对头孢类和喹诺酮类药物敏感,而对强力霉素、氟苯尼考和阿莫西林等耐药.进一步通过荚膜多糖血清分型和多位点序列分型技术对病原菌进行分子特征分析,结果表明2株病原菌荚膜多糖血清型均为致病性Ia型;2株病原菌多位点序列分型均被鉴定为新的序列型ZST-1,与序列型为ST-261的亲缘关系最近,此新型ZST-1在glnA位点发生变异,提交序列到MLST数据库获得新的等位基因编号81(登录ID:BIGSdb_ 20151110 081850_13025_35759).

  18. Isolated patellofemoral osteoarthritis.

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


    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 discu

  19. Isolated patellofemoral osteoarthritis.

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


    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

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

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


    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

  1. 罗非鱼源无乳链球菌S-核糖基高半胱氨酸酶基因(luxS)的克隆及其推导蛋白的三维结构预测%Cloning of Streptococcus agalactiae luxS gene from tilapia and 3D structure prediction of deduced protein

    马艳平; 李嘉彬; 郝乐; 刘振兴; 冯国清; 周结珊; 柯浩


    We have amplified, cloned and determined the sequence of Streptococcus agalactiae luxS gene from Tilapia sp. by PCR. The characteristics of the deduced luxS protein were predicted by ExPAsy software; the 3D structures of luxS and the deduced protein were established and analyzed by SWISS-Model and SwisS-PDBviewer software, respectively. The results indicate that the deduced luxS protein contains conserved active center and Zn2+ binding site, which may affect biofilm formation and regulate virulence factor. The Ramachandram plot shows that the structure of modeled luxS protein is reasonable.%利用PCR技术对罗非鱼源无乳链球菌(Streptococcus agalactiae)S-核糖基高半胱氨酸酶(luxS)基因全长DNA进行了扩增、克隆和序列测定,采用ExPAsy软件包预测了推导蛋白的特性,利用SwisS-Model服务器建立了luxS 三维结构,利用SwisS-PDBviewer软件进行了蛋白质三维结构的分析.预测结果显示,罗非鱼源无乳链球菌luxS推导蛋白包括保守的酶活性中心和锌结合位点,具有影响生物被膜形成、毒力因子调控等特性功能;经拉氏构象图(Ramachandran plot)分析,所构建的luxS的空间结构合理.

  2. Mutation and premating isolation.

    Woodruff, R C; Thompson, J N


    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.

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



    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

  4. 某院妇产科住院患者的无乳链球菌感染情况及耐药性研究%Research of infectious situation and drug resistance of Streptococcus agalactiae in patients of the department of gynecology and obstetrics

    刘泽滨; 聂署萍; 陆学东; 林立鹏; 邓凤娴


    目的:研究本院产科、妇科住院患者中的无乳链球菌感染情况,尤其在首胎产妇和二胎产妇中的无乳链球菌感染情况,并对培养出的阳性菌株进行药敏试验,了解其耐药情况。方法将阴道拭子标本接种于血平板上,用Vitek2 compact全自动细菌鉴定及药敏分析系统对异常菌落进行鉴定,并对培养出的无乳链球菌进行药敏分析。结果培养产科孕妇阴道拭子标本共730份,其中无乳链球菌培养阳性为12例,阳性率为1.64%;培养妇科患者阴道拭子标本共583份,其中无乳链球菌培养阳性为6例,阳性率为1.03%。首胎孕妇为505例,无乳链球菌培养阳性为8例,阳性率1.59%;二胎孕妇为227例,无乳链球菌培养阳性为4例,阳性率1.76%。首胎孕妇与二胎孕妇的无乳链球菌感染率进行比较,差异无统计学意义( P>0.05)。所有无乳链球菌阳性菌株对头孢菌素类抗菌药物有较高的敏感性,未出现头孢哌酮、头孢呋辛钠、头孢曲松的耐药菌株,敏感率为100%。无乳链球菌阳性菌株对环丙沙星和左旋氧氟沙星的耐药率较高,分别为38.89%、27.78%。结论产科患者的无乳链球菌感染率较妇科患者的高,临床上仍需加强对孕产妇的无乳链球菌感染监测。无乳链球菌培养阳性的菌株对头孢菌素类抗菌药物有较高敏感性,对喹诺酮类抗菌药物有较高的耐药性。%Objective To explore the infectious situation and drug resistance of Streptococcus agalactiae in patients of the de‐partment of gynecology and obstetrics ,especially in the first fetal pregnant women and second fetal pregnant women .Methods The vaginal swab specimens were inoculated on blood plates ,and then the abnormal bacterial colonies were identified by compact Vitek2 automatic bacteria identification and drug sensitivity analysis system .Results The total number of

  5. Prevalence of antimicrobial resistance in bacteria isolated from central nervous system specimens as reported by U.S. hospital laboratories from 2000 to 2002

    Karlowsky James A


    Full Text Available Abstract Background Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy. The widespread emergence of resistance among bacterial species is a cause for concern. Current antibacterial susceptibility data among central nervous system (CNS pathogens is important to define current prevalence of resistance. Methods Antimicrobial susceptibility of pathogens isolated from CNS specimens was analyzed using The Surveillance Database (TSN® USA Database which gathers routine antibiotic susceptibility data from >300 US hospital laboratories. A total of 6029 organisms derived from CNS specimen sources during 2000–2002, were isolated and susceptibility tested. Results Staphylococcus aureus (23.7% and Streptococcus pneumoniae (11.0% were the most common gram-positive pathogens. Gram-negative species comprised approximately 25% of isolates. The modal patient age was 1 or S. aureus from cerebrospinal fluid (CSF and brain abscesses were 29.9–32.9%. Penicillin resistance rates were 16.6% for S. pneumoniae, 5.3% for viridans group streptococci, and 0% for S. agalactiae. For CSF isolates, ceftriaxone resistance was S. pneumoniae (3.5%, E. coli (0.6%, Klebsiella pneumoniae (2.8%, Serratia marcescens (5.6%, Enterobacter cloacae (25.0%, Haemophilus influenzae (0%. Listeria monocytogenes and N. meningitidis are not routinely susceptibility tested. Conclusions Resistance is commonly detected, albeit still at relatively low levels for key drugs classes such as third-generation cephalosporins. This data demonstrates the need to consider predominant resistance phenotypes when choosing empiric therapies to treat CNS infections.

  6. Isolated Galaxies and Isolated Satellite Systems

    Ann, H B; Choi, Yun-Young


    We search for isolated galaxies using a volume-limited sample of galaxies with 0.02r_{vir,nei} and \\rho <\\bar{\\rho} well segregates the CIG galaxies. We confirm the morphology conformity between the host and their satellites, which suggests importance of hydrodynamic interaction among galaxies within their virial radii in galaxy evolution.

  7. Alternativ isolering i bygninger

    Andersen, T.; Skibstrup Eriksen, S.

    Denne SBI-meddelelse beskriver resultaterne af en undersøgelse af bygninger, hvor der er anvendt alternativ isolering. Undersøgelsen omfatter termografering og fugtmåling af udvendige bygningsdele isoleret med alternativ isolering. Endvidere rapporteres resultaterne af målinger af indholdet af tu...

  8. Population structure and characterization of viridans group streptococci (VGS) including Streptococcus pneumoniae isolated from adult patients with cystic fibrosis (CF).

    Maeda, Yasunori; Elborn, J Stuart; Parkins, Michael D; Reihill, James; Goldsmith, Colin E; Coulter, Wilson A; Mason, Charlene; Millar, B Cherie; Dooley, James S G; Lowery, Colm J; Ennis, Madeleine; Rendall, Jacqueline C; Moore, John E


    A study was undertaken to examine the population structure of viridans group streptococci (VGS) in the sputum of adult patients with cystic fibrosis (CF). Freshly expectorated sputa (n=58) from 45 adult CF patients were examined by selective conventional culture on Mitis-Salivarius agar and yielded 190 isolates of VGS. Sequence analyses of the rpnB and 16-23S rRNA ITS genes identified these isolates to belong to 12 species of VGS and included S. anginosus, S. australis, S. cristatus, S. gordonii, S. infantis, S. mitis, S. mutans, S. oralis, S. parasanguinis, S. pneumoniae, S. salivarius and S. sanguinis. The most frequently VGS organism isolated was S. salivarius (47/190; 24.7%), followed by S. mitis (36/190; 19%), S. sanguinis (25/190; 13.2%), S. oralis (20/190; 11.0%), S. pneumoniae (19/190; 10.0%), S. parasanguinis (16/190; 8.4%), S. infantis (11/190; 5.8%), S. gordonii (7/190; 3.7%), S. anginosus (4/190; 2.1%), S. cristatus (2/190; 1.1%), S. australis (1/190; 0.5%), S. mutans (1/190; 0.5%) and S. agalactiae (1/190; 0.5%). All, but four, patients harboured at least one VGS species, which ranged from one to five streptococcal species, with a mean of 2.85 species per patient. There was no clonality at the subspecies level employing ERIC RAPD PCR. Antibiotic susceptibility was determined by Minimum Inhibitory Concentration (MIC) testing against penicillin, erythromycin and ciprofloxacin. Overall, resistance to penicillin with all VGS was 73/190 (38.4%) and 167/190 (87.9%) for erythromycin. With regard to ciprofloxacin, 27/190 (14.2%) were fully resistant, whilst a further 21/190 (11.1%) showed intermediate resistance, which equated to approximately three quarters (74.7%) of isolates being fully sensitive to this agent. In addition, as a comparator control population, we examined antibiotic susceptibility, as above, in a non-CF population comprising 12 individuals (50 VGS isolates), who were not receiving chronic antibiotics. In comparison, 8% and 38% of VGS

  9. Isolated Vascular Vertigo


    Strokes in the distribution of the posterior circulation may present with vertigo, imbalance, and nystagmus. Although the vertigo due to a posterior circulation stroke is usually associated with other neurologic symptoms or signs, small infarcts involving the cerebellum or brainstem can develop vertigo without other localizing symptoms. Approximately 11% of the patients with an isolated cerebellar infarction present with isolated vertigo, nystagmus, and postural unsteadiness mimicking acute peripheral vestibular disorders. The head impulse test can differentiate acute isolated vertigo associated with cerebellar strokes (particularly within the territory of the posterior inferior cerebellar artery) from more benign disorders involving the inner ear. Acute audiovestibular loss may herald impending infarction in the territory of anterior inferior cerebellar artery. Appropriate bedside evaluation is superior to MRIs for detecting central vascular vertigo syndromes. This article reviews the keys to diagnosis of acute isolated vertigo syndrome due to posterior circulation strokes involving the brainstem and cerebellum. PMID:25328871

  10. Base isolation: Fresh insight

    Shustov, V.


    The objective of the research is a further development of the engineering concept of seismic isolation. Neglecting the transient stage of seismic loading results in a widespread misjudgement: The force of resistance associated with velocity is mostly conceived as a source of damping vibrations, though it is an active force at the same time, during an earthquake type excitation. For very pliant systems such as base isolated structures with relatively low bearing stiffness and with artificially added heavy damping mechanism, the so called `damping`` force may occur even the main pushing force at an earthquake. Thus, one of the two basic pillars of the common seismic isolation philosophy, namely, the doctrine of usefulness and necessity of a strong damping mechanism, is turning out to be a self-deception, sometimes even jeopardizing the safety of structures and discrediting the very idea of seismic isolation. There is a way out: breaking with damping dependancy.

  11. Alternativ isolering i bygninger

    Hansen, Ernst Jan de Place


    Resume af rapport om målinger på alternative isoleringsmaterialer i bygninger, udgivet af Statens Byggeforskningsinstitut (SBI meddelelse 128) under Energistyrelsens udviklingsprogram "Miljø- og arbejdsmiljøvenlig isolering"...

  12. Isolated sleep paralysis

    Sawant, Neena S.; Parkar, Shubhangi R.; Tambe, Ravindra


    Sleep paralysis (SP) is a cardinal symptom of narcolepsy. However, little is available in the literature about isolated sleep paralysis. This report discusses the case of a patient with isolated sleep paralysis who progressed from mild to severe SP over 8 years. He also restarted drinking alcohol to be able to fall asleep and allay his anxiety symptoms. The patient was taught relaxation techniques and he showed complete remission of the symptoms of SP on follow up after 8 months.

  13. Isolated sleep paralysis

    Sawant, Neena S.; Shubhangi R Parkar; Tambe, Ravindra


    Sleep paralysis (SP) is a cardinal symptom of narcolepsy. However, little is available in the literature about isolated sleep paralysis. This report discusses the case of a patient with isolated sleep paralysis who progressed from mild to severe SP over 8 years. He also restarted drinking alcohol to be able to fall asleep and allay his anxiety symptoms. The patient was taught relaxation techniques and he showed complete remission of the symptoms of SP on follow up after 8 months.

  14. Isolated patellofemoral osteoarthritis

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


    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 discussion of treatment options. METHODS: A systematic computerized database search (Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, MEDLINE (PubMed), and EMBASE...

  15. 新生儿无乳链球菌败血症合并脑膜炎12例临床分析%Clinical and prognostic analysis of sepsis caused by Streptococcus agalactiae combined with purulent meningitis in 12 ;neonates

    杨欢欢; 李菁


    目的:探讨无乳链球菌(GBS)败血症合并脑膜炎新生儿的临床表现、治疗及预后情况。方法回顾性分析2012年1月至2015年5月收治的12例无乳链球菌脑膜炎新生儿的临床资料。结果12例患儿中,男女各6例,均为足月儿,早发型4例、晚发型8例,顺产7例、剖宫产5例,母亲均无明显感染病史,亦未行常规GBS筛查。新生儿血降钙素原均升高。头颅磁共振(MRI)可见脑膜强化,5例硬膜下积液,3例脑实质受累。1例单用青霉素,1例应用美罗培南之后降阶梯单用青霉素效果良好,大部分患儿需联合用药。随访20~29个月,1例语言发育落后,2例运动发育落后,其余9例发育正常。结论新生儿无乳链球菌败血症合并脑膜炎病情危重,需及时应用敏感抗生素,必要时联合用药。建议完善产前筛查。%Objective To investigate the clinical manifestations, treatment and prognosis of sepsis caused by Streptococcus agalactiae combined with purulent meningitis in neonates. Methods Clinical data of 12 neonates with purulent meningitis caused by Streptococcus agalactiae were analyzed retrospectively from January 2012 to May 2015. Results In 12 full term neonates (6 males and 6 females) , there were 4 early onset cases and 8 late onset cases;7 cases of vaginal delivery and 5 cases of cesarean section. All of the mothers had no history of infections, and also no routine screening for group B streptococcus. The serum levels of procalcitonin were elevated in all neonates. Brain magnetic resonance imaging results indicated abnormal meningeal enhancement, subdural effusion in 5 cases and brain parenchyma involvement in 3 cases. One case was treated with penicillin only. One case was treated with meropenem ifrst and then penicillin only with good efifcacy. Most of the neonates need drug combination therapy. During 20 ~ 29 months of following-up, one case had language development delays, 2

  16. Isolated Splenic Hydatid Disease

    Alper Dilli


    Full Text Available Hydatid disease (HD continues to be a significant health problem in areas where animal husbandry is common but no proper veterinary control exists. The involvement of the spleen in HD is rare, and isolated splenic involvement is even less common. In this case report, we present isolated splenic HD in a 26-year-old female with complaint of abdominal pain, and we discuss some of the clinical aspects of HD. Evaluation of the patient with ultrasonography, computed tomography, and magnetic resonance imaging revealed the presence of an isolated splenic HD as a multivesicular cystic mass located near splenic hilus, measuring 12×11 cm. No other organ or system involvement could be demonstrated.

  17. Isolated Fallopian Tube Torsion

    S. Kardakis


    Full Text Available Isolated torsion of the Fallopian tube is a rare gynecological cause of acute lower abdominal pain, and diagnosis is difficult. There are no pathognomonic symptoms; clinical, imaging, or laboratory findings. A preoperative ultrasound showing tubular adnexal masses of heterogeneous echogenicity with cystic component is often present. Diagnosis can rarely be made before operation, and laparoscopy is necessary to establish the diagnosis. Unfortunately, surgery often is performed too late for tube conservation. Isolated Fallopian tube torsion should be suspected in case of acute pelvic pain, and prompt intervention is necessary.

  18. Isolation of Photons

    Wielers, M


    Photon identification in ATLAS is based mainly on the shower shapes in the calorimeters, conversion reconstruction and a track veto. In this note the additional gamma/jet separation power is evaluated if isolation is required around the shower as additional criteria.

  19. Motion of isolated bodies

    Choquet-Bruhat, Yvonne [Academie des Sciences, 23 Quai de Conti, 75270 Paris Cedex 06 (France); Friedrich, Helmut [Max-Planck-Institut fuer Gravitationsphysik, Am Muehlenberg 1, 14476 Golm (Germany)


    It is shown that sufficiently smooth initial data for the Einstein-dust or the Einstein-Maxwell-dust equations with non-negative density of compact support develop into solutions representing isolated bodies in the sense that the matter field has spatially compact support and is embedded in an exterior vacuum solution.

  20. Motion of Isolated bodies

    Choquet-Bruhat, Y; Choquet-Bruhat, Yvonne; Friedrich, Helmut


    It is shown that sufficiently smooth initial data for the Einstein-dust or the Einstein-Maxwell-dust equations with non-negative density of compact support develop into solutions representing isolated bodies in the sense that the matter field has spatially compact support and is embedded in an exterior vacuum solution.

  1. Biological Isolation Garment


    A spinoff of astronaut's biological garment will allow hospital patients who are highly vulnerable to infection to leave their sterile habitats for several hours, carrying their germ free environment with them. Garments can be used in any of some 200 hospitals where isolation rooms are installed to treat leukemia.



    In this paper, some results obtained on the formation of isolated molecules of composition SnOx in silver and SnFx in copper-are reviewed. Hyperfine interaction and ion beam interaction techniques were used for the identification of these molecules.



    In this paper, some results obtained on the formation of isolated molecules of composition SnOx in silver and SnFx in copper-are reviewed. Hyperfine interaction and ion beam interaction techniques were used for the identification of these molecules.

  4. Isolated lingual leishmaniasis

    Habibzadeh F


    Full Text Available Cutaneous leishmaniasis is endemic in Fars Province, southern Iran. However, mucosal leishmaniasis is extremely uncommon. Herein, we report a patient with isolated lingual leishmaniasis in an immunocompetent 40-year-old man. The lesion was totally excised. The patient was cured completely and is doing well after four years of follow-up, with no medical treatment

  5. Isolated jejunal varices.

    Bhagwat S


    Full Text Available Isolated jejunal varices are an uncommon manifestation of portal hypertension. A one and a half year old boy presented with recurrent, massive gastrointestinal bleeding from jejunal varices. The bleeding site was identified at exploratory laparotomy. Jejunal resection and anastomosis resulted in complete resolution of the bleeding and there has been no recurrent bleeding over an eight month follow-up period.

  6. Proteoglycan isolation and analysis

    Woods, A; Couchman, J R


    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...

  7. Isolating Triggered Star Formation

    Barton, Elizabeth J.; Arnold, Jacob A.; /UC, Irvine; Zentner, Andrew R.; /KICP, Chicago /Chicago U., EFI; Bullock, James S.; /UC, Irvine; Wechsler, Risa H.; /KIPAC, Menlo


    Galaxy pairs provide a potentially powerful means of studying triggered star formation from galaxy interactions. We use a large cosmological N-body simulation coupled with a well-tested semi-analytic substructure model to demonstrate that the majority of galaxies in close pairs reside within cluster or group-size halos and therefore represent a biased population, poorly suited for direct comparison to 'field' galaxies. Thus, the frequent observation that some types of galaxies in pairs have redder colors than 'field' galaxies is primarily a selection effect. We use our simulations to devise a means to select galaxy pairs that are isolated in their dark matter halos with respect to other massive subhalos (N= 2 halos) and to select a control sample of isolated galaxies (N= 1 halos) for comparison. We then apply these selection criteria to a volume-limited subset of the 2dF Galaxy Redshift Survey with M{sub B,j} {le} -19 and obtain the first clean measure of the typical fraction of galaxies affected by triggered star formation and the average elevation in the star formation rate. We find that 24% (30.5 %) of these L* and sub-L* galaxies in isolated 50 (30) h{sup -1} kpc pairs exhibit star formation that is boosted by a factor of {approx}> 5 above their average past value, while only 10% of isolated galaxies in the control sample show this level of enhancement. Thus, 14% (20 %) of the galaxies in these close pairs show clear triggered star formation. Our orbit models suggest that 12% (16%) of 50 (30) h{sup -1} kpc close pairs that are isolated according to our definition have had a close ({le} 30 h{sup -1} kpc) pass within the last Gyr. Thus, the data are broadly consistent with a scenario in which most or all close passes of isolated pairs result in triggered star formation. The isolation criteria we develop provide a means to constrain star formation and feedback prescriptions in hydrodynamic simulations and a very general method of understanding

  8. Chemical composition and antimicrobial activity of the essential oil of Syzygium aromaticum flower bud (Clove) against fish systemic bacteria isolated from aquaculture sites

    Seongwei LEE; Musa NAJIAH; Wee WENDY; Musa NADIRAH


    This paper describes the chemical composition and antimicrobial activity of the essential oil of Syzygium aromaticum against Vibrio spp. (n=6), Edwardsiella spp. (n=21), Aeromonas spp. (n=2), Escherichia coli (n=2), Flavobacterium spp. (n=1), Salmonella spp. (n=2), Streptococcus spp. (n=1) and Pseudomonas spp. (n=1) isolated from aquaculture sites as well as seven reference strains of bacteria, namely, Escherichia coli (ATCC 25922), Citrobacter freundii (ATCC 8090), Aeromonas hydrophila (ATCC 49140), Pseudomonas aeruginosa (ATCC 35032), Streptococcus agalactiae (ATCC13813), Edwardsiella tarda (ATCC 15947) and Yersinia enterocolitica (ATCC 23715). Nowadays, most antibiotics are no longer effective in controlling diseases in aquaculture, especially fish systemic bacterial diseases, due to increasing incidences of antibiotic resistance among pathogenic bacteria. Furthermore, many countries have banned antibiotics in aquaculture use due to public health concerns and environmental hazards. Therefore, this study was carried out to evaluate the potential of the essential oil of S. aromaticum as an alternate commercial antibiotic to antimicrobial agents against fish systemic bacteria in aquaculture. The essential oil of S. aromaticum was prepared using a steam distillation method, and the chemical composition was analysed using Gas chromatography-mass spectroscopy (GC-MS). Minimum inhibitory concentration (MIC) values of the essential oils against the tested bacteria were determined using the broth two fold micro dilution method, with kanamycin and eugenol as positive controls. The MIC values of the essential oil of S. aromaticum ranged from 0.015 μg·mL-1 to 0.062 μg·mL-1 against the tested bacterial isolates. A total of nine chemical compounds were detected in the essential oil, with eugenol (49.0%) and caryophyllene (7.5%) being the major compounds. The results of the present study indicate that the essential oil of S. aromati- cum shows a huge potential to

  9. Prediction of DNA-binding Sites in Streptococcus agalactiae GlnR Factor and Construction of glnR Mutant Gene%无乳链球菌GlnR因子DNA结合位点预测及其突变基因的构建

    夏杰; 易弋; 王佳; 杨军; 黄稀; 黄翠姬; 伍时华


    利用生物信息学的方法对无乳链球菌(Streptococcus agalactiae)GlnR因子与DNA结合的位点进行预测,筛选出GlnR因子中与DNA结合的关键位点的氨基酸残基R47和R48.然后利用重叠PCR的方法将glnR基因上的第139~144位上的碱基CGCCGC突变为GCGGCG,使其编码的2个精氨酸残基突变为丙氨酸残基,将突变基因片段TA克隆至PMD 18-T载体上进行测序验证.测序结果表明,利用重叠PCR成功突变了这6个碱基,为后续研究GlnR因子的突变位点对DNA结合的影响提供了参考.

  10. A Nested PCR-based Method for Detection of 16S rRNA of Stretococcus agalactiae in Milk and Its Application%巢式PCR检测无乳链球菌16SrRNA方法的建立及应用

    贾玉萍; 周东顺; 万仁忠; 刘文强; 胡敬东; 赵宏坤


    无乳链球菌(Streptococcus agalactiae)引起的奶牛乳房炎主要表现为隐性型,需要通过实验室检查才能确诊.研究建立了一种特异、敏感和快速的巢式PCR方法用于检测奶样中的无乳链球菌.利用链球菌属(Streptococcus)16SrRNA基因的保守区设计通用引物作为链球菌属的阳性控制,在保守区内的可变区设计无乳链球菌特异的引物.研究结果表明,通过奶样中无乳链球菌的增菌培养,简便、快速提取DNA和特异性PCR反应,可以检测到奶样中1 CFU/mL的无乳链球菌,可以用作牛群中无乳链球菌感染的早期流行病学调查.

  11. Offset trench isolation

    Roth, S. S.; Cooper, K. J.; Ray, W.; Kirsch, H. C.; Grove, Charlotte


    Feasibility of a new, recessed isolation technique that utilizes an offset, shallow trench in combination with thermal oxidation for achieving near zero final encroachment with excellent planarity is demonstrated. Etch of the shallow trench is offset from the original hardmask by an oxide sidewall spacer. After trench etch, HF is used to remove the hardmask oxide and sidewall spacers and to form a cavity which is self-aligned to the nitride edge. Exposed silicon regions are then reoxidized and encapsulated with polysilicon. Field oxide is then grown. The final field oxide profile exhibits steep sidewall angles without inducing substrate defects as evidenced by low diode leakage. Other isolation sensitive device parametrics such as gate oxide quality and metal oxide semiconductor field effect transistor threshold voltage stability are presented and exhibit good characteristics.

  12. Idiopathic isolated orbicularis weakness

    MacVie, O P; Majid, M A; Husssin, H M; Ung, T; Manners, R M; Ormerod, I; Pawade, J; Harrad, R A


    Purpose Orbicularis weakness is commonly associated with seventh nerve palsy or neuromuscular and myopathic conditions such as myotonic dystrophy and myasethenia gravis. We report four cases of idiopathic isolated orbicularis weakness. Methods All four cases were female and the presenting symptoms of ocular irritation and epiphora had been present for over 7 years in three patients. All patients had lagophthalmos and three had ectropion. Three patients underwent full investigations which excluded known causes of orbicularis weakness. Two patients underwent oribularis oculi muscle biopsy and histological confirmation of orbicularis atrophy. Results All patients underwent surgery to specifically address the orbicularis weakness with satisfactory outcomes and alleviation of symptoms in all cases. Isolated orbicularis weakness may be a relatively common entity that is frequently overlooked. Conclusion Early recognition of this condition may lead to better management and prevent patients undergoing unnecessary surgical procedures. PMID:22322997

  13. Isolated Pulmonary Valve Endocarditis

    Mohammad Ali Hatamizadeh


    Full Text Available Infective endocarditis is one of the most severe complications of parenteral drug abuse. The outstanding clinical feature of infective endocarditis in intravenous drug abusers is the high incidence of right-sided valve infection, and the tricuspid valve is involved in 60% to 70% of the cases. We herein report a case of isolated pulmonic valve infective endocarditis with a native pulmonary valve.

  14. Isolated unilateral disk edema

    Varner P


    Paul VarnerJohn J Pershing VAMC, Poplar Bluff, MO, USAAbstract: Isolated unilateral disk edema is a familiar clinical presentation with myriad associations. Related, non-consensus terminology is a barrier to understanding a common pathogenesis. Mechanisms for the development of disk edema are reviewed, and a new framework for clinical differentiation of medical associations is presented.Keywords: disk edema, axoplasmic flow, clinical multiplier, optic neuritis, ischemic optic neuropathy, papi...

  15. Isolated unilateral disk edema

    Varner P


    Full Text Available Paul VarnerJohn J Pershing VAMC, Poplar Bluff, MO, USAAbstract: Isolated unilateral disk edema is a familiar clinical presentation with myriad associations. Related, non-consensus terminology is a barrier to understanding a common pathogenesis. Mechanisms for the development of disk edema are reviewed, and a new framework for clinical differentiation of medical associations is presented.Keywords: disk edema, axoplasmic flow, clinical multiplier, optic neuritis, ischemic optic neuropathy, papilledema

  16. Isolating Triggered Star Formation

    Barton, Elizabeth J; Zentner, Andrew R; Bullock, James S; Wechsler, Risa H


    Galaxy pairs provide a potentially powerful means of studying triggered star formation from galaxy interactions. We use a large cosmological N-body simulation coupled with a well-tested semi-analytic substructure model to demonstrate that the majority of galaxies in close pairs reside within cluster or group-size halos and therefore represent a biased population, poorly suited for direct comparison to ``field'' galaxies. Thus, the frequent observation that some types of galaxies in pairs have redder colors than ``field'' galaxies is primarily a selection effect. We select galaxy pairs that are isolated in their dark matter halos with respect to other massive subhalos (N=2 halos) and a control sample of isolated galaxies (N=1 halos) for comparison. We then apply these selection criteria to a volume-limited subset of the 2dF Galaxy Redshift Survey with M_Bj ~ 5 above their average past value, while only 10% of isolated galaxies in the control sample show this level of enhancement. Thus, 14% (20 %) of the galaxi...

  17. Isolation by distance, web service

    Bohonak Andrew J; Jensen Jeffrey L; Kelley Scott T


    Abstract Background The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user-friendly. Results We have created Isolation by Distance Web Service (IBDWS) a user-friendly web interface for determining patterns of isolation by distance. Using this site, ...

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

    Gita Eslami


    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

  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

    Jinhu eHuang


    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.


    Svetlana Shumarova


    Full Text Available Background: Isolated adrenal gland metastases are not frequently finding. The aim of the present retrospective study was to estimate clinical and pathological parameters that could be used to predict survival after adrenalectomy. Material and methods: A total of 34 patients with adrenal gland masses suspected to be metastases was included in this study. The group of patients with isolated metastases was19 (56% and a group of patients with adrenal adenomas -15 (44%. Results: The sample of patients consisted of 18 (53% men and 16 (47% women from 40 to 81 years old with a mean (±SD age of 61.6±10.3 years, presented with adrenal mass suspected to be metastases. Nineteen (56% of them had demonstrated metastases and 16 (84% metachronous with median overall survival (OS 54.6 (range 43-66 months. Median OS in the group with metastases was 22.6 months. Lung carcinoma was the most common primary tumour metastasizing in the adrenal gland – 58% of all metastases with a disease-free interval (DFI of 13 months. It was presented by shorter median survival than the rest primary tumour types (37.8 vs. 96.7months; log-rank test, p=0.028. In the multivariate Cox’s hazard analysis of the surgical technique was found to be an independent prognostic factor (p=0.047, together with lung carcinoma vs. renal cell carcinoma (p=0.045. Conclusion: Adrenalectomy due to isolated metastases in the adrenal glands showed the median overall survival of 22.6 months. Shorter survival periods were associated with lung carcinoma, DFI<12months, conversion to open surgery, synchronous metastases, but not with age, tumour size or resection status.


    Gowri Sankar


    Full Text Available ABSTRACT: Xanthomas are often a manifestation of underlying lipid abnormalities.A 17 - year - old female presented to our hospital with pain and swelling over t he left knee joint.Routine investigations and lipid profile were normal. Dermatological examination and Psychiatric examination were done and found to be normal. Histopathology from the lesion revealed macrophages and Touton type of giant cells. We present a case of normolipemic isolated xanthoma, which not only is an an uncommon occurrence but also needs a regular follow up because a high association with reticuloendothelial malignancies has also been reported.

  2. Journey from isolation

    Wim A. Dreyer


    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. Diabetes in population isolates

    Grarup, Niels; Moltke, Ida; Albrechtsen, Anders


    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...

  4. Isolation of RNP granules

    Jønson, Lars; Nielsen, Finn Cilius; Christiansen, Jan


    be regarded as a supramolecular assembly of RNA and protein, probably representing several overlapping post-transcriptional operons. The present protocol describes how RNP granules may be isolated by the transgenic expression of a 3X FLAG version of an RNA-binding protein under tetracycline control via...... the tetracycline receptor/operator complex. In this way, inclusion of an appropriate tetracycline concentration ensures expression of the tagged version at the endogenous level, and the 3X FLAG tag is a convenient "handle" for the subsequent immunoprecipitation by immobilized anti-FLAG antibody....

  5. [Isolated Ledderhose fibromatosis plantaris].

    Runkel, N; Göhring, U; Friedl, W; Roeren, T


    Plantar fibromatosis or Ledderhose syndrome has rarely been discussed in the medical literature. This clinical entity includes nodular Dupuytren-like indurations of the plantar aponeurosis. We present a case of a young man with isolated disease of his right foot and describe the clinical and pathomorphological features of this disease as well as its characteristic findings at magnetic resonance tomography. The therapy of choice is a subtotal resection of the plantar aponeurosis to prevent irreversible contractions of the toes. The prognosis following fascietomy is good.

  6. Isolation of Mouse Neutrophils.

    Swamydas, Muthulekha; Luo, Yi; Dorf, Martin E; Lionakis, Michail S


    Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation-based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in UNIT. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments.

  7. Schizosaccharomyces isolation method

    Benito Santiago


    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. 无乳链球菌sodA基因的克隆、序列分析及原核表达载体构建%Cloning and Bioinformatics Analysis of sodA Gene from Streptococcus agalactiae and Constructing pET28a-sodA Plasmid

    王松艳; 蔡双虎; 鲁义善; 简纪常; 吴灶和


    无乳链球菌(Streptococcus agalactiae)是引起我国南方罗非鱼链球菌病的主要病原之一,SodA是无乳链球菌重要的毒力因子之一.根据链球菌sodA基因保守区设计引物,采用PCR扩增sodA基因部分序列,反向PCR和巢式PCR扩增其侧翼序列,成功获得无乳链球菌sodA基因.无乳链球菌sodA基因序列全长为699 bp,含开放阅读框609 bp,可编码202个氨基酸,演绎的SodA蛋白主要由α螺旋和不规则卷曲构成.BLAST分析表明,该蛋白氨基酸序列与犬链球菌(S.canis)及副乳房链球菌(S.parauberis)相应蛋白的氨基酸序列相似性分别高达80%和79%.将sodA基因克隆至原核表达载体pET28a上成功构建了重组质粒pET28a-sodA,导入Escherichia coli BL21 (DE3),诱导表达的重组蛋白质分子质量约为24 ku,主要以包涵体形式存在于表达菌中.

  9. 多重PCR检测原料乳中沙门氏菌、无乳链球菌和金黄色葡萄球菌的研究%Study on the Detection of Salmonella, Streptococcus Agalactiae and Staphylococcus Aureus in Raw Milk by Multiplex PCR

    刘继超; 姜铁民; 陈历俊; 赵长新


    目的:建立快速检测沙门氏菌、无乳链球菌和金黄色葡萄球菌的多重PCR方法.方法:根据沙门氏菌(Salmonella)组氨酸转运操纵子基因、无乳链球菌(streptococcus agalactiae)的STRA-Agl-23-1D基因和金黄色葡萄球菌(Staphylococcus aureus)的耐热核酸酶肌nuc基因分别设计引物,进行PCR扩增及反应条件的优化,建立这3种菌的多重PCR检测方法.结果:通过建立多重PCR方法,3对引物同时特异性地扩增出495 bp、360 bp和279 bp目的片段;3种菌的检测限:沙门氏菌1.9×102cfu/ml、无乳链球菌2.0×102cfu/mL、金黄色葡萄球菌2.9×102 cfu/mL;3种菌DNA含量的(最低)检出限分别为3.86、25.5、3.47pg.结论:本文建立的多重PCR检测方法,简单、快速、灵敏度高,具有很好的应用前景.

  10. Isolated hepatosplenic tuberculosis

    Vinay Kumar; Durgatosh Pandey


    BACKGROUND:Tuberculosis rarely involves the liver and spleen and when it does so, it is usually associated with disseminated disease. METHOD:We report a patient with isolated tuberculosis of the liver and spleen. RESULTS:A 50-year-old man presented with weakness and loss of weight for two months. CT scan of the abdomen showed multiple small hypodense lesions in the liver and spleen. Image-guided biopsy of liver lesion was inconclusive as it showed normal hepatocytes with interspersed lymphocytes. On laparotomy, the liver and spleen were studded with multiple small nodules. A 5-cm wedge of the liver was resected. Histopathological examination conifrmed the diagnosis of tubercular hepatitis. A detailed work-up failed to identify any other focus of tuberculosis. CONCLUSION:Although rare, tuberculosis should also be considered in the differential diagnosis for multiple nodules in the liver and spleen, along with lymphoproliferative diseases and metastatic lesions.

  11. Clinically isolated laryngeal sarcoidosis

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


    Laryngeal sarcoidosis is rare (0.5% of patients with sarcoidosis), the pathogenesis is unknown and the optimal treatment remains a matter of debate. We undertook this study to elucidate possible pathogenic factors in clinically isolated laryngeal sarcoidosis and to describe results...... of supraglottoplastic surgery. From 1995 to 2009, we identified six patients with histologically proven sarcoidosis of the larynx treated at Rigshospitalet. All patients were subjected to a panel of blood tests and MR scan of the head and neck. All patients had dyspnoea at admission, and five were subjected...... successful to maintain normal breathing. None of the many parameters examined--some previously having been found to be abnormal in sarcoidosis--were abnormal in the present cohort. We are therefore unable to elucidate the pathogenesis. The combined surgical approach re-established normal airway function...

  12. Isolated sulfite oxidase deficiency.

    Relinque, B; Bardallo, L; Granero, M; Jiménez, P J; Luna, S


    Sulfite oxidase deficiency is an uncommon metabolic disease. Only few cases of its isolated form have been reported in the literature. We report a case of severe neonatal onset. A newborn baby of 41 weeks gestational age, weighted at birth of 3240 grams and had an Apgar score of 6-10-10. Fifty-three hours after being born, the baby started with seizures that were refractory to antiepileptic treatment. Brain function was monitored using a-EEG. Laboratory and imaging tests were performed. All of them were consistent with sulfite oxidase deficiency. The diagnosis was confirmed by genetic testing. We highlight the importance of this disease as part of the differential diagnosis of seizures during the neonatal period, as well as the importance of the therapeutic support based on dietary restrictions. It's also remarkable the possibility of prenatal diagnosis by quantifying enzyme activity and it's also possible carrying out DNA mutational analysis.

  13. Patients experience of source isolation

    Johansen, Kamilla; Pedersen, Didde; Kragbak, Nina


    Patients´ experience of source isolation - a literature review Title: Patients experience of source isolation – a literature review Authors: Nina Kragbak*, Didde Pedersen*, Kamilla Johansen* and Peter E. Jensen** *students, **lecturer, bachelor in science and Ph.D., at VIA University College......, 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...

  14. Protein loss during nuclear isolation


    Cryomicrodissection makes possible the measurement of the entire in vivo protein content of the amphibian oocyte nucleus and provides a heretofore missing baseline for estimating protein loss during nuclear isolation by other methods. When oocyte nuclei are isolated into an aqueous medium, they lose 95% of their protein with a half-time of 250 s. This result implies an even more rapid loss of protein from aqueously isolated nuclei of ordinary-size cells.

  15. A hybrid base isolation system

    Hart, G.C. [Univ. of California, Los Angeles, CA (United States); Lobo, R.F.; Srinivasan, M. [Hart Consultant Group, Santa Monica, CA (United States); Asher, J.W. [kpff Engineers, Santa Monica, CA (United States)


    This paper proposes a new analysis procedure for hybrid base isolation buildings when considering the displacement response of a base isolated building to wind loads. The system is considered hybrid because of the presence of viscous dampers in the building above the isolator level. The proposed analysis approach incorporates a detailed site specific wind study combined with a dynamic nonlinear analysis of the building response.

  16. Isolation and the parish ministry

    Irvine, Andrew R.


    The purpose of this thesis was to examine the concept of isolation as it occurs within the profession of ministry. Isolation, for the purpose of this thesis, is defined social-psychologically. Within the field research isolation is considered as evidenced professionally, socially and spiritually. This study utilized as its sample base 200 hundred Church of Scotland ministers (15% of total population) which provided 159 usable responses to an extensive mail survey. The mai...

  17. Isolated Pelvic Hyperthermochemotherapeutic Perfusion -An Experimental Study on Isolating Efficacy


    Hyperthermochemotherapeutic perfusion model through isolated pelvic vessels was developed to evaluate the leakage of hyperthermia and drugs (such as adriamycin) from the isolated pelvic circulation to systemic circulation and its associated side/toxic effects. The isolated pelvic circulation was perfused through a femoral artery catheter with hyperthermic (48 ℃ to 55 ℃) adriamycin solution (50 μg/ml) for 30 min. The efflux was drained through a femoral vein catheter. And the pelvic temperature was kept at the level of 43±0.5 ℃. The temperature of pelvic circulation was kept at 4 ℃ to 5 ℃ greater than the systemic/core temperature. The adriamycin concentration of pelvic efflux was 12 to 46 folds of that of systemic serum. The difference between them was very significant (P<0.001). As the perfusion pressure was increased, which kept lower than the mean systemic artery pressure, the leakage of the adriamycin from the isolated pelvic circulation to systemic circulation was increased, but there was no significant difference between them (P>0.05). During isolated perfusion, the systemic blood dynamics remained stable and there were no organic injuries on the important organs. It was suggested that the isolating efficacy of the modality of isolated pelvic hyperthermochemotherapeutic perfusion through vessels was rather high. The hyperthermia and drugs could be effectively limited in the isolated pelvic region with minor side effects on the systemic circulation and important organs.

  18. Geometry of isolated horizons

    Flandera, Aleš


    While the formalism of isolated horizons is known for some time, only quite recently the near horizon solution of Einstein's equations has been found in the Bondi-like coordinates by Krishnan in 2012. In this framework, the space-time is regarded as the characteristic initial value problem with the initial data given on the horizon and another null hypersurface. It is not clear, however, what initial data reproduce the simplest physically relevant black hole solution, namely that of Kerr-Newman which describes stationary, axisymmetric black hole with charge. Moreover, Krishnan's construction employs the non-twisting null geodesic congruence and the tetrad which is parallelly propagated along this congruence. While the existence of such tetrad can be easily established in general, its explicit form can be very difficult to find and, in fact it has not been provided for the Kerr-Newman metric. The goal of this thesis was to fill this gap and provide a full description of the Kerr-Newman metric in the framework ...

  19. Isolated subtalar arthrodesis.

    Yildirim, Timur; Sofu, Hakan; Çamurcu, Yalkin; Özcan, Çağri; Öner, Ali; Şahin, Vedat


    The aim of this study is to review the results of isolated subtalar arthrodesis in adults and to make a comparative analysis of the clinical outcomes between the patients with posttraumatic subtalar arthritis and the ones with other etiologic factors, and to evaluate the effects of grafting. This study included 19 men and 12 women. The mean postoperative follow-up was 36.8 months. The mean AOFAS hindfoot score improved from a mean of 46 preoperatively to a mean of 77.3 postoperatively. Thirty-one of 33 arthrodeses achieved bony union at a mean time of 15.7 weeks. The mean increase in the talocalcaneal height was 3.8 mm in the feet operated without grafting, whereas it was 8.1 mm in the feet for which grafting was performed. Improvement in talocalcaneal angle was significantly better in the feet operated with bone grafting. The feet with posttraumatic subtalar arthritis were more prone to Reflex Sympathetic Dystrophy.

  20. Seismic isolation for Advanced LIGO

    Abbott, R; Allen, G; Cowley, S; Daw, E; Debra, D; Giaime, J; Hammond, G; Hammond, M; Hardham, C; How, J; Hua, W; Johnson, W; Lantz, B; Mason, K; Mittleman, R; Nichol, J; Richman, S; Rollins, J; Shoemaker, D; Stapfer, G; Stebbins, R


    The baseline design concept for a seismic isolation component of the proposed 'Advanced LIGO' detector upgrade has been developed with proof-of-principle experiments and computer models. It consists of a two-stage in-vacuum active isolation platform that is supported by an external hydraulic actuation stage. Construction is underway for prototype testing of a full-scale preliminary design.

  1. Improved Active Vibration Isolation Systems


    The control force, feedback gain, and actuator stroke of several active vibration isolation systems were analyzed based on a single-layer active vibration isolation system. The analysis shows that the feedback gain and actuator stroke cannot be selected independently and the active isolation system design must make a compromise between the feedback gain and actuator stroke. The performance of active isolation systems can be improved by the joint vibration reduction using an active vibration isolation system with an adaptive dynamic vibration absorber. The results show that the joint vibration reduction method can successfully avoid the compromise between the feedback gain and actuator stroke. The control force and the object vibration amplitude are also greatly reduced.

  2. Dynamic isolation technologies in negative pressure isolation wards

    Xu, Zhonglin


    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.

  3. Isolated Proofs of Knowledge and Isolated Zero Knowledge

    Damgård, Ivan Bjerre; Nielsen, Jesper Buus; Wichs, Daniel


    . 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...

  4. Helicopter gearbox isolation using periodically layered fluidic isolators

    Szefi, Joseph Thomas


    In rotorcraft transmissions, vibration generation by meshing gear pairs is a significant source of vibration and cabin noise. This high-frequency gearbox noise is primarily transmitted to the fuselage through rigid connections, which do not appreciably attenuate vibratory energy. The high-frequency vibrations typically include discrete gear-meshing frequencies in the range of 500--2000 Hz, and are often considered irritating and can reduce pilot effectiveness and passenger comfort. Periodically-layered isolators were identified as potential passive attenuators of these high frequency vibrations. Layered isolators exhibit transmissibility "stop bands," or frequency ranges in which there is very low transmissibility. An axisymmetric model was developed to accurately predict the location of these stop bands for isolators in compression. A Ritz approximation method was used to model the axisymmetric elastic behavior of layered cylindrical isolators. This model of layered isolators was validated with experiments. The physical design constraints of the proposed helicopter gearbox isolators were then estimated. Namely, constraints associated with isolator mass, axial stiffness, geometry, and elastomeric fatigue were determined. The passive performance limits of layered isolators were then determined using a design optimization methodology employing a simulated annealing algorithm. The results suggest that layered isolators cannot always meet frequency targets given a certain set of design constraints. Many passive and active design enhancements were considered to address this problem, and the use of embedded inertial amplifiers was found to exhibit a combination of advantageous effects. The first benefit was a lowering of the beginning stop band frequency, and thus a widening of the original stop band. The second was a tuned absorber effect, where the elastomer layer stiffness and the amplified tuned mass combined to act as a vibration absorber within the stop band. The

  5. Isolation of rat adrenocortical mitochondria

    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: [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)


    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.

  6. Isolated Echinococcosis of cervical region

    Khare, Pratima; Kala, Pooja; Gupta, Renu; Chauhan, Nidhi


    .... 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...

  7. Atomically thin nonreciprocal optical isolation

    Lin, Xiao; Wang, Zuojia; Gao, Fei; Zhang, Baile; Chen, Hongsheng


    Optical isolators will play a critical role in next-generation photonic circuits, but their on-chip integration requires miniaturization with suitable nonreciprocal photonic materials. Here, we theoretically demonstrate the thinnest possible and polarization-selective nonreciprocal isolation for circularly polarized waves by using graphene monolayer under an external magnetic field. The underlying mechanism is that graphene electron velocity can be largely different for the incident wave propagating in opposite directions at cyclotron frequency, making graphene highly conductive and reflective in one propagation direction while transparent in the opposite propagation direction under an external magnetic field. When some practical loss is introduced, nonreciprocal isolation with graphene monolayer still possesses good performance in a broad bandwidth. Our work shows the first study on the extreme limit of thickness for optical isolation and provides theoretical guidance in future practical applications. PMID:24569672

  8. Waste Isolation Pilot Plant Overview

    Weaver, Douglas James [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)


    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.

  9. Pneumothorax complicating isolated clavicle fracture.

    Hani, Redouane; Ennaciri, Badr; Jeddi, Idriss; El Bardouni, Ahmed; Mahfoud, Mustapha; Berrada, Mohamed Saleh


    Isolated clavicle fractures are among the commonest of traumatic fractures in the emergency department. Complications of isolated clavicle fractures are rare. Pneumothorax has been described as a complication of a fractured clavicle only rarely in English literature. In all the reported cases, the pneumothorax was treated by a thoracostomy and the clavicle fracture was treated conservatively. In our case, the pneumothorax required a chest drain insertion and the clavicle fracture was treated surgically with good result.

  10. Base isolation of fluid containers

    Kim, S. [Cygna Group Inc./ICF Kaiser International, Oakland, CA (United States)


    Fluid containers often constitute critical internal equipment in power plants. However, due to possible structure-equipment interaction effect they are particularly vulnerable during strong earthquake events. An effective technique for protecting fluid containers is base isolation. By deflecting the possible seismic input energy into the superstructure, base isolation can substantially reduce seismic demand on the containers, making it more cost effective than equivalent conventional design.


    HABASHI, Akbar Sheidaei


    The effect of weak shear rigidity is important in the analysis of elastomeric bearings used for base isolation of buildings. In particular, the buckling load is low and the P-Δ effect is significant as compared to standard columns neglecting shear deformations. In this paper, a simple mechanical model accounting for both shear and flexural deformations is proposed to treat the P-Δ effect of elastomeric isolation bearings. With adequate choice of the model parameters, the simplified model agre...

  12. Neuroimmunological Responses to Social Isolation

    Ramirez DDS, MSc, PhD, Karol


    Objective social isolation and perceived social isolation are psychosocial stressors that may impair the normal functioning of the neuroimmune system. Chronic activation of the neuro-immuno-endocrine communication and the consequent loss of homeostasis may lead to the appearance of pathologies and associated mood disorders. For example, alterations in the hypothalamic-pituitary- adrenal axis and sympathetic nervous system dynamics may account for the observed predisposition to inflammatory di...

  13. Market study: Biological isolation garment


    The biological isolation garment was originally designed for Apollo astronauts to wear upon their return to earth from the moon to avoid the possibility of their contaminating the environment. The concept has been adapted for medical use to protect certain patients from environmental contamination and the risk of infection. The nature and size of the anticipated market are examined with certain findings and conclusions relative to clinical acceptability and potential commercial viability of the biological isolation garment.

  14. Isolation, biochemical and molecular characterization of 2 ...

    Isolation, biochemical and molecular characterization of ... Abstract. Pure cultures of 2-chlorophenol degrading bacteria were isolated from a natural enrichment that may be adapted to ... three isolates out of six yielded positive PCR products.

  15. Progress in Exosome Isolation Techniques

    Li, Pin; Kaslan, Melisa; Lee, Sze Han; Yao, Justin; Gao, Zhiqiang


    Exosomes are one type of membrane vesicles secreted into extracellular space by most types of cells. In addition to performing many biological functions particularly in cell-cell communication, cumulative evidence has suggested that several biological entities in exosomes like proteins and microRNAs are closely associated with the pathogenesis of most human malignancies and they may serve as invaluable biomarkers for disease diagnosis, prognosis, and therapy. This provides a commanding impetus and growing demands for simple, efficient, and affordable techniques to isolate exosomes. Capitalizing on the physicochemical and biochemical properties of exosomes, a number of techniques have been developed for the isolation of exosomes. This article summarizes the advances in exosome isolation techniques with an emphasis on their isolation mechanism, performance, challenges, and prospects. We hope that this article will provide an overview of exosome isolation techniques, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations of exosomes from a wide range of biological matrices. PMID:28255367

  16. Geomorphology of Minnesota - Isolated Landform Structures

    Minnesota Department of Natural Resources — Geomorphology of Minnesota - Isolated Landform Structures are essentially cartographic arcs representing isolated glacial features that were mapped in conjunction...

  17. Base isolation system and verificational experiment of base isolated building

    Takeuchi, Mikio; Harada, Osamu; Aoyagi, Sakae; Matsuda, Taiji


    With the objective of rationalization of the earthquake resistant design and the economical design based thereupon, many base isolation systems have been proposed and its research, development and application have been made in recent years. In order to disseminate the system, it is necessary to accumulate the data obtained from vibration tests and earthquake observations and verify the reliability of the system. From this viewpoint, the Central Research Institute of Electric power Industry and Okumura Corporation did the following experiments with a base isolated building as the object: 1) static power application experiments, 2) shaking experiments, 3) free vibration experiments, 4) regular slight vibration observations and 5) earthquake response observations (continuing). This article reports the outline of the base isolation system and the base isolated building concerned as well as the results of the verification experiments 1) through 3) above. From the results of these verification experiments, the basic vibration characteristics of the base isolation system consisting of laminated rubber and plastic damper were revealed and its functions were able to be verified. Especially during the free vibration experiments, the initial displacement even up to a maximum of 10cm was applied to the portion between the foundation and the structure and this displacement corresponds to the responded amplitude in case of the earthquake of seismic intensity of the 6th degree. It is planned to continue the verification further. (18 figs, 3 tabs, 3 photos, 6 refs)

  18. 响应面法优化丹参中抗无乳链球菌活性成分的提取工艺%Optimization of the Extraction Conditions of Active Components for Inhibition of Streptococcus agalactiae from Salvia miltiorrhiza Bge. Using Response Surface Methodology

    邓恒为; 郭伟良; 孙晓飞; 谢珍玉; 王世锋; 胡文婷; 周永灿


    In recent yearsStreptococcus agalactiae(SA) has become the most harmful pathogen to cultured tilapia in Hainan and many southern provinces of China.Salvia miltiorrhizaBge is highly resistant to SA, but it contains complex components and which component is responsible for the resistance to SA has not been reported. Here we investigated the effects of extraction conditions on the yield of anti-SA components inS. miltiorrhizaBge and optimized the extraction method. We first developed the agar diffusion method for the quantitative analysis of extracted anti-SA components; then we used the yields of anti-SA components fromSalvia miltiorrhizaBge as the responsive values to study the effects of extraction conditions on the yield of the anti-SA components, including the extraction solvent, extraction times, the ratio of liquid to solid, infiltration time, temperature and extraction time in single-factor experiments; finally we further optimized the ratio of liquid to solid, infiltration time, temperature and extraction time using the four-factor-three-level Box-Behnken experimental design with response surface method. The results of the agar diffusion showed that the standard equation for the determination of anti-SA components fromS. miltiorrhizaBge wasY= 4.842X+ 16.707 (R2= 0.9873). The results of the single-factor experiments showed that the most suitable extraction solvent and times was 20% ethanol aqueous solution for 1 time. The results of the Box-Behnken experimental design with response surface method showed that the infiltration time, the extraction temperature, and the ratio of liquid to solid had extremely vital effects on the responsive values. The optimum extraction conditions were as follows: the ratio of liquid to solid was 37.8:1.0 (ml:g), the infiltration time was 104 min, the extraction temperature was 86.7℃ and the extraction time was 182 min. The predicted maximum yield of anti-SA components was 25.95%, under the RSM model-generated optimal extraction

  19. Entropy of Isolated Horizons revisited

    Basu, Rudranil; Majumdar, Parthasarathi


    The decade-old formulation of the isolated horizon classically and within loop quantum gravity, and the extraction of the microcanonical entropy of such a horizon from this formulation, is reviewed, in view of recent renewed interest. There are two main approaches to this problem: one employs an SU(2) Chern-Simons theory describing the isolated horizon degrees of freedom, while the other uses a reduced U(1) Chern-Simons theory obtained from the SU(2) theory, with appropriate constraints imposed on the spectrum of boundary states `living' on the horizon. It is shown that both these ways lead to the same infinite series asymptotic in horizon area for the microcanonical entropy of an isolated horizon. The leading area term is followed by an unambiguous correction term logarithmic in area with a coefficient $-\\frac32$, with subleading corrections dropping off as inverse powers of the area.


    Smirnov Vladimir Alexandrovich


    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.

  1. Isolated Echinococcosis of cervical region

    Pratima Khare


    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.

  2. Solo doctors and ethical isolation.

    Cooper, R J


    This paper uses the case of solo doctors to explore whether working in relative isolation from one's peers may be detrimental to ethical decision-making. Drawing upon the relevance of communication and interaction for ethical decision-making in the ethical theories of Habermas, Mead and Gadamer, it is argued that doctors benefit from ethical discussion with their peers and that solo practice may make this more difficult. The paper identifies a paucity of empirical research related to solo practice and ethics but draws upon more general medical ethics research and a study that identified ethical isolation among community pharmacists to support the theoretical claims made. The paper concludes by using the literary analogy of Soderberg's Doctor Glas to illustrate the issues raised and how ethical decision-making in relative isolation may be problematical.

  3. Isolation and use of rafts.

    Brown, Deborah A


    This unit describes methods for isolating and analyzing rafts by detergent insolubility. To distinguish these rafts from raft-like membranes isolated by other methods, they are referred to here as detergent-resistant membranes (DRMs). DRMs can be isolated by flotation on sucrose density gradients or by pelleting after detergent extraction. DRM proteins can be analyzed by SDS-PAGE and immunoblotting. Additionally, radiolabeled DRM proteins can be analyzed, and lipids can be quantitated by high-performance thin layer chromatography. Support protocols needed for the lipid analysis are also provided. Finally, protocols for raft disruption by cholesterol removal and measuring the kinetics of such removal are included together with a method that reverses the cholesterol removal (cholesterol repletion).

  4. Bacteria isolated from amoebae/bacteria consortium

    Tyndall, Richard L.


    New protozoan derived microbial consortia and method for their isolation are provided. Consortia and bacteria isolated therefrom are useful for treating wastes such as trichloroethylene and trinitrotoluene. Consortia, bacteria isolated therefrom, and dispersants isolated therefrom are useful for dispersing hydrocarbons such as oil, creosote, wax, and grease.

  5. Active Fault Isolation in MIMO Systems

    Niemann, Hans Henrik; Poulsen, Niels Kjølstad


    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 iso...

  6. Completely Isolated? Health Information Seeking among Social Isolates

    Askelson, Natoshia M.; Campo, Shelly; Carter, Knute D.


    To better target messages it is important to determine where people seek their health information. Interpersonal networks are a common way most people gather health information, but some people have limited networks. Using data from the 2004 General Social Survey (N = 984), we compared social isolates and nonisolates in their health…

  7. Completely Isolated? Health Information Seeking among Social Isolates

    Askelson, Natoshia M.; Campo, Shelly; Carter, Knute D.


    To better target messages it is important to determine where people seek their health information. Interpersonal networks are a common way most people gather health information, but some people have limited networks. Using data from the 2004 General Social Survey (N = 984), we compared social isolates and nonisolates in their health…

  8. Is an isolated cleft lip an isolated anomaly?

    Deelder, J. D.; Breugem, C. C.; de Vries, Iris; de Bruin, M.; van der Molen, A. B. Mink; van der Horst, C. M. A. M.

    Introduction: It is well known that patients with cleft lip/palate or cleft palate can have associated anomalies. However, there is a relative paucity of information about the possible anomalies associated with an isolated cleft lip. A recent study (Vallino et al., 2008) showed that children with

  9. Is an isolated cleft lip an isolated anomaly?

    Deelder, J. D.; Breugem, C. C.; de Vries, Iris; de Bruin, M.; van der Molen, A. B. Mink; van der Horst, C. M. A. M.


    Introduction: It is well known that patients with cleft lip/palate or cleft palate can have associated anomalies. However, there is a relative paucity of information about the possible anomalies associated with an isolated cleft lip. A recent study (Vallino et al., 2008) showed that children with cl

  10. Optical Isolator Utilizing Surface Plasmons

    Shinji Yuasa


    Full Text Available Feasibility of usage of surface plasmons in a new design of an integrated optical isolator has been studied. In the case of surface plasmons propagating at a boundary between a transition metal and a double-layer dielectric, there is a significant difference of optical loss for surface plasmons propagating in opposite directions. Utilizing this structure, it is feasible to fabricate a competitive plasmonic isolator, which benefits from a broad wavelength operational bandwidth and a good technological compatibility for integration into the Photonic Integrated Circuits (PIC. The linear dispersion relation was derived for plasmons propagating in a multilayer magneto-optical slab.

  11. Isolation and Cultivation of Anaerobes

    Aragao Börner, Rosa


    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...... microorganisms is still a promising venture, and conventional methodologies as well as considerations and modifications are presented here. An insight into new methodologies and devices as well as a discussion on future perspectives for the cultivation of anaerobes may open the prospects of the exploitation...

  12. Isolation contactor state control system

    Bissontz, Jay E.


    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.

  13. Spontaneous isolated celiac artery dissection

    Tuba Cimilli Ozturk


    Full Text Available Dyspepsia with mild, stabbing epigastric discomfort without history of trauma is a very common symptom that emergency physicians see in their daily practice. Vascular emergencies, mostly the aortic dissection and aneurysm, are always described in the differential diagnosis with persistent symptoms. Isolated celiac artery dissection occurring spontaneously is a very rare diagnosis. The involvement of branch vessels is generally observed and patients show various clinical signs and symptoms according to the involved branch vessel. Here we are presenting a case with spontaneous isolated celiac artery dissection, without any branch vessel involvement or visceral damage, detected by computed tomography scans taken on admission.

  14. Isolated complete corpus callosal agenesis

    Jaiganesh S, Venkateshwaran A, Naresh Kumar C, Rajasekhar KV


    Full Text Available Isolated complete corpus callosal agenesis is a rare entity. Usually this condition will be an associated finding in other syndromes. 3 month old male child came with complaints of deformed foot on both sides, not having a social smile and neck holding. Patient referred to the Radiology department for MRI brain which showed complete absence of corpus callosum, widely separated and parallely placed lateral ventricles, colpocephaly, high riding of 3rd ventricle and absence of cingulate gyrus and radial arrangement of gyri along the interhemispheric fissure. Hence it was reported as isolated complete corpus callosal agenesis and this article describes the Embryogenesis, anatomy, developmental anomalies and its clinical manifestations & prognosis.

  15. [Isolated giant hydatid in kidney].

    Ozgör, Faruk; Erbin, Akif; Berberoğlu, Ahmet Yalçın; Binbay, Murat; Sarılar, Omer; Müslümanoğlu, Ahmet Yaser


    Cyst hydatid of the kidney is parasitic condition caused by Echinococcus granulosus and identified in many countries, especially associated with sheep farming. Echinococcal larvae enter the bloodstream using the digestive system and invade any organs in the human body. The urinary system is the third most common area affected by parasitic infection after liver and lungs, but isolated renal involvement is a very rare situation, even in endemic areas. İn our case, we aimed to report a 57-year-old female patient with an 18-centimeter isolated renal cyst hydatid treated by retroperitoneal nephrectomy. The diagnosis was based on imaging findings and confirmed by histopathologically.

  16. Isolation of Leptospira from blood culture bottles.

    Girault, Dominique; Soupé-Gilbert, Marie-Estelle; Geroult, Sophie; Colot, Julien; Goarant, Cyrille


    With the increasing use of real-time PCR techniques, Leptospira isolation has mostly been abandoned for the diagnosis of human leptospirosis. However, there is a great value of collecting Leptospira isolates to better understand the epidemiology of this complex zoonosis and to provide the researchers with different isolates. In this study, we have successfully isolated different Leptospira strains from BacT/Alert aerobic blood culture bottles and suggest that this privileged biological material offers an opportunity to isolate leptospires.

  17. Effectiveness of Base Isolation Technique and Influence of Isolator Characteristics on Response of a Base Isolated Building

    Sunita Tolani


    Full Text Available This study concerns with the seismic response comparison of a fixed base building with a base isolated building and parametric study of a base isolated building. The structural system considered for analysis is a three storey reinforced concrete building, which is idealized as a shear type building with one lateral degree of freedom at each floor level. The isolation systems considered for this study are Laminated Rubber bearing (LRB, Lead Rubber Bearing (N-Z bearing and Friction Pendulum System (FPS. The response of fixed base building and of base isolated building is compared in terms of maximum top floor acceleration, interstorey drift, maximum floor displacements and base shear. For parametric study important isolation system parameters considered are: (i isolation time period, isolator damping for LRB; (ii isolator yield strength, isolation time period, isolator damping for N-Z bearing and (iii isolation time period, friction coefficient for FPS. It is found that base isolation technique is very effective in reducing seismic response of structure and isolation system parameters significantly influence the earthquake response of a base isolated structure.

  18. Providencia stuartii Isolates from Greece

    Oikonomou, Olga; Liakopoulos, Apostolos; Phee, Lynette M.; Betts, Jonathan; Mevius, Dik; Wareham, David W.


    Providencia stuartii has emerged as an important nosocomial pathogen. We describe an outbreak due to a multidrug-resistant strain over a 4-month period in a critical care unit in Athens. Molecular typing revealed each of the isolates to be clonally related with coresistance to cephalosporins, car


    The major historical use of zein, the predominant corn protein, has been in the fibers market. In order to evaluate new formulations of zein in the fibers market a simple means of isolating and generating fibers is necessary. We have evaluated the ability to electrospin zein from numerous solvents...

  20. Isolated superior mesenteric artery dissection

    Lalitha Palle


    Full Text Available Isolated superior mesenteric artery (SMA dissection without involvement of the aorta and the SMA origin is unusual. We present a case of an elderly gentleman who had chronic abdominal pain, worse after meals. CT angiography, performed on a 64-slice CT scanner, revealed SMA dissection with a thrombus. A large artery of Drummond was also seen. The patient was managed conservatively.

  1. Mitochondria: isolation, structure and function.

    Picard, Martin; Taivassalo, Tanja; Gouspillou, Gilles; Hepple, Russell T


    Mitochondria are complex organelles constantly undergoing processes of fusion and fission, processes that not only modulate their morphology, but also their function. Yet the assessment of mitochondrial function in skeletal muscle often involves mechanical isolation of the mitochondria, a process which disrupts their normally heterogeneous branching structure and yields relatively homogeneous spherical organelles. Alternatively, methods have been used where the sarcolemma is permeabilized and mitochondrial morphology is preserved, but both methods face the downside that they remove potential influences of the intracellular milieu on mitochondrial function. Importantly, recent evidence shows that the fragmented mitochondrial morphology resulting from routine mitochondrial isolation procedures used with skeletal muscle alters key indices of function in a manner qualitatively similar to mitochondria undergoing fission in vivo. Although these results warrant caution when interpreting data obtained with mitochondria isolated from skeletal muscle, they also suggest that isolated mitochondrial preparations might present a useful way of interrogating the stress resistance of mitochondria. More importantly, these new findings underscore the empirical value of studying mitochondrial function in minimally disruptive experimental preparations. In this review, we briefly discuss several considerations and hypotheses emerging from this work.

  2. Isolation by distance, web service

    Bohonak Andrew J


    Full Text Available Abstract Background The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user-friendly. Results We have created Isolation by Distance Web Service (IBDWS a user-friendly web interface for determining patterns of isolation by distance. Using this site, population geneticists can perform a variety of powerful statistical tests including Mantel tests, Reduced Major Axis (RMA regression analysis, as well as calculate FST between all pairs of populations and perform basic summary statistics (e.g., heterozygosity. All statistical results, including publication-quality scatter plots in Postscript format, are returned rapidly to the user and can be easily downloaded. Conclusion IBDWS population genetics analysis software is hosted at and documentation is available at The source code has been made available on Source Forge at

  3. Vapor-barrier Vacuum Isolation System

    Weinstein, Leonard M. (Inventor); Taminger, Karen M. (Inventor)


    A system includes a collimated beam source within a vacuum chamber, a condensable barrier gas, cooling material, a pump, and isolation chambers cooled by the cooling material to condense the barrier gas. Pressure levels of each isolation chamber are substantially greater than in the vacuum chamber. Coaxially-aligned orifices connect a working chamber, the isolation chambers, and the vacuum chamber. The pump evacuates uncondensed barrier gas. The barrier gas blocks entry of atmospheric vapor from the working chamber into the isolation chambers, and undergoes supersonic flow expansion upon entering each isolation chamber. A method includes connecting the isolation chambers to the vacuum chamber, directing vapor to a boundary with the working chamber, and supersonically expanding the vapor as it enters the isolation chambers via the orifices. The vapor condenses in each isolation chamber using the cooling material, and uncondensed vapor is pumped out of the isolation chambers via the pump.

  4. Construction and expression of prokaryotic expression vector for LrrG-Sip fusion gene of Streptococcus agalactiae in tilapia%罗非鱼无乳链球菌LrrG-Sip融合基因原核表达载体的构建及表达

    曾祖聪; 曹建萌; 卢迈新; 可小丽; 刘志刚; 高风英; 朱华平


    LrrG( leucine-rich repeat protein from GBS)and Sip( surface immunogenic protein),which are two kinds of surface anti-gen proteins from Streptococcus agalactiae in tilapia,have good immunogenicity. To obtain the LrrG-Sip fusion protein via coalescing surface antigen protein LrrG and Sip of S. agalctiae in tilapia,we cloned Sip and LrrG genes into vector pCold Ⅱ one by one using double enzyme method of gene splicing technology,and constructed a prokaryotic expression vector pCold Ⅱ-LrrG-Sip. The recombi-nant plasmid was transformed into E. coli BL21(DE3),and the result indicated that 9 h,15 ℃,0. 5 mmol·L-1IPTG were the opti-mum inducing conditions under which fusion protein was most soluble and abundant. Western blotting test showed that the LrrG-Sip fu-sion protein was about 160 kDa,consistent with the prediction(162 kDa),which suggested the prokaryotic expression vector pColdⅡ-LrrG-Sip was constructed successfully and laid the foundation for developing subunit vaccines for S. agalctiae in tilapia.%LrrG和表面免疫原性蛋白( Sip)是无乳链球菌( Streptococcus agalactiae)的2种表面蛋白,具有良好的免疫原性。为获得罗非鱼无乳链球菌表面蛋白LrrG和Sip蛋白的融合蛋白,该试验采用基因拼接技术中的双酶切法分2步逐个将Sip和LrrG基因插入pCold Ⅱ载体中,构建原核表达载体pCold Ⅱ-LrrG-Sip。将成功构建的融合基因原核表达载体转化感受态细胞BL21(DE3),进行诱导表达条件的优化。结果显示,15℃、IPTG 0.5 mmol·L-1诱导9 h,目的蛋白呈可溶状态的表达量最高。Western Blot检测结果显示LrrG-Sip融合蛋白大小与预测一致(162 kDa),说明成功构建了融合基因,为罗非鱼源无乳链球菌亚单位疫苗的研制奠定了基础。

  5. NASDA's activities on vibration isolation technology


    The National Space Development Agency's (NASDA) activities in providing various vibration isolation technologies for the Space Station Mission are covered in viewgraph form. Technologies covered include an active vibration isolation system for extra sensitive missions in the low frequency range, a passive damping system consisting of a damping rack for the reduction of resonance amplification, and an isolator for vibration isolation from low frequencies. Information is given in viewgraph form on the active vibration isolation concept, voice coil type electromagnetic suspension, a profile of an active vibration isolation system, a three degree of freedom ground experiment, and acceleration feedback.

  6. Sympathicotomy for isolated facial blushing

    Licht, Peter Bjørn; Pilegaard, Hans K; Ladegaard, Lars


    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...... in all social and mental domains in both groups. Overall, 85% of the patients had an excellent or satisfactory result, with no significant difference between the R2 procedure and the R2-R3 procedure. Mild recurrence of facial blushing occurred in 30% of patients within the first year. One patient...

  7. Isolated urachal malakoplakia mimicking malignancy

    Saisriharsha Pakalapati


    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.

  8. Pediatric isolated bilateral iliac aneurysm.

    Chithra, R; Sundar, R Ajai; Velladuraichi, B; Sritharan, N; Amalorpavanathan, J; Vidyasagaran, T


    Aneurysms are rare in children. Isolated iliac artery aneurysms are very rare, especially bilateral aneurysms. Pediatric aneurysms are usually secondary to connective tissue disorders, arteritis, or mycotic causes. We present a case of a 3-year-old child with bilateral idiopathic common iliac aneurysms that were successfully repaired with autogenous vein grafts. Copyright © 2013 Society for Vascular Surgery. Published by Mosby, Inc. All rights reserved.

  9. Isolated complete corpus callosal agenesis

    Jaiganesh S, Venkateshwaran A, Naresh Kumar C, Rajasekhar KV


    Isolated complete corpus callosal agenesis is a rare entity. Usually this condition will be an associated finding in other syndromes. 3 month old male child came with complaints of deformed foot on both sides, not having a social smile and neck holding. Patient referred to the Radiology department for MRI brain which showed complete absence of corpus callosum, widely separated and parallely placed lateral ventricles, colpocephaly, high riding of 3rd ventricle and absence of cingulate gyrus an...

  10. Transformerless dc-Isolated Converter

    Rippel, Wally E.


    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.

  11. Isolation of intraflagellar transport trains.

    Mencarelli, Caterina; Mitchell, Aaron; Leoncini, Roberto; Rosenbaum, Joel; Lupetti, Pietro


    The intraflagellar transport (IFT) system was first identified in situ by electron microscopy in thin sections of plastic-embedded flagella as linear arrays of electrondense particles, located between the B tubules of the outer doublets and the flagellar membrane. These arrays of particles are referred to as IFT trains. Upon membrane rupture, IFT trains are thought to easily dissociate to yield soluble IFT particles, which are commonly purified through sucrose gradients as ∼16-17S complexes. The latters easily dissociate into two subcomplexes, named A and B. We report here the isolation, visualization, and identification by immunolabeling of flexible strings of IFT particles, which are structurally similar to in situ IFT trains and appear to be formed by both complex A and complex B polypeptides. Moreover, the particles forming isolated IFT trains are structurally similar to the individual particles found in the ∼17S gradient peak. Our results provide the first direct evidence that ∼17S particles do indeed compose the IFT trains. The paper also represents the first isolation of the IFT trains, and opens new possibilities for higher resolution studies on their structure and how particles are attached to each other to form the particle trains.

  12. Immunogenic properties of Streptococcus agalactiae FbsA fragments.

    Salvatore Papasergi

    Full Text Available Several species of Gram-positive bacteria can avidly bind soluble and surface-associated fibrinogen (Fng, a property that is considered important in the pathogenesis of human infections. To gain insights into the mechanism by which group B Streptococcus (GBS, a frequent neonatal pathogen, interacts with Fng, we have screened two phage displayed genomic GBS libraries. All of the Fng-binding phage clones contained inserts encoding fragments of FbsA, a protein displaying multiple repeats. Since the functional role of this protein is only partially understood, representative fragments were recombinantly expressed and analyzed for Fng binding affinity and ability to induce immune protection against GBS infection. Maternal immunization with 6pGST, a fragment containing five repeats, significantly protected mouse pups against lethal GBS challenge and these protective effects could be recapitulated by administration of anti-6pGST serum from adult animals. Notably, a monoclonal antibody that was capable of neutralizing Fng binding by 6pGST, but not a non-neutralizing antibody, could significantly protect pups against lethal GBS challenge. These data suggest that FbsA-Fng interaction promotes GBS pathogenesis and that blocking such interaction is a viable strategy to prevent or treat GBS infections.

  13. Isolation of Cellulolytic Actinomycetes from Marine Sediments

    Veiga, Manuel; Esparis, Azucena; Fabregas, Jaime


    The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity.

  14. Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

    Pringle, M; Landén, A; Franklin, A


    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.

  15. Isolation, production, purification, assay and characterization of ...

    Isolation, production, purification, assay and characterization of fibrinolytic ... are isolated from Bacillus subtilis, β-haemolytic Streptococci and urine sample. ... recombinant E.coli containing short fragment genomic DNA of Pseudomonas sp.

  16. Mycoplasma contamination of Chlamydia pneumoniae isolates

    Huniche, BS; Jensen, Lise Torp; Birkelund, Svend


    We examined 6 C. pneumonia isolates from The American Type Culture Collection (ATCC) and 2 Finnish isolates for Mycoplasma contamination. Three of the ATCC isolates and both of the Finnish isolates were Mycoplasma-contaminated. The contaminants were characterized by means of growth in BEa and BEg...... 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...

  17. Isolation of haloorganic groundwater humic substances

    Krog, M.; Grøn, C.


    Humic substances were isolated from groundwater according to a revised method designed to avoid organohalogen artefacts. The prepared humic substances exhibited lower halogen contents than humic substances isolated according to the conventionally used method. Excessive oxidation or hydrolysis...

  18. Genetics Home Reference: isolated lissencephaly sequence

    ... Facebook Share on Twitter Your Guide to Understanding Genetic Conditions Search MENU Toggle navigation Home Page Search ... Conditions Genes Chromosomes & mtDNA Resources Help Me Understand Genetics Home Health Conditions isolated lissencephaly sequence isolated lissencephaly ...

  19. Isolation, characterization and antimicrobial activity of Streptomyces ...



    Dec 18, 2013 ... series on all media. The results showed that glycerol yeast extract and starch casein were the best .... MATERIALS AND METHODS. Collection of .... Genomic DNA was isolated from the isolates using a bacterial genomic DNA ...

  20. Are Isolated Indigenous Populations Headed toward Extinction?

    Walker, Robert S; Kesler, Dylan C; Hill, Kim R


    .... 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...

  1. Isolated Malignant Melanoma Metastasis to the Pancreas

    Anne K. Larsen, MD


    Full Text Available 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. common bacterial isolates from infected eyes abstract


    The common bacterial isolates and their antibiotics susceptibility were studied in 298 bacterial ... Bacteria were isolated most on the eye infections of the conjunctiva ... practitioners to avoid development of resistance from indiscriminate use.

  3. Isolated malignant melanoma metastasis to the pancreas

    Larsen, Anne K; Krag, Christen; Geertsen, Poul


    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.......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....

  4. Molecular characterization of Danish Cryptosporidium parvum isolates

    Enemark, Heidi L.; Ahrens, Peter; Juel, Cynthia Dawn


    was characterized as C. meleagridis. The porcine Cryptosporidium isolates (N = 4) revealed a pattern which was genetically distinct from human and bovine isolates. Cryptosporidium in a hedgehog (Erinaceus europaeus L.) was identified for the first time. By microsatellite sequencing the hedgehog isolate showed...

  5. Fungi isolated in school buildings

    Elżbieta Ejdys


    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.

  6. Thermodynamic laws in isolated systems.

    Hilbert, Stefan; Hänggi, Peter; Dunkel, Jörn


    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.

  7. [Neuroanatomy of Isolated Body Lateropulsion].

    Nakazato, Yoshihiko; Tamura, Naotoshi; Ikeda, Kei; Tanaka, Ai; Yamamoto, Toshimasa


    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.

  8. Isoflavonoids isolated from Cuban propolis.

    Piccinelli, Anna Lisa; Campo Fernandez, Mercedes; Cuesta-Rubio, Osmany; Márquez Hernández, Ingrid; De Simone, Francesco; Rastrelli, Luca


    Chemical investigation of a red-type Cuban propolis sample has led to the isolation of 11 isoflavonoids (2 isoflavones, 3 isoflavans, and 6 pterocarpans), together with gallic acid, isoliquiritigenin, and (-)-liquiritigenin. Structural determination, including the absolute stereochemistry, was accomplished by spectroscopic analysis, particularly CD and 2D NMR techniques. The fragmentation behavior of pterocarpans was studied by electrospray ionization (ESI) tandem mass spectrometry (MS/MS) using an ion-trap analyzer, and a generalized fragmentation pathway, useful in the identification and structural characterization of pterocarpans, is proposed. Isoflavonoids are reported for the first time from propolis samples.

  9. Simplified design procedure for base isolation system

    Takayama, Mineo; Tada, Hideyuki [Fukuoka Univ. (Japan). Faculty of Engineering


    This paper presents a simplified design procedure which incorporates an existing response prediction method for base isolated buildings into design methods for isolation devices (rubber bearings and hysteresis dampers). The procedure enables a designer to easily identify the relationship between tile seismic behavior of base-isolated buildings and the characteristics of isolation devices. The prediction method, proposed by Prof. Akiyama, is based on energy balance between the total input seismic energy and the energy absorbed by the isolation devices. The method is very accurate. The design methods for devices were developed by authors based on experimental and finite element analysis results.

  10. From rumors to genetic isolates

    Eduardo E. Castilla


    Full Text Available Here we propose a registration process for population genetic isolates, usually geographic clusters of genetic disorders, based on the systematic search of rumors, defined as any type of account regardless of its reliability. Systematically ascertained rumors are recorded, and validated through a progressive process of pre-established steps. This paper outlines the conceptual basis for this approach and presents the preliminary results from a rumor-based nationwide registry of genetically isolated populations, named CENISO (Censo Nacional de Isolados, operating in Brazil since 2009. During the first four years of its existence (2009-2013, a total of 191 Rumors were registered and validated, resulting in a prevalence rate of one per million inhabitants of Brazil. When the five statutory geographic regions of Brazil were considered, more Rumors were registered for the Northeast (2.11; 1.74-2.54 per 10(6 than for the remaining four regions, North, Center-West, Southeast, and South, which did not differ among themselves. About half (86/191 of the recorded rumors were proven to be geographic clusters; of these disorders, 58 were autosomal recessive, 17 autosomal dominant, 5 X-linked, 3 multifactorial, and one environmental (thalidomide embryopathy.

  11. Blackbody Radiation from Isolated Neptunes

    Ginzburg, Sivan; Loeb, Abraham


    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 which slows down their cooling. Therefore, a measurement of the blackbody temperature, $T_{\\rm eff}\\sim 50\\textrm{K}$, at which a Neptune emits can be used to estimate the mass of its atmosphere, $M_{\\rm atm}$. Explicitly, we find the relation $T_{\\r...

  12. Isolation of technogenic magnetic particles

    Catinon, Mickaël, E-mail: [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Ayrault, Sophie, E-mail: [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Boudouma, Omar, E-mail: [Service du MEB, UFR928, Université Pierre et Marie Curie, 75252 Paris VI (France); Bordier, Louise, E-mail: [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Agnello, Gregory, E-mail: [Evinrude, Espace St Germain, 38200 Vienne (France); Reynaud, Stéphane, E-mail: [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Tissut, Michel, E-mail: [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France)


    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.

  13. Braking Index of Isolated Pulsars

    Hamil, Oliver Q; Urbanec, Martin; Urbancova, Gabriela


    Isolated pulsars are rotating neutron stars with accurately measured angular velocities $\\Omega$, and their time derivatives that show unambiguously that the pulsars are slowing down. The commonly accepted view is that it arises through emission of magnetic dipole radiation (MDR) from a rotating magnetized body. The calculated energy loss by a rotating pulsar with a constant moment of inertia is assumed proportional to a model dependent power of $\\Omega$. This relation leads to the power law $\\dot{\\Omega}$ = -K $\\Omega^{\\rm n}$ where $n$ is called the braking index. The MDR model predicts $n$ exactly equal to 3. Selected observations of isolated pulsars provide rather precise values of $n$, individually accurate to a few percent or better, in the range 1$ <$ n $ < $ 2.8, which is consistently less than the predictions of the MDR model. In spite of an extensive investigation of various modifications of the MDR model, no satisfactory explanation of observation has been found yet. The aim of this work is t...

  14. 山东潍坊市部分奶牛场奶牛乳房炎病原菌的分离鉴定及耐药性检测%Isolation and Identiifcation of Pathogenic Bacteria in Cows with Mastitis in Weifang and the Drug Resistance Test

    薛梅; 朱俊平


    为了解潍坊市当前奶牛乳房炎的发病以及病原菌的种类和耐药性情况,为该地区奶牛乳房炎的防治提供参考,采集潍坊市6个奶牛场临床型和隐性型乳房炎奶样各100份,进行了细菌分离和鉴定。结果细菌检出率为91.50%(183/200),细菌混合感染奶样占检出细菌奶样的84.70%(155/183);共分离到265株细菌,其中链球菌(无乳链球菌、停乳链球菌、乳房链球菌)占38.11%,金黄色葡萄球菌占34.34%,大肠杆菌占16.98%,这些是引起奶牛乳房炎的主要病原菌。将这些病原菌进行药物敏感试验,结果显示,敏感率高的药物依次是头孢喹肟(80.59%)、恩诺沙星(68.35%)、头孢噻呋(68.35%)和氟苯尼考(67.51%),耐药率高的药物依次是青霉素(37.97%)、链霉素(26.16%)和四环素(10.55%)。%In order to understand the cow mastitis situation,kinds of pathogenic bacteria and drug resistance status, provide references for prevention in Weifang,100 milk samples were collected from 6 dairy cattle farms each with clin-ical or recessive mastitis in Weifang for pathogenic bacteria isolation and identification,resulting in bacteria detection rate of 91.50%. Milk samples with mixed bacteria infection reached 84.70%. 265 bacteria strains were isolated,includ-ing 38.11%Streptococcus strains(S. agalactiae,S. dysgalactiae and S. uberis),34.34%Staphylococcus aureus and 16.98%E. coli,forming the dominant pathogenic bacteria of dairy cow mastitis in Weifang. The drug sensitivity test of the dominant mastitis bacteria strains in Weifang indicated that the high sensitive drugs were cefquinome (80.59%), enrofloxacin(68.35%),ceftiofur(68.35%)and florfenicol(67.51%)and the high resistant drugs were penicillin(37.97%), streptomycin(26.16%)and tetracycline(10.55%).

  15. Molecular characterization of bacteria isolated from the Kingdom of ...



    Nov 6, 2012 ... ... bacterial isolation. Bacterial isolates were evaluated for their antimicrobial activities against pathogenic fungi which cause ... Molecular techniques based on genomic DNA have ... Isolation of total DNA from isolated strains.

  16. Reovirus, isolated from SARS patients

    DUAN Qing; SONG Lihua; GAN Yonghua; TAN Hua; JIN Baofeng; LI Huiyan; ZUO Tingting; CHEN Dehui; ZHANG Xuemin; ZHU Hong; YANG Yi; LI Weihua; ZHOU Yusen; HE Jun; HE Kun; ZHANG Haojie; ZHOU Tao


    Beijing has been severely affected by SARS, and SARS-associated coronavirus has been confirmed as its cause. However, clinical and experimental evidence implicates the possibility of co-infection. In this report, reovirus was isolated from throat swabs of SARS patients, including the first case in Beijing andher mother. Identification with the electron microscopy revealed the characteristic features of reovirus. 24 of 38 samples from other SARS cases were found to have serologic responses to the reovirus. Primers designed for reovirus have amplified several fragments of DNA, one of which was sequenced (S2 gene fragment), which indicates it as a unique reovirus (orthoreovirus). Preliminary animal experiment showed that inoculation of the reovirus in mice caused death with atypical pneumonia. Nevertheless, the association of reovirus with SARS outbreak requires to be further investigated.

  17. Multiple isolated cutaneous plexiform schwannomas

    Enas A. S. Attia


    Full Text Available Plexiform schwannoma is a rare neurogenic tumor, arising from skin and subcutaneous tissue. The presence of multiple schwannomas suggests a possible association with neurofibromatosis type 2 (NF2. A 50-year old male patient presented with multiple papulo-nodular cutaneous lesions on both arms and forearms. Histopathological examination revealed a dermal multinodular pattern of well-circumscribed masses of closely packed cells, with peripheral myxoid tissue, well-encapsulated in a thin collagenous capsule. S-100 immunohistochemical staining was diffusely and strongly positive. Neuron-specific enolase was positive, confirming a neural tissue tumor. An audiogram and Magnetic Resonance Imaging (MRI of cerebro-pontine angle showed no detected abnormality, excluding acoustic neuroma. Thus, we present a case of multiple bilateral isolated cutaneous plexiform schwannomas, not associated with NF2. Multiple plexiform schwannomas is a very rare entity, distinct from neurofibromatosis (NF, and being confined to the dermis is even more rarely reported.

  18. Propellant isolation shutoff valve program

    Merritt, F. L.


    An analysis and design effort directed to advancing the state-of-the-art of space storable isolation valves for control of flow of the propellants liquid fluorine/hydrazine and Flox/monomethylhydrazine is discussed. Emphasis is on achieving zero liquid leakage and capability of withstanding missions up to 10 years in interplanetary space. Included is a study of all-metal poppet sealing theory, an evaluation of candidate seal configurations, a valve actuator trade-off study and design description of a pneumo-thermally actuated soft metal poppet seal valve. The concepts and analysis leading to the soft seal approach are documented. A theoretical evaluation of seal leakage versus seal loading, related finishes and yield strengths of various materials is provided. Application of a confined soft aluminum seal loaded to 2 to 3 times yield strength is recommended. Use of either an electro-mechanical or pneumatic actuator appears to be feasible for the application.

  19. Bacterial isolates degrading aliphatic polycarbonates.

    Suyama, T; Hosoya, H; Tokiwa, Y


    Bacteria that degrade an aliphatic polycarbonate, poly(hexamethylene carbonate), were isolated from river water in Ibaraki. Prefecture, Japan, after enrichment in liquid medium containing poly(hexamethylene carbonate) suspensions as carbon source, and dilution to single cells. Four of the strains, 35L, WFF52, 61A and 61B2, degraded poly(hexamethylene carbonate) on agar plate containing suspended poly(hexamethylene carbonate). Degradation of poly(hexamethylene carbonate) was confirmed by gel permeation chromatography. Besides poly(hexamethylene carbonate), the strains were found to degrade poly(tetramethylene carbonate). The strains were characterized morphologically, physiologically, and by 16S rDNA sequence analysis. Strains 35L and WFF52 were tentatively identified as Pseudomonas sp. and Variovorax sp., respectively, while strains 61A and 61B2 constitute an unidentified branch within the beta subclass of the Proteobacteria.

  20. An isolated nasolacrimal duct osteoma.

    Kim, Joo Yeon; Kwon, Jae Hwan


    Osteomas of the nose and paranasal sinus are common benign tumors that can extend to surrounding structures and result in orbital or intracranial involvement. Presenting symptoms include facial pain, headache, cerebral symptoms, ocular symptoms, and so on, depending on the location and size of the tumor. They commonly occur within the frontal, ethmoid, maxillary, and sphenoid sinuses; however, there are rare cases of reported osteomas in the nasal cavity, turbinate, or orbit. Our case report describes a patient with nasolacrimal duct osteoma who presented with ipsilateral ocular pain, epiphora, and medial canthal swelling. We performed intranasal dacryocystorhinostomy using a nasal endoscope and removed the lacrimal duct osteoma. This report describes symptoms and management of an isolated nasolacrimal duct stone with a review of the literature.