Agalactiae Isolates Research Articles

Antibiotic Resistance and Presence of Persister Cells in the Biofilm-like Environments in Streptococcus agalactiae

Objectives: This study investigated antibiotic resistance and presence of persister cells in Streptococcus agalactiae strains belonging to capsular types Ia/ST-103, III/ST-17, and V/ST-26 in biofilm-like environments. Results: S. agalactiae strains were susceptible to penicillin, clindamycin, and erythromycin. Resistance genes were associated with tetM (80%), tetO (20%), ermB (80%), and linB (40%). Persister cells were detected in bacterial strains exposed to high concentrations of penicillin, clindamycin, and erythromycin. S. agalactiae capsular type III/ST-17 exhibited the highest percentage of persister cells in response to penicillin and clindamycin, while type Ia/ST-103 presented the lowest percentages of persister cells for all antimicrobials tested. Additionally, persister cells were also detected at lower levels for erythromycin, regardless of capsular type or sequence type. Further, all S. agalactiae isolates presented efflux pump activity in ethidium bromide-refractory cell assays. LIVE/DEAD fluorescence microscopy confirmed the presence of >85% viable persister cells after antibiotic treatment. Conclusions: These findings suggest that persister cells play a key role in the persistence of S. agalactiae during antibiotic therapy, interfering with the treatment of invasive infections. Monitoring persister formation is crucial for developing strategies to combat recurrent infections caused by this pathogen.

First report of Streptococcus agalactiae isolated from a healthy captive sichuan golden snub-nosed monkey (Rhinopithecus roxellana) in China

Streptococcus agalactiae (S. agalactiae) is an opportunistic pathogen, and to date, studies have mainly focused on S. agalactiae strains isolated from humans, dairy cows, and fish. We reported one S. agalactiae strain, named CFFB, which was isolated from a healthy Sichuan golden snub-nosed monkey. Classical bacteriological approaches, as well as, next-generation sequencing, comparative genomics, and mice challenge test were used to characterize this strain. CFFB was identified as serotype III, ST19 combination which is a common type found in human strains. Phylogenetic analysis showed that the genome of CFFB was closely related to human clinical isolates, rather far away from animal strains. In total, CFFB contained fewer virulence-associated genes and antibiotic resistance genes than human isolates that were close to CFFB in evolutionary relationships. In the mice challenge test, CFFB had a relative weak virulence that just caused death in 33 % of ICR mice at a dose of 108 CFU by intraperitoneal injection, and CFFB was reisolated from the cardiac blood of the dead mice. Meanwhile, two intact prophages (prophage 1 and 2) were identified in the CFFB genome and shared high similarities with phage Javan52 and Javan29 which from human S. agalactiae isolate Gottschalk 1002A and RBH03, respectively. Moreover, the type II-A CRISPR-Cas system was detected in the CFFB genome, and the spacers from CFFB were the same to the streptococci isolates from human. These results suggest that CFFB isolated from healthy Sichuan golden snub-nosed monkeys may have its origin in human S. agalactiae. Our results suggested some genomic similarities between the S. agalactiae colonized in Sichuan golden snub-nosed monkey and those in infected humans.

Molecular characterization, antimicrobial resistance and invasion of epithelial cells by Streptococcus agalactiae strains isolated from colonized pregnant women and newborns in Rio de Janeiro, Brazil.

To evaluate the prevalence, molecular characteristics, antimicrobial susceptibility, and epithelial invasion of Streptococcus agalactiae strains isolated from pregnant women and newborns in Rio de Janeiro, Brazil. A total of 67 S. agalactiae isolates, 48 isolates from pregnant women and 19 from neonates, were analyzed. Capsular type Ia and V were predominant (35.8%/each). The multilocus sequence typing analysis revealed the presence of 19 STs grouped into 6 clonal complexes with prevalence of CC17/40.3% and CC23/34.3%. The lmb and iag virulence genes were found in 100% of isolates. Four S. agalactiae strains, belonging to CC17/ST1249 and CC23/ST23, were able to adhere to A549 respiratory epithelial cells. Antimicrobial resistance was verified mainly to tetracycline (85%), erythromycin (70.8%), and clindamycin (58.3%). Four S. agalactiae isolates were multidrug resistant. The resistance genes tested were found in 92.5% of isolates for tetM, 58.2% for ermB, 28.4% for mefAE, and 10.4% for tetO. The study showed a high prevalence of virulence and antimicrobial genes in S. agalactiae strains isolated from pregnant women and newborns, supporting the idea that continued surveillance is necessary to identify risk factors and perform long-term follow-up in pregnant women and neonates in Rio de Janeiro.

Extraction of high-molecular-weight DNA from Streptococcus spp. for nanopore sequencing in resource-limited settings.

The long-read sequencing platform MinION, developed by Oxford Nanopore Technologies, enables the sequencing of bacterial genomes in resource-limited settings, such as field conditions or low- and middle-income countries. For this purpose, protocols for extracting high-molecular-weight DNA using nonhazardous, inexpensive reagents and equipment are needed, and some methods have been developed for gram-negative bacteria. However, we found that without modification, these protocols are unsuitable for gram-positive Streptococcus spp., a major threat to fish farming and food security in low- and middle-income countries. Multiple approaches were evaluated, and the most effective was an extraction method using lysozyme, sodium dodecyl sulfate, and proteinase K for lysis of bacterial cells and magnetic beads for DNA recovery. We optimized the method to consistently achieve sufficient yields of pure high-molecular-weight DNA with minimal reagents and time and developed a version of the protocol which can be performed without a centrifuge or electrical power. The suitability of the method was verified by MinION sequencing and assembly of 12 genomes of epidemiologically diverse fish-pathogenic Streptococcus iniae and Streptococcus agalactiae isolates. The combination of effective high-molecular-weight DNA extraction and MinION sequencing enabled the discovery of a naturally occurring 15 kb low-copy number mobilizable plasmid in S. iniae, which we name pSI1. We expect that our resource-limited settings-adapted protocol for high-molecular-weight DNA extraction could be implemented successfully for similarly recalcitrant-to-lysis gram-positive bacteria, and it represents a method of choice for MinION-based disease diagnostics in low- and middle-income countries.

Virulence Determinants of Buffalo Mastitis Originated Streptococcus agalactiae Isolates

Streptococci as a cause of mastitis have become the major concern to the dairy industry worldwide due to huge economic losses. Streptococcus agalactiae is a major contagious mastitis pathogen and continues to be a major cause of mastitis in dairy cattle and buffaloes. The aim of the study was to investigate the virulence determinants of S. agalactiae strains isolated from buffalo milk. Within the scope of the study, 24 S. agalactiae isolates from buffalo mastitis were examined. Biofilm production of isolates was investigated phenotypically by CRA method. It was observed that 18 (75%) isolates were positive for biofilm production. The presence of hylB, fnbB, scpB and spb1 virulence genes in S. agalactiae isolates were investigated by PCR. It was determined that 19 (79.17%) of the isolates were positive for scpB and 6 (25%) for fnbB virulence genes. None of the isolates were found to contain hylB and spb1 virulence genes. The antibiotic resistance profiles of the isolates among kanamycin, ampicillin, enrofloxacin, erythromycin, tetracycline, trimethoprim-sulfamethoxazole antibiotic discs were determined by Kirby Bauer Disc Diffusion Method. Resistance were evaluated as % 41.7, % 45.9, % 25, % 12.5, % 20.9, and % 33.3, respectively. RAPD-PCR patterns of all isolates were determined using the ERIC-2 primer. The dendrograms of the RAPD patterns were plotted with the UPGMA method. It was determined that the isolates showed similarity between 59-95%. In conclusion, the research confirms the prevalence of various virulence genes in S. agalactiae isolated from buffalo mastitis. Further studies are therefore necessary to determine the molecular epidemiology and variability of S. agalactiae isolated from buffaloes, with the aim of improving mastitis control programs with regard to S. agalactiae.

Identification of antibiotic induced persister cells in Streptococcus agalactiae.

Antibiotic persistence is a phenomenon, where a small fraction of a bacterial population expresses a phenotypic variation that allows them to survive antibiotic treatment, which is lethal to the rest of the population. These cells are called persisters cells, and their occurrence has been associated with recurrent disease. Streptococcus agalactiae is a human pathobiont, able to cause invasive infections, and recurrent infections have been reported to occur in both newborns and adults. In this study, we demonstrated that S. agalactiae NEM316 can form persister cells when exposed to antibiotics from different classes. The frequency of persister cell formation was dependent on bacterial growth phase and the class of antibiotics. The ability to form persister cells in response to penicillin was shown to be a general trait among different clinical S. agalactiae isolates, independent of sero- and sequence-type. Taken together, this study shows the existence of antibiotic tolerant S. agalactiae persister cells, which may explain why this bacterial species frequently persists after treatment of invasive infection and can be associated with recurrent disease.

KHÁNG KHÁNG SINH CỦA Streptococcus agalactiae GÂY BỆNH TRÊN CÁ RÔ PHI (Oreochromis sp.) Ở MIỀN BẮC VIỆT NAM

Antimicrobial resistance (AMR) is a threat to animal and human health globally. Antimicrobials are used in aquaculture to control different bacterial diseases. However, inappropriate use of antimicrobials can accelerate the emergence of AMR. This study was conducted to determine antimicrobial resistance in Streptococcus agalactiae, which affects and causes diseases with high mortality in tilapia. A total of 26 S. agalactiae isolates were streaked from kidney/brain/liver samples of diseased farmed tilapia from 2017 to 2021 at Hai Duong, Bac Ninh, Hoa Binh, Yen Bai provinces. The sensitivity of the isolates to 7 antimicrobials was established by disc diffusion (Oxiod). The results showed that levels of resistance vary from S. agalactiae to DOX (34.4%), RIF (64.1%), TET (45.0%), ERY (64.8%), TMPSMX (70%), FLO (35.3%), and AMP (75%). The resistance rate to DOX was significantly different (p<0.05) compared to ERY. S. agalactiae has the highest resistance to 2 types of antibiotics at 19.2%, followed by resistance to 3 types (15.4%), and resistance to 6 and 7 types of antibiotics at a rate of 11.5%. The resistance in 1 type, 4 types, and 5 types of antibiotics was observed lowest at 7.7%. Vietnamese tilapia farmers need guidelines for effective antimicrobial treatment and wwider take up of vaccine or sustainable farming technology to prevent infections caused by S. agalactiae and reduce emergence of AMR

Antimicrobial resistance of Streptococcus dysgalactiae, Streptococcus agalactiae, and Streptococcus canis in quarter milk samples from Bavaria, Southern Germany, between 2012 and 2022

The objective of this study was to analyze the in vitro antimicrobial resistance (AMR) of Streptococcus (Sc.) dysgalactiae, Sc. agalactiae, and Sc. canis over a 10-year period from 2012 to 2022 against the most commonly used antimicrobial agents. For this purpose, all quarter milk samples (QMS) submitted to the milk laboratory of the Bavarian Animal Health Service (TGD) were analyzed. Each QMS was tested using the California Mastitis Test (CMT) and categorized as negative (N), subclinical (S), or clinical (C) mastitis if the milk character was abnormal. Samples with Sc. dysgalactiae, Sc. agalactiae, or Sc. canis were included and a subset of isolates were further tested for in vitro antimicrobial resistance by breakpoint analysis with broth microdilution. Sc. dysgalactiae (61%, n = 65,750) was the most abundant pathogen among those 3 species, followed by Sc. agalactiae (28%, n = 30,486), and Sc. canis (11%, n = 11,336). All 3 species showed the highest resistance to the same 4 antimicrobial agents: erythromycin, marbofloxacin, pirlimycin, and cefalexin/kanamycin with varying degrees of resistance. Throughout the study period, Sc. dysgalactiae, Sc. agalactiae, and Sc. canis were largely susceptible to the remaining antimicrobial agents tested (penicillin, amoxicillin-clavulanate, oxacillin, cefazolin, cefoperazone, cefquinome). Only less than 14% of isolates of Sc. dysgalactiae and Sc. canis were resistant against any of the antimicrobials tested. Sc. agalactiae was the species with the highest percentage of resistant isolates. While the percentage of resistant isolates from Sc. canis and Sc. dysgalactiae decreased, the percentage of resistant Sc. agalactiae isolates increased since 2017. In summary, most isolates were not resistant to the most commonly used antimicrobial agents for mastitis therapy, including β-lactam antibiotics and penicillin should remain the first-choice therapy against streptococcal mastitis.

Serotype distribution, virulence and antibiotic resistance of Streptococcus agalactiae isolated from cultured tilapia Oreochromis niloticus in Lake Volta, Ghana.

Streptococcus agalactiae infection is one of the major factors limiting the expansion of tilapia farming globally. In this study, we investigated the serotype distribution, virulence and antimicrobial resistance of S. agalactiae isolates from tilapia farmed in Lake Volta, Ghana. Isolates from 300 moribund fish were characterised by Gram staining, MALDI-TOF/MS and 16S rRNA sequencing. Serotype identification was based on multiplex polymerase chain reaction (PCR) amplification of the capsular polysaccharide genes. Detection of virulence genes (cfb, fbsA and cspA) and histopathology were used to infer the pathogenicity of the isolates. The susceptibility of isolates to antibiotics was tested using the Kirby-Bauer disk diffusion assay. All 32 isolates identified as S. agalactiae were of serotype Ia. This was notably different from isolates previously collected from the farms in 2017, which belonged to serotype Ib, suggesting a possible serotype replacement. The prevalence of the pathogen was related to the scale of farm operation, with large-scale farms showing higher S. agalactiae positivity. Data from histopathological analysis and PCR amplification of targeted virulence genes confirmed the virulence potential and ability of the isolates to cause systemic infection in tilapia. Except for gentamicin, the majority of the isolates were less resistant to the tested antibiotics. All isolates were fully sensitive to oxytetracycline, erythromycin, florfenicol, enrofloxacin, ampicillin and amoxicillin. This study has improved our understanding of the specific S. agalactiae serotypes circulating in Lake Volta and demonstrates the need for continuous monitoring to guide the use of antimicrobials and vaccines against streptococcal infections in Ghanaian aquaculture systems.

Capsular gene distribution and RAPD typing of Streptococcus agalactiae isolated from pregnant women

Streptococcus agalactiae has different virulence factors, from which the capsule has the most significant role in the pathogenesis of this organism. We aimed to investigate the distribution of more prevalent capsular genes among different Random Amplified Polymorphic DNA (RAPD) types of S. agalactiae isolated from pregnant women. A total of 106 isolates were collected from 420 vaginal and rectal swabs obtained from pregnant women. The specimens were transferred using Todd Hewitt Broth and were cultured on a blood agar containing antibiotics. The S. agalactiae isolates were identified by the standard microbiological and biochemical tests. The genomic DNAs of S. agalactiae isolates were extracted using an extraction kit. Then, the PCR method was used to detection of the capsular genes. Moreover, The RAPD PCR was used to genotyping of the isolates. The colonization rate of the pregnant women was 25.23%, and there was a statistically significant correlation between the weeks of gestation and the probability of colonization (p-value < 0.05). Also, 31 (29.24%) and 18 (16.98%) pregnant women had a history of abortion and membrane rupture, respectively. In addition, 20 (18.86%), 32 (30.18%), 4 (3.77%), and 6 (5.66%) isolates carried genes encoding capsular types Ia, Ib, III, and V, respectively. None isolates had the type II capsular gene, and other 44 isolates were non-typeable. Nine clones (clusters) of S. agalactiae were observed in the present study with 70% similarity, and 53 different types were identified among the isolates. Except for capsular types III and V that belonged to clones 3, 5, 7, and 9, other capsular types were detected in different RAPD types. We found that the capsular types Ib and Ia were predominant among pregnant women in this area, indicating their significance for vaccine designation. Also, our isolates showed a lower genotypic diversity in RAPD typing. This may be due to the same sources of most isolates.

Streptococcus agalactiae Associated with "Dark Body" Disease on Snakeskin Gourami Farmed in the Mekong Delta, Vietnam

Streptococcosis, due to Streptococcus agalactiae, has become a serious challenge for aquaculture around the world. Therefore, the main purpose of this work was to confirm the etiological agent that caused "dark body" disease in snakeskin gourami (Trichogaster pectoralis) cultured in the Mekong Delta, Vietnam. Infected fish displayed clinical signs, including anorexia, lethargic swimming on the water surface, corneal opacity, and hemorrhage in the base of the fin. Remarkably, abnormal black coloration on the body and serious hemorrhage at the base of the pectoral fin, and on the body were found in infected fish. In total, 75 bacterial strains were isolated from different diseased fish farms. Based on morphological and physiological characteristics, the API 20 Strep test, and 16S rRNA sequence analysis, the result illustrated that the bacterial isolates were identified as S. agalactiae. Additionally, antibiotic sensitivity testing revealed that all the S. agalactiae isolates were sensitive to amoxicillin, rifampicin, clarithromycin, erythromycin, doxycycline, cephalexin, novobiocin, and sulfamethoxazole-trimethoprim. Interestingly, S. agalactiae strains were only resistant to gentamycin in this study. Two strains, SRT41 and SRT43, carried out an experimental challenge with concentrations from 104 to 107 CFU/ml in healthy snakeskin gourami, and the LD50 values of the isolates, SRĐT41 and SRĐT43, were determined to be 2.15 × 105 and 3.59 × 103 CFU/ml, respectively, after 7 days. To our knowledge, this is the first report of S. agalactiae derived from intensively cultured snakeskin gourami in the Mekong Delta, Vietnam.

Genetic diversity and biofilm formation ofinvasive and noninvasive Streptococcus agalactiae isolates: Emergence of hypervirulentCC19 strains in Tehran, Iran.

The prevalence of Streptococcus agalactiae infections in adult populations is increasing. The current study aimed to characterize the genetic features of S. agalactiae strains responsible for different infections. Across-sectional study was performed on 65 S. agalactiae strains (30 invasive and 35 noninvasive) isolated from non-pregnant women. All S. agalactiae isolates were confirmed by atr and dltS PCR assays. Antibiotic susceptibility patterns were determined using the disk diffusion method. Biofilm production was investigated by microtiter plate assay. PCR was done to detect resistance determinants. Isolates were characterized using the multilocus sequence typing (MLST) method. cMLSB, iMLSB, and M phenotypes accounted for 47.7%, 30.8%, and 6.2%, respectively. MDR was detected in 15.4% of noninvasive and 44.6% of invasive isolates. MtP assay indicated that 80% of isolates were biofilm producers. Biofilm formation was common among noninvasive compared with invasive strains (94.3% versus 66.7%). tet (M) (46.2%) and erm (B) (69.2%) were the most prevalent tetracycline and macrolide-resistance genes. The most prevalent serotype was type III (50.8%), followed by Ia (18.4%), II (15.4%), V (12.3%), and IV (3.1%). The frequency of serotype III among biofilm producer strains (81.8%) was found to be significantly higher than that of non-producer isolates (18.2%) (P < 0.05). S. agalactiae was resolved within four clonal complexes, including CC19 (46.2%; in both invasive and noninvasive), followed by CC23 (30.8%; only noninvasive isolates), CC1 (15.4%; only noninvasive isolates) and CC17 (7.6%; only invasive isolates). Themain sequence types (STs) found were ST19 (27.7%), ST17 (7.7%), ST27 (6.2%), and ST28 (4.6%) linked with invasive infections and ST23 (18.4%), ST933 (12.3%), ST644 (9.2%), ST19 (7.7%), ST1 (6.2%) found in noninvasive infections. The high prevalence of CC19 and CC23 clones among S. agalactiae strains reflects the emergence of these lineages as successful clones in Iran.

Proteomics and metabolomics analyses of Streptococcus agalactiae isolates from human and animal sources

Streptococcus agalactiae (S. agalactiae), group B Streptococcus (GBS), a major cause of infection in a wide variety of diseases, have been compared in different human and animal sources. We aimed to compare the bacterial proteome and metabolome profiles of human and animal S. agalactiae strains to delineate biological interactions relevant to infection. With the innovative advancement in mass spectrometry, a comparative result between both strains provided a solid impression of different responses to the host. For instance, stress-related proteins (Asp23/Gls24 family envelope stress response protein and heat shock protein 70), which play a role in the survival of GBS under extreme environmental conditions or during treatment, are highly expressed in human and animal strains. One human strain contains ꞵ-lactamase (serine hydrolase) and biofilm regulatory protein (lytR), which are important virulence regulators and potential targets for the design of novel antimicrobials. Another human strain contains the aminoglycosides-resistance bifunctional AAC/APH (A0A0U2QMQ5) protein, which confers resistance to almost all clinically used aminoglycosides. Fifteen different metabolites were annotated between the two groups. L-aspartic acid, ureidopropionic acid, adenosine monophosphate, L-tryptophan, and guanosine monophosphate were annotated at higher levels in human strains. Butyric acid, fumaric acid, isoleucine, leucine, and hippuric acid have been found in both human and animal strains. Certain metabolites were uniquely expressed in animal strains, with fold changes greater than 2. For example, putrescine modulates biofilm formation. Overall, this study provides biological insights into the substantial possible bacterial response reflected in its macromolecular production, either at the proteomic or metabolomic level.

207. The increased frequency and clinical characteristics of Streptococcus agalactiae serotype VIII from blood isolates at two tertiary care hospital in South Korea over 14 years

Abstract Background Invasive group B Streptococcus (GBS) infections are increasing among non-pregnant adults with underlying diseases. Previous studies have shown that the proportion of GBS serotype of pregnant women were differed from geographic regions. However, studies on the serotype of invasive GBS infection in non-pregnant adults are sparse. This study aimed to evaluate epidemiology and clinical characteristics of strains caused GBS bloodstream infection according to serotypes. Methods S. agalactiae blood isolates were collected between March 2007 and December 2021, and their clinical data were obtained from two tertiary hospital of South Korea. Species identification was performed using MALDI-TOF MS and serotyping test was performed using commercial latex particle agglutination kit. Antimicrobial susceptibility tests are conducted using automated equipment. Results A total of 130 patients with GBS bacteremia were identified and among them 125 isolates were serotypeable. The median age were 66 years and 46% were male. Serotype VIII was most dominant (31.5%) and Ib, III, Ia, V were as follows (20.8%, 13.8%, 12.3%, 11.5% retrospectively). The proportion of serotype VIII increased from 27.3% to 53.3% from 2015 to 2021. Most common source of GBS bloodstream infection were primary bacteremia (34.4%), followed by bone and joint infections (16.8%) and skin and soft tissue infections (13.6%). Cardiovascular disease (79.2%) was most frequently reported in GBS bacteremia patients, followed by malignancy (44.6%) and diabetes mellitus (40%). The 30-day mortality rate was 12.3% and there was no difference in 30-day mortality according to serotypes (log rank test P=0.39). In logistic regression analysis, high SOFA score was significantly associated with 30-day mortality (odd ratio 1.28, 95% C.I. 1.28-2.53, P&amp;lt; 0.01). All isolated GBS were susceptible to penicillin and resistances were identified against clindamycin, erythromycin and levofloxacin as 30%, 26.9%, 24.6% of the isolates, respectively. Conclusion We found the most common capsular serotype was VIII (31.5%) in Korea and the proportion of type VIII increasing over time. It is necessary to consider GBS vaccine in adults with underlying disease and its composition for serotype VIII according to epidemiologic characteristics. Disclosures All Authors: No reported disclosures

Genotypic and antimicrobial susceptibility of Streptococcus agalactiae causing bovine mastitis in the central region of Thailand.

Streptococcus agalactiae is a highly contagious pathogen that causes bovine mastitis, leading to significant economic losses. This study aimed to (1) identify and characterize S. agalactiae strains responsible for bovine mastitis by examining their phenotypic and genotypic characteristics in Thai dairy-intensive farming areas and (2) determine their susceptibility profiles to antimicrobial agents. In total, 100 S. agalactiae isolates obtained from clinical and subclinical mastitis cases from 13 dairy herds located in the central region of Thailand were examined. To confirm the identity of the bacterial pathogens, conventional microbiological procedures recommended by the National Mastitis Council (NMC) and the VITEK® 2 system were employed. All 100 isolates were successfully identified as S. agalactiae using the NMC procedure, whereas 94 isolates were identified as S. agalactiae using the VITEK® 2 system. Finally, the S. agalactiae-specific gene dlt S was identified in all the examined isolates using polymerase chain reaction. Capsular polysaccharide (CPS) typing revealed that all strains belonged to CPS type Ia. Multilocus sequence typing identified 33 selected isolates as sequence type 103. Random amplified polymorphic DNA (RAPD) typing yielded 43 RAPD types, with 6 RAPD clusters identified. These results demonstrated a high level of genetic diversity among S. agalactiae within the studied herds. RAPD analysis suggested that specific S. agalactiae strains could persist in dairy farms for 2-12 months. Furthermore, antimicrobial susceptibility testing was performed using the broth microdilution method. Most strains demonstrated susceptibility to ampicillin, penicillin, penicillin/novobiocin, cephalothin, oxacillin, ceftiofur, and erythromycin. This study revealed the phenotypic and genotypic characteristics of S. agalactiae isolates responsible for bovine mastitis in the central region of Thailand. The rapid identification of S. agalactiae and application of molecular typing methods can provide valuable epidemiological information regarding S. agalactiae causing mastitis in dairy farms. The antimicrobial susceptibility of S. agalactiae indicates that antimicrobial treatment for control and eradication could be a successful protocol. Our findings revealed that a single clonal strain of S. agalactiae affected the 13 studied farms. Further research is needed to explore the feasibility of vaccine development and application.

Incidence of virulence determinants among Streptococcus agalactiae isolated from pregnant women and association with their serotypes

ABSTRACT Colonization of pregnant women by Streptococcus agalactiae (S. agalactiae) is the main reason for intrauterine infection or transmission during parturition. This study investigated the prevalence, antimicrobial susceptibility, serotypes, virulence traits of S. agalactiae in pregnant women across three hospitals in Dakahlia Governorate, Egypt. Isolates were identified by culturing onto Hichrome Sterp B selective medium, latex agglutination and biochemical tests. The antimicrobial susceptibility was determined by Kirby-Bauer disc diffusion method. Capsular serotypes were determined by latex agglutination test. Hemolysin production and biofilm formation were assessed quantitively. Virulence genes were detected by polymerase chain reaction (PCR). Among 290 samples, 31.7% were S. agalactiae. All isolates were sensitive to penicillin G, ampicillin, cefotaxime, and vancomycin, but notably resistant to tetracycline (83.6%), erythromycin (68.4%), and clindamycin (54.3%). Serotype III (26.08%) was most common. Strong biofilm was produced by 72.8% of isolates. High hemolytic activity was seen (80.4%). Prevalent virulence genes included fsbA (97.8%), fbsB (96.7%), and scpB (96.7%). rib and alp4 were frequent surface protein genes (60.8% and 57.6%, respectively). Serotypes were significantly correlated to clindamycin resistance, biofilm production (Ia, III, VI serotypes), hemolytic activity (serotype V), and several virulence genes and surface proteins. Our study found a high prevalence of S. agalactiae isolates in pregnant women with diverse resistance patterns, high virulence indicating potential for increased pathogenicity.

In Vitro, In Vivo, and In Silico Activities of Ginkgolic Acid C15:1 against Streptococcus agalactiae Clinical Isolates.

Streptococcus agalactiae is the major cause of invasive neonatal infections and is a recognized pathogen associated with various diseases in nonpregnant adults. The emergence and spread of antibiotic-resistant S. agalactiae necessitate the development of a novel antibacterial agent. Here, the potential antibacterial activities and mechanisms of ginkgolic acid C15:1 (GA (15:1)) from Ginkgo biloba against clinical S. agalactiae are characterized. The MIC50 and MIC90 values for GA (15:1) against 72 clinical S. agalactiae isolates were 6.25 and 12.5 μM, respectively. GA (15:1) showed a strong bactericidal effect against both planktonic bacteria and bacteria embedded in biofilms as well as significant effectiveness in suppressing the growth of S. agalactiae biofilms. Moreover, GA (15:1) possesses intracellular antibacterial activity and could significantly decrease the bacterial burden in the intraperitoneal infection model of S. agalactiae. Mechanistic studies showed that GA (15:1) triggers membrane damage of S. agalactiae through a unique dual-targeting mechanism of action (MoA). First, GA (15:1) targets phospholipids in the bacterial cytoplasmic membrane. Second, by using mass-spectrometry-based drug affinity responsive target stability (DARTS) and molecular docking, lipoprotein signaling peptidase II (lspA) was identified as a target protein of GA (15:1), whose role is crucial for maintaining bacterial membrane depolarization and permeabilization. Our findings suggest a potential therapeutic strategy for developing GA (15:1) to combat S. agalactiae infections.

Identification of inducible clindamycin resistance gene in Streptococcus agalactiae isolates colonizing pregnant women in Suez Canal University Hospitals

Identification of inducible clindamycin resistance gene in Streptococcus agalactiae isolates colonizing pregnant women in Suez Canal University Hospitals

Streptococcosis on Nile tilapia (Oreochromis niloticus) in Indonesian freshwater aquaculture

This review aimed to provide the current status of streptococcosis in freshwater aquaculture in Indonesia. So far, this disease has been mostly reported in tilapia aquaculture, and there have been no serious reports of this disease in other species of freshwater aquaculture. The disease is mostly encountered during the dry season or the transition period from the dry season to the rainy season or vice versa. Initially, the disease was mostly reported in tilapia larger than 250 grams, but in its development, it was also reported in smaller fish. The Ministry of Marine Affairs and Fisheries (MMAF) has classified this disease as a major disease in tilapia in Indonesia since 2014. Based on the characteristics of Streptococcus agalactiae isolates collected from diseased tilapia from various regions in Indonesia, two biotypes were found consisting of ?-hemolytic S. agalactiae (biotype I) and non-hemolytic S. agalactiae (biotype II). Biotype I cause disease with acute mortality patterns, while biotype II causes disease with chronic and persistent mortality patterns. Based on susceptibility to S. agalactiae during artificial infection testing, it is known that all tilapia strains that have been released to the public for aquaculture purposes are susceptible to infection with the bacteria; although each strain has a different level of susceptibility. Fish health management strategies that are based on preventive measures and relevant to sustainable aquaculture programs should be based on ecological-biological approaches such as the implementation of biosecurity systems, the use of vaccines for endemic diseases, probiotics, essential herbs, and the development of more disease-resistant populations.Keywords: health management, streptococcosis, Streptococcus agalactiae, tilapia

Variable resistance to zinc intoxication among Streptococcus agalactiae reveals a novel IS1381 insertion element within the zinc efflux transporter gene czcD.

Streptococcus agalactiae, also known as group B Streptococcus, is an important human and animal pathogen. Zinc (Zn) is an essential trace element for normal bacterial physiology but intoxicates bacteria at high concentrations. Molecular systems for Zn detoxification exist in S. agalactiae, however the degree to which Zn detoxification may vary among different S. agalactiae isolates is not clear. We measured resistance to Zn intoxication in a diverse collection of clinical isolates of S. agalactiae by comparing the growth of the bacteria in defined conditions of Zn stress. We found significant differences in the ability of different S. agalactiae isolates to resist Zn intoxication; some strains such as S. agalactiae 18RS21 were able to survive and grow at 3.8-fold higher levels of Zn stress compared to other reference strains such as BM110 (6.4mM vs 1.68mM Zn as inhibitory, respectively). We performed in silico analysis of the available genomes of the S. agalactiae isolates used in this study to examine the sequence of czcD, which encodes an efflux protein for Zn that supports resistance in S. agalactiae. Interestingly, this revealed the presence of a mobile insertion sequence (IS) element, termed IS1381, in the 5' region of czcD in S. agalactiae strain 834, which was hyper-resistant to Zn intoxication. Interrogating a wider collection of S. agalactiae genomes revealed identical placement of IS1381 in czcD in other isolates from the clonal-complex-19 (CC19) 19 lineage. Collectively, these results show a resistance spectrum among S. agalactiae isolates enables survival in varying degrees of Zn stress, and this phenotypic variability has implications for understanding bacterial survival in metal stress.