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Background: Neonatal sepsis, often attributed to Group B Streptococcus (GBS) infection, poses a critical health risk to infants, demanding rapid and accurate diagnostic approaches. Existing diagnostic approaches are dependent on traditional culture methods, a process that requires substantial time and has the potential to delay crucial therapeutic assessments. Methods: This study introduces an innovative Loop-Mediated Isothermal Amplification (LAMP) assay for the early on-site detection of GBS infection from neonatal sepsis blood samples. To develop a LAMP assay, the primers are designed for the selective targeting of a highly conserved segment within the cfb gene encoding the CAMP factor in Streptococcus agalactiae ensuring high specificity. Results: Rigorous optimization of reaction conditions, including temperature and incubation time, enhances the efficiency of the LAMP assay, enabling rapid and reliable GBS detection within a short timeframe. The diagnostic efficacy of the LAMP assay was evaluated using spiked blood samples by eliminating the DNA extraction step. The simplified colorimetric LAMP assay has the capability to detect S. agalactiae in a neonatal blood sample containing 2 CFU/mL during sepsis. Additionally, the LAMP assay effectively detected S. agalactiae in both the standard and spiked blood samples, with no detectable interference with blood. Conclusion: This optimised LAMP assay emerges as a promising tool for early GBS detection, offering a rapid and accurate on-site solution that has the potential to inform timely interventions and improve outcomes in neonatal sepsis cases. [ABSTRACT FROM AUTHOR]

ABSTRACT The tkt (transketolase) gene is one of the seven gene fragments used in the multilocus sequence typing (MLST) system for Streptococcus agalactiae. We discovered that the tkt_134 allele is derived from a homologous gene (which we designate tktX) that is not present in all S. agalactiae; all known strains that contain a match to the tkt_134 allele also contain a gene sequence that is much closer in sequence identity to the other non-tkt_134 alleles (i.e., the canonical tkt gene) in the database. Based on these data, the tkt_134 allele has been removed from the MLST database as of September 2021, and all sequence types containing tkt_134 have also been removed.IMPORTANCEMultilocus sequence typing (MLST) databases are a common good and remain important for research, medical, and epidemiological purposes. This remains true even in the context of widespread whole-genome sequencing. We discovered a contaminating allele of the tkt gene in the S. agalactiae MLST database that led to unstable, ambiguous, or erroneous MLST assignment. The allele has since been removed from the public database based on the results presented in this manuscript.

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The pathogen S. agalactiae, categorized as Lancefield group B streptococci (GBS), is a Gram-positive bacterium, encapsulated, anaerobic facultative bacillus, which occurs in pairs or short chains. It was responsible for more than 319,000 cases of infections in neonates. Early and late streptococci, in the course of an illness or medical condition, are infections that have high mortality and morbidity rates, with an incidence of 2.0 to 5.0 per 1000 live births in North America, Europe, and Australia. The infection of newborns usually happens through transmission from infected mothers during and after childbirth, with GBS being among the critical bacterial infections in the Brazilian epidemiological picture. S. agalactiae strains isolated from different regions of Brazil, indicate a great diversity among the isolated serotypes but with a higher incidence of serotype Ia. Publications report that, in pregnant women in Brazil, the prevalence of serotype Ia has been constant between 2002 and 2018, followed by serotypes II and IV. Serotypes Ib, III, and V also appear, but with some fluctuations in different regions of the country. The pathogen which causes invasive disease are usually surrounded by a polysaccharide capsule and is its major virulence factor, being that the key antigen in protective response triggered by polysaccharides and, more recently, protein-polysaccharide conjugate vaccines. The development of antibiotic resistance by these microorganisms to different antibiotics has been the guideline in leading therapeutic options against these infections including the use of prophylactic vaccines in pregnant women, immunosuppressed or older people. Such prophylactic alternatives can be constructed from capsular polysaccharides.Based on the experience in the production and purification of Neisseria meningitidis polysaccharides for vaccines, Bio-Manguinhos developed a process for the production, purification and characterization of S. agalactiae Ia polysaccharides. In the present study, the capsular polysaccharide from the strain isolated in clinical material of a public hospital, in Rio de Janeiro – Brazil, was obtained from simple batch cultures, with purification process achieving about 20 mg g−1 in yield for cell dry weight (CDW) and ca. of 100 % of purity. Hence, to prove that the CPS of GBS Ia can be consistently produced, purified, and characterized by NMR, HRGC-MS, and SEC-MALS – from such strain – to be used in vaccine productions against S. agalactiae.

Introduction: Group B streptococcus (GBS) is recognized as a significant cause of neonatal infections worldwide. The study investigated the burden of early-onset neonatal disease and assessed the cost of routine GBS screening in a hospital of the eastern region of Saudi Arabia. Materials and Methods: Pregnancy-related information and laboratory-related data for pregnant women for the period of 2016-2017 were used to determine the colonization rate of GBS. The efficiency of intrapartum antibiotic prophylaxis (IAP) was examined among pregnant women carrying GBS and delivered at the hospital. The cost of GBS routine screening was estimated based on the cost of rectovaginal swab enrichment in BBLTM LIM broth and confirmation by polymerase chain reaction. The burden of early-onset disease and rates of resistance to antibiotics among vaginal and invasive neonatal isolates were investigated for seven years. Results: Screening of 2,476 samples from 1,162 pregnant women in two years showed a GBS colonization rate of 23% (270/1162). Of the 270 GBS-positive women, only 98 (37%) delivered at the hospital, of whom 42% received IAP. Cesarean delivery was associated with significantly higher rates of IAP administration (odds ratio 5.2; p value 0.001). The annual incidence of early-onset disease increased from a baseline level of 0.5 in 2012 to 3.9 per 1,000 births in 2018. These included 23 cases of invasive infections with three fatal cases caused by meningitis or sepsis (3/23, 13%). Group B streptococcus isolates remained uniformly sensitive to penicillin and vancomycin but exhibited resistance to clindamycin in 35.2%. The annual estimated costs of routine screening would be $110,880 ($41 per delivery) and $161,280 ($60 per delivery) for the BD MAX™ system and GeneXpert® System, respectively. Conclusion: While the incidence of early-onset GBS disease was increasing, improved compliance with the IAP is needed to inform the cost-effectiveness of routine GBS screening.

Antimicrobial therapy is frequently associated with the emergence of resistant bacteria with a high rate of morbidity and mortality worldwide. The present study was aimed at investigating the impact of a neodymium-doped yttrium aluminum (Nd:YAG) laser, and a static magnetic field (SMF) on cellular growth and DNA alteration in some clinical bacterial isolates. Samples from cutaneous wounds were collected by sterile cotton swabs from three elderly women admitted to Tikrit Teaching Hospital, Tikrit City, Iraq. Isolation and identification of Streptococcus agalactiae, Staphylococcus aureus, and Pseudomonas aeruginosa were carried out using cultural characteristics, microscopy, and biochemical tests. Three broth cultures were prepared for each of the test isolates. The first broth culture served as untreated control, the second was exposed to an Nd:YAG laser and the third was exposed to SMF. Colony counting was done on all the samples. DNA was extracted from the test bacteria and used to perform the RAPD-PCR assay. In contrast to the untreated control, the results showed that Nd:YAG laser radiation was more effective than SMF at inhibiting the cellular growth of the test isolates. Also, the radiation caused DNA alteration, which was established by decreased microbial growth, as well as the development of new bands and the loss of original bands. According to the findings of this study, the Nd:YAG laser is a promising technique for influencing the healing of infected cutaneous wounds. RAPD-PCR is also a useful biomarker assay for assessing the biological impact of laser radiation and SMF on bacteria.

Intramammary infection (IMI) from the environment and infected quarters can cause co-infection. The objective of this study was to determine the ability of coagulase-negative staphylococci (CNS) to survive in the same environment as Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli as major pathogens. In total, 15 and 242 CNS strains were used in Experiment I and Experiment II, respectively. Both experiments were separated into three conditions: culture with CNS 24 h before (PRIOR), after (AFTER), and at the same time (EQUAL). The lack of a clear zone, regardless of size, was determined to be the key to the survival of both. The CNS species' percentages of survival against major pathogens were tested using Fisher's exact test. Differences in the percentages of survival were evident among the CNS species in all conditions. For the PRIOR condition, all CNS mostly survived when living with major strains; however, S. chromogenes could degrade S. agalactiae. Although most CNS strains were degraded in the AFTER and EQUAL conditions, some strains of S. hominis and S. simulans could resist S. aureus and S. agalactiae. In conclusion, some specific strains of CNS are able to survive in an environment with major pathogens. Research into the survival strains may indicate that the concept of novel bacteria with bacteriolytic capabilities might be possible as a novel mastitis treatment. [ABSTRACT FROM AUTHOR]

Abstract Background Mastitis is an inflammation of the mammary glands caused by a microbial infection. The common bacteria causing this infection in dairy farms are Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. The aptamer is a new biosensor platform for detecting pathogens; however, its use for simultaneous detection of S. aureus, S. agalactiae, and E. coli bacteria has not been reported. This study’s objective is to isolate and characterize polyclonal DNA aptamer with broad reactivity to the mastitis bacteria S. aureus, S. agalactiae, and E. coli using a sequential toggle cell-SELEX. Methods and results The DNA aptamer pool from SELEX 15 was inserted into the pGEM-T easy plasmid. Furthermore, the transformant clones were selected by PCR colony, plasmid isolation, and sequencing. Six DNA aptamers, consisting of S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20 with a constant region and the right size of 81 bp were derived from the sequencing analysis. The secondary structure of the DNA was predicted using Mfold software. The DNA was analyzed with binding characteristics, including binding capacity and affinity (Kd), using qPCR. The results indicated aptamer S15K15 has the highest binding ability into S. agalactiae, while S15K13 performed binding capacity most to E. coli EPEC 4, and S15K3 has the highest capacity of binding to S. aureus BPA-12. Conclusion Aptamer S15K3 has the best binding characteristics on all three bacterial targets.

Background: Mastitis is an inflammation of the mammary glands caused by a microbial infection. The common bacteria causing this infection in dairy farms are Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. The aptamer is a new biosensor platform for detecting pathogens; however, its use for simultaneous detection of S. aureus, S. agalactiae, and E. coli bacteria has not been reported. This study’s objective is to isolate and characterize polyclonal DNA aptamer with broad reactivity to the mastitis bacteria S. aureus, S. agalactiae, and E. coli using a sequential toggle cell-SELEX. Methods and results: The DNA aptamer pool from SELEX 15 was inserted into the pGEM-T easy plasmid. Furthermore, the transformant clones were selected by PCR colony, plasmid isolation, and sequencing. Six DNA aptamers, consisting of S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20 with a constant region and the right size of 81 bp were derived from the sequencing analysis. The secondary structure of the DNA was predicted using Mfold software. The DNA was analyzed with binding characteristics, including binding capacity and affinity (Kd), using qPCR. The results indicated aptamer S15K15 has the highest binding ability into S. agalactiae, while S15K13 performed binding capacity most to E. coli EPEC 4, and S15K3 has the highest capacity of binding to S. aureus BPA-12. Conclusion: Aptamer S15K3 has the best binding characteristics on all three bacterial targets. [ABSTRACT FROM AUTHOR]

Mastitis disease causes significant economic losses in dairy farms by reducing milk production, increasing production costs, and reducing milk quality. Streptococcus agalactiae continues to be a major cause of mastitis in dairy cattle. To date, there has been no approved multi-epitope vaccine against this pathogen in the market. In the present study, an efficient multi-epitope vaccine against S. agalactiae, the causative agent of mastitis, was designed using various immonoinformtics approaches. Potential epitopes were selected from Sip protein to improve vaccine immunogenicity. The designed vaccine is more antigenic in nature. Then, linkers and profilin adjuvant were added to enhance the immunity of vaccines. The designed vaccine was evaluated in terms of molecular weight, PI, immunogenicity, Toxicity, and allergenicity. Prediction of three-dimensional (3 D) structure of multi-epitope vaccine, followed by refinement and validation, was conducted to obtain a high-quality 3 D structure of the designed multi-epitope vaccine. The designed vaccine was then subjected to molecular docking with Toll-like receptor 11 (TLR11) receptor to evaluate its binding efficiency followed by dynamic simulation for stable interaction. In silico cloning approach was carried out to improve the expression of the vaccine construct. These analyses indicate that the designed multi-epitope vaccine may produce particular immune responses against S. agalactiae and may be further helpful to control mastitis after in vitro and in vivo immunological assays. [ABSTRACT FROM AUTHOR]

Objectives: Streptococcus agalactiae is a zoonotic human and animal pathogen that causes global economic losses in aquaculture and fatal outcomes in Tilapia. This study aimed to identify S. agalactiae isolated from different fish sources intended for human consumption phenotypically and genotypically and to characterize the virulence-associated genes fbsA (fibrinogen-binding protein FbsA), cfb (CAMP factor), and pbp1A/ponA (penicillin-binding protein 1A). Materials and Methods: Three hundred Nile Tilapia fish (Oreochromis niloticus) were collected from different farms and retail shops in Dakahlia and Damietta, Egypt, during the summer of 2020. The samples were examined using routine phenotypic methods, then characterized using polymerase chain reaction (PCR) targeting S. agalactiae-specific dltS gene. All S. agalactiae isolates were examined for the susceptibility to ten antimicrobial agents by the disc diffusion method. The virulence-associated genes (fbsA, cfb, and pbp1A/ponA) were characterized using multiplex-PCR. Results: Streptococcus agalactiae was detected in 7% (n = 21/300) samples. The isolates showed high resistance against ampicillin and erythromycin (20/21; 95%) for each. The most predominant antibiotypes through isolates were P, CN, SXT, CRO, TE, CTX, E, AMP, at 10.5% for each antibiotype. A total of 19 (90.5%) of S. agalactiae isolates showed multi-drug resistance (MDR), and those were recovered from market Tilapia fish. The virulence-associated genes (fbsA, cfb, and pbp1A/ ponA) were identified in the S. agalactiae as 100%, 76%, and 52%, respectively. Conclusions: The MDR S. agalactiae detected in raw Tilapia fish pose a significant health hazard to consumers due to their zoonotic characteristics. [J Adv Vet Anim Res 2022; 9(1.000): 95-103]

Antimicrobial therapy is frequently associated with the emergence of resistant bacteria with a high rate of morbidity and mortality worldwide. The present study was aimed at investigating the impact of a neodymium-doped yttrium aluminum (Nd:YAG) laser, and a static magnetic field (SMF) on cellular growth and DNA alteration in some clinical bacterial isolates. Samples from cutaneous wounds were collected by sterile cotton swabs from three elderly women admitted to Tikrit Teaching Hospital, Tikrit City, Iraq. Isolation and identification of Streptococcus agalactiae, Staphylococcus aureus, and Pseudomonas aeruginosa were carried out using cultural characteristics, microscopy, and biochemical tests. Three broth cultures were prepared for each of the test isolates. The first broth culture served as untreated control, the second was exposed to an Nd:YAG laser and the third was exposed to SMF. Colony counting was done on all the samples. DNA was extracted from the test bacteria and used to perform the RAPD-PCR assay. In contrast to the untreated control, the results showed that Nd:YAG laser radiation was more effective than SMF at inhibiting the cellular growth of the test isolates. Also, the radiation caused DNA alteration, which was established by decreased microbial growth, as well as the development of new bands and the loss of original bands. According to the findings of this study, the Nd:YAG laser is a promising technique for influencing the healing of infected cutaneous wounds. RAPD-PCR is also a useful biomarker assay for assessing the biological impact of laser radiation and SMF on bacteria. [ABSTRACT FROM AUTHOR]

In this study, a novel interleukin-1-family member (nIL-1Fm) was studied from the Nile tilapia (Oreochromis niloticus), a widely cultured fish species in brackish water and freshwater in Thailand and worldwide. The complete sequence of the Nile tilapia nIL-1Fm comprised 2474 bp, encoding 350 amino acids. The genome organization of nIL-1Fm had seven exons and six introns, which were conserved in teleost fish. During tissue distribution analysis, the nIL-1Fm was extremely increased in the head kidney, tailfin and trunk kidney, which had the highest levels, followed by the spleen. Additionally, nIL-1Fm was significantly up-regulated under PAMP stimulation, whereas the stimulation of LPS, PHA and poly I:C could significantly induce nIL-1Fm expression in vitro. Moreover, nIL-1Fm transcription was significantly induced when infected with Streptococcus agalactiae in the spleen, liver and intestine. Taking together, our results indicated that nIL-1Fm can expand our knowledge of the IL-1 cytokine family in fish immune response. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae is a well-known pathogen in humans and food-producing animals. Therefore, this bacterium is a paradigmatic example of a pathogen to be controlled by a One Health approach. Indeed, the zoonotic and reverse-zoonotic potential of the bacteria, the prevalence of Group B Streptococci (GBS) diseases in both human and animal domains, and the threatening global situation on GBS antibiotic resistance make these bacteria an important target for control programs. An epidemiological analysis using a public database containing sequences of S. agalactiae from all over the world was conducted to evaluate the frequency and evolution of antibiotic resistance genes in those isolates. The database we considered (NCBI pathogen detection isolate browser—NPDIB) is maintained on a voluntary basis. Therefore, it does not follow strict epidemiological criteria. However, it may be considered representative of the bacterial population related to human diseases. The results showed that the number of reported sequences increased largely in the last four years, and about 50% are of European origin. The frequency data and the cluster analysis showed that the AMR genes increased in frequency in recent years and suggest the importance of verifying the application of prudent protocols for antimicrobials in areas with an increasing frequency of GBS infections both in human and veterinary medicine. [ABSTRACT FROM AUTHOR]

IgY is an immunoglobulin primarily found in the serum and egg yolk of birds, amphibians, and reptiles. Recent years, IgY is considered to have a good application prospect in the immunodiagnostics and passive immunotherapy of aquatic diseases. In this study, we prepared a specific IgY against Streptococcus agalactiae in tilapia after immunizing the hens for 4 times. The result of ELISA detection showed that the IgY titers in water-soluble fraction (WSF) after 6 weeks of immunization reached 1:51200 and last for 4 weeks. Western blot (WB) analysis data showed that the specific IgY could recognize the target band, the specific IgY showed a concentration-dependent inhibitory effect on the growth of S. agalactiae, altered cell wall structure and aggluted of S. agalactiae. The quantitative reverse transcription PCR (qRT-PCR) analysis data suggested that the specific IgY downregulated the expression of pro-inflammatory factors (IL-8, TNF-α), upregulated the anti-inflammatory factors (IL-10, TGF-β). In addition, the histopathological results showed that the specific IgY significantly decreased the pathological manifestations, dramatically improved the survival rates of tilapia in injection, feeding, and immersion experiments. Collectively, our findings demonstrated that the broad potential of specific IgY for the prevention and treatment of S. agalactiae infection in tilapia., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The tkt (transketolase) gene is one of the seven gene fragments used in the multilocus sequence typing (MLST) system for Streptococcus agalactiae . We discovered that the tkt_134 allele is derived from a homologous gene (which we designate tktX ) that is not present in all S. agalactiae ; all known strains that contain a match to the tkt_134 allele also contain a gene sequence that is much closer in sequence identity to the other non-tkt_134 alleles (i.e., the canonical tkt gene) in the database. Based on these data, the tkt_134 allele has been removed from the MLST database as of September 2021, and all sequence types containing tkt_134 have also been removed.IMPORTANCEMultilocus sequence typing (MLST) databases are a common good and remain important for research, medical, and epidemiological purposes. This remains true even in the context of widespread whole-genome sequencing. We discovered a contaminating allele of the tkt gene in the S. agalactiae MLST database that led to unstable, ambiguous, or erroneous MLST assignment. The allele has since been removed from the public database based on the results presented in this manuscript., Competing Interests: The authors declare no conflict of interest.

Background: Mastitis is one of the most expensive diseases in the dairy industry. It causes heavy monetary losses by decreasing milk production and treatment cost. Streptococcus agalactiae, the cause of contagious bovine mastitis, possesses various virulence factors that contribute to pathogenicity. Aims: The main aim of the study was to evaluate the distribution of virulence genes of S. agalactiae. Methods: In the current work, 98 Streptococcus species were isolated from 320 milk samples, collected from Veterinary Clinical Complex, Junagadh. Out of the isolates, 42 S. agalactiae isolates were used for virulence genes detection. Results: All Streptococcus spp. were confirmed at genus level by targeting tuf gene, and S. agalactiae was identified at species level by targeting 16S rRNA gene. For virulence gene detection, scpB, cfb, and cylE genes were targeted. Of 42 S. agalactiae isolates, 15, 16, and 10 isolates possessed scpB, cfb, and cylE genes, respectively. Conclusion: This study aids us to know virulence characteristics and mechanisms responsible for the development of new strains in mastitis epidemiology in response to prevention and control strategies. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae causes sepsis and meningitis in neonates, presenting substantial clinical challenges. Type VII secretion system (T7SS), an important secretion system identified in Mycobacterium sp. and Gram-positive bacteria, was recently characterized in S. agalactiae and considered to contribute to its virulence and pathogenesis. In the present study, 128 complete genomic sequences of S. agalactiae were retrieved from GenBank to build a public dataset, and their sequences, capsular types, and clonal complexes were determined. Polymerase chain reaction (PCR) screening and genomic sequencing were conducted in an additional clinical dataset. STs and capsular types were determined using PCR. Eleven different types of T7SS were detected with similarities in gene order but differences in gene content. Strains with incomplete T7SS or lack of T7SS were also identified. Deletion, insertion, and segmentation of T7SS might be related to insertion sequences. The genetic environment of T7SS in S. agalactiae was also investigated and different patterns were identified downstream the T7SS, which were related to the diversity of T7SS putative effectors. The T7SS demonstrated possible sequence type (ST)-dependent diversity in both datasets. This work elucidated detailed genetic characteristics of T7SS and its genetic environment in S. agalactiae and further identified its possible ST-dependent diversity, which gave a clue of its mode of transmission. Further investigations are required to elucidate the underlying mechanisms and its functions. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae causes sepsis and meningitis in neonates, presenting substantial clinical challenges. Type VII secretion system (T7SS), an important secretion system identified in Mycobacterium sp. and Gram-positive bacteria, was recently characterized in S. agalactiae and considered to contribute to its virulence and pathogenesis. In the present study, 128 complete genomic sequences of S. agalactiae were retrieved from GenBank to build a public dataset, and their sequences, capsular types, and clonal complexes were determined. Polymerase chain reaction (PCR) screening and genomic sequencing were conducted in an additional clinical dataset. STs and capsular types were determined using PCR. Eleven different types of T7SS were detected with similarities in gene order but differences in gene content. Strains with incomplete T7SS or lack of T7SS were also identified. Deletion, insertion, and segmentation of T7SS might be related to insertion sequences. The genetic environment of T7SS in S. agalactiae was also investigated and different patterns were identified downstream the T7SS, which were related to the diversity of T7SS putative effectors. The T7SS demonstrated possible sequence type (ST)-dependent diversity in both datasets. This work elucidated detailed genetic characteristics of T7SS and its genetic environment in S. agalactiae and further identified its possible ST-dependent diversity, which gave a clue of its mode of transmission. Further investigations are required to elucidate the underlying mechanisms and its functions.

Intramammary infection (IMI) from the environment and infected quarters can cause co-infection. The objective of this study was to determine the ability of coagulase-negative staphylococci (CNS) to survive in the same environment as Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli as major pathogens. In total, 15 and 242 CNS strains were used in Experiment I and Experiment II, respectively. Both experiments were separated into three conditions: culture with CNS 24 h before (PRIOR), after (AFTER), and at the same time (EQUAL). The lack of a clear zone, regardless of size, was determined to be the key to the survival of both. The CNS species’ percentages of survival against major pathogens were tested using Fisher’s exact test. Differences in the percentages of survival were evident among the CNS species in all conditions. For the PRIOR condition, all CNS mostly survived when living with major strains; however, S. chromogenes could degrade S. agalactiae. Although most CNS strains were degraded in the AFTER and EQUAL conditions, some strains of S. hominis and S. simulans could resist S. aureus and S. agalactiae. In conclusion, some specific strains of CNS are able to survive in an environment with major pathogens. Research into the survival strains may indicate that the concept of novel bacteria with bacteriolytic capabilities might be possible as a novel mastitis treatment.

This study aimed to investigate the potential causes of the summer mortality syndrome affecting the earthen pond cultured Nile tilapia, Oreochromis niloticus in some Egyptian farms. A total of 100 moribund O. niloticus showing non-specific clinical signs of septicemia and mortality (30- 65%) were collected from five farms at Tolombat 7, Kafr El-Sheikh, Egypt, during June 2019. All the fish farms share the same water source and perform some bad management practices. Inadequate water quality measures were noticed in all the investigated farms. Molecular detection of tilapia lake virus (TiLV) using PCR confirmed the non-existence of the TiLV in all examined specimens. A total of 19 pathogenic bacterial isolates were obtained from moribund O. niloticus: Streptococcus agalactiae (14), Aeromonas hydrophila (3), and Vibrio cholera (2). The predominance of Streptococcus agalactiae infections points to its intense involvement in such mortalities. S. agalactiae was confirmed by 16s rRNA gene sequencing and phylogenetic analysis. The PCR detection of virulence genes showed existence of Hyl, cylE, scpB and camp genes in all S. agalactiae strains. Isolates of S. agalactiae showed variable susceptibility to the tested antibiotics with high resistance against ampicillin, amoxicillin and gentamycin. Infections were associated with several pathological alterations in naturally infected fish. The pathogenicity of S. agalactiae was confirmed in O. niloticus (90 % cumulative mortality). The data in the present study proved that the tilapia summer mortality syndrome is multifactorial. Thus, good managemental practices and water quality measures are recommended to minimize fish mortality. [ABSTRACT FROM AUTHOR]

Objectives: Streptococcus agalactiae is a zoonotic human and animal pathogen that causes global economic losses in aquaculture and fatal outcomes in Tilapia. This study aimed to identify S. agalactiae isolated from different fish sources intended for human consumption phenotypically and genotypically and to characterize the virulence-associated genes fbsA (fibrinogen-binding protein FbsA), cfb (CAMP factor), and pbp1A/ponA (penicillin-binding protein 1A). Materials and Methods: Three hundred Nile Tilapia fish (Oreochromis niloticus) were collected from different farms and retail shops in Dakahlia and Damietta, Egypt, during the summer of 2020. The samples were examined using routine phenotypic methods, then characterized using polymerase chain reaction (PCR) targeting S. agalactiae-specific dltS gene. All S. agalactiae isolates were examined for the susceptibility to ten antimicrobial agents by the disc diffusion method. The virulence-associated genes (fbsA, cfb, and pbp1A/ponA) were characterized using multiplex-PCR. Results: Streptococcus agalactiae was detected in 7% (n = 21/300) samples. The isolates showed high resistance against ampicillin and erythromycin (20/21; 95%) for each. The most predominant antibiotypes through isolates were P, CN, SXT, CRO, TE, CTX, E, AMP, at 10.5% for each antibiotype. A total of 19 (90.5%) of S. agalactiae isolates showed multi-drug resistance (MDR), and those were recovered from market Tilapia fish. The virulence-associated genes (fbsA, cfb, and pbp1A/ ponA) were identified in the S. agalactiae as 100%, 76%, and 52%, respectively. Conclusions: The MDR S. agalactiae detected in raw Tilapia fish pose a significant health hazard to consumers due to their zoonotic characteristics. [ABSTRACT FROM AUTHOR]

The class B scavenger receptors (scarb) are members of important pattern recognition receptors which play key roles for host immune response to pathogen infection. In the present study, two scarb member 2 paralogous (Onscarb2a and Onscarb2c) were identified and characterized in Nile tilapia. Onscarb2a was identified with an open reading frame (ORF) of 1593 bp which encoded a polypeptide of 530 amino acids, and Onscarb2c was consisting of an ORF of 1446 bp which encoded a polypeptide of 461 amino acids. Both Onscarb2a and Onscarb2c share high sequence identity/similarity with their counterparts from other teleost species. Multiple sequence alignment and phylogenetic tree analyses show that scarb2c from teleost fish is more closely related to other vertebrate scarb2s. Gene expression studies revealed that both scarb2a and scarb2c were highly expressed in liver, with the lowest expression in muscle in healthy tilapia. After challenges with Streptococcus agalactiae and Aeromonas hydrophila, both Onscarb2a and Onscarb2c were markedly up-regulated in liver and head kidney. In addition, the up-regulation of Onscarb2a and Onscarb2c were also detected in head kidney monocytes/macrophages and hepatocytes. In conclusion, this study revealed that Onscarb2a and Onscarb2c might play important roles in the innate immune responses to bacterial invasion in Nile tilapia.

Pyogenic ventriculitis is a disorder characterized by inflammation of the cerebral ventricular lining secondary to infection within the ventricular system. Very few cases of primary pyogenic ventriculitis have been reported among adults. We present a case report of a 74-year-old female with a history of hypertension and diabetes mellitus who presented with Group B Streptococcus (GBS) primary pyogenic ventriculitis. She was successfully treated with intravenous (IV) antibiotics. To our knowledge, this is the only case of adult Streptococcus  agalactiae primary pyogenic ventriculitis., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Sangam et al.)

Streptococcus agalactiae is a well-known pathogen in humans and food-producing animals. Therefore, this bacterium is a paradigmatic example of a pathogen to be controlled by a One Health approach. Indeed, the zoonotic and reverse-zoonotic potential of the bacteria, the prevalence of Group B Streptococci (GBS) diseases in both human and animal domains, and the threatening global situation on GBS antibiotic resistance make these bacteria an important target for control programs. An epidemiological analysis using a public database containing sequences of S. agalactiae from all over the world was conducted to evaluate the frequency and evolution of antibiotic resistance genes in those isolates. The database we considered (NCBI pathogen detection isolate browser—NPDIB) is maintained on a voluntary basis. Therefore, it does not follow strict epidemiological criteria. However, it may be considered representative of the bacterial population related to human diseases. The results showed that the number of reported sequences increased largely in the last four years, and about 50% are of European origin. The frequency data and the cluster analysis showed that the AMR genes increased in frequency in recent years and suggest the importance of verifying the application of prudent protocols for antimicrobials in areas with an increasing frequency of GBS infections both in human and veterinary medicine.

In this study, a novel interleukin-1-family member (nIL-1Fm) was studied from the Nile tilapia (Oreochromis niloticus), a widely cultured fish species in brackish water and freshwater in Thailand and worldwide. The complete sequence of the Nile tilapia nIL-1Fm comprised 2474 bp, encoding 350 amino acids. The genome organization of nIL-1Fm had seven exons and six introns, which were conserved in teleost fish. During tissue distribution analysis, the nIL-1Fm was extremely increased in the head kidney, tailfin and trunk kidney, which had the highest levels, followed by the spleen. Additionally, nIL-1Fm was significantly up-regulated under PAMP stimulation, whereas the stimulation of LPS, PHA and poly I:C could significantly induce nIL-1Fm expression in vitro. Moreover, nIL-1Fm transcription was significantly induced when infected with Streptococcus agalactiae in the spleen, liver and intestine. Taking together, our results indicated that nIL-1Fm can expand our knowledge of the IL-1 cytokine family in fish immune response.

Group B Streptococcus (GBS) is a leading cause of neonatal meningitis, pneumonia, and sepsis. The biggest contributing factor of neonatal infections is due to vertical transmission from maternal colonisation of GBS in the genitourinary tract. Multiple serotype colonisation is often not investigated in epidemiological studies, but it is an important consideration for serotype-based vaccine development and implementation to ensure less abundant serotypes are not under-represented. In this study, we show that RAPD PCR is a quick tool useful in screening the presence of genetically different strains using multiple colony picks from a single patient swab. We observed a maximum of five different GBS strains colonising a single patient at a specific time. • Co-colonisation of GBS strains is present in pregnant women and infants. • GBS serotype acquisition and loss is a common occurrence during colonisation. • RAPD PCR can be used as a screening tool to identify genetically distinct strains. • Assay may be useful to complement serotyping before sending GBS isolates for further characterisations. [ABSTRACT FROM AUTHOR]

Group B Streptococcus (GBS) infection in newborns during childbirth can result in death. We described a method to detect GBS from vaginal swabs in pregnant women, without prior enrichment, using real-time polymerase chain reaction (qPCR), and compared its results to the culture method. The qPCR outperforms culture method. [Display omitted] • qPCR has great sensitivity for GBS detection in pregnant women; • qPCR can improve the strategies against GBS infection; • qPCR should be the gold standard for GBS detection. [ABSTRACT FROM AUTHOR]