Streptococcus Agalactiae Strains Research Articles

Macrolide and lincosamide resistance of Streptococcus agalactiae in pregnant women in Poland

Knowing about the antibiotic resistance, serotypes, and virulence-associated genes of Group B Streptococcus for epidemiological and vaccine development is very important. We have determined antimicrobial susceptibility patterns, serotype, and virulence profiles. The antibiotic susceptibility was assessed for a total of 421 Streptococcus agalactiae strains, isolated from pregnant women and neonates. Then, 89 erythromycin and/or clindamycin-resistant strains (82 isolates obtained from pregnant women and seven isolates derived from neonates) were assessed in detail. PCR techniques were used to identify the studied strains, perform serotyping, and assess genes encoding selected virulence factors. Phenotypic and genotypic methods determined the mechanisms of resistance. All tested strains were sensitive to penicillin and levofloxacin. The constitutive MLSB mechanism (78.2%), inducible MLSB mechanism (14.9%), and M phenotype (6.9%) were identified in the macrolide-resistant strains. It was found that macrolide resistance is strongly associated with the presence of the ermB gene and serotype V. FbsA, fbsB, fbsC, scpB, and lmb formed the most recurring pattern of genes among the nine surface proteins whose genes were analysed. A minority (7.9%) of the GBS isolates exhibited resistance to lincosamides and macrolides, or either, including those that comprised the hypervirulent clone ST-17. The representative antibiotic resistance pattern consisted of erythromycin, clindamycin, and tetracycline resistance (71.9%). An increase in the fraction of strains resistant to macrolides and lincosamides indicates the need for monitoring both the susceptibility of these strains and the presence of the ST-17 clone.

Analysis of molecular characteristics of CAMP-negative Streptococcus agalactiae strains.

Streptococcus agalactiae can produce CAMP factor, which can promote the β-hemolysin activity of Staphylococcus aureus, forming an arrow-shaped hemolysis enhancement zone at the intersection of the two bacterial species on a blood agar plate. This characteristic feature of Streptococcus agalactiae has led to the widespread use of the CAMP test as an identification method. Vaginal/rectal swabs, collected from women at 35-37 weeks of pregnancy, were first inoculated into a selective enrichment broth media, then subcultured onto GBS chromogenic agar and 5% sheep blood agar sequentially. The VITEK-2 automatic identification system and MALDI-TOF MS were initially employed for identification, followed by the CAMP test. CAMP-negative strains underwent 16S rDNA and cfb gene sequence analysis, as well as bacterial multilocus sequence typing. A total of 190 strains were isolated, with 15 identified as CAMP-negative. Further 16S rDNA gene sequence analysis confirmed that all 15 strains were Streptococcus agalactiae. The MLST typing assay revealed that these 15 strains were of the ST862 type. The cfb gene was amplified and electrophoresed, but no specific fragments were found, indicating that these strains lack the CAMP factor due to cfb gene deletion. Antibiotic susceptibility tests demonstrated no resistance to penicillin, ampicillin, vancomycin and linezolid among the GBS strains. However, there are significant differences in resistance rates to tetracycline. This study found that 7.9% of GBS strains isolated from the vagina/rectum of pregnant women were CAMP-negative, suggesting that the CAMP test method or primers targeting the cfb gene should not be used as the sole presumptive test for GBS identification.

Microbiological monitoring of antibiotic resistance of strains of Streptococcus agalactiae among pregnant women

Group B Streptococcus (GBS) is the causative agent in 2–7% of all urinary tract infections (UTI), including asymptomatic bacteriuria (AB), cystitis, and pyelonephritis. We used the bacteriological quantitative method of sowing urine samples of pregnant women on Columbia agar with 5% lamb blood), Strepto B chromogenic agar and Todd Hewitt broth, identification of GBS strains with determination of sensitivity to bacitracin, hippurate hydrolysis test and CAMP test. Susceptibility to antimicrobial drugs was determined by the disk-diffusion method according to recommendations of the European Committee on Antimicrobial Susceptibility Testing. For the period of 2021, out of 3,356 urine samples of pregnant women, there were 149 samples of the investigated biomaterial with a positive result for GBS (resultancy – 4.4%). It was established that among the studied contingent, 41 strains of GBS were isolated in the first trimester of pregnancy (27.5%), in the second trimester – 59 strains (39.5%), in the third trimester – 49 strains (33.0%). It is established that out of the 149 strains of GBS, 38 strains (25.0%) were resistant to norfloxacin, 45 strains (30.0%) to erythromycin, 41 strains (28.0%) to clindamycin,125 strains (84.0%) to tetracycline, 20 strains (13.0%) to levofloxacin, 3 strains (2.0%) to nitrofurantoin. All strains of Streptococcus agalactiae were found to be sensitive to benzylpenicillin. The number of resistant strains of SGB identified from the urine of pregnant women was 47 strains. All resistant strains were resistant to at least three groups of antibacterial drugs, except beta-lactams. The results of the scientific research will allow us to obtain dynamic data on the antibiotic resistance of GBS strains in order to study the rate of development of antibiotic sensitivity of this microorganism. In the future, the research data can be used by scientists when reviewing protocols for the treatment of urinary tract infections in pregnant women.

Serotypes and antibiotic susceptibility of Streptococcus agalactiae strains isolated from invasive neonatal infections

The purpose of the study: to determine the serotypes and antibiotic susceptibility of strains of streptococcus agalactiae isolated from invasive neonatal infections. Material and methods: Fifty-five strains of Streptococcus agalactiae were selected isolated from invasive cerebrospinal fluid (CSF) and blood infections. The strains were confirmed by the Api 20Strep galleries and tested against the following antibiotics: penicillin G, ampicillin, erythromycin, clindamycin, tetracycline, ofloacin, levofloacin, gentamycin 500 μg, quinpristin-dalfopristin, chloramphenicol, vancomycin, interpretation was carried out according to the recommendations of the CLSI 2020 and kanamycin 1000 μg was interpreted according to the Ca-SFM standards. The serotyping of the strains was carried out according to the protocol of Monica Impéri Results: All strains were sensitive to penicillin G and ampicillin as well as vancomycin and fluoroquinolone; 92.7% are resistant to tetracyclines, 34.5% to erythromycin, 32.7% to clindamycin, 27.3% to chloramphenicol and 25.5% to kanamycin. The distribution of serotypes was as follows: serotype III is the majority with 56.4% followed by serotype Ia with 21.8% then serotype V with 14.5%. Conclusion: The strains of Streptococcus agalactiae remain sensitive to betalactamines, resistance to macrolides was around 35%, whereas that of aminoglycosides was 25%.Serotype III is the main serotype in invasive strains

Anti-biofilm Activity of Human Milk Oligosaccharides in Clinical Strains of Streptococcus agalactiae with Diverse Capsular and Sequence Types.

Group B Streptococcus (GBS) is an encapsulated Gram-positive bacterial pathogen that causes severe perinatal infections. Human milk oligosaccharides (HMOs) are short-chain sugars that have recently been shown to possess antimicrobial and anti-biofilm activity against a variety of bacterial pathogens, including GBS. We have expanded these studies to demonstrate that HMOs can inhibit and dismantle biofilm in both invasive and colonizing strains of GBS. A cohort of 30 diverse strains of GBS were analyzed for susceptibility to HMO-dependent biofilm inhibition or destruction. HMOs were significantly effective at inhibiting biofilm in capsular-type- and sequence-type-specific fashion, with significant efficacy in CpsIb, CpsII, CpsIII, CpsV, and CpsVI strains as well as ST-1, ST-12, ST-19, and ST-23 strains. Interestingly, CpsIa as well as ST-7 and ST-17 were not susceptible to the anti-biofilm activity of HMOs, underscoring the strain-specific effects of these important antimicrobial molecules against the perinatal pathogen Streptococcus agalactiae.

Comparative genome analysis of Streptococcus strains to identify virulent genes causing neonatal meningitis

To determine Streptococcus agalactiae genes responsible for causing neonatal meningitis. Streptococcus agalactiae strain 2603V/R is causative agent of neonatal meningitis, maternal infection and sepsis in young children. World health organisation reported high burden of new born death caused by this bacterium. Streptococcus agalactiae colonizing epithelial cells of vagina and endothelial cells have high resistance to available antibiotic drugs which makes it essential to determine new drug targets. To compare the genome of selected strain with the non-pathogenic strains of streptococcus and identify the virulent and antibiotic resistant genes for adaptation in host environment. The whole genome of human pathogen Streptococcus agalactiae strain 2603V/R was analysed and compared with Streptococcus dysgalactiae strains using visualization and annotation tools. Genomic islands, mobile genetic elements, virulent and resistant genes were studied. Genetically pathogenic strain is most similar to Streptococcus dysgalactiae subsp. equisimilis strain NCTC 7136. Comparative analysis revealed the importance of capsular polysaccharides and surface proteins responsible for avoiding immune system attachment to host epithelial cells and virulent behaviour. High number of genes coding for antibiotics resistance may provide a competitive advantage for survival of pathogenic Streptococcus agalactiae strain 2603V/R in its niche. The comparative analysis of pathogenic strain Streptococcus agalactiae with non-pathogenic strains of Streptococcus dysgalactiae provided new insights in pathogenicity that could aid in recognization for new regions and genes for development of new drug development strategies considering presence of high number of resistance genes.

Potential of Pm11 antimicrobial peptide against bovine mastitis pathogens

To investigate an alternative treatment for bovine mastitis by using Pm11 antimicrobial peptide. 5 bovine mastitis pathogens that were previously isolated from cows affected by either clinical or subclinical mastitis. The current study introduces Pm11 antimicrobial peptide as an alternative treatment for bovine mastitis. The antibacterial activity of Pm11 was tested against Escherichia coli strain SCM1249, Klebsiella spp strain SCM1282, Staphylococcus aureus strain CM967, Streptococcus agalactiae strain SCM1084, and Streptococcus uberis strain SCM1310 using minimum bactericidal concentrations (MBCs) and time-kill kinetics. The pathogens' morphological changes were demonstrated using a scanning electron microscope (SEM). The cytotoxicity of Pm11 was assessed using the minimum hemolytic concentration assay. MBCs ranged from 2.5 to 10 μM and IC50 ranged from 0.32 to 2.07 μM. Time-kill kinetics at MBC demonstrated that Pm11 reduced viable cell counts of S agalactiae strain SCM1084 and S uberis strain SCM1310 from 105 to 0 CFU/mL within 1 h. E coli strain SCM1249 and S aureus strain CM967 were reduced from 105 to 0 CFU/mL within 4 h. The average Pm11-induced hemolytic activity was < 10% for all Pm11 concentrations tested except at the maximum concentration tested (160 μM: 10.19 ± 2.29%). Based on SEM, Pm11 induced morphological and cellular changes in S aureus and E coli. Pm11 antimicrobial peptide demonstrated in vitro antibacterial activity against the common bovine mastitis pathogens E coli, S aureus, S agalactiae, and S uberis, except Klebsiella spp, and should be further investigated in vivo.

Clonal Complex 12 Serotype Ib Streptococcus agalactiae Strain Causing Complicated Sepsis in Neonates: Clinical Features and Genetic Characteristics.

Streptococcus agalactiae (group B Streptococcus [GBS]) is well known to cause serious diseases in infants. A serotype Ib GBS strain has recently emerged and become prevalent in Southeast Asia. We aimed to investigate the clinical and genetic characteristics of this strain. All neonates with invasive GBS diseases from a tertiary-level medical center in Taiwan between 2003 and 2020 were analyzed. The capsule serotyping, multilocus sequence typing, and antimicrobial resistance analyses were performed on all the invasive GBS isolates, and whole-genome sequencing (WGS) was performed specifically on the type Ib GBS strain. A total of 188 neonates with invasive GBS disease during the study period were identified. The type Ib GBS strain accounted for 7.4% (n = 14) of neonatal GBS invasive diseases. Almost all type Ib GBS isolates belonged to sequence type 12 (13/14, 92.9%) and clonal complex 12. Neonates with type Ib GBS disease had a significantly higher rate of complicated sepsis (10/14, 71.4%; P < 0.05) and sepsis-attributable mortality (6/14, 42.9%; P < 0.05). Additionally, type Ib GBS isolates had significantly higher rates of resistance to erythromycin and clindamycin (both 100%; P < 0.05) than other GBS serotypes. WGS revealed the presence of an ~75-kb integrative and conjugative element, ICESag37, comprising multiple antibiotic resistance and virulence genes, and PI-1 plus PI-2a were noted in all type Ib serotype 12 (ST12) GBS isolates; these isolates may be responsible for its high invasiveness and antimicrobial resistance rates. The genomic characteristics of the type Ib clonal complex 12 (CC12) GBS strain may account for the high illness severity associated with this strain and its antibiotic resistance. Continuous monitoring and advanced strategies to control the spread of type Ib CC12 GBS should be considered. IMPORTANCE A type Ib ST12 GBS strain is not a common isolate in neonatal invasive diseases and has been ignored for a long time. However, the recent literature and our data showed that such a GBS strain is associated with a significantly higher risk of severe sepsis, higher illness severity, and a significantly higher rate of sepsis-attributable mortality. This study found a novel gene cluster, including the presence of ICESag37 and specific pilus genes, carrying multiple antimicrobial resistance and virulence genes, which may be responsible for the clinical characteristics. Because of the higher mortality and severity of illness, we concluded that continuous monitoring of the type Ib ST12 GBS strain is warranted in the future.

Attenuated Streptococcus agalactiae WC1535 ∆Sia perturbs the gut microbiota of Oreochromis niloticus, massively colonizes the intestine, and induces intestinal mucosal immunity after intraperitoneal inoculation.

We previously developed and assessed the effectiveness of the attenuated Streptococcus agalactiae (Group B Streptococcus, GBS) strain WC1535 ∆Sia (with neuA-D gene cluster deletion) vaccine in tilapia (Oreochromis niloticus). In this study, we characterized the bacterial communities of the tilapia intestines by 16S rRNA high-throughput sequencing and assessed the serum antibody response, expression of immune-related genes, and histological changes following formalin-killed GBS vaccine (FKV) and the live attenuated vaccine ∆Sia (LAV). Results showed that FKV and LAV induced robust systemic and intestinal mucosal immune responses in tilapia without causing obvious pathological changes in the hindgut, spleen, and head kidney but exerted different effects on intestinal bacterial communities. The richness or diversity of the intestinal bacterial community of FKV tilapia showed no significant changes compared with that of the control fish (p > 0.05) at either day 21 post-initial vaccination (21 dpiv) or day 35 (day 14 after the second immunization) (35 dpiv). The community composition of FKV tilapia and controls was significantly similar, although the relative abundance of some genera was significantly altered. Relative to control fish, the gut ecosystem of LAV tilapia was significantly disturbed with a substantial increase in community diversity at 21 dpiv (p < 0.05) and a significant decrease at 35 dpiv in fish with high serum antibody response (ΔSia35H) (p < 0.05). However, there was no significant difference between ΔSia35H and ΔSia35L (low serum antibody response) fish (p > 0.05). Moreover, the community composition of LAV tilapia at 21 dpiv or 35 dpiv was considerably different from that of the controls. Particularly, GBS ∆Sia was found to be abundant in the intestine at 21 and 35 dpiv. This result suggested that the parenteral administration of the LAV (∆Sia) may also have the effect of oral vaccination in addition to the immune effect of injection vaccination. In addition, a significant correlation was found between the expression of immune-related genes and certain bacterial species in the intestinal mucosal flora. Our findings will contribute to a better understanding of the effects of inactivated and attenuated vaccines on gut microbiota and their relationship with the immune response.

Correlation Study of Alcoholic Clove extract against Streptococcus agalactiae virulence factors isolated from Neonatal patients

Plant extracts promise natural sources of antivirulence chemicals thanks to their broad availability and various phytochemical antivirulence components. The main antivirulence components of plants are essential oils. 130 isolates were obtained from neonates (≤1 year old) from February, 2020 to February, 2021. All studied isolates were diagnosed by traditional methods like biochemical tests, Vitek-2 system, and 16srRNA using conventional PCR. Profile of antibiotics resistance ratio for Streptococcus agalactiae were Penicillin 20%, Clindamycin 68%, Chloramphenicol 46%, Tetracycline 82%, Erythromycin 100%, Imipenem 12%, and Ciprofloxacillin 26.47%. All genes (Cyl E, Spb1, and bibA) were detected by conventional PCR and the molecular frequency showed 64% of strains positive for Cyl E gene and 56% of strains harbored bibA genes, while 48% of Streptococcus agalactiae contain Spb1 gene. This research showed the coexistence of bla Cyl E, Spb 1 and bibA genes in 48% strains of Streptococcus agalactiae. Extraction of clove extract was carried out using methanol and it analysed using GC-MS to detect active compounds, such as Eugenol (60.10%). The effect of clove extract against biofilm formation, haemolysin, and protease activity was significant (p-value &lt; 0.01).

CRISPR Contributes to Adhesion, Invasion, and Biofilm Formation in Streptococcus agalactiae by Repressing Capsular Polysaccharide Production.

ABSTRACTThe clustered regularly interspaced palindromic repeat (CRISPR)-associated (Cas) system functions classically as a prokaryotic defense system against invading mobile genetic elements, such as phages, plasmids, and viruses. Our previous study revealed that CRISPR deletion caused increased transcription of capsular polysaccharide (CPS) synthesis-related genes and severely attenuated virulence in the hypervirulent piscine Streptococcus agalactiae strain GD201008-001. Here, we found that CRISPR deficiency resulted in reduced adhesion, invasion, and biofilm formation abilities in this strain by upregulating the production of CPS. However, enhanced CPS production was not responsible for the attenuated phenotype of the ΔCRISPR mutant. RNA degradation assays indicated that inhibited transcription of the cps operon by CRISPR RNA (crRNA) was not due to the base pairing of the crRNA with the cps mRNA but to the repression of the promoter activity of cpsA, which is a putative transcriptional regulator of the capsule locus.IMPORTANCE Beyond protection from invading nucleic acids, CRISPR-Cas systems have been shown to have an important role in regulating bacterial endogenous genes. In this study, we demonstrate that crRNA inhibits the transcription of the cps operon by repressing the activity of promoter PcpsA, leading to increases in the abilities of adhesion, invasion, and biofilm formation in S. agalactiae. This study highlights the regulatory role of crRNA in bacterial physiology and provides a new explanation for the mechanism of crRNA-mediated endogenous gene regulation in S. agalactiae.

Small Things Matter: The 11.6-kDa TraB Protein is Crucial for Antibiotic Resistance Transfer Among Enterococci.

Conjugative transfer is the most important means for spreading antibiotic resistance genes. It is used by Gram-positive and Gram-negative bacteria, and archaea as well. Conjugative transfer is mediated by molecular membrane-spanning nanomachines, so called Type 4 Secretion Systems (T4SS). The T4SS of the broad-host-range inc18-plasmid pIP501 is organized in a single operon encoding 15 putative transfer proteins. pIP501 was originally isolated from a clinical Streptococcus agalactiae strain but is mainly found in Enterococci. In this study, we demonstrate that the small transmembrane protein TraB is essential for pIP501 transfer. Complementation of a markerless pIP501∆traB knockout by traB lacking its secretion signal sequence did not fully restore conjugative transfer. Pull-downs with Strep-tagged TraB demonstrated interactions of TraB with the putative mating pair formation proteins, TraF, TraH, TraK, TraM, and with the lytic transglycosylase TraG. As TraB is the only putative mating pair formation complex protein containing a secretion signal sequence, we speculate on its role as T4SS recruitment factor. Moreover, structural features of TraB and TraB orthologs are presented, making an essential role of TraB-like proteins in antibiotic resistance transfer among Firmicutes likely.

Bacteriocin Producing Streptococcus agalactiae Strains Isolated from Bovine Mastitis in Brazil.

Antibiotic resistance is one of the biggest health challenges of our time. We are now facing a post-antibiotic era in which microbial infections, currently treatable, could become fatal. In this scenario, antimicrobial peptides such as bacteriocins represent an alternative solution to traditional antibiotics because they are produced by many organisms and can inhibit bacteria, fungi, and/or viruses. Herein, we assessed the antimicrobial activity and biotechnological potential of 54 Streptococcus agalactiae strains isolated from bovine mastitis. Deferred plate antagonism assays revealed an inhibition spectrum focused on species of the genus Streptococcus—namely, S. pyogenes, S. agalactiae, S. porcinus, and S. uberis. Three genomes were successfully sequenced, allowing for their taxonomic confirmation via a multilocus sequence analysis (MLSA). Virulence potential and antibiotic resistance assessments showed that strain LGMAI_St_08 is slightly more pathogenic than the others. Moreover, the mreA gene was identified in the three strains. This gene is associated with resistance against erythromycin, azithromycin, and spiramycin. Assessments for secondary metabolites and antimicrobial peptides detected the bacteriocin zoocin A. Finally, comparative genomics evidenced high similarity among the genomes, with more significant similarity between the LGMAI_St_11 and LGMAI_St_14 strains. Thus, the current study shows promising antimicrobial and biotechnological potential for the Streptococcus agalactiae strains.

Identifying large-scale recombination and capsular switching events in Streptococcus agalactiae strains causing disease in adults in the UK between 2014 and 2015.

Cases of invasive group B streptococcal infection in the adult UK population have steadily increased over recent years, with the most common serotypes being V, III and Ia, but less is known of the genetic background of these strains. We have carried out in-depth analysis of the whole-genome sequences of 193 clinically important group B Streptococcus (GBS) isolates (184 were from invasive infection, 8 were from non-invasive infection and 1 had no information on isolation site) isolated from adults and submitted to the National Reference Laboratory at the UK Health Security Agency between January 2014 and December 2015. We have determined that capsular serotypes III (26.9%), Ia (26.4%) and V (15.0%) were most commonly identified, with slight differences in gender and age distribution. Most isolates (n=182) grouped to five clonal complexes (CCs), CC1, CC8/CC10, CC17, CC19 and CC23, with common associations between specific serotypes and virulence genes. Additionally, we have identified large recombination events mediating potential capsular switching events between sequence type (ST)1 serotype V and serotypes Ib (n=2 isolates), II (n=2 isolates) and VI (n=2 isolates); between ST19 serotype III and serotype V (n=5 isolates); and between CC17 serotype III and serotype IV (n=1 isolate). The high genetic diversity of disease-causing isolates and multiple recombination events reported in this study highlight the need for routine surveillance of the circulating disease-causing GBS strains. This information is crucial to better understand the global spread of GBS serotypes and genotypes, and will form the baseline information for any future GBS vaccine research in the UK and worldwide.

Molecular determination, serotyping, antibiotic profile and virulence factors of group B Streptococcus isolated from invasive patients at Arabcare Hospital Laboratory, Palestine

BackgroundStreptococcus agalactiae (group B Streptococcus) is beta-hemolytic, catalase negative, gram-positive cocci, recognized as main bacterial pathogen causing infections in newborns, infants, adults, and elderly people around the world. The aim of this study is to investigate group B Streptococcus samples recovered from invasive patients and determine serotype, virulent genes, and antibiotic-resistant profile of Streptococcus agalactiae in Palestine. MethodsA total of 95 group B Streptococcus strains were isolated from neonates, infants, pregnant and non-pregnant women and males at Arabcare Hospital Laboratory, Palestine, between the period of June 2018 and September 2020. Species identification was carried out through cultivation and conventional biochemical tests. A conventional Polymerase Chain Reaction (cPCR) was used to determine the 5 serotypes and virulent genes of the Streptococcus agalactiae strains. The antibiotic resistance test of group B Streptococcus was evaluated using Kirby-Bauer disk susceptibility. Sequencing and BLAST analysis were used to determine the relationship of the isolates in this study to worldwide isolates. ResultsSerotype III (35%) was the major group B Streptococcus strains serotype causing invasive infections in neonates, infants, pregnant and nonpregnant women, and males, followed by serotypes V (19%), Ia, and II (15%), Ib (6%), respectively. All our isolates encoding for surface protein virulent factors, including a highly virulent gene (HvgA) were mostly found in strains isolated from pregnant women (12%). These group B Streptococcus strains exhibited a high rate of resistance to clindamycin (26%). The overall percentage of levofloxacin resistance was 11%, while vancomycin and ampicillin showed higher resistance, at 14.7 and 16% respectively. In addition, the phylogenetic relationship dendrogram illustrates that Streptococcus agalactiae isolated from an invasive patient (newborn) in Palestine was similar to strains found in China and Japan. ConclusionsThe outcomes of this study demonstrate that resistant group B Streptococcus strains are common in Palestine, therefore, evidence-based infection prevention and antibiotic stewardship efforts are necessary.

Genomic Characterization of Serotype III/ST-17 Group B Streptococcus Strains with Antimicrobial Resistance Using Whole Genome Sequencing.

Background: Antibiotic-resistant type III/ST-17 Streptococcus agalactiae (group B Streptococcus, GBS) strain is predominant in neonatal invasive GBS diseases. We aimed to investigate the antibiotic resistance profiles and genetic characteristics of type III/ST-17 GBS strains. Methods: A total of 681 non-duplicate GBS isolates were typed (MLST, capsular types) and their antibiotic resistances were performed. Several molecular methods (WGS, PCR, sequencing and sequence analysis) were used to determine the genetic context of antibiotic resistant genes and pili genes. Results: The antibiotic resistant rates were significantly higher in type Ib (90.1%) and type III (71.1%) GBS isolates. WGS revealed that the loss of PI-1 genes and absence of ISSag5 was found in antibiotic-resistant III/ST-17 GBS isolates, which is replaced by a ~75-kb integrative and conjugative element, ICESag37, comprising multiple antibiotic resistance and virulence genes. Among 190 serotype III GBS isolates, the most common pilus island was PI-2b (58.4%) alone, which was found in 81.3% of the III/ST-17 GBS isolates. Loss of PI-1 and ISSag5 was significantly associated with antibiotic resistance (95.5% vs. 27.8%, p < 0.001). The presence of ICESag37 was found in 83.6% of all III/ST-17 GBS isolates and 99.1% (105/106) of the antibiotic-resistant III/ST-17 GBS isolates. Conclusions: Loss of PI-1 and ISSag5, which is replaced by ICESag37 carrying multiple antibiotic resistance genes, accounts for the high antibiotic resistance rate in III/ST-17 GBS isolates. The emerging clonal expansion of this hypervirulent strain with antibiotic resistance after acquisition of ICESag37 highlights the urgent need for continuous surveillance of GBS infections.

Analysis of Susceptibility to the Antimicrobial and Anti-Biofilm Activity of Human Milk Lactoferrin in Clinical Strains of Streptococcus agalactiae With Diverse Capsular and Sequence Types.

Group B Streptococcus (GBS) is one of the leading infection-related causes of adverse maternal and neonatal outcomes. This includes chorioamnionitis, which leads to preterm ruptures of membranes and can ultimately result in preterm or stillbirth. Infection can also lead to maternal and neonatal sepsis that may contribute to mortality. Currently, treatment for GBS infection include a bolus of intrapartum antibiotic prophylaxis to mothers testing positive for GBS colonization during late pregnancy. Lactoferrin is an antimicrobial peptide expressed in human breast milk, mucosal epithelia, and secondary granules of neutrophils. We previously demonstrated that lactoferrin possesses antimicrobial and antibiofilm properties against several strains of GBS. This is largely due to the ability of lactoferrin to bind and sequester iron. We expanded upon that study by assessing the effects of purified human breast milk lactoferrin against a panel of phenotypically and genetically diverse isolates of GBS. Of the 25 GBS isolates screened, lactoferrin reduced bacterial growth in 14 and biofilm formation in 21 strains. Stratifying the data, we observed that colonizing strains were more susceptible to the growth inhibition activity of lactoferrin than invasive isolates at lactoferrin concentrations between 250-750 µg/mL. Treatment with 750 µg/mL of lactoferrin resulted in differences in bacterial growth and biofilm formation between discrete sequence types. Differences in bacterial growth were also observed between capsular serotypes 1a and III. Maternally isolated strains were more susceptible to lactoferrin with respect to bacterial growth, but not biofilm formation, compared to neonatal sepsis isolates. Finally, high biofilm forming GBS strains were more impacted by lactoferrin across all isolates tested. Taken together, this study demonstrates that lactoferrin possesses antimicrobial and antibiofilm properties against a wide range of GBS isolates, with maternally isolated colonizing strains being the most susceptible.

Features of acute purulent lymphadenites and maxillofacial adenophlegmons course in children

Objective. The purpose of the research was to study the features of the course of acute purulent lymphadenites and adenophlegmons of the maxillofacial region in children, to describe the species composition of pathogens, to determine their sensitivity to antibiotics of various groups.&#x0D; Materials and methods. The medical histories of 147 patients were analyzed; microbiological studies were carried out (microscopic examination of clinical specimens with gram stain, cultural method, determination of sensitivity of isolated pathogen to antimicrobial drugs).&#x0D; Results. Most often, acute purulent lymphadenites and adenophlegmons were observed in the submandibular tissue space (40.1 %) and in the neck (38.1 %). The main causative agents of the inflammatory process were Staphylococcus aureus (38.1 %) and Streptococcus agalactiae (BHSA) (14.3 %). Staphylococcus aureus strains had the highest sensitivity to beta-lactam antibiotics in 83 %, to macrolides in 60.4 %, to fluoroquinolones in 56.6 %; 3.8 % showed resistance to lincomycin, 1.9 % to beta-lactams. The strains of Streptococcus agalactiae (BHSA) were 100 % sensitive to beta-lactams, 73.7 % to macrolides, they were resistant to lincomycin and macrolides (10.5 % each).&#x0D; Conclusions. The data obtained are of great practical importance, since they allow to correctly orient doctors in the choice of rational antibiotic therapy.

Novel molecular diagnosis of Cyl E, Spb 1 and bib A virulence genes of Streptococcus agalactiae from pregnant women

Group B Streptococcus is a gram-positive coccal bacterium which produces an arrow zone of blood hemolysis on blood agar. Group B Streptococcus is the most common cause of invasive bacterial illness in neonates, including sepsis, meningitis, septicemia, and pneumonia. For the first time, such a study of Streptococcus agalactiae has been conducted in hospitals of Anbar Governorate .All Streptococcus agalactiae identified by conventianal methods , automated methods using Vitek-2 system , and molecular methods by conventional PCR. All isolates gave a positive result as 16S rRNA. Distribution of resistance ratio for Streptococcus agalactiae were Penicillin 20% , Clindamycin 68% , Chloramphenicol % 46 , Tetracycline 82% , Erythromycin % 100 ,Imipenem % 12 , and Ciprofloxacillin 26.47%. The Cyl E and bibA genes were detected by conventional PCR and the result showed (72%) strains positive, but in case of bla Cyl E gene (53.48%) of strains harbored bla bibA genes , while 48.83 of Streptococcus agalactiae contain Spb1 gene . This approach showed the coexistence of bla Cyl E , Spb 1 and bibA genes in 48.83% strains of Streptococcus agalactiae.

Antimicrobial, modulatory, and antibiofilm activity of tt-farnesol on bacterial and fungal strains of importance to human health

In this study, we analyzed the antimicrobial, antibiofilm, and modulatory activities of trans–trans-farnesol (tt-farnesol). The minimum inhibitory concentration (MIC) of this sesquiterpene was evaluated against 31 Gram-positive and Gram-negative bacterial strains and 4 species of the genus Candida. Furthermore, we examined its inhibitory action on biofilm production as well as antibiotic modulation. Only Gram-positive species presented susceptibility to tt-farnesol (MIC ranging from 8 µg/mL to 128 µg/mL). No synergistic or antagonistic effects were observed between tt-farnesol (1/4 and 1/8 of MIC) and first-choice antibiotics against multidrug resistant strains. However, the modulatory action of tt-farnesol (1/2 and 1/4 of the MIC) decreased 8 × MIC of non-inhibitory β-lactam antibiotic against a Methicillin-resistant strain. In the antibiofilm assay, tt-farnesol inhibited biofilm formation, especially in Methicillin-resistant Staphylococcus aureus (MRSA) strains, at concentrations ranging from 2 μg/mL to 128 μg/mL. Additionally, in the molecular docking study, the tt-farnesol molecule demonstrated a remarkable binding affinity with important proteins involved in the biofilm production, such as IcaA and Srt proteins. The antimicrobial action of tt-farnesol on Streptococcus pyogenes and Streptococcus agalactiae strains was evaluated for the first time, presenting an MIC of 16 µg/mL for both strains. Our findings reveal the antibacterial, antibiofilm, and modulatory potential of tt-farnesol to aid in the fight against infectious processes.