Detection Of Virulence Genes Research Articles

Detection of Virulence Factors and Antibiotic Resistance among Klebsiella pneumoniae Isolates from Iran.

The current study assessed the detection of virulence genes and drug resistance among Klebsiella pneumoniae isolates from Iran. During 2018 to 2020, 52 K. pneumoniae isolates were obtained from patients at Iran hospitals. By disk diffusion method, the antimicrobial susceptibility of K. pneumoniae isolates was assessed, and ESBL-producing K. pneumoniae isolates were detected by CDDT method. PCR analysis was done to detect virulence genes (iucB, iutA, iroN, kfu, allS, fimH, ybtS, mrkD, and entB); ESBL-encoding genes (bla TEM, bla PER, bla CTX-M, bla VEB, and bla SHV); and class D (bla OXA-48), class B (bla VIM, bla NDM, and bla IMP), and class A (bla KPC and bla GES) carbapenemase genes. Among all isolates, 84.6%, 13.5%, and 1.9% isolates were multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR), respectively. Also, 84.6% were ESBL-producing and 71.2%, 53.8%, 40.4%, and 9.6% of all isolates were carrying bla TEM, bla SHV, bla CTX-M, and bla OXA-48 genes, respectively. Six isolates (11.5%) were positive for bla NDM gene. In contrast, no isolates were positive for the presence of bla KPC, bla IMP, and bla VIM. Virulence factor genes including iucB, iutA, iroN, kfu, allS, fimH, ybtS, mrkD, and entB were carried by 24%, 46.2%, 25%, 11.5%, 17.3%, 86.5%, 75%, 88.5%, and 100% isolates, respectively. This study evaluated the distribution and prevalence of virulence factor genes among K. pneumoniae isolates. The treatment of these infections is challenging due to the existence of particular virulence factors and the rise of antibiotic resistance. Therefore, the current study accentuates the necessity of finding new and efficient solutions for stopping the increase of antibiotic resistance.

Genetic relationship, antibiotic resistance pattern and virulence factors of Klebsiella pneunoniae strains isolated from meningitis patients

Backgroud: Meningitis due to Klebsiella pneunoniae is increasingly reported from several Asian countries. Microbiological characteristic of K. pneumoniae strains causing meningitis is not yet explored in Iran. The aim of this study was to demonstrate antibiotic resistance pattern and virulence factors, as well as the genetic relationship of K. pneumoniae strains isolated from meningitis patients. Materials and Mehtods: Eight K. pneumoniae isolates were collected from hospitalized patients at Imam Khomeini hospital, Tehran-Iran from 14 May 2018 to 15 Mar 2020. The antibiotic resistance pattern was determined by disc diffusion method. Antibiotic resistance genes and virulence-associated genes were traced by Polymerase Chain Reaction. Repetitive Extragenic Palindromic Polymerase Chain Reaction was used for evaluation of the genetic relationships among isolates. Findings: Six out of eight isolates were resistant to almost all the 15 antibiotics tested. These six isolates harbored triple antibiotic resistance genes: blaOXA-48, blaSHV, and aac (6’)-Ib. Five of these isolates co-harbored aac (3)-IIa gene. Besides, blaTEM and blaCTXM-1 were detected in five and three isolates respectively. blaNDM-1 was identified in one isolate. All isolates harbored acrAB, ompK36, and tolC resistance genes. None of the isolates were related to K1 or K2 capsular serotypes. The most commonly detected virulence genes were entB (100%), mrkD (100%), ybtS (62.5%) and kfu (25%). rep-PCR fingerprinting discriminated seven isolates into three clusters. Conclusion: Simulations presence of the genes coding for Extended Spectrum Beta Lactamases, Carbapenemases, and Aminoglycoside Modifying Enzymes narrows therapeutic alternatives and imposes a heavy load on the public health system. Antibiotic susceptibility test is recommended however, Carbapenems can still be considered as first-line medications for preliminary empirical treatment of K. pneumoniae meningitis before obtaining susceptibility test results. Low genetic diversity suggests the circulation of certain K. pneumoniae clones in hospital which highlights the establishing of effective infection observation and prevention program.

Molecular Characterization of Enterococcus faecalis and Enterococcus faecium Isolated from a Meat Source in Shahrekord Local Markets, Iran.

Enterococcus faecalis (E. faecalis) and E. faecium (E. faecium) are commensals of the gastrointestinal biota of humans and animals and are considered opportunistic pathogens. This study aimed to improve the knowledge about E. faecalis and E. faecium isolated from meat. To this end, 104 meat samples were collected from sheep, goats, cattle, and calves from Shahrekord local markets in Iran. Presumptive colonies of E. faecalis and E. faecium were isolated from the samples and subjected to identification tests. Antimicrobial susceptibility was determined using the Kirby-Bauer disc diffusion method, and polymerase chain reaction (PCR) was performed to detect some virulence genes. Furthermore, randomly amplified polymorphic DNA typing and repetitive element sequence-based PCR fingerprinting were used to show the clonal relatedness of the isolates. The results revealed that enterococci were present in 90 out of 104 samples. Specifically, E. faecalis and E. faecium were the commonly isolated species, with the predominance of E. faecalis, which exhibited high resistance to streptomycin (95%) but was susceptible to vancomycin (85.6%). Virulence genes detection showed that ccf and cpd genes were the most prevalent genes in both species. In addition, the molecular typing method indicated that the isolates belonged to separate subgroups. This study shows the contamination of meat products by potential pathogens and resistant enterococci. There is a need to implement regular surveillance to monitor the emergence of antimicrobial-resistant E. faecalis and E. faeciumin food, particularly in meat production.

Carbapenem-resistant hypermucoviscous Klebsiella pneumoniae clinical isolates from a tertiary hospital in China: Antimicrobial susceptibility, resistance phenotype, epidemiological characteristics, microbial virulence, and risk factors.

Hypervirulent and multidrug-resistant Klebsiella pneumoniae poses a significant threat to public health. We aimed to determine the common carbapenemase genotypes and the carriage patterns, main antibiotic resistance mechanisms, and in vitro susceptibility of clinical isolates of carbapenem-resistant K. pneumoniae (CRKP) to ceftazidime/avibactam (CZA) for the reasonable selection of antimicrobial agents and determine whether hypermucoviscous (HMV) phenotype and virulence-associated genes are key factors for CRKP colonization and persistence. Antibiotics susceptibility of clinical CRKP isolates and carbapenemase types were detected. CRKP isolates were identified as hypermucoviscous K. pneumoniae (HMKP) using the string test, and detection of virulence gene was performed using capsular serotyping. The bla KPC-2, bla NDM, bla IMP, and/or bla OXA-48-like were detected in 96.4% (402/417) of the isolates, and the bla KPC-2 (64.7%, 260/402) was significantly higher (P<0.05) than those of bla NDM (25.1%), bla OXA-48-like (10.4%), and bla IMP (4.2%). Carriage of a single carbapenemase gene was observed in 96.3% of the isolates, making it the dominant antibiotic resistance genotype carriage pattern (P < 0.05). Approximately 3.7% of the isolates carried two or more carbapenemase genotypes, with bla KPC-2 + bla NDM and bla NDM + bla IMP being the dominant multiple antibiotic resistance genotype. In addition, 43 CRKP isolates were identified as HMKP, with a prevalence of 10.3% and 2.7% among CRKP and all K. pneumoniae isolates, respectively. Most clinical CRKP isolates were isolated from elderly patients, and carbapenemase production was the main mechanism of drug resistance. Tigecycline and polymyxin B exhibited exceptional antimicrobial activity against CRKP isolates in vitro. Furthermore, bla KPC-2, bla NDM, and bla OXA-48-like were the main carbapenemase genes carried by the CRKP isolates. CZA demonstrated excellent antimicrobial activity against isolates carrying the single bla KPC-2 or bla OXA-48-like genotype. Capsular serotype K2 was the main capsular serotype of the carbapenem-resistant HMKP isolates. Survival rates of Galleria mellonella injected with K. pneumoniae 1-7 were 20.0, 16.7, 6.7, 23.3, 16.7, 3.3, and 13.3, respectively. Therefore, worldwide surveillance of these novel CRKP isolates and carbapenem-resistant HMKP isolates as well as the implementation of stricter control measures are needed to prevent further dissemination in hospital settings.

Sewage from a secondary hospital in Ribeirão Preto, southeastern Brazil: a source of multidrug-resistant Enterobacteriaceae.

Antimicrobial resistance is one of the severe threats to global health. Hospital sewage can serve as a reservoir for multi-resistant bacteria and promote the spread of antimicrobial resistance. This study aimed to investigate the antimicrobial susceptibility and the pathogenic potential of Enterobacteriaceae isolated from the sewage of a secondary hospital in Ribeirão Preto, a city in southeastern Brazil. The strains were isolated by membrane filtration and identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF). The antimicrobial susceptibility profile was performed by disk diffusion. Polymerase chain reaction (PCR) assays were used to detect virulence genes among the strains. Twenty-eight isolates were obtained, with Klebsiella pneumoniae being the predominant species (71.4%, n = 20). All isolates were classified as multidrug-resistant, including four isolates that were non-susceptible to at least 50% of the tested antibiotics. All isolates were also non-susceptible to cefuroxime and sulfonamides antibiotics; however, they were susceptible to norfloxacin, ofloxacin, amikacin, gentamicin, netilmicin, ertapenem, cefazolin, cefaclor, and cefotetan. The virulence genes ycfM, fimH, mrkD, kfu, and entB were detected in several isolates. Our study showed that even in a secondary hospital, without the routine of major surgeries and intensive care admissions, the hospital sewage can harbor a high percentage of multidrug-resistant bacteria with pathogenic potential. This leads to the worrying risk of public health and environmental contamination.

Staphylococcus coagulans possesses many virulence factors of Staph. aureus and Staph. pseudintermedius.

To understand the Staphylococcus coagulans prevalence in causing skin infections in dogs and detection of various virulence genes in Staph. coagulans isolates. Staph. coagulans was isolated from pus swabs collected from dogs with skin infection and identified by detecting thermonuclease, coagulase, and urease genes. The presence of methicillin-resistant gene (mecA) was performed by PCR. Antimicrobial susceptibility test was carried out by disc diffusion method. In total, 38 Staph. coagulans clinical isolates and 42 Staph. coagulans genomes available in NCBI database were screened for 19 virulence genes by PCR and in silico prediction, respectively. A prevalence of 13.8% (38/275) of Staph. coagulans dog skin infection was observed and 15.8% (6/38) of Staph. coagulans isolates carried mecA gene. Many Staph. coagulans isolates were susceptible to all tested antimicrobials. Twenty nine per cent isolates were resistant to ciprofloxacin. Genes encoding leukotoxins, DNase, exfoliative toxin, superantigen-like exotoxin, immunoglobulin-binding proteins, fibrinogen-binding proteins, autolysin, and rod shape-determining protein were detected in almost all the Staph. coagulans clinical isolates and genomes from NCBI database, whereas anti-adhesin plasma-sensitive protein genes were present in relatively lesser number of Staph. coagulans clinical isolates and genomes from NCBI database. Staph. coagulans possesses many virulence factors that are present in other coagulase-positive staphylococci, such as Staph. aureus and Staph. pseudintermedius. The presence of two bi-component leukotoxin genes in tandem with other virulence factor genes in a single pathogenic island in the Staph. coagulans genomes explained their eminence in the virulence of Staph. coagulans causing infections. Staph. coagulans was classified as a separate species in the year 2020 and primarily causes skin infections in dogs. Identification of this species is not included in any of the automated bacterial identification systems. Hence, many veterinary laboratories do not have a strategy to identify this bacterium. This study will help in the identification of Staph. coagulans in veterinary laboratories by PCR apart from detecting various virulence factors present in this pathogen. The existence of many virulence factors and prevalence in different animals in varied geographical locations suggest that Staph. coagulans is an important coagulase-positive staphylococcal pathogen in animals.

Prevalence and genetic diversity of Aeromonas veronii isolated from aquaculture systems in the Poyang Lake area, China.

The area around Poyang Lake is the main aquaculture area in Jiangxi Province, China, and an important base for the supply of freshwater aquatic products. Aquaculture in the Poyang Lake area is severely threatened by diseases caused by bacterial pathogens, and Aeromonas veronii has been the main pathogen in recent years. In this paper, ERIC-PCR genotyping, virulence gene and antimicrobial resistance gene detection, and drug susceptibility tests were carried out on 46 A. veronii isolates obtained from aquaculture systems in the Poyang Lake area from 2016 to 2020. The results showed that the A. veronii strains in the Poyang Lake area had high genetic diversity, and 46 strains produced 36 ERIC genotypes. There were no geographical and temporal differences in the cluster analysis results and no dominant clones. All 13 virulence genes tested were detected, and all isolates had harbored 2 or more virulence genes, with a maximum of 12 virulence genes detected. Among the 22 antimicrobial resistance genes selected, 15 were detected; 97.8% of the isolates contained 2 or more antimicrobial resistance genes, with a maximum of 9 antimicrobial resistance genes. Drug susceptibility tests showed that some strains were resistant to a variety of traditionally effective drugs for Aeromomas, such as enrofloxacin and florfenicol. This study provides a reference for exploring the impact of aquaculture in the Poyang Lake area on public health.

Prevalence of Helicobacter pylori Virulence Genes and Their Association with Chronic Gastritis in Beijing, China

Helicobacter pylori is closely related to chronic gastritis. The aim of the study was to investigate the correlation between H. pylori virulence genes and chronic gastritis in order to determine the pathogenic role of H. pylori virulence genes in chronic gastritis. Gastric mucosal tissues were obtained from 142 patients with chronic gastritis at three Beijing hospitals. The presence of virulence genes was determined by polymerase chain reaction (PCR) from H. pylori DNA. Multilocus sequence typing (MLST) and a phylogenetic tree were performed to characterize the overall genetic diversity. 91 new sequence types were identified by MLST in this study, and all strains showed high genetic diversity. The H. pylori isolates were divided into three types: hspEAsia strains (61 strains), hpEurope strains (15 strains), and mixed strains (16 strains). Some virulence genes were found to be significantly different between strains. The highest positive rates were found for dupA in chronic atrophic gastritis (AG), iceA1 in chronic non-atrophic gastritis with erosions, and iceA2 in chronic non-atrophic gastritis. The presence of dupA was found to be inversely related to the risk of AG. The H. pylori strains display high genetic diversity. Some virulence genes were found to be significantly different between diseases. The detection of various virulence genes is critical for screening high-risk populations for precancerous lesions and for the early prevention and control of gastric cancer.

Distribution of emm, superantigen and other virulence genotypes and detection of phylogenetic relationships in group A streptococcal isolates.

Group A streptococci are important pathogens with various virulence factors, such as M protein, superantigens, hemolysins, deoxyribonuclease, and proteases. The aims of this study are to investigate the detection of emm genotypes and other virulence genes, such as SAgs, DNase, protease, antibiotic resistance, and phylogenetic relationships in GAS strains isolated from clinical samples.Test strains were obtained from Çukurova University Balcalı Hospital and regional hospitals in Adana province. The M proteins were detected by sequence analysis of emm genes. SAgs and other virulence gene profiles were determined using the Multiplex-PCR method. The antibiotic susceptibility of the isolates was performed by the disc diffusion method and evaluated according to CLSI criteria. The PFGE method was used to determine the clonal relationship between the strains.The emm gene was positive in 86 isolates. The most common emm genotypes were emm28 (22%), emm1 (18.6%), emm12 (13.9%), and emm3 (11.6%). Also, the most common virulence genes were speG (58.1%), speC (56.9%), sdaB (53.4%), and mac (53.4%). The rates of resistance to erythromycin, clindamycin, levofloxacin, ciprofloxacin and telithromycin were 19.8%, 16.3%, 4.7%, 3.5%, and 3.5%, respectively.As a result, additional regional studies on the detection and prevalence of GAS virulence factors in Turkey are required. We believe that this study will provide valuable information for epidemiological studies on emm sequences, Sags, and other virulence factors of Streptococcus pyogenes in Turkey.

An emerging Panton–Valentine leukocidin-positive CC5-meticillin-resistant Staphylococcus aureus-IVc clone recovered from hospital and community settings over a 17-year period from 12 countries investigated by whole-genome sequencing

A novel Panton-Valentine leukocidin (PVL)-positive meticillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC)5-MRSA-IVc ('Sri Lankan' clone) was recently described from Sri Lanka. Similar isolates caused a recent Irish hospital outbreak. To investigate the international dissemination and diversity of PVL-positive CC5-MRSA-IVc isolates from hospital and community settings using whole-genome sequencing (WGS). Core-genome single nucleotide polymorphism (cgSNP) analysis, core-genome multi-locus sequence typing (cgMLST) and microarray-based detection of antimicrobial-resistance and virulence genes were used to investigate PVL-positive CC5-MRSA-IVc (N= 214 including 46 'Sri Lankan' clone) from hospital and community settings in 12 countries over 17 years. Comparators included 29 PVL-positive and 23 PVL-negative CC5/ST5-MRSA-I/II/IVa/IVc/IVg/V. Maximum-likelihood cgSNP analysis grouped 209/214 (97.7%) CC5-MRSA-IVc into Clade I; average of 110 cgSNPs between isolates. Clade III contained the five remaining CC5-MRSA-IVc; average of 92 cgSNPs between isolates. Clade II contained seven PVL-positive CC5-MRSA-IVa comparators, whereas the remaining 45 comparators formed an outlier group. Minimum-spanning cgMLST analysis revealed a comparably low average of 57 allelic differences between all CC5/ST5-MRSA-IVc. All 214 CC5/ST5-MRSA-IVc were identified as 'Sri Lankan' clone, predominantly spa type t002 (186/214) with low population diversity and harboured a similar range of virulence genes and variable antimicrobial-resistance genes. All 214 Sri Lankan clone isolates and Clade II comparators harboured a 9616-bp chromosomal PVL-encoding phage remnant, suggesting both arose from a PVL-positive meticillin-susceptible ancestor. Over half of Sri Lankan clone isolates were from infections (142/214), and where detailed metadata were available (168/214), most were community associated (85/168). Stable chromosomal retention of pvl may facilitate Sri-Lankan clone dissemination.

Detection of optrA and poxtA genes in linezolid-resistant Enterococcus isolates from fur animals in China.

The emergence of linezolid-resistant (LR) enterococci found in food of animal origin arouses attention, but little is known about LR enterococci in fur animals. A total of 342 Enterococcus faecalis and 265 E. faecium strains isolated from fur animals in China from 2015 to 2017 were investigated to determine if LR enterococci (≥16 μg ml-1 ) are present. Overall, two E. faecalis and 12 E. faecium among these isolates were resistant to linezolid. In addition, all LR isolates were classified as multidrug-resistant isolates. We further explore the resistance genes of the LR enterococci, four E. faecalis and two E. faecium isolates contained optrA gene. Two of them co-harboured optrA and poxtA genes. We detected virulence genes in LR enterococci were the following: asa1, cylA, esp, gelE and hyl, among which the highest carrying rate gene was asa1. Besides, all of the LR enterococci we tested had the biofilm-forming ability. It is worth noting that we detected a novel ST type ST2010 from E. faecium 82-2. These data show LR enterococci exist in fur animals and have unique characteristics.

Assessment of the molecular epidemiology and genetic multiplicity of Listeria monocytogenes recovered from ready-to-eat foods following the South African listeriosis outbreak

Following the recent listeriosis outbreak in South Africa, this study was carried out to assess the safety level of various common ready-to-eat foods (RTE) obtained from supermarkets and grocery stores in major towns and cities within the Amathole, Chris Hani and Sarah Baartman Districts Municipalities, Eastern Cape Province, South Africa. A sum of 239 food samples was collected from these locations, and Listeria monocytogenes (Lm) was isolated in line with the recommended techniques by the International Organization for Standardization EN ISO 11290:2017 parts 1 and 2. Identification of the pathogen and detection of various associated virulence genes was done using Polymerase Chain Reaction (PCR) techniques. From the RTE food samples processed, Lm was detected in 107 (44.77%) of the samples. Russian sausage was the most contaminated (78.57%), followed by sliced polony (61.90%), muffins (58.33%), polony (52.63%), and pies (52.38%), while all vetkoek samples examined were negative for Lm. Although the prevalence of Lm in the food samples was very high, concentrations were generally < 100 CFU/g. Strains of Lm recovered from the RTE foods were predominantly epidemiological strains belonging to serotypes 1/2a, 1/2b and 4b. The prevalence of 10 virulence genes including the inlA, InlC, inlJ, plcA, hlyA, plcB, prfA, mpl, inlB, and actA were detected among Lm isolates. Most of the isolates (69.07%) demonstrated the potential for biofilm formation and were categorized as weak (14.95%), moderate (13.40%) and strong (40.72) biofilm formers. Furthermore, molecular typing revealed high levels of genetic diversity among Lm isolates. The findings of this investigation suggested that the presence of Lm in the RTE foods may constitute potential threats to the food sector and could pose public health hazards to consumers, particularly the high-risk group of the population. We, therefore, recommend that adequate food monitoring for safety and proper regulation enforcement in the food sector must be ensured to avoid any future listeriosis outbreak that could be linked to RTE foods in South Africa.

Escherichia coli in different animal feces: phylotypes and virulence genes.

In this study, it was aimed to determine the phylogroups of Escherichia coli isolates from horse, cat, dog, sheep, cattle, and chicken feces samples and to investigate some important virulence genes of the isolates. For this purpose, a total of 600 feces samples, 100 from each animal species, were used as material. For the isolation of E.coli, feces samples were directly inoculated on MacConkey agar. The identification of the isolates was performed via phenotypic tests and species-specific multiplex Polymerase Chain Reaction (mPCR) method. PCR methods were used to phylotype E.coli isolates and to investigate virulence genes (bfpA, eaeA, LT, ST, Stx1, and Stx2). Of the total 600 E.coli isolates recovered in this study, 120 (20%), 269 (44.8%), 58 (9.7%), 19 (3.2%), 35 (5.8%), 56 (9.3%), 31 (5.2%), and 12 (2%) were identified as phylogroup A, B1, B2, C, D, E, F, and Escherichia clade I, respectively. While the virulence gene was detected in 149 (24.8%) E.coli isolates, no virulence gene was detected in 451 (75.2%) isolates. According to the analysis results, the most determined virulence gene was Stx1, while the least determined virulence gene was LT. In conclusion, in this study, when both the animal species and the number of E.coli isolates examined are considered, the data obtained are of great importance in epidemiological terms. However, the detection of virulence genes in 13.5% among phylogroup A, B1, and C isolates with commensal characteristics suggest that these isolates may show pathogenic characteristics with the virulence genes they contain.

Detection of Virulence Genes of Proteus Mirabilis Isolated from Clinical Samples of some Hospitals in Nasiriyah City

The study aimed to detect some virulence genes of Proteus mirabilis isolates, isolation and diagnosis by biochemical tests and Vitek-2 system . The research was conducted in Nasiriyah City/Dhi-Qar, Iraq, in the period between April 2021 and January 2022, and 70 urine and 30 wound samples were obtained from patients admitted to some Hospitals . Isolates diagnosis was made based on morphological and cultural characteristics of blood agar, MacConkey agar, as well as conventional biochemical tests and Vitek-2 identification system. Depending on these criteria, out of 100 samples bacterial growth appeared in only 80 (80.0%) samples, while no bacterial growth appeared in the remaining 20 (20.0%). only 20 (20%) isolates were obtained from P. mirabilis. chosen for genetic testing in order to detect the virulence factors (15) isolates from Proteus mirabilis, such as colonization factor antigen (CFA), extracellular protease, swarming activity, and urease, were identified. The results showed that all isolates (100%) possessed UreC gene, ZapA gene, flaA gene and mrp gene. Keywords: Proteus mirabilis, UTIs, UreC, ZapA, mrp and flaA

A First Report of Molecular Typing, Virulence Traits, and Phenotypic and Genotypic Resistance Patterns of Newly Emerging XDR and MDR Aeromonas veronii in Mugil seheli.

Aeromonas veronii is associated with substantial economic losses in the fish industry and with food-borne illness in humans. This study aimed to determine the prevalence, antibiogram profiles, sequence analysis, virulence and antimicrobial resistance genes, and pathogenicity of A. veronii recovered from Mugil seheli. A total of 80 fish were randomly gathered from various private farms in Suez Province, Egypt. Subsequently, samples were subjected to clinical, post-mortem, and bacteriological examinations. The retrieved isolates were tested for sequence analysis, antibiogram profile, pathogenicity, and PCR detection of virulence and resistance genes. The prevalence of A. veronii in the examined M. seheli was 22.5 % (18/80). The phylogenetic analyses revealed that the tested A. veronii strains shared high genetic similarity with other A. veronii strains from India, UK, and China. Using PCR it was revealed that the retrieved A. veronii isolates harbored the aerA, alt, ser, ompAII, act, ahp, and nuc virulence genes with prevalence of 100%, 82.9%, 61.7%, 55.3%, 44.7%, 36.17%, and 29.8%, respectively. Our findings revealed that 29.8% (14/47) of the retrieved A. veronii strains were XDR to nine antimicrobial classes and carried blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 resistance genes. Likewise, 19.1% (9/47) of the obtained A. veronii strains were MDR to eight classes and possessed blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 genes. The pathogenicity testing indicated that the mortality rates positively correlated with the prevalence of virulence-determinant genes. To our knowledge, this is the first report to reveal the occurrence of XDR and MDR A. veronii in M. seheli, an emergence that represents a risk to public health. Emerging XDR and MDR A. veronii in M. seheli frequently harbored aerA, alt, ser, ompAII, and act virulence genes, and blaTEM, sul1, tetA, blaCTX-M, blaSHV, and aadA1 resistance genes.

Prevalence of some Helicobacter pylori virulence genes such vacA, oipA and dupA in Iraqi patients

Development of H. pylori-associated diseases is determined by a number of virulence factors. So, numerous molecular studies performed about the frequency of its virulence genes in infected patients. To evaluate the prevalence of some H. pylori virulence genes such vacA, oipA and dupA in some Iraqi infected patients. A total of 105 patients suffering from gastritis, clinical manifestations of dyspepsia or burning, vomiting, bloating and others symptoms, with age ranged 17-85 years who attended the Gastroenterology and Hepaetology Teaching Centre, Baghdad – Iraq. They were diagnosed through physicians according to endoscopic findings. Further biopsy samples from different places of stomach were collected by gastroenterologists from each patient, used for histopathological examination and genetic detection of virulence genes (vacA, dupA and oipA) by PCR technique. The result showed that histological study pointed, 78(74.2%) positive for H.pylori, then PCR technique applied on these positive samples to detect the presence of virulence genes which found, VacA gene was the most prevalent 73(93.5%) of positive samples, followed by oipA gene 42(53.8%) and dupA 25(32.05%). The study revealed a significant association between virulence genes and development of certain form of gastric infections. So, detection of the H. pylori virulence genes in gastric biopsy samples can be provides a clear prognosis of clinical outcomes.

Genetic Diversity, Antibiotic Resistance, and Pathogenicity of Aeromonas Species from Food Products in Shanghai, China

ObjectiveAeromonas has recently been recognized as an emerging human pathogen. Aeromonas-associated diarrhea is a phenomenon occurring worldwide. This study was designed to determine the prevalence, genetic diversity, antibiotic resistance, and pathogenicity of Aeromonas strains isolated from food products in Shanghai. MethodsAeromonas isolates (n = 79) collected from food samples were analyzed using concatenated gyrB-cpn60 sequencing. The antibiotic resistance of these isolates was determined using antimicrobial susceptibility testing. Pathogenicity was assessed using β-hemolytic, extracellular protease, virulence gene detection, C. elegans liquid toxicity (LT), and cytotoxicity assays. ResultsEight different species were identified among the 79 isolates. The most prevalent Aeromonas species were A. veronii [62 (78.5%)], A. caviae [6 (7.6%)], A. dhakensis [3 (3.8%)], and A. salmonicida [3 (3.8%)]. The Aeromonas isolates were divided into 73 sequence types (STs), of which 65 were novel. The isolates were hemolytic (45.6%) and protease-positive (81.0%). The most prevalent virulence genes were act (73.4%), fla (69.6%), aexT (36.7%), and ascV (30.4%). The results of C. elegans LT and cytotoxicity assays revealed that A. dhakensis and A. hydrophila were more virulent than A. veronii, A. caviae, and A. bivalvium. Antibiotic resistance genes [tetE, blaTEM, tetA, qnrS, aac(6)-Ib, mcr-1, and mcr-3] were detected in the isolates. The multidrug-resistance rate of the Aeromonas isolates was 11.4%, and 93.7% of the Aeromonas isolates were resistant to cefazolin. ConclusionThe taxonomy, antibiotic resistance, and pathogenicity of different Aeromonas species varied. The Aeromonas isolates A. dhakensis and A. hydrophila were highly pathogenic, indicating that food-derived Aeromonas isolates are potential risks for public health and food safety. The monitoring of food quality and safety will result in better prevention and treatment strategies to control diarrhea illnesses in China.

Prevalence, Antimicrobial Resistance, and Molecular Description of Pseudomonas aeruginosa Isolated from Meat and Meat Products

Resistant and virulent Pseudomonas aeruginosa (P. aeruginosa) bacteria are measured as the major cause of food spoilage and food-borne diseases. This survey assesses the prevalence, antibiotic resistance properties, and virulence factors distribution in P. aeruginosa bacteria isolated from meat and meat products. A total of 370 raw, frozen, and imported bovine meat samples and diverse types of meat product samples were collected from Alborz province, Iran. P. aeruginosa bacteria were identified by culture. Disk diffusion was used to assess the antibiotic resistance of bacteria. Furthermore, the PCR was used to assess the virulence and antibiotic resistance genes. Twenty nine out of 370 (7.83%) samples were contaminated with P. aeruginosa. Imported frozen bovine meat (20%) harbored the highest distribution, while sausage (2%) harbored the lowest. High resistance rates were observed toward ampicillin (89.65%), penicillin (86.20%), tetracycline (82.75%), cefoxitin (37.93%), gentamicin (34.48%), and clindamycin (31.03%). The most commonly detected antibiotic resistance genes were blaDHA (93.10%), blaCTX-M (83.65%), and blaSHV (48.27%). BlaDHA (93.10%), blaCTX-M (83.65%), and blaSHV (48.27%) were the most frequently detected resistance genes. The most commonly detected virulence genes were exoS (75.86%), lasA (68.96%), exoU (58.62%), lasB (51.72%), plcH (48.27%), and algD (44.82%). Meat and meat product samples may be sources of P. aeruginosa, which show an important threat to their consumption. Nevertheless, additional inquiries are obligatory to find supplementary epidemiological properties of P. aeruginosa in meat and meat product samples.

Identification of faecal Escherichia coli isolates with similar patterns of virulence and antimicrobial resistance genes in dogs and their owners.

The presence of antimicrobial resistance and virulence genes in Escherichia coli allows them to survive and cause infections. The close contact between humans and pets can reinforce the risk of transmitting resistant and virulent bacteria between them. This study aims to compare the patterns of the presence of tetracycline and streptomycin resistance genes, as well as important virulence genes in E. coli isolated from faeces of healthy dogs and their owners. Polymerase chain reactions were performed for detection of antimicrobial resistance (tetA, tetB, tetC, tetD, strA and strB) and virulence (fimH, iss, sitA and malX) genes in 144 faecal E. coli isolates from 28 dog-owner pairs and 16 humans who did not keep any pets as controls. Among the investigated antimicrobial resistance and virulence genes, tetA (52.1%) and fimH (86.8%) genes had the highest prevalence. No statistically significant difference was found between the prevalence of antimicrobial resistance and virulence genes in isolates of dogs and their owners. In total, 46.4% of dog-owner pairs had the same patterns of presence or absence of six antimicrobial resistance genes, 50.0% had the same patterns of presence or absence of four virulence genes and 25.0% had the same patterns of presence or absence of all 10 tested genes. The presence of antimicrobial-resistant virulent E. coli in humans and pets may predispose them to infections that are hard to cure with conventional antibiotics. Notable frequency of dogs' and their owners' E. coli isolates with similar patterns of antimicrobial resistance and virulence genes may indicate the possibility of sharing virulent antimicrobial resistant E. coli between them.

Molecular identification of virulence genes of Pseudomonas aeruginosa isolated from fish (Cyprinus carpio) in Mosul city

Depending on the nature of the fish's environment, they are susceptible to many pathogens, including bacterial causes, so the goals of the current study are isolation and molecular identification of Pseudomonas aeruginosa with its prevalence rate and detected virulence genes from fresh common carp fish. The swabs were taken from the gills, skin, intestine and muscles of 75 fish samples from variable localities from Mosul city during the period September to December in the year 2021. The prevalence percentage of bacteria was 26.66% which was confirmed by traditional microbiological tests which included (phenotype culture, microscopically features and API-test) and molecular identification. The isolates formed 42.5, 37.5, 15, 5% from gills, skin, intestine and muscles, respectively. The molecular results of forty isolates determine that Pseudomonas aeruginosa have rpoB 100%, and virulence genes oprL, toxA, and algD, which are express the outer membrane protein, exotoxin A and alginate respectively occur as 97.5% for the oprL gene and 100% of both toxA and algD genes.