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Abstract The health of calves has a significant impact on the production of cows and livestock. Some desert plants have pharmacological importance, as they can be used to reduce antibiotic resistance. Our hypothesis is designed to detect Virulent- Multidrug-Resistant and Extended- spectrum Beta- lactamase Enterobacteriaceae (Virulent-MDR-ESBL Enterobacteriaceae and to determine whether Moringa oleifera has antibacterial activity against the detected isolates. A total of 39 Enterobacteriaceae isolates from 28 diarrheic samples were collected from calves aged between 20 days and 20 months from 3 different flocks in North Sinai, Sahl-Eltina region, Egypt. E.coli 46% (18/39), O157 13% (5/39), Klebsiella pneumoniae 41% (16/39). MDR members accounted for 87%, while ESBL isolates accounted for 43%. The antibacterial activity is represented by microdilution. Minimum inhibition concentration (MIC) for the methanol extract of Moringa oleifera ranged from 2.5,5,10, and 25mg/ ml among E.coli isolates, and O157 was susceptible to (2.5mg/ ml), Klebsiella pneumoniae isolates were susceptible to (5-50mg/ ml). Analysis of the methanol extract revealed that ferulic acid was the dominant phenolic compound with a concentration of 29,832 parts per million (ppm). In silico docking study expected the active site of ferulic acid to act on the tyrosine bacterial enzyme through Pi-alkyl, Pi-anion, Carbon hydrogen bonds, and extra ionic attractive interactions with copper ions which can stabilize ferulic acid inside the targeted pocket Diverse virulent gene profiles were observed in E. coli. The Shiga toxin-producing Escherichia coli (STEC) was reported in 83% of the isolated E. coli, while the DNA gyrase (gyrA) was harbored in 100% of Klebsiella pneumoniae isolates. Various profiles of antibiotic resistance genes for both E. coli and Klebsiella pneumoniae isolates were distinguished. blaTEM genes were detected in 99% of E. coli and 100% of Klebsiella pneumoniae. Sequence analysis for E. coli strain DRC-North Sinai-Eg was placed in accession numbers (OP955786) for the Shiga toxin 2 gene (Stx2A), (OP997748) and (OP997749) for the Adhesion to host cell gene (Eae). For the hemolysine gene (hylA), the accession number was (OP946183). Klebsiella pneumoniae strain DRC-North Sinai-Eg was placed in (OP946180) for (gyrA). This study has proven the broad range of Moringa oliefera's antibacterial effects in vitro against the virulent-MDR- ESBL E. coli and Klebsiella pneumoniae isolated from North Sinai calves diarrhea. These are congruent with the disability effect on bacterial tyrosinase enzyme through docking study therefore, we recommend the usage of this desert plant as a prospective feed additive, we endorse this as an antibacterial new insight natural source and for the medication of considered pathogens with zoonotic impacts. Graphical Abstract

Background: The relationship between gastroesophageal reflux disease (GERD) and Helicobacter pylori infection is a complicated topic. This study aimed to evaluate the association between H. pylori infection and GERD and identify the associated risk factors in North India. Methods: All individuals who were referred for an endoscopy because they had dyspepsia were included in a cross-sectional study. Using endoscopic assessment, GERD was diagnosed. As a comparison group, patients with normal esophageal function were chosen. H. pylori was cultured and genotyped to assess the virulent genes such as cagA, vacA, and iceA. The relationship of H. pylori infection with GERD and its associated risk factors were estimated with Chi-square analysis, odds ratios, and 95% confidence intervals using multiple logistic regression analysis. Results: In total, 420 patients were included in this study, of whom 335 (79.8%) had GERD. The mean age was 44.01 ± 16.52 years. Overall, 143 (34%) patients were positive for H. pylori infection. cagA-negative H. pylori isolates were significantly associated with the GERD group. Variables such as gender, age, alcohol, smoking, body mass index (BMI), smoking, and hiatus hernia in patients had no significant association with H. pylori infection. However, increased age, gender, Frequency Scale for the Symptoms of GERD (FSSG) score, BMI, and presence of hiatus hernia were significantly associated with the GERD group. Conclusion: In our results, there was no association between H. pylori infection and GERD. The risk factors for GERD were age, gender, FSSG score, BMI, and hiatus hernia. Further studies are recommended.

The health of calves has a significant impact on the production of cows and livestock. Some desert plants have pharmacological importance, as they can be used to reduce antibiotic resistance. Our hypothesis is designed to detect Virulent- Multidrug-Resistant and Extended- spectrum Beta- lactamase Enterobacteriaceae (Virulent-MDR-ESBL Enterobacteriaceae and to determine whether Moringa oleifera has antibacterial activity against the detected isolates. A total of 39 Enterobacteriaceae isolates from 28 diarrheic samples were collected from calves aged between 20 days and 20 months from 3 different flocks in North Sinai, Sahl-Eltina region, Egypt. E.coli 46% (18/39), O157 13% (5/39), Klebsiella pneumoniae 41% (16/39). MDR members accounted for 87%, while ESBL isolates accounted for 43%. The antibacterial activity is represented by microdilution. Minimum inhibition concentration (MIC) for the methanol extract of Moringa oleifera ranged from 2.5,5,10, and 25mg/ ml among E.coli isolates, and O157 was susceptible to (2.5mg/ ml), Klebsiella pneumoniae isolates were susceptible to (5-50mg/ ml). Analysis of the methanol extract revealed that ferulic acid was the dominant phenolic compound with a concentration of 29,832 parts per million (ppm). In silico docking study expected the active site of ferulic acid to act on the tyrosine bacterial enzyme through Pi-alkyl, Pi-anion, Carbon hydrogen bonds, and extra ionic attractive interactions with copper ions which can stabilize ferulic acid inside the targeted pocket Diverse virulent gene profiles were observed in E. coli. The Shiga toxin-producing Escherichia coli (STEC) was reported in 83% of the isolated E. coli, while the DNA gyrase (gyrA) was harbored in 100% of Klebsiella pneumoniae isolates. Various profiles of antibiotic resistance genes for both E. coli and Klebsiella pneumoniae isolates were distinguished. blaTEM genes were detected in 99% of E. coli and 100% of Klebsiella pneumoniae. Sequence analysis for E. coli strain DRC-North Sinai-Eg was placed in accession numbers (OP955786) for the Shiga toxin 2 gene (Stx2A), (OP997748) and (OP997749) for the Adhesion to host cell gene (Eae). For the hemolysine gene (hylA), the accession number was (OP946183). Klebsiellapneumoniae strain DRC-North Sinai-Eg was placed in (OP946180) for (gyrA). This study has proven the broad range of Moringa oliefera's antibacterial effects in vitro against the virulent-MDR- ESBL E. coli and Klebsiella pneumoniae isolated from North Sinai calves diarrhea. These are congruent with the disability effect on bacterial tyrosinase enzyme through docking study therefore, we recommend the usage of this desert plant as a prospective feed additive, we endorse this as an antibacterial new insight natural source and for the medication of considered pathogens with zoonotic impacts. [ABSTRACT FROM AUTHOR]

Background: The relationship between gastroesophageal reflux disease (GERD) and Helicobacter pylori infection is a complicated topic. This study aimed to evaluate the association between H. pylori infection and GERD and identify the associated risk factors in North India. Methods: All individuals who were referred for an endoscopy because they had dyspepsia were included in a cross-sectional study. Using endoscopic assessment, GERD was diagnosed. As a comparison group, patients with normal esophageal function were chosen. H. pylori was cultured and genotyped to assess the virulent genes such as cagA, vacA, and iceA. The relationship of H. pylori infection with GERD and its associated risk factors were estimated with Chi‑square analysis, odds ratios, and 95% confidence intervals using multiple logistic regression analysis. Results: In total, 420 patients were included in this study, of whom 335 (79.8%) had GERD. The mean age was 44.01 ± 16.52 years. Overall, 143 (34%) patients were positive for H. pylori infection. cagA-negative H. pylori isolates were significantly associated with the GERD group. Variables such as gender, age, alcohol, smoking, body mass index (BMI), smoking, and hiatus hernia in patients had no significant association with H. pylori infection. However, increased age, gender, Frequency Scale for the Symptoms of GERD (FSSG) score, BMI, and presence of hiatus hernia were significantly associated with the GERD group. Conclusion: In our results, there was no association between H. pylori infection and GERD. The risk factors for GERD were age, gender, FSSG score, BMI, and hiatus hernia. Further studies are recommended. [ABSTRACT FROM AUTHOR]

The Present study was undertaken to assess the prevalence of B. cereus in milk and some milk Products and its survival in yoghurt and Damietta cheese. A total of 100 samples of raw milk, Pasteurized milk, Damietta cheese and yoghurt (25 for each) were collected from supermarkets in Assiut city and were examined for isolation of B. cereus. It was isolated from 24, 32, 68 and 20% of the tested samples, respectively. All the isolated strains were confirmed positive for B. cereus except one strain for each raw milk and Damietta cheese. 14.7%, 85.3, 50.0 and 76.5% of the confirmed strains were carried Ces, Nhe, Hbl and cytK genes. Concerning its survival in yoghurt, there was a significant difference between the effect of yoghurt and acidophilus yoghurt on B. cereus growth, where the organism remained viable to the 9th day in yoghurt and failed to be isolated at the 5th day in yoghurt with probiotics. In case of Damietta cheese, 2 and 5% salt were more favorable for the pathogen growth, while 10% salt was inconvenient for its growth. In addition, the inhibitory effect of thyme essential oil was more obvious than the effect of rosemary EO, where the bacterial population was decreased to a count of 2.04 and 2.46 log cfu/ g. at the 2nd week in case of addition of 1.65% thyme EO and could not be detected at the 4th week, whereas the organism could survive to the 6th week in case of rosemary. In conclusion, strict hygienic measures must be implemented during the manufacture of dairy products and addition of biopreservatives to control B. cereus growth is substantial. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is an opportunistic pathogen that causes invasive infections in humans. In recent years, increasing studies have focused on the prevalence of S. aureus infections in adults; however, the epidemiology and molecular characteristics of S. aureus from Chinese pediatric patients remain unknown. The present study examined the population structure, antimicrobial resistance, and virulent factors of methicillin-resistant and -susceptible S. aureus isolated from Chinese pediatric patients from one medical center in eastern China. A total of 81 cases were screened with positive S. aureus infections among 864 pediatric patients between 2016 and 2022 in eastern China. Molecular analysis showed that ST22 (28.4%) and ST59 (13.6%) were the most typical strains, and associations between different clonal complex (CC) types/serotype types (ST) and the age of pediatric patients were observed in this study. CC398 was the predominant type in neonates under 1 month of age, while CC22 was mainly found in term-infant (under 1 year of age) and toddlers (over 1 year of age). Additionally, 17 S. aureus isolates were resistant to at least three antimicrobials and majority of them belonged to CC59. The blaZ gene was found in 59 isolates and mecA gene was present in 26 strains identified as methicillin-resistant. Numerous virulent factors were detected in S. aureus isolated from present pediatric patients. Remarkably, lukF-PV and lukS-PV were dominantly carried by CC22, tsst-1 genes were detected in CC188, CC7, and CC15, while exfoliative toxin genes were found only in CC121. Only 41.98% of the S. aureus isolates possessed scn gene, indicating that the sources of infections in pediatric patients may include both human-to-human transmissions as well as environmental and nosocomial infections. Together, the present study provided a phylogenetic and genotypic comparison of S. aureus from Chinese pediatric patients in Suzhou city. Our results suggested that the colonization of multi-drug resistant isolates of S. aureus may raise concern among pediatric patients, at least from the present medical center in eastern China. [ABSTRACT FROM AUTHOR]

The emergence of antibiotic-resistant microorganisms is a significant concern in global health. Antibiotic resistance is attributed to various virulent factors and genetic elements. This study investigated the virulence factors of Staphylococcus aureus to create an mRNA-based vaccine that could help prevent antibiotic resistance. Distinct strains of the bacteria were selected for molecular identification of virulence genes, such as spa, fmhA, lukD, and hla-D, which were performed utilizing PCR techniques. DNA extraction from samples of Staphylococcus aureus was conducted using the Cetyl Trimethyl Ammonium Bromide (CTAB) method, which was confirmed and visualized using a gel doc; 16S rRNA was utilized to identify the bacterial strains, and primers of spa, lukD, fmhA, and hla-D genes were employed to identify the specific genes. Sequencing was carried out at Applied Bioscience International (ABI) in Malaysia. Phylogenetic analysis and alignment of the strains were subsequently constructed. We also performed an in silico analysis of the spa, fmhA, lukD, and hla-D genes to generate an antigen-specific vaccine. The virulence genes were translated into proteins, and a chimera was created using various linkers. The mRNA vaccine candidate was produced utilizing 18 epitopes, linkers, and an adjuvant, known as RpfE, to target the immune system. Testing determined that this design covered 90% of the population conservancy. An in silico immunological vaccine simulation was conducted to verify the hypothesis, including validating and predicting secondary and tertiary structures and molecular dynamics simulations to evaluate the vaccine's long-term viability. This vaccine design may be further evaluated through in vivo and in vitro testing to assess its efficacy. [ABSTRACT FROM AUTHOR]

Yuqi Hao,1 Yong’ang Jiang,1 Hafiz Muhammad Ishaq,2 Wenke Liu,1 Huajie Zhao,1 Mingyong Wang,3 Fan Yang1 1Xinxiang Key Laboratory of Pathogenic Biology, Department of Pathogenic Biology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, People’s Republic of China; 2Faculty of Veterinary and Animal Sciences, Muhammad Nawaz Shareef University of Agriculture, Multan, Pakistan; 3Xinxiang Key Laboratory of Immunoregulation and Molecular Diagnostics, School of Laboratory Medicine, Xinxiang Medical University,, Xinxiang, People’s Republic of ChinaCorrespondence: Fan Yang; Mingyong Wang, Email yangf77@163.com; wmy118@126.comBackground: In clinical practice, Klebsiella pneumoniae (K. pneumoniae) is a common opportunistic pathogen responsible for nosocomial infection. This study aimed to analyze the trend of antimicrobial susceptibility and virulent characteristics of K. pneumoniae isolated from sputum. In clinics, data of the current study will help in the clinical treatment of K. pneumoniae infection.Results: The current research showed the resistance rates of the 20 K. pneumoniae isolates against 13 antibiotics ranged from 15.0% to 80.0%. The detection rate of extended spectrum β-lactamases (ESBLs) was up to 55%, while blaSHV was the most prevalent ESBLs genes. Four strains (25.0%) of K. pneumoniae presented hypermucoviscous phenotype (HMV). Moreover, 18 strains (90.0%) showed the stronger biofilm-forming ability. wzi, wabG, fimH, mrkD were the most prevalent virulence genes in current research. Ten strains were found capsule typing and the higher genetic diversity of colonizing K. pneumoniae in this region. K19 exhibited a strong positive correlation with imipenem resistance, while K1 showed strong correlations with magA. Furthermore, HMV phenotype showed significantly negative correlations with multidrug-resistant.Conclusion: In the hospital, the antibiotic resistance of K. pneumoniae (isolated from sputum samples) has a serious concern. Additionally, strains of K. pneumoniae show the higher genetic diversity.Keywords: Klebsiella pneumoniae, antimicrobial resistance, resistant genes, virulent genes, biofilm-forming

Diarrheagenic Escherichia coli (DEC) are the leading cause of infectious diarrhea and pose a significant global, regional, and national burden of disease. This study aimed to investigate the prevalence of six DEC pathotypes in children with diarrhea and determine their antibiotic resistance patterns. Samples from 107 diarrheagenic children were collected and processed for Escherichia coli (E. coli). Single-plex PCR was used to detect target virulence genes as well as characterize and categorize DEC pathotypes. Antibiotic resistance patterns were determined by the Kirby–Bauer disk diffusion method. E. coli was detected in 79 diarrheal stool samples, accounting for 73.8% of the samples collected. Additionally, 49.4% (39 out of 79) of the isolates harbored various typical virulence factors. Results revealed six pathotypes of virulence: enterotoxigenic E. coli (ETEC) (53.8%), enteropathogenic E. coli (EPEC) (12.8%), enteroaggregative E. coli (EAEC) (10.3%), Heteropathotypes (7.8%), Shiga toxin-producing E. coli (STEC), and enterohemorrhagic E. coli (EHEC) (7.7% each). The isolates exhibited high antibiotic resistance against trimethoprim/sulfamethoxazole (82.1%), amoxicillin (79.5%), ampicillin (74.4%), gentamicin (69.2%), and streptomycin (64.1%). An overall occurrence of 84.6% of multiple-drug resistance was observed in the isolates, with resistance ranging from three to four antibiotic classes. Our findings revealed a high level of pathogenic E. coli that were highly resistant to multiple categories of antibiotics among children in the Awi zone. These findings highlight the potential role of pathogenic E. coli in childhood diarrhea in tropical low-resource settings and underscore the need for continued research on the characteristics of pathogenic and antibiotic-resistant strains.

Salmonellosis is one of the most important bacterial diseases in human and animals. Rapid diagnosis and sub sequence accurate treatment of Salmonella carriers help reduce the salmonellosis in human and livestock animals. In this study, 420 fecal samples were taken during year 2019 from buffalo in the Urmia, Khoy and Piranshahr regions in west Azerbaijan province, Iran. Samplings were carried out in different seasons. Presence of Salmonella invasion genes (FimA, Stn and InvA) were evaluated by polymerase chain reaction. The bacterial culture and biochemical tests were performed on feces samples for isolation of bacterium Salmonella; however, all samples were negative in culture method. PCR findings showed that, 50 (11.90%) fecal samples were positive to the genes. The analysis of results showed that frequency of salmonellosis outbreak in different parts of west Azerbaijan province followed a similar pattern and the incidence of salmonellosis according to forecast in the warm seasons (spring and summer) was more than in cold seasons (autumn and winter). The prevalence of Salmonella in buffalo's feces based on warm and cold seasons were 32 (64.00%) and 18 (36.00%), respectively. The results showed significant difference between cold and warm season in the prevalence of salmonellosis. Therefore, the application of molecular technics is essential for the prevention and treatment of salmonellosis. The results also showed that specificity of PCR method was better than culture method for detection of Salmonella in feces sample. [ABSTRACT FROM AUTHOR]

The recent outbreak of the emerging pathogenic races of Fusarium wilt and early blight causing pathogens capable of infecting numerous commercial tomato varieties/cultivars highlights the need for robust disease resistance in crop plants. To breed plants with increased and robust disease resistance using quantitative disease resistance genes and resistance gene pyramids is being used to combat Fusarium wilt and early blight resistance, but broader acceptance of these methods is required to maintain resistance effectiveness. A greater understanding of the molecular basis of plant disease resistance at host levels offers further possibilities for Fusarium wilt and early blight resistance using omics approaches, such as genomics, transcriptomics, proteomics, and metabolomics. These omics technologies are delivering us many candidate genes that might lead to increased disease resistance through genetic engineering. There are several strategies for manipulating these genes, which can come from plants, pathogens, or other organisms. However, truly durable tomato cultivars with Fusarium wilt and early blight resistance appear a doubtful prospect in the face of frequently evolving virulent populations of associated pathogens. This article discusses recent developments and future perspectives for improving tomato plants resistant to Fusarium wilt and early blight.

In this study, the antibacterial effects of para-coumaric acid (PCA), as an intermediate product in the phenylpropanoid biosynthesis pathway, on the virulence of Aeromonas hydrophila in both in vivo and in vitro conditions were assayed. First, the minimum inhibitory concentration (MIC) of PCA against A. hydrophila was determined. The existence of ascV and aopB virulence genes as essential components of the Type III secretion system (T3SS) of A. hydrophila was determined using RT-PCR analysis. Then, the effect of sub-MIC concentrations of PCA on the expression of ascV and aopB genes of bacteria was investigated using qRT-PCR. In the in vivo phase of the study, 135 juvenile common carp (weighing 30.2 ± 4.53 g) were equally divided into three groups in triplicates. Group 1 was fed with un-supplemented food, whereas groups 2 and 3 were fed with food supplemented with PCA or oxytetracycline (OTC), respectively, for 5 days. Following that, all groups were challenged with A. hydrophila, and mortality rates, the presence of A. hydrophila in surviving fish, and the expression of the ascV and aopB genes in the fish head kidney were assessed. Our results revealed that the expression of ascV and aopB genes was significantly suppressed after exposure to sub-MICs of PCA compared to control in both in vivo and in vitro assays (p < 0.05). Although the PCA-treated group showed higher mortality (= 36.7%) compared to the OTC group following bacterial challenge (= 23.3%), the expression of ascV and aopB genes in isolated bacteria in the PCA group was significantly lower than that in the OTC and control groups (p < 0.05). This is the first report of the in vitro and in vivo effects of PCA on decreasing the virulence properties of A. hydrophila in fish. It can be concluded that PCA has an inhibitory effect on the virulence and pathogenicity of A. hydrophila and may be effective in the control of Aeromonas infections in fish farms. [ABSTRACT FROM AUTHOR]

The poultry industry in developing countries still faces a significant threat from fowl typhoid, a disease caused by Salmonella Gallinarum that has been well contained in more economically developed countries. In addition to the virulence exhibited by large virulence plasmid (85 kb), Salmonella Pathogenicity Island 2 in S. Gallinarum plays a key role in mediating disease through its type III secretion systems (TTSS). The TTSS secrete effector protein across the Salmonella containing vacuoles and mediate the internalization of bacteria by modulating vesicular passage. In this study, candidate virulent ssaU gene (~1 kb) encoding type III secretion system was successfully deleted from indigenously isolated S. Gallinarum genome through homology-directed repair using CRISPR/Cas9 and lambda recombination systems. CRISPR/Cas9-based genome editing of poultry-derived Salmonella Gallinarum has not been previously reported, which might be linked to a lack of efficiency in its genetic tools. This is the first study which demonstrates a complete CRISPR/Cas9-based gene deletion from this bacterial genome. More importantly, a poultry experimental model was employed to assess the virulence potential of this mutant strain (ΔssaU_SG18) which was unable to produce any mortality in the experimentally challenged birds as compared to the wild type strain. No effect on weight gain was observed whereas bacteria were unable to colonize the intestine and liver in our challenge model. This in vivo loss of virulence in mutant strain provides an excellent functionality of this system to be useful in live vaccine development against this resistant and patho genic bacteria. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) is one of the most prevalent bacterial pathogens recovered from diabetic foot infections (DFIs). Most S. aureus isolates exhibit methicillin resistance, so treatment is recommended with antimicrobials active against methicillin-resistant S. aureus (MRSA) in patients who have risk factors associated with MRSA infections. The main goal of this study was to see if proteomics and molecular methods could be effective in identifying and distinguishing MRSA recovered from DFIs. Since MRSA is highly resistant to β-lactam antibiotics and usually does not respond to other antimicrobial drugs, we evaluated the resistance of MRSA isolates against different antibiotics. The standard procedures were followed for a culture of 250 skin swabs collected from diabetic foot patients. The phenotypic characteristics of 48 suspected S. aureus cultures were determined via microscopic examination, Gram staining, a coagulase test, a BBL™ Staphyloslide™ Latex test, a Staph ID 32 API system, and a Vitek 2 Compact system. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to examine the protein profile of all isolates, and real-time PCR was then used to identify mecA and PVL virulence genes. S aureus isolates were tested using the Vitek 2 Compact for antimicrobial susceptibility using Gram-positive cards (GP71). Among the 48 bacterial isolates tested, 45 (93.75%), 42 (87.5%), and 46 (95.83%) were positive in tube coagulase, the Staph ID 32 API system, and the Vitek 2 Compact system, respectively. We correctly identified all suspected S. aureus isolates (100%) via MALDI-TOF MS with a score value ≥2.00 and differentiated them into 22/48 MRSA (45.83%) and 26/48 MSSA (54.17%) isolates. A higher peak intensity at masses of 5530 Da, 6580 Da, 6710 Da, and 6820 Da was detected in MRSA, but not in MSSA. All MRSA isolates tested positive for the mecA gene, while all isolates tested negative for the PVL gene. The antibiotic susceptibility results showed that 22 (100%), 20 (90.91%), 19 (86.36%), 18 (81.82%), 17 (77.27%), 15 (68.18%), 13 (59.1%), and 12 (54.55%) MRSA strains were resistant to cefoxitin, daptomycin, erythromycin, benzylpenicillin, ciprofloxacin, oxacillin, and clindamycin, respectively. In contrast, all MRSA strains were extremely susceptible (100%) to linezolid, nitrofurantoin, quinupristin–dalfopristin, tigecycline, and vancomycin. Moreover, 20 (90.91%), 18 (81.82%), and 17 (77.27%) of the MRSA strains exhibited high sensitivity against rifampin, trimethoprim–sulfamethoxazole, and gentamicin, respectively. In DFIs, MALDI-TOF MS is a powerful and accurate method of identifying and distinguishing both MRSA and MSSA isolates. A high level of antimicrobial resistance was found in MRSA isolates, and antibiotic therapy based on antibiotic susceptibility patterns is essential for a successful outcome. [ABSTRACT FROM AUTHOR]

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Verotoxigenic E.coli (VTEC) is one of the emerging pathogens worldwide. The aim of this study was to investigate the prevalence of VTEC among One hundred from different dairy products samples (Kareish cheese and plain yogurt) were collected randomly from street-vendors, supermarkets and small dairies at Alexandria City, Egypt. (50 of each). The Prevalence of Verotocytotoxigenic Escherichia coli in examined locally manufactured dairy products samples using Sorbitol MacConky agar give rise for 24% (6/25), 0% (0/25) and 12% (3/25), 12% (3/25) positive samples from street-vendor and supermarket Kareish cheese, and Supermarket Plain yogurt and Small dairies Plain yogurt, respectively while, its prevalence based on molecular identification revealed that only 8% (2/25) positive samples from street-vendor Kareish cheese. E.coli isolated from street-vendor Kareish cheese could be serologically identified into E.coli O26, O55 and O127, while that isolated from plain yoghurt samples into O55 and O86a. All screened E.coli strains had a determinant gene (phoA) while, stx1 and stx2 genes could be detected only in of selected street-vended Kareish cheese isolates., meanwhile both virulence genes could not be detected in all selected small dairies plain yoghurt isolates. On contrary, virulence gene eaeA could be detected in all test E.Coli isolates from street-vendor Kareish cheese and small dairies plain yoghurt. Prevalence of Verotoxigenic E.coli (VTEC) poses public health threats to consumers. Food safety concept should be implemented for all steps through manufacture to produce safe and good quality dairy products. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is an opportunistic pathogen that causes invasive infections in humans. In recent years, increasing studies have focused on the prevalence of S. aureus infections in adults; however, the epidemiology and molecular characteristics of S. aureus from Chinese pediatric patients remain unknown. The present study examined the population structure, antimicrobial resistance, and virulent factors of methicillin-resistant and -susceptible S. aureus isolated from Chinese pediatric patients from one medical center in eastern China. A total of 81 cases were screened with positive S. aureus infections among 864 pediatric patients between 2016 and 2022 in eastern China. Molecular analysis showed that ST22 (28.4%) and ST59 (13.6%) were the most typical strains, and associations between different clonal complex (CC) types/serotype types (ST) and the age of pediatric patients were observed in this study. CC398 was the predominant type in neonates under 1 month of age, while CC22 was mainly found in term-infant (under 1 year of age) and toddlers (over 1 year of age). Additionally, 17 S. aureus isolates were resistant to at least three antimicrobials and majority of them belonged to CC59. The blaZ gene was found in 59 isolates and mecA gene was present in 26 strains identified as methicillin-resistant. Numerous virulent factors were detected in S. aureus isolated from present pediatric patients. Remarkably, lukF-PV and lukS-PV were dominantly carried by CC22, tsst-1 genes were detected in CC188, CC7, and CC15, while exfoliative toxin genes were found only in CC121. Only 41.98% of the S. aureus isolates possessed scn gene, indicating that the sources of infections in pediatric patients may include both human-to-human transmissions as well as environmental and nosocomial infections. Together, the present study provided a phylogenetic and genotypic comparison of S. aureus from Chinese pediatric patients in Suzhou city. Our results suggested that the colonization of multi-drug resistant isolates of S. aureus may raise concern among pediatric patients, at least from the present medical center in eastern China.

Background and Objectives: Pathogenic diseases resulting from microbial contamination of food have been widely distributed in many parts of the world. Among these, Escherichia coli is one of the most important foodborne pathogenic bacteria. Diarrhea is one of the major causes of children’s death in developing countries, with approximately 2 million deaths annually. The current study aimed to determine the frequency of diarrheagenic E. coli pathotypes such as Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroaggregative E. coli (EAEC), and Shiga toxin-producing E. coli (STEC) in Brassica oleracea cultivars in order to provide information on the assessment of diarrheagenic E. coli pathogenesis risk. Materials and Methods: 100 samples of vegetables were collected in Tehran, including cabbage, cauliflower, broccoli and Brussels sprouts. After homogenizing samples, enrichment was done in the EC broth medium. Five colonies of pure culture were used for DNA extraction. Pathotypes were identified by PCR using virulence genes. Results: The results showed that the prevalence of diarrheagenic E. coli strains was 7%. The EPEC prevalence was 3%, All EPEC isolates were atypical. The ETEC frequency was 3%, And the EAEC prevalence was 1%. Conclusion: These findings indicated that Brassica oleracea cultivars could be consideredas a source of contamination with diarrhea-causing E. coli strains.

Background and Objectives: Pathogenic diseases resulting from microbial contamination of food have been widely distributed in many parts of the world. Among these, Escherichia coli is one of the most important foodborne pathogenic bacteria. Diarrhea is one of the major causes of children's death in developing countries, with approximately 2 million deaths annually. The current study aimed to determine the frequency of diarrheagenic E. coli pathotypes such as Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroaggregative E. coli (EAEC), and Shiga toxin-producing E. coli (STEC) in Brassica oleracea cultivars in order to provide information on the assessment of diarrheagenic E. coli pathogenesis risk. Materials and Methods: 100 samples of vegetables were collected in Tehran, including cabbage, cauliflower, broccoli and Brussels sprouts. After homogenizing samples, enrichment was done in the EC broth medium. Five colonies of pure culture were used for DNA extraction. Pathotypes were identified by PCR using virulence genes. Results: The results showed that the prevalence of diarrheagenic E. coli strains was 7%. The EPEC prevalence was 3%, All EPEC isolates were atypical. The ETEC frequency was 3%, And the EAEC prevalence was 1%. Conclusion: These findingsindicated that Brassica oleracea cultivars could be consideredas a source of contamination with diarrhea-causing E. coli strains. [ABSTRACT FROM AUTHOR]

Dumpsites are reported as a major source of human exposure to pathogenic microorganisms. In the current study, we investigated the prevalence of Vibrio sp. and Escherichia coli from market dumpsites in selected local government areas in Edo-South Senatorial district. Bacterial isolates were isolated using standard microbiological techniques, and further identified by their morphological, biochemical and 16 S rRNA sequence. The antibiotic susceptibility test and multidrug resistance profile were done by the agar disc diffusion method. DNA extraction was done by boiling method and the presence of virulent genes (Ctx, TcpAEl tor, Zot, flicH7, Lt and Stx) in the isolates were detected by polymerase chain reaction as well as agarose gel electrophoresis. Vibrio cholerae and Escherichia coli were prevalent in soil and leachates from market dumpsites in selected local government areas in Edo-South Senatorial district. All the isolates were multidrug resistant, while a total of 31 isolates (26 Escherichia coli and 5 Vibrio sp.) had at least, one of the virulent genes (Lt, FliCH7, UidA, CtxA and TcpAEl tor) of interest. This study shows that Escherichia coli and Vibrio sp. isolated from market dumpsites are multidrug resistant, and possesses virulent genes, thus, making them a threat to public health. [ABSTRACT FROM AUTHOR]

The emergence of antibiotic-resistant microorganisms is a significant concern in global health. Antibiotic resistance is attributed to various virulent factors and genetic elements. This study investigated the virulence factors of Staphylococcus aureus to create an mRNA-based vaccine that could help prevent antibiotic resistance. Distinct strains of the bacteria were selected for molecular identification of virulence genes, such as spa, fmhA, lukD, and hla-D, which were performed utilizing PCR techniques. DNA extraction from samples of Staphylococcus aureus was conducted using the Cetyl Trimethyl Ammonium Bromide (CTAB) method, which was confirmed and visualized using a gel doc; 16S rRNA was utilized to identify the bacterial strains, and primers of spa, lukD, fmhA, and hla-D genes were employed to identify the specific genes. Sequencing was carried out at Applied Bioscience International (ABI) in Malaysia. Phylogenetic analysis and alignment of the strains were subsequently constructed. We also performed an in silico analysis of the spa, fmhA, lukD, and hla-D genes to generate an antigen-specific vaccine. The virulence genes were translated into proteins, and a chimera was created using various linkers. The mRNA vaccine candidate was produced utilizing 18 epitopes, linkers, and an adjuvant, known as RpfE, to target the immune system. Testing determined that this design covered 90% of the population conservancy. An in silico immunological vaccine simulation was conducted to verify the hypothesis, including validating and predicting secondary and tertiary structures and molecular dynamics simulations to evaluate the vaccine’s long-term viability. This vaccine design may be further evaluated through in vivo and in vitro testing to assess its efficacy.

Abstract Objectives This review fills the paucity of information on K. pneumoniae as a nosocomial pathogen by providing pooled data on epidemiological risk factors, resistant trends and profiles and resistant and virulent genes of this organism in Asia. Methods Exhaustive search was conducted using PubMed, Web of Science, and Google scholar for most studies addressing the prevalence, risk factors, drug resistant-mediated genes and/or virulent factors of K. pneumoniae in Asia. Data extracted for meta-analysis were analyzed using comprehensive meta-analysis version 3. Trends data for the isolation rate and resistance rates were entered into Excel spread sheet and the results were presented in graphs. Results The prevalence rate of drug resistance in K. pneumoniae were; amikacin (40.8%) [95% CI 31.9–50.4], aztreonam (73.3%) [95% CI 59.9–83.4], ceftazidime (75.7%) [95% CI 65.4–83.6], ciprofloxacin (59.8%) [95% CI 48.6–70.1], colistin (2.9%) [95% CI 1.8–4.4], cefotaxime (79.2%) [95% CI 68.0–87.2], cefepime (72.6) [95% CI 57.7–83.8] and imipenem (65.6%) [95% CI 30.8–89.0]. TEM (39.5%) [95% CI 15.4–70.1], SHV-11 (41.8%) [95% CI 16.2–72.6] and KPC-2 (14.6%) [95% CI 6.0–31.4] were some of the resistance mediated genes observed in this study. The most virulent factors utilized by K. pneumoniae are; hypermucoviscous phenotype and mucoviscosity-related genes, genes for biosynthesis of lipopolysaccharide, iron uptake and transport genes and finally, adhesive genes. Conclusion It can be concluded that, antimicrobial resistant in K. pneumoniae is a clear and present danger in Asia which needs strong surveillance to curb this menace. It is very important for public healthcare departments to monitor and report changes in antimicrobial-resistant isolates.

Shiga toxin-producing Escherichia coli (STEC) is the pathogenic E. coli causing disease in humans via the consumption or handling of animal food products. The high prevalence of these organisms in ruminants has been widely reported. Among STECs, O157 is one of the most lethal serotypes causing serious disease in humans. The present study investigated the prevalence of sorbitol non-fermenting STECs in goats reared in the lower region of southern Thailand and described the virulent factors carried by those isolates. Sorbitol non-fermenting (SNF)-STECs were found in 57 out of 646 goats (8.82%; 95% CI 6.75% to 11.28%). Molecular identification revealed that 0.77% of SNF-STEC isolates were the O157 serotype. Shiga toxin genes (stx1 and stx2) and other virulent genes (i.e., eaeA, ehxA, and saa) were detected by molecular techniques. The presence of stx1 (75.44%) was significantly higher than that of stx2 (22.81%), whereas 1.75% of the total isolates carried both stx1 and stx2. Most of the isolates carried ehxA for 75.44%, followed by saa (42.11%) and eaeA (12.28%). In addition, 21.05% of STEC isolates did not carry any eaeA, ehxA, or saa. The first investigation on SNF-STECs in goat was conducted in the lower region of southern Thailand. The present study revealed that goats could be one of the potential carriers of SNF-STECs in the observing area.

The poultry industry in developing countries still faces a significant threat from fowl typhoid, a disease caused by Salmonella Gallinarum that has been well contained in more economically developed countries. In addition to the virulence exhibited by large virulence plasmid (85 kb), Salmonella Pathogenicity Island 2 in S. Gallinarum plays a key role in mediating disease through its type III secretion systems (TTSS). The TTSS secrete effector protein across the Salmonella containing vacuoles and mediate the internalization of bacteria by modulating vesicular passage. In this study, candidate virulent ssaU gene (~1 kb) encoding type III secretion system was successfully deleted from indigenously isolated S. Gallinarum genome through homology-directed repair using CRISPR/Cas9 and lambda recombination systems. CRISPR/Cas9-based genome editing of poultry-derived Salmonella Gallinarum has not been previously reported, which might be linked to a lack of efficiency in its genetic tools. This is the first study which demonstrates a complete CRISPR/Cas9-based gene deletion from this bacterial genome. More importantly, a poultry experimental model was employed to assess the virulence potential of this mutant strain (ΔssaU_SG18) which was unable to produce any mortality in the experimentally challenged birds as compared to the wild type strain. No effect on weight gain was observed whereas bacteria were unable to colonize the intestine and liver in our challenge model. This in vivo loss of virulence in mutant strain provides an excellent functionality of this system to be useful in live vaccine development against this resistant and patho genic bacteria.

Staphylococcus aureus (S. aureus) is one of the most prevalent bacterial pathogens recovered from diabetic foot infections (DFIs). Most S. aureus isolates exhibit methicillin resistance, so treatment is recommended with antimicrobials active against methicillin-resistant S. aureus (MRSA) in patients who have risk factors associated with MRSA infections. The main goal of this study was to see if proteomics and molecular methods could be effective in identifying and distinguishing MRSA recovered from DFIs. Since MRSA is highly resistant to β-lactam antibiotics and usually does not respond to other antimicrobial drugs, we evaluated the resistance of MRSA isolates against different antibiotics. The standard procedures were followed for a culture of 250 skin swabs collected from diabetic foot patients. The phenotypic characteristics of 48 suspected S. aureus cultures were determined via microscopic examination, Gram staining, a coagulase test, a BBL™ Staphyloslide™ Latex test, a Staph ID 32 API system, and a Vitek 2 Compact system. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to examine the protein profile of all isolates, and real-time PCR was then used to identify mecA and PVL virulence genes. S aureus isolates were tested using the Vitek 2 Compact for antimicrobial susceptibility using Gram-positive cards (GP71). Among the 48 bacterial isolates tested, 45 (93.75%), 42 (87.5%), and 46 (95.83%) were positive in tube coagulase, the Staph ID 32 API system, and the Vitek 2 Compact system, respectively. We correctly identified all suspected S. aureus isolates (100%) via MALDI-TOF MS with a score value ≥2.00 and differentiated them into 22/48 MRSA (45.83%) and 26/48 MSSA (54.17%) isolates. A higher peak intensity at masses of 5530 Da, 6580 Da, 6710 Da, and 6820 Da was detected in MRSA, but not in MSSA. All MRSA isolates tested positive for the mecA gene, while all isolates tested negative for the PVL gene. The antibiotic susceptibility results showed that 22 (100%), 20 (90.91%), 19 (86.36%), 18 (81.82%), 17 (77.27%), 15 (68.18%), 13 (59.1%), and 12 (54.55%) MRSA strains were resistant to cefoxitin, daptomycin, erythromycin, benzylpenicillin, ciprofloxacin, oxacillin, and clindamycin, respectively. In contrast, all MRSA strains were extremely susceptible (100%) to linezolid, nitrofurantoin, quinupristin–dalfopristin, tigecycline, and vancomycin. Moreover, 20 (90.91%), 18 (81.82%), and 17 (77.27%) of the MRSA strains exhibited high sensitivity against rifampin, trimethoprim–sulfamethoxazole, and gentamicin, respectively. In DFIs, MALDI-TOF MS is a powerful and accurate method of identifying and distinguishing both MRSA and MSSA isolates. A high level of antimicrobial resistance was found in MRSA isolates, and antibiotic therapy based on antibiotic susceptibility patterns is essential for a successful outcome.

Purpose: Salmonella is a significant pathogen affecting wider range of animals and one of the main causes of diarroeal diseases leading to millions of human cases globally. Pathogenicity islands of Salmonella (SPI) are imperative in invasion of host cell and pathogenesis within the cell. It is found that SPI from 1 to 5 are present in every serovars of S. enterica and these are laterally acquired virulence regions. Research Method: This study investigated the presence of some virulence genes that belong to the SPI1, SPI2, SPI3 and SPI5 within 23 Salmonella isolates from broiler chicken meat in Sri Lanka using PCR method and further some virulence genes (gene encoding the invasion-associated protein, structural gene for attachment and invasion, regulatory gene for invasion and like nucleoid structuring gene) were quantified using RT- PCR. Findings: The study revealed the presence of Pathogenicity Island 1 genes such as hilA, invH, invF, invA and hns in all the isolates. Among the SPI 2 genes investigated, all the isolates of Salmonella showed the presence of ssaO, ssaQ, ssaP and ssaS (genes encoding for T3SS apparatus proteins), sscB and sscA (gene for secretion system chaperon proteins), sseF, sseD (genes coding for secreted effector proteins). In all isolates, SPI 3 genes, responsible in magenisum transport (mgtB and mgtC) were found. The rhuM gene (that supposed to code for a cytoplasmic protein) was absent in four isolates whereas the gene cigR (which predicted as codes for membrane protein) was absent in two isolates. The SPI 5 genes pipB was present in all the isolates except in one isolate whereas sopB was absent in one isolate. Originality/Value: This study found that some SPI genes are conserved in most of the Salmonella isolates from fresh broiler chicken meat in Sri Lanka and the expression of different virulence genes vary with the isolate. This molecular basic will pave the way to explore more on local isolates of Salmonella, detection methods and control methods. [ABSTRACT FROM AUTHOR]

The lesser-known traditionally processed meat products such as beef kargyong, pork kargyong, satchu, and khyopeh are popular food items in the Himalayan state of Sikkim in India. The present study aimed to assess the microbiological safety of traditional meat products by identifying the potential spoilage or pathogenic bacteria, detecting the enterotoxins, and screening the antibiotic susceptibility patterns. The pH and moisture contents of the meat products varied from 5.3 to 5.9 and from 1.5 to 18%, respectively. The microbial loads of aerobic bacteria were 105 to 107 cfu/g, Staphylococcus 103 to 106 cfu/g, Bacillus 104 to 106 cfu/g, and total coliform 102 to 107 cfu/g, respectively. Based on 16S rRNA gene sequencing, the bacterial species isolated from traditionally processed meat products were Staphylococcus piscifermentans, Citrobacter freundii, Enterococcus faecalis, Salmonella enterica, Staphylococcus aureus, Citrobacter werkmanii, Klebsiella pneumoniae, Macrococcus caseolyticus, Klebsiella aerogenes, Staphylococcus saprophyticus, Pseudocitrobacter anthropi, Citrobacter europaeus, Shigella sonnei, Escherichia fergusonii, Klebsiella grimontii, Burkholderia cepacia, and Bacillus cereus. The enzyme-linked immunosorbent assay (ELISA) tests detected Salmonella spp. and enterotoxins produced by B. cereus well as Staphylococcus in a few tested samples. However, the PCR method did not detect the virulence genes of B. cereus and Salmonella in the isolates. Virulence gene (sea) was detected in S. piscifermentans BSLST44 and S. piscifermentans BULST54 isolated from beef kargyong and in S. aureus PSST53 isolated from pork kargyong. No enterotoxins were detected in khyopeh samples. The antibiotic sensitivity test showed that all bacterial strains were susceptible toward gentamicin, cotrimoxazole, norfloxacin, and trimethoprim. Gram-positive bacteria showed 100% sensitivity against clindamycin and erythromycin; however, 50% of the resistance pattern was observed against oxacillin followed by penicillin (33%) and ampicillin (27%).

Escherichia coli remain significant problem caused urinary tract infection; Results indicated a higher urinary tract infection in women compared with men across all age groups. Phylogenetic analysis showed that majority of uropathogenic isolated E. coli belong to phylogroup B2 followed by D. The isolates showed the existence of the first type of fimbriae, maximum P fimbriae positive isolates 74/94 (80.43%) were widely correlating with common UTI. colony adhesion factor (CAFÐ) represented in 40 (43.47%) and 5 (5.43%) colony adhesion factor (CAFØ). Six major clusters (A-F) were identified depending on antibiograms typing. B2 is the most phylogenetic type showed wide range of resistant from 1 to 12 resistant to antibiotic of remaining strains. 14 isolates (15.21%) were detected as ESâLs producers and 30 isolates (32.60%) of them were AmpC b-lactamases producers. prevalence of virulent genes occurred in 51 papC (55.43%), 66 fim H (71.73%) and SfaDE detected in low occurrence 21(22.82). These results emphasize the low or moderate resistant to antibiotics focused in isolates with high genetic content .These results indicated that the greater the resistance to antibiotics the less the genetic expression of the virulence factors, this confirms the reverse relationship. The genes required for uropathogenicity in a single isolate may not reflect virulence in another isolate. Pathogenicity is a multi-factor characteristic, the result of adhesion-related genes which interact in separate set in various genetic backgrounds.

The study of biodiversity of Pyricularia oryzae Cav. in the rice-growing areas of the south of Russia using the methods of PCR On the basis of molecular-genetic approaches there has been conducted the study of biodiversity of phyto pathogenic fungus Pyricularia oryzae Cav. in the south of Russia. We have conducted the monitoring and selected the isolates of the pathogen from the infected herbarium material, collected from the fields of six rice-growing areas of the Krasnodar Kray with various degree of harmfulness of rice blast pathogen. Their DNA-passports have been made. The studied isolates of the blast pathogen have been characterized in respect of their morphological traits. The phyto-pathological test with the application of varieties-differentiators of rice has established quantitative and qualitative composition of the virulent genes in fungus strains. The study has found the genes of pathogen tolerance, greatly efficient in the south of Russia, which can be recommended for the breeding programs for growing rice varieties resistant to rice blast.

The lesser-known traditionally processed meat products such as beef kargyong , pork kargyong , satchu , and khyopeh are popular food items in the Himalayan state of Sikkim in India. The present study aimed to assess the microbiological safety of traditional meat products by identifying the potential spoilage or pathogenic bacteria, detecting the enterotoxins, and screening the antibiotic susceptibility patterns. The pH and moisture contents of the meat products varied from 5.3 to 5.9 and from 1.5 to 18%, respectively. The microbial loads of aerobic bacteria were 105 to 107 cfu/g, Staphylococcus 103 to 106 cfu/g, Bacillus 104 to 106 cfu/g, and total coliform 102 to 107 cfu/g, respectively. Based on 16S rRNA gene sequencing, the bacterial species isolated from traditionally processed meat products were Staphylococcus piscifermentans , Citrobacter freundii , Enterococcus faecalis , Salmonella enterica , Staphylococcus aureus , Citrobacter werkmanii , Klebsiella pneumoniae , Macrococcus caseolyticus , Klebsiella aerogenes , Staphylococcus saprophyticus , Pseudocitrobacter anthropi , Citrobacter europaeus , Shigella sonnei , Escherichia fergusonii , Klebsiella grimontii , Burkholderia cepacia , and Bacillus cereus. The enzyme-linked immunosorbent assay (ELISA) tests detected Salmonella spp. and enterotoxins produced by B. cereus well as Staphylococcus in a few tested samples. However, the PCR method did not detect the virulence genes of B. cereus and Salmonella in the isolates. Virulence gene (sea) was detected in S. piscifermentans BSLST44 and S. piscifermentans BULST54 isolated from beef kargyong and in S. aureus PSST53 isolated from pork kargyong. No enterotoxins were detected in khyopeh samples. The antibiotic sensitivity test showed that all bacterial strains were susceptible toward gentamicin, cotrimoxazole, norfloxacin, and trimethoprim. Gram-positive bacteria showed 100% sensitivity against clindamycin and erythromycin; however, 50% of the resistance pattern was observed against oxacillin followed by penicillin (33%) and ampicillin (27%). [ABSTRACT FROM AUTHOR]

Microbial contamination of soft cheese may originate from various sources as contaminated equipment's or from the workers themselves during cheese production. Our study was aimed to characterize Bacillus cereus organisms using culture-based and "PCR" techniques among 285 selected samples along Damietta and Kareish cheese producing line in a small cheese plant at Alexandria governorate which including raw milk, cheese, whey, rennet, water, and swabs from all contacts surfaces. Overall, of 10.90 % among raw milk, cheese, whey, rennet, water, and swabs from all contacts surfaces were positive for Bacillus species in Damietta and Kareish cheese producing line. Incidence of Bacillus cereus in receiving raw milk, curd, whey, fresh cheese samples were 6.66, 13.33 ,0 and 13.33%, respectively for traditional Damietta cheese plant. Bacillus cereus organisms was found only within 6.66 % in receiving raw milk, pasteurized milk and fresh cheese samples for traditional Kareish cheese plant. Hbl, Nhe and Cytk genes were detected in Bacillus cereus isolated from curd and Damietta cheese. Nhe gene was detected in Bacillus cereus isolated from raw milk, pasteurized milk and Kareish cheese While, Ces gene was detected only in Bacillus cereus isolated from fresh Kareish cheese samples. Prevalence of Eneterotoxigenic virulent B. cereus poses public health threats to consumers. HACCP should be implemented for all steps through manufacture in order to produce safe and good quality dairy products. [ABSTRACT FROM AUTHOR]

Aims: The occurrence of multiple pathogenic Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa are important nosocomial and hazardous infection clinically challenge worldwide. Thus, the aim of this study was to screen for the virulent genes profiles to ascertain their prevalence in swiftlets in Borneo. Methodology and results: The Enterococci, E. coli and P. aeruginosa bacteria were isolated from the swiftlets' faeces and air inside swiftlet houses, which located in the Southern, Central and Northern regions of Borneo. The isolates were identified to the species level by 16S rRNA sequencing assay. Specific primers were designed for detection of the potential virulence genes in E. faecalis (ace, AS, efaA and gelE), E. coli (stx) and P. aeruginosa (oprL) by PCR assay. A total of 38 Enterococci, 26 of E. coli and 2 of P. aeruginosa fecal and airborne bacteria were identified. Sixty-seven percent of E. faecalis isolates were detected positive for four virulence genes, 27% possessed three (AS, efaA, gelE) genes and 6% possessed two (ace, AS) genes. There were no stx genes detected among all the E. coli isolates. The oprL gene was detected in all the P. aeruginosa isolates. Conclusion, significance and impact of study: Virulence genes are important in the pathogenesis of both clinical and avian infections which considered to be a serious public health threat. The high incidence of virulence genes detection in E. faecalis and P. aeruginosa indicates these genes were widely disseminated among the bacteria found in swiftlet houses, suggesting the important issues in the pathogenesis of infections and diseases which may cause potential health risks to humans. [ABSTRACT FROM AUTHOR]

[Display omitted] • UV 185+254 radiation instantly deactivated ARGs and VGs in indoor bioaerosol. • Removal of target genes was up to 2 orders of magnitude lower than host bacteria. • Thicker cell wall and plasmid carrier prominently hindered ARGs and VGs destruction. • UV 185+254 exhibited superior temperature adaptability and energy efficiency. • Filled in the gap of airborne ARGs and VGs removal in full-scale ventilation ducts. The energy-efficient inactivation of airborne antibiotic resistance genes (ARGs) and virulence genes was seldom investigated in full-scale ventilation systems. This study examined the potential of UV disinfection to inactivate two types of chromosome- or plasmid-encoded ARGs (mecA and amp) and one chromosomal virulence gene (tst1) within two host bacterial species: Staphylococcus aureus and Escherichia coli. UV 185+254 disinfection exhibited dual damages (i.e., physical by 254 nm UV and oxidative by the produced ozone) on those target genes, resulting in relatively strong adaptability to temperature and low electrical energy per order in specific gene removal (down to 1 kJ m−3). It was found that the disinfection efficiency of target genes was up to 2 orders of magnitude lower than that of the host bacteria. The target genes with more bases and higher weighted bipyrimidine content were more susceptible to UV 185+254 treatment. Thicker cell walls and plasmid carriers prominently hindered UV 185+254 deactivation, requiring larger radiation power to enhance the gene removal efficiency in bioaerosols. [ABSTRACT FROM AUTHOR]

Objectives: This review fills the paucity of information on K. pneumoniae as a nosocomial pathogen by providing pooled data on epidemiological risk factors, resistant trends and profiles and resistant and virulent genes of this organism in Asia. Methods: Exhaustive search was conducted using PubMed, Web of Science, and Google scholar for most studies addressing the prevalence, risk factors, drug resistant-mediated genes and/or virulent factors of K. pneumoniae in Asia. Data extracted for meta-analysis were analyzed using comprehensive meta-analysis version 3. Trends data for the isolation rate and resistance rates were entered into Excel spread sheet and the results were presented in graphs. Results: The prevalence rate of drug resistance in K. pneumoniae were; amikacin (40.8%) [95% CI 31.9–50.4], aztreonam (73.3%) [95% CI 59.9–83.4], ceftazidime (75.7%) [95% CI 65.4–83.6], ciprofloxacin (59.8%) [95% CI 48.6–70.1], colistin (2.9%) [95% CI 1.8–4.4], cefotaxime (79.2%) [95% CI 68.0–87.2], cefepime (72.6) [95% CI 57.7–83.8] and imipenem (65.6%) [95% CI 30.8–89.0]. TEM (39.5%) [95% CI 15.4–70.1], SHV-11 (41.8%) [95% CI 16.2–72.6] and KPC-2 (14.6%) [95% CI 6.0–31.4] were some of the resistance mediated genes observed in this study. The most virulent factors utilized by K. pneumoniae are; hypermucoviscous phenotype and mucoviscosity-related genes, genes for biosynthesis of lipopolysaccharide, iron uptake and transport genes and finally, adhesive genes. Conclusion: It can be concluded that, antimicrobial resistant in K. pneumoniae is a clear and present danger in Asia which needs strong surveillance to curb this menace. It is very important for public healthcare departments to monitor and report changes in antimicrobial-resistant isolates. [ABSTRACT FROM AUTHOR]

Purpose: The aquaculture sector is a major contributor to the economic and nutritional security for a number of countries. India's total seafood exports for the year 2017–2018 accounted for US$ Million 7082. One of the major setbacks in this sector is the frequent outbreaks of diseases often due to bacterial pathogens. Vibriosis is one of the major diseases caused by bacteria of Vibrio spp., causing significant economic loss to the aquaculture sector. The objective of this study was to understand the genetic composition of Vibrio spp. Methods: Thirty-five complete genomes were downloaded from GenBank comprising seven vibrio species, namely, Vibrio alginolyticus, V. anguillarum, V. campbellii, V. harveyi, V. furnissii, V. parahaemolyticus, and V. vulnificus. Pan-genome analysis was carried out with coding sequences (CDS) generated from all the Vibrio genomes. In addition, genomes were mined for genes coding for toxin-antitoxin systems, antibiotic resistance, genomic islands, and virulence factors. Results: Results revealed an open pan-genome comprising of 2004 core, 8249 accessory, and 6780 unique genes. Downstream analysis of genomes and the identified unique genes resulted in 312 antibiotic resistance genes, 430 genes coding for toxin and antitoxin systems along with 4802, and 4825 putative virulent genes from genomic island regions and unique gene sets, respectively. Conclusion: Pan-genome and other downstream analytical procedures followed in this study have the potential to predict strain-specific genes and their association with habitat and pathogenicity. [ABSTRACT FROM AUTHOR]

A total of 100 migratory birds were collected from Dakahlia and Damietta Governorates. All samples were subjected to clinical and postmortem (P.M) examination followed by bacteriological examination for detection of Avian Pathogenic E. coli infection. Out of 100 samples, 25 samples were distinguished as positive for Escherichia coli. Accurately, 4 E. coli serogroupes were identified in 25 positive samples and the most predominant serogroup were O125 (76 %), O126 (8%), O158 (8%), O86 (4%) and untypable E. coli (4 %). Studying antibiotic resistance pattern of E. coli isolates revealed that 64% of the isolates demonstrated multidrug resistance pattern. The highest resistance rates were recorded against Ciprofloxacin (88%) followed by Gentamycin (80%), Streptomycin (68%), Norfloxacin (40%), Colistin (16%) and Doxycycline (4%). Conventional PCR was done for detection of virulence genes (fimH, tsh, eaeA, stx1and stx2) in 10 isolates and the results revealed that; 100%, 60%, 40%, 0% and 0% were positive for these virulence genes, respectively. By studying the effect of one isolate of O125 No. 78 in commercial chicken (cobb) and ducks (pekin) we found that, more sever signs, pathological lesion and 100% mortality rate in cobb chicken than pekin ducks which had less signs nor pathological lesion with 10% mortality rate only. [ABSTRACT FROM AUTHOR]

A total of 150 fish samples, including 75 samples each of Tilapia nilotica and Mugil cephalus collected from different fish markets distributed in Mansoura city, Egypt were examined bacteriologically to determine the prevalence, antimicrobial susceptibility and the virulence of Aeromonas hydrophila regarding to aerolysin and haemolysin toxins using PCR technique. Phenotypically, presumptive A. hydrophila could be detected in 43/75 (57.33%) and 38/75 (50.67%) of the examined Tilapia nilotica and Mugil cephalus, respectively, with an overall incidence of 81/150 (54%). The results of PCR test based on using A. hydrophila 16S rRNA gene demonstrated that 32/75 (42.67%) and 29/75 (38.67%) of the examined Tilapia nilotica and Mugil cephalus, respectively were contaminated with A. hydrophila, with an overall incidence of 61/150 (40.67%). The aerA gene was detected in all (61) A. hydrophila isolates, while hlyA gene couldn't detected in any of the identified strains. The results of antimicrobial susceptibility test showed that all (100%) isolated A. hydrophila strain were resistant to both ampicillin and carbenicillin, while chloramphenicol and amikacin showed the same percentage but account for sensitivity, whereas higher percentage of sensitivity was detected against ciprofloxacin (92.59%), followed by cefoxitin (87.65%), cefotaxime (80.24%), trimethoprim/sulfamethoxazole (79%), tetracycline (74%). The result declared that examined fish samples were markedly contaminated with virulent and multidrug strains of A. hydrophila which may be of a potential health risk. [ABSTRACT FROM AUTHOR]

The study evaluated the safety and colonisation properties of four lactic acid bacteria (LAB), by determining their cell hydrophobicity and aggregation abilities. In addition, the presence of virulence and resistance genes was assayed in these probiotic candidates. Lactobacillus reuteri ZJ625, Lactobacillus reuteri VB4, Lactobacillus salivarius ZJ614 and Streptococcus salivarius NBRC13956 were tested for cell surface hydrophobicity abilities against xylene, chloroform and ethyl acetate. The isolates were also tested for auto-aggregation and co-aggregation abilities; the optical densities of cell growth were measured after 1, 2, 3 and 4 h of experimental set-up. DNA was extracted from all the four isolates and amplified using PCR with specific primers to detect virulence genes of adhesion collagen protein (ace) and aggregation substances (agg and asa); also, resistance genes of Vancomycin vanA, Vancomycin vanC1 and Vancomycin vanC2/3 were assayed in the four isolates. The isolates showed high hydrophobicity to all solvents: xylene (78–84%), chloroform (68–75%) and ethyl acetate (52–60%). High auto- and co-aggregations ranging from 60 to 70% and from 45 to 56% respectively were observed in the isolates after 4 h of incubation at 37 °C. Some of the tested isolates showed the presence of virulence and resistance genes; however, this does not indicate that these genes are unsafe because their transmission and expression abilities are unknown. Therefore, in this study, the isolates studied are considered safe for use as future probiotics, as revealed from results presented, which generally represents the scanned safety evaluations of the isolates as promising probiotics. [ABSTRACT FROM AUTHOR]

A total of 100 diseased broiler chickens of average age 22-28 days old were collected from different farms located in Dakahlia Governorate. All samples were subjected to clinical and postmortem (P.M) examination as well as bacteriological examination for detection of infection. Thus, E. coli was recovered from examined organs of 37 chickens with an incidence rate of 37 % (37 out of 100). Accuratly, 10 E. coli serogroupes were identified in 28 positive samples and the most predominant serogroup were O125 16%, O91 14%, O1 11%, O26 K60 11% & O86 K64 8 & O128 5% & O55 K59 3% & O166 3% & O103 3% and O144 3% and untypable E. coli (24 %). Studying antibiogram resistance pattern of E. coli isolates revealed that all isolates demonstrated multidrug resistence pattern. The highest resistance rates were recorded against Lincomycin (100%) and lowest are Colistin (7.14%) and other tested antibiotics showed variations regarding their resistance patterns. The most predominant serotypes of this study have the ability to form biofilm on the inner wall of the glass. PCR was done for detection of resistant genes (blaTEM and tetA(A)) and virulence genes (csgD, adrA, eaeA and fimH) in the 10 tested isolated serotypes and the results revelead that, 100% and 90% &100%, 90%, 90% and 100% were positive for resistant genes and virulence genes, respectively and investigate the relationship between them was discovered. By studying the effect of Olive Leaf Extract on multidrug resistant isolates, results were revealed that all tested serotypes are sensitive to Olive Leaf Extract (10% conc.). [ABSTRACT FROM AUTHOR]

Background and Objectives: Non-typhoidal Salmonellosis, a zoonotic infection associated with acute gastroenteritis is caused by non-typhoidal salmonellae (NTS). The study was carried out to determine the prevalence of NTS serovars and their antimicrobial resistance along with the presence of the virulence gene (invA gene) in poultry samples. Materials and Methods: This is a prospective cross-sectional study carried out at the Enteric Diseases Division, Kasturba Medical College, Manipal, South India from January 2016– December 2017. Poultry samples were collected randomly from two local poultry farms in Udupi district and processed following CDC standard protocol. Results: From the 396 poultry meat samples, intestinal contents and faecal samples collected, 58 NTS serovars were isolated showing a prevalence of 14.64%. Salmonella Infantis, 43.1%, 25/58 was the commonest serovar. Resistance to ciprofloxacin 72.41%, ampicillin 32.8%, gentamicin 17.24%, cotrimoxazole 29.31% and amoxicillin-clavulanic acid 6.9% was observed. The invA gene was detected in 43 NTS isolates (74.13%). Conclusion: Poultry sources are recognized as a significant cause for non-typhoidal salmonellosis. Therefore, hygienic measures should be initiated to reduce the contamination of meat and poultry products with virulent strains of Salmonella that are of public health significance.

The prevalent Staphylococcus aureus clones and antibiotic susceptibility profiles are known to change dynamically and geographically; however, recent S. aureus strains causing infections in women and children in China have not been characterized. In this study, we analyzed the molecular epidemiology and antimicrobial resistance of S. aureus isolated from patients in four centers for women and children in Guangzhou, China. In total, 131 S. aureus isolates (100 from children and 31 from women) were analyzed by spa typing, multi-locus sequence typing, virulence gene and antimicrobial resistance profiling, staphylococcal chromosomal cassette mec typing, and mutation analyses of rpoB. A total of 58 spa types, 27 sequence types (STs), and 10 clonal complexes (CCs) were identified. While CC59 (ST59-IV, 48.8%; ST338-III, 35.7%) and CC45 (ST45-IV, 100%) were the major clones (84.4%) among MRSA isolates, CC5 (ST188, 24.3%; ST1, 21.6%) and CC398 (ST398, 70%) were the major ones (70.1%) among MSSA isolates. ST338-MRSA-III mostly found in pus but hardly in respiratory tract samples while ST45-MRSA-IV was on the opposite, even though they both found in blood and cerebrospinal fluid sample frequently. Staphylococcal enterotoxin genes seb-seq-sek were strongly associated with ST59 and ST338, while sec was associated with ST45, ST121, ST22, and ST30. All ST338, ST1232, and SCCmec III isolates carried lukF/S-PV genes. A total of 80% of ST338 isolates were resistant to erythromycin, clindamycin, and tetracycline. All ST45 isolates exhibited intermediate or complete resistance to rifampicin. In total, 481 HIS/ASN mutations in rpoB were found in rifampicin-resistant or intermediate-resistant isolates. ST338-III and ST45-IV emerged as two of three major clones in MRSA isolates from women and children in Guangzhou, China, though ST59-MRSA-IV remained the most prevalent MRSA clone. Clonal distribution of S. aureus varied, depending on the specimen source. Virulence genes and antibiograms were closely associated with the clonal lineage. These results clarified the molecular epidemiology of S. aureus from women and children in Guangzhou, China, and provide critical information for the control and treatment of S. aureus infections.

Background and Objectives: Non-typhoidal Salmonellosis, a zoonotic infection associated with acute gastroenteritis is caused by non-typhoidal salmonellae (NTS). The study was carried out to determine the prevalence of NTS serovars and their antimicrobial resistance along with the presence of the virulence gene (invA gene) in poultry samples. Materials and Methods: This is a prospective cross-sectional study carried out at the Enteric Diseases Division, Kasturba Medical College, Manipal, South India from January 2016– December 2017. Poultry samples were collected randomly from two local poultry farms in Udupi district and processed following CDC standard protocol. Results: From the 396 poultry meat samples, intestinal contents and faecal samples collected, 58 NTS serovars were isolated showing a prevalence of 14.64%. Salmonella Infantis, 43.1%, 25/58 was the commonest serovar. Resistance to ciprofloxacin 72.41%, ampicillin 32.8%, gentamicin 17.24%, cotrimoxazole 29.31% and amoxicillin-clavulanic acid 6.9% was observed. The invA gene was detected in 43 NTS isolates (74.13%). Conclusion: Poultry sources are recognized as a significant cause for non-typhoidal salmonellosis. Therefore, hygienic measures should be initiated to reduce the contamination of meat and poultry products with virulent strains of Salmonella that are of public health significance. [ABSTRACT FROM AUTHOR]

Aeromonas spp., is an important opportunistic pathogen responsible for great economic losses in aquaculture. In this paper, we report our study of an Aeromonas infection that occurred in cultured sheatfish Silurus glanis in May 2016 in a farm in Tianjin city. Typical external symptoms included petechial skin hemorrhages, ascitic distension of the abdomen, redness and swelling of the anus. The isolate SAV-165 from the diseased fish was identified as A. veronii biovar sobria by biochemical properties and molecular techniques. Experimental challenge showed that a 50% lethal dose (LD50) of SAV-165 was 1.59 × 106 CFU/ml in zebrafish Danio rerio. The antimicrobial susceptibility test indicated that the isolate was susceptible to aminoglycosides and chloramphenicol, but resistant to penicillins and sulphonamides. Histopathological examination showed fatty changes and swelling of hepatocytes in the liver, congestion in the submucosa of intestine and the renal veins of the kidney. This is the first report of A. veronii biovar sobria infection in S. glanis in China. [ABSTRACT FROM AUTHOR]

Escherichia coli remain significant problem caused urinary tract infection; Results indicated a higher urinary tract infection in women compared with men across all age groups. Phylogenetic analysis showed that majority of uropathogenic isolated E. coli belong to phylogroup B2 followed by D. The isolates showed the existence of the first type of fimbriae, maximum P fimbriae positive isolates 74/94 (80.43%) were widely correlating with common UTI. colony adhesion factor (CAFÐ) represented in 40 (43.47%) and 5 (5.43%) colony adhesion factor (CAFØ). Six major clusters (A-F) were identified depending on antibiograms typing. B2 is the most phylogenetic type showed wide range of resistant from 1 to 12 resistant to antibiotic of remaining strains. 14 isolates (15.21%) were detected as ESâLs producers and 30 isolates (32.60%) of them were AmpC â-lactamases producers. prevalence of virulent genes occurred in 51 papC (55.43%), 66 fim H (71.73%) and SfaDE detected in low occurrence 21(22.82) . These results emphasize the low or moderate resistant to antibiotics focused in isolates with high genetic content .These results indicated that the greater the resistance to antibiotics the less the genetic expression of the virulence factors, this confirms the reverse relationship. The genes required for uropathogenicity in a single isolate may not reflect virulence in another isolate. Pathogenicity is a multi-factor characteristic, the result of adhesion-related genes which interact in separate set in various genetic backgrounds. [ABSTRACT FROM AUTHOR]

Streptococcus mutans is a primary etiological agent of dental caries. Farnesol, as a potential antimicrobial agent, inhibits the development of S. mutans biofilm. In this study, we hypothesized that farnesol inhibits caries development in vitro and interferes with biofilm formation by regulating virulence-associated gene expression. The inhibitory effects of farnesol to S. mutans biofilms on enamel surfaces were investigated by determining micro-hardness and calcium measurements. Additionally, the morphological changes of S. mutans biofilms were compared using field emission scanning electron microscopy and confocal laser scanning microscopy, and the vitality and oxygen sensitivity of S. mutans biofilms were compared using MTT assays. To investigate the molecular mechanisms of farnesol's effects, expressions of possible target genes luxS, brpA, ffh, recA, nth, and smx were analyzed using reverse-transcription polymerase chain reaction (PCR) and quantitative PCR. Farnesol-treated groups exhibited significantly higher micro-hardness on the enamel surface and lower calcium concentration of the supernatants as compared to the-untreated control. Microscopy revealed that a thinner film with less extracellular matrix formed in the farnesol-treated groups. As compared to the-untreated control, farnesol inhibited biofilm formation by 26.4% with 500 μmol/L and by 37.1% with 1,000 μmol/L (P < 0.05). Last, decreased transcription levels of luxS, brpA, ffh, recA, nth, and smx genes were expressed in farnesol-treated biofilms. In vitro farnesol inhibits caries development and S. mutans biofilm formation. The regulation of luxS, brpA, ffh, recA, nth, and smx genes may contribute to the inhibitory effects of farnesol. [ABSTRACT FROM AUTHOR]

This Study was carried out on 200 random samples of chicken meat products represented by chicken luncheon, chicken burger, chicken sausage and chicken shawerma (50 of each). Samples were randomly collected from different supermarkets and retailers of different sanitation levels at Mansoura city, El Dakahlia Province, Egypt and bacteriologically analyzed to assess the prevalence of Staph. aureus, E. coli and S. spp. and their enterotoxigenic virulence genes using PCR in some chicken meat products intended for direct consumption. The obtained results revealed that the prevalence of Staph. aureus in examined chicken luncheon, chicken burger, chicken sausage and chicken shawerma were 6%, 2%, 2% and 2%., respectively. While E. coli were 2%, 4%, 0% and 2% in examined samples respectively and S. spp. was isolated by 2% from shawerma only. The isolated S. typhimurium harbor invA and stn genes. The isolated E. coli showed presence of shiga toxin genes (stx1 and stx2). The examined coagulase positive Staph. aureus showed the presence of different enterotoxin genes sea, seb, sec, sed and see. Thus it is necessary to adopt a regime of good, safe and healthy production of such chicken meat products with cleaning and disinfection and hygienic packaging in order to ensure safe products for consumers. [ABSTRACT FROM AUTHOR]