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A total of sixty-two isolates were tested to assess the presence of the irp2 gene in different isolates of Klebsiella pneumoniae. The isolates discussed in this study were obtained from patients who had acquired illnesses either within the hospital premises or in the surrounding vicinity. These isolates were sourced from three hospitals located in Baghdad, namely Al-Imam Ali, Al-Zaafaranya, and Ibin-Albady hospitals. One interesting thing about Klebsiella pneumoniae is that it makes siderophores, especially yersiniabactin. This is because of a gene that controls this trait. The application of DNA sequencing methodologies has facilitated the identification of the irp2 gene in 44% of Klebsiella pneumoniae. According to amino acid sequences and differentiation of nucleotide, the current work reports findings on the identification of the K. pneumoniae irp2 gene isolates collected from patients in Iraq. This event represents the initial recorded occurrence of such detection. The presence of this gene is considered an unconventional human pathogen. The aim is to explore the correlation between genetic analysis and the diagnosis of genetic variation by examining isolates documented in the global GenBank database (LC791754.1, LC791755.1, LC791756.1, LC791757.1, LC791758.1, LC791759.1, LC791760.1). Additionally, it seeks to provide insights into the magnitude of genetic variation observed within these isolates. [ABSTRACT FROM AUTHOR]

Gallinarum disease caused by Salmonella gallinarum is one of the most challenging diseases in the poultry industry. In this study, the level of Salmonella gallinarum contamination of industrial chicken flocks in northwestern Iran, antibiotic resistance and frequency of some virulence genes (msgA, spiA, pagC) were investigated and evaluated. All the samples taken from the herds with suspected clinical symptoms of salmonellosis from the clinics of East Azarbaijan, West Azarbaijan, Ardabil and Zanjan provinces were examined and confirmed by microbial culture, chemical analysis, PCR and serological methods. After confirming the diagnosis of involvement with Salmonella gallinarum, antibiotic resistance was evaluated by disking method and the abundance of virulence genes in each isolate was checked. Of the 25 isolates of Salmonella gallinarum in this study, the highest microbial resistance was reported to tetracycline (76%), oxytetracycline (72%) and the highest sensitivity to amikacin (80%) and Fozbek (72%). 100% of the isolates were resistant to at least 6 types of antibiotics. In the study of the isolates, 8% had the msgA gene, 28% had the spiA gene, and 8% had the pagC gene, of which two isolates had all three virulence genes. [ABSTRACT FROM AUTHOR]

To investigate the clinical features of endogenous Klebsiella pneumoniae endophthalmitis (EKPE) and the molecular characteristics of pathogenic K. pneumoniae. A retrospective study was conducted between January 2014 and January 2021. Clinical data were extracted. K. pneumoniae isolates cultured from EKPE patients' specimens were characterized by antimicrobial susceptibility testing, multilocus sequence typing, capsular serotyping, and virulence gene profiles. Virulence phenotypes were confirmed by mouse lethality assay. Thirty-nine patients (47 eyes) were diagnosed with EKPE. Administration of systemic antibiotics and eye treatment within 24 hours of ocular symptom onset was statistically associated with better visual outcomes (p =.007). Sixteen K. pneumoniae isolates were collected, three of which were carbapenem-resistant and avirulent. Only salmochelin encoding gene was harbored by all isolates. Systemic and eye treatment administered within 24 hours from the onset improves visual prognosis. Besides hypervirulent strains, avirulent carbapenem-resistant K. pneumoniae can cause EKPE. [ABSTRACT FROM AUTHOR]

Background: Macrophomina phaseolina is a pathogen that causes an opportunistic disease that spreads by soil and seeds and affects more than 500 different plant species, like fruits, trees, and row crops. Mycotoxins, such as phaseolinic acid, and phaseolinone, are produced by M. phaseolina isolates in previous investigations; however, the production of these mycotoxins seems to vary depending on the host and the region. Methods and results: In this study, Macrophomina phaseolina strain 3 A was isolated from rotten cassava tuber and identified using the analysis of the sequences of the internal transcribed spacer region. The isolate was inoculated on a fresh healthy cassava tuber at 25 °C and tuber-rotting potential was monitored for 4 weeks. Virulence genes MPH_06603, MPH_06955, and MPH_01521 were determined with designed primers, and secondary metabolites were characterized by FTIR and GCMS. The rotten tuber effect was observed from the 2nd week of the experiment with severe tuber rot and weight reduction. The PCR showed the presence of MPH_06603 virulence gene. The GCMS showed N-Methylpivalamide (115.0 m/z), Butane, 1,4-dimethoxy- (119.0 m/z), and 5-Hydroxymethylfurfural (126.0 m/z) were the predominant metabolites produced by the pathogen. The compounds in the metabolites inhibit CYP3A4 enzymes, cause eye irritation, and Human Ether-a-go-go-related gene inhibition. Conclusion: This study revealed that M. phaseolina was responsible for the cassava tuber rot which leads to a lower yield of farm produce. The metabolites produced are toxic and unsafe for human consumption. It is suggested that farmers should destroy any cassava affected by this pathogen to prevent its toxic effects on humans and animals. [ABSTRACT FROM AUTHOR]

Background: Enterobacter cloacae complex (ECC) including different species are isolated from different human clinical samples. ECC is armed by many different virulence genes (VGs) and they were also classified among ESKAPE group by WHO recently. The present study was designed to find probable association between VGs and antibiotic susceptibility in different ECC species. Methods: Forty-five Enterobacter isolates that were harvested from different clinical samples were classified in four different species. Seven VGs were screened by PCR technique and antibiotic susceptibility assessment was performed by disk-diffusion assay. Result: Four Enterobacter species; Enterobacter cloacae (33.3%), Enterobacter hormaechei (55.6%), Enterobacter kobei (6.7%) and Enterobacter roggenkampii (4.4%) were detected. Minimum antibiotic resistance was against carbapenem agents and amikacin even in MDR isolates. 33.3% and 13.3% of isolates were MDR and XDR respectively. The rpoS (97.8%) and csgD (11.1%) showed maximum and minimum frequency respectively. Blood sample isolated were highly virulent but less resistant in comparison to the other sample isolates. The csgA, csgD and iutA genes were associated with cefepime sensitivity. Conclusion: The fepA showed a predictory role for differentiating of E. hormaechei from other species. More evolved iron acquisition system in E. hormaechei was hypothesized. The fepA gene introduced as a suitable target for designing novel anti-virulence/antibiotic agents against E. hormaechei. Complementary studies on other VGs and ARGs and with bigger study population is recommended. [ABSTRACT FROM AUTHOR]

Wild birds as source of pathogenic microorganisms infecting livestock and humans are an interesting topic that has received increased attention in recent years. Here we study occurrence of pathogenic bacteria in birds in north-east Iran with focus on to identifying virulence gene in wild and domestic birds. Wild birds were trapped and sampled in different localities in northeast Iran from April to September in 2018 and 2019. From 184 birds representing 32 species of wild passerine birds, potentially pathogenic bacteria were isolated from 171 samples (92.9). Escherichia coli was the most frequently isolated bacterium with a prevalence of 70.1%, followed by Enterobacter spp. (53.8%) and Salmonella spp (3.2%) isolated from 129, 99 and 6 wild birds, respectively. In total 79.6% of the isolates carried virulence factors. A higher frequency of virulence genes was detected in domestic birds (88.6%) than in wild birds (76%). The results showed that birds in north-east Iran can serve as a potential source of bacterial pathogens and virulence factors that may cause disease in both humans and birds. Furthermore, our results introduce new host birds as source of pathogenic microorganisms that can carry and disseminate them in the environment. [ABSTRACT FROM AUTHOR]

The present study aimed to isolate and identify Methicillin-Resistant DOIStaphylococcus aureus MRSA from the nasal cavity of healthy cows and buffaloes and their breeders in Nineveh Governorate and detect some virulence factors by using molecular methods. A total of 150 samples of cotton swabs were collected randomly from different areas of Nineveh governorate. The samples were cotton swabs from the nasal passages of healthy cattle, buffaloes, and their breeders (50 swabs of each type). All the samples were subjected to culture and molecular testing. The results showed the highest isolation percentage of S. aureus from cattle followed by breeders, then buffaloes, at 54%, 40%, and 32%, respectively. The total isolation percentage of MRSA was 65.1%. The highest percentage was in buffaloes, followed by breeders and cattle, at 93.75%, 70%, and 44.44%, respectively. Out of 41 isolates from cattle, buffaloes, and their breeders, the virulence genes clfA, clfB, and coa were detected in MRSA at rates of 100%, 80.49%, and 65.85%, respectively. The current study concluded that cattle and buffalo are considered carriers and potential transmitters of MRSA, which makes them risk factors for infection in humans, especially those who are in direct contact with animals. Together, these findings also highlight the need to prevent the transmission of zoonotic pathogens to humans via occupational exposure or consumption of contaminated animal products. [ABSTRACT FROM AUTHOR]

Abstract Background Enterobacter cloacae complex (ECC) including different species are isolated from different human clinical samples. ECC is armed by many different virulence genes (VGs) and they were also classified among ESKAPE group by WHO recently. The present study was designed to find probable association between VGs and antibiotic susceptibility in different ECC species. Methods Forty-five Enterobacter isolates that were harvested from different clinical samples were classified in four different species. Seven VGs were screened by PCR technique and antibiotic susceptibility assessment was performed by disk-diffusion assay. Result Four Enterobacter species; Enterobacter cloacae (33.3%), Enterobacter hormaechei (55.6%), Enterobacter kobei (6.7%) and Enterobacter roggenkampii (4.4%) were detected. Minimum antibiotic resistance was against carbapenem agents and amikacin even in MDR isolates. 33.3% and 13.3% of isolates were MDR and XDR respectively. The rpoS (97.8%) and csgD (11.1%) showed maximum and minimum frequency respectively. Blood sample isolated were highly virulent but less resistant in comparison to the other sample isolates. The csgA, csgD and iutA genes were associated with cefepime sensitivity. Conclusion The fepA showed a predictory role for differentiating of E. hormaechei from other species. More evolved iron acquisition system in E. hormaechei was hypothesized. The fepA gene introduced as a suitable target for designing novel anti-virulence/antibiotic agents against E. hormaechei. Complementary studies on other VGs and ARGs and with bigger study population is recommended.

The present study aimed to isolate and identify Methicillin-Resistant Staphylococcus aureus MRSA from the nasal cavity of healthy cows and buffaloes and their breeders in Nineveh Governorate and detect some virulence factors by using molecular methods. A total of 150 samples of cotton swabs were collected randomly from different areas of Nineveh governorate. The samples were cotton swabs from the nasal passages of healthy cattle, buffaloes, and their breeders (50 swabs of each type). All the samples were subjected to culture and molecular testing. The results showed the highest isolation percentage of S. aureus from cattle followed by breeders, then buffaloes, at 54%, 40%, and 32%, respectively. The total isolation percentage of MRSA was 65.1%. The highest percentage was in buffaloes, followed by breeders and cattle, at 93.75%, 70%, and 44.44%, respectively. Out of 41 isolates from cattle, buffaloes, and their breeders, the virulence genes clfA, clfB, and coa were detected in MRSA at rates of 100%, 80.49%, and 65.85%, respectively. The current study concluded that cattle and buffalo are considered carriers and potential transmitters of MRSA, which makes them risk factors for infection in humans, especially those who are in direct contact with animals. Together, these findings also highlight the need to prevent the transmission of zoonotic pathogens to humans via occupational exposure or consumption of contaminated animal products.

Xiaojuan Ji,1,* Wenjie Zhu,1,* Hongfei Lu,2 Zhiwei Wu,1 Huaqing Chen,1 Chunli Lin,1 Zhaolin Zeng,1 Cong You,1 Longnian Li1 1Department of Dermatology, First Affiliated Hospital of Gannan Medical University, Joint Organization of Jiangxi Clinical Medicine Research Center for Dermatology, Ganzhou, People’s Republic of China; 2Clinical Laboratory, First Affiliated Hospital of Gannan Medical University, Ganzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Cong You; Longnian Li, Department of Dermatology, First Affiliated Hospital of Gannan Medical University, Ganzhou, People’s Republic of China, Email youcong1987@tmu.edu.cn; li_longnian@foxmail.comObjective: To analyze the antibiotic resistance profile, virulence genes, and molecular typing of Staphylococcus aureus (S. aureus) strains isolated in skin and soft tissue infections at the First Affiliated Hospital, Gannan Medical University, to better understand the molecular epidemiological characteristics of S. aureus.Methods: In 2023, 65 S. aureus strains were isolated from patients with skin and soft tissue infections. Strain identification and susceptibility tests were performed using VITEK 2 and gram-positive bacteria identification cards. DNA was extracted using a DNA extraction kit, and all genes were amplified using polymerase chain reaction. Multilocus sequence typing (MLST) was used for molecular typing.Results: In this study, of the 65 S. aureus strains were tested for their susceptibility to 16 antibiotics, the highest resistance rate to penicillin G was 95.4%. None of the staphylococcal isolates showed resistance to ceftaroline, daptomycin, linezolid, tigecycline, teicoplanin, or vancomycin. fnbA was the most prevalent virulence gene (100%) in S. aureus strains isolated in skin and soft tissue infections, followed by arcA (98.5%). Statistical analyses showed that the resistance rates of methicillin-resistant S. aureus isolates to various antibiotics were significantly higher than those of methicillin-susceptible S. aureus isolates. Fifty sequence types (STs), including 44 new ones, were identified by MLST.Conclusion: In this study, the high resistance rate to penicillin G and the high carrying rate of virulence gene fnbA and arcA of S.aureus were determine, and 44 new STs were identified, which may be associated with the geographical location of southern Jiangxi and local trends in antibiotic use. The study of the clonal lineage and evolutionary relationships of S. aureus in these regions may help in understanding the molecular epidemiology and provide the experimental basis for pathogenic bacteria prevention and treatment.Keywords: Staphylococcus aureus, skin and soft tissue infections, antibiotic resistance, multilocus sequence typing, virulence gene

Abstract Background Salmonella, an important foodborne pathogen, was estimated to be responsible for 95.1 million cases and 50,771 deaths worldwide. Sixteen serovars were responsible for approximately 80% of Salmonella infections in humans in China, and infections caused by a few uncommon serovars have been reported in recent years, though not with S. Welikade. This study reports the first clinical case caused by S. Welikade in China and places Chinese S. Welikade isolates in the context of global isolates via genomic analysis. For comparison, S. Welikade isolates were also screened in the Chinese Local Surveillance System for Salmonella (CLSSS). The minimum inhibitory concentrations (MICs) of 28 antimicrobial agents were determined using the broth microdilution method. The isolates were sequenced on an Illumina platform to identify antimicrobial resistance genes, virulence genes, and phylogenetic relationships. Results The S. Welikade isolate (Sal097) was isolated from a two-year-old boy with acute gastroenteritis in 2021. Along with the other two isolates found in CLSSS, the three Chinese isolates were susceptible to all the examined antimicrobial agents, and their sequence types (STs) were ST5123 (n = 2) and ST3774 (n = 1). Single nucleotide polymorphism (SNP)-based phylogenetic analysis revealed that global S. Welikade strains can be divided into four groups, and these three Chinese isolates were assigned to B (n = 2; Sal097 and XXB1016) and C (n = 1; XXB700). In Group B, the two Chinese ST5123 isolates were closely clustered with three UK ST5123 isolates. In Group C, the Chinese isolate was closely related to the other 12 ST3774 isolates. The number of virulence genes in the S. Welikade isolates ranged from 59 to 152. The galF gene was only present in Group A, the pipB2 gene was only absent from Group A, the avrA gene was only absent from Group B, and the allB, sseK1, sspH2, STM0287, and tlde1 were found only within Group C and D isolates. There were 15 loci unique to the Sal097 isolate. Conclusion This study is the first to characterize and investigate clinical S. Welikade isolates in China. Responsible for a pediatric case of gastroenteritis in 2021, the clinical isolate harbored no antimicrobial resistance and belonged to phylogenetic Group B of global S. Welikade genomes.

Objective: To investigate the frequency of exoU and exoY genes in patients with Pseudomonas aeruginosa infection. Methods: In this study, 100 clinical isolates of Pseudomonas aeruginosa were collected from patients hospitalized in educational-therapeutic hospitals and were identified using standard microbiological tests. Then, the antibiotic resistance pattern of the isolates was determined by the disk agar diffusion method. The bacterial DNAs were extracted by the alkaline lysis method. Finally, the presence of exoU and exoY genes was evaluated by the PCR test. Results: In this study, 47%, 72%, 29%, 39%, 40%, and 44% of the isolates were non-susceptible to piperacillin, aztreonam, ceftazidime, imipenem, tobramycin, and ciprofloxacin, respectively. In addition, 95% and 93% of the clinical isolates carried the exoU and exoY genes. Blood and fecal isolates had both virulence genes, while only one wound isolate had neither genes. Meanwhile, all urinary isolates contained the exoY gene and only one isolate lacked the exoU gene. Also, 88 isolates simultaneously had both exoU and exoY genes. Conclusions: High prevalence of exoU and exoY genes in this region indicates a significant role of type III secretion system in pathogenesis of Pseudomonas aeruginosa. The type III secretion system may be a suitable target to reduce the pathogenicity of this bacterium.

【Objective】The study aimed to isolate Escherichia coli without virulence genes and antibiotic resistance from pig intestines and to analyze their growth performance and genetic evolution, with a view to laying a foundation for the development of microecologics or oral vaccines for pigs.【Method】Fecal samples of 11 and 15 healthy adult pigs were collected from two pig farms in Guangdong Province, respectively. First, E. coli strains were isolated by McConkey plate and identified by 16S rRNA gene sequencing. Then, PCR was used to verify whether the selected E. coli contained 13 common virulence genes of pig pathogenic E. coli. Antimicrobial susceptibility tests were performed with 7 classes (14 kinds) of antimicrobial agents. Finally, the growth curves of selected E. coli strains were analyzed, and a phylogenetic tree was constructed based on the extracted genome sequences.【Result】Among 260 suspected E. coli strains, 206 strains were identified as E. coli by 16S rRNA gene sequencing analysis. 107 strains of E. coli did not contain 13 common virulence genes, of which 25 strains were sensitive to 7 classes (14 kinds) of antibiotics. Among the 25 E. coli strains, 23 non-pathogenic and non-antibiotic resistant E. coli strains (numbered as 2-9, 3-2, 3-4, 5-1, 6-1, 6-2, 6-9, 6-10, 8-2, 8-9, 10-5, B-4, B-6, B-7, B-10, D-10, E-2, E-10, J-1, J-4, K-6, L-6 and O-9) showed similar growth performance to E. coli Nissle 1917 and MG1655, and 2 E. coli strains (numbered as 6-4 and 5-2) showed higher growth rates than those of E. coli Nissle 1917 and MG1655. Among them, 10 strains (numbered as E-10, E-2, 6-4, 6-9, 8-2, O-9, 3-2, 6-2, J-1, K-6) had good acid tolerance and may be suitable for long-term colonization in the gastrointestinal tract of pigs. The phylogenetic tree analysis based on SNP core genome showed that 9 strains (numbered as 3-2, 6-10, 10-5, 6-1, 8-9, 5-2, L-6, O-9 and K-6) of the 25 strains were in the same group at the beginning of the phylogenetic tree, which were most likely to belong to the ancestral group of pig native E. coli.【Conclusion】Based on the comprehensive analysis of antibiotic sensitivity, growth performance, genome-wide evolutionary tree and gastric environment tolerance, one strain (numbered as O-9) was dominant in these four aspects. The growth kinetics of this strain is similar to or even better than that of E. coli Nissle 1977 and MG1655, which means that O-9 strain of E. coli isolates was likely to be high-quality pig native E. coli strain, with great research potential.

Acinetobacter baumannii is a non-fermentative Gram-negative bacterium that can cause nosocomial infections in critically ill patients. Carbapenem-resistant A. baumannii (CRAB) has spread rapidly in clinical settings and has become a key concern. The main objective of this study was to identify the distribution of integrons and biofilm-formation-related virulence genes in CRAB isolates. A total of 269 A. baumannii isolates (219 isolates of CRAB and 50 isolates of carbapenem-sensitive A. baumannii (CSAB)) were collected. Carbapenemase genes (blaKPC, blaVIM, blaIMP, blaNDM, and blaOXA-23-like) and biofilm-formation-related virulence genes (abal, bfms, bap, and cusE) were screened with PCR. Class 1 integron was screened with PCR, and common promoters and gene cassette arrays were determined with restriction pattern analysis combined with primer walking sequencing. Whole-genome sequencing was conducted, and data were analyzed for a blaOXA-23-like-negative isolate. All 219 CRAB isolates were negative for blaKPC, blaVIM, blaIMP, and blaNDM, while blaOXA-23-like was detected in 218 isolates. The detection rates for abal, bfms, bap, and cusE in 219 CRAB were 93.15%, 63.93%, 88.13%, and 77.63%, respectively. Class 1 integron was detected in 75 CRAB (34.25%) and in 3 CSAB. The single gene cassette array aacA4-catB8-aadA1 with relatively strong PcH2 promoter was detected in class 1 integrons. The blaOXA-23-like-negative CRAB isolate was revealed to be a new sequence type (Oxford 3272, Pasteur 2520) carrying blaOXA-72, blaOXA-259, and blaADC-26. In conclusion, blaOXA-23-like was the main reason for CRAB’s resistance to carbapenems. A new (Oxford 3272, Pasteur 2520) CRAB sequence type carrying the blaOXA-72, blaOXA-259, and blaADC-26 was reported.

Methicillin-resistant strains of Staphylococcus aureus (MRSA) can harm public health as they can cause widespread food poisoning and resistance to multiple antibiotics. Hence, our objective was to examine the prevalence of S. aureus and MRSA among fish shops, utensils, and workers' hands in Mosul City. We were able to find the nuc and mecA genes in S. aureus and MRSA isolates, respectively, via the PCR approach. In March and April of 2023, one hundred samples randomly selected from fish and various other store surfaces in Mosul were taken for this inquiry. S. aureus isolates the nuc gene, which can be identified in 19% of the samples. The worker hand samples had eight of the top isolates, in contrast to the fish's three isolates out of twenty. Additionally, because the mecA gene existed in 12 of the 19 S. aureus isolates (63.2%), methicillin resistance was shown. The hand samples of the workers had a higher percentage of methicillin-resistant S. aureus isolates 75% (6/8) than other samples. According to the study, a significant amount of S. aureus was showing in fish and associated utensils, revealing that the fishing industry was not suitably subsequent food safety laws. Based on this study, a significant amount of S. aureus was discovered in fish and associated utensils, revealing that the fishing industry was not appropriately following food safety laws.

Background: Salmonella, an important foodborne pathogen, was estimated to be responsible for 95.1 million cases and 50,771 deaths worldwide. Sixteen serovars were responsible for approximately 80% of Salmonella infections in humans in China, and infections caused by a few uncommon serovars have been reported in recent years, though not with S. Welikade. This study reports the first clinical case caused by S. Welikade in China and places Chinese S. Welikade isolates in the context of global isolates via genomic analysis. For comparison, S. Welikade isolates were also screened in the Chinese Local Surveillance System for Salmonella (CLSSS). The minimum inhibitory concentrations (MICs) of 28 antimicrobial agents were determined using the broth microdilution method. The isolates were sequenced on an Illumina platform to identify antimicrobial resistance genes, virulence genes, and phylogenetic relationships. Results: The S. Welikade isolate (Sal097) was isolated from a two-year-old boy with acute gastroenteritis in 2021. Along with the other two isolates found in CLSSS, the three Chinese isolates were susceptible to all the examined antimicrobial agents, and their sequence types (STs) were ST5123 (n = 2) and ST3774 (n = 1). Single nucleotide polymorphism (SNP)-based phylogenetic analysis revealed that global S. Welikade strains can be divided into four groups, and these three Chinese isolates were assigned to B (n = 2; Sal097 and XXB1016) and C (n = 1; XXB700). In Group B, the two Chinese ST5123 isolates were closely clustered with three UK ST5123 isolates. In Group C, the Chinese isolate was closely related to the other 12 ST3774 isolates. The number of virulence genes in the S. Welikade isolates ranged from 59 to 152. The galF gene was only present in Group A, the pipB2 gene was only absent from Group A, the avrA gene was only absent from Group B, and the allB, sseK1, sspH2, STM0287, and tlde1 were found only within Group C and D isolates. There were 15 loci unique to the Sal097 isolate. Conclusion: This study is the first to characterize and investigate clinical S. Welikade isolates in China. Responsible for a pediatric case of gastroenteritis in 2021, the clinical isolate harbored no antimicrobial resistance and belonged to phylogenetic Group B of global S. Welikade genomes. [ABSTRACT FROM AUTHOR]

Drug-resistant virulent bacteria and antibiotic residues can enter sewage and aquatic ecosystems from urban areas, hospitals, animal farms and slaughterhouse waste. Even the influx of treated wastewater into water and soil can be a potential risk to public health. In this study, 51 strains of Escherichia coli (E. coli) were isolated from 70 samples of slaughterhouse wastewater and urban sewage treatment plant influent in Shiraz, Iran. The most frequent and least frequent extended-spectrum beta-lactamase, tetracycline resistance genes and virulence genes in these isolates were blaTEM (19.6%) and blaCTX−M−1 (2%), tetA (72.5%) and tetD (0%), and bcsA and crl (92.2%) and sfa/focDE (0%). The highest and lowest frequencies of antibiotic resistance were observed for amoxicillin (70.6%) and nitrofurantoin (0%) disks, respectively. None of the urban sewage isolates were resistant to cefotaxime, ceftazidime, ceftriaxone, nitrofurantoin, or gentamicin, while only nitrofurantoin resistance was not observed among the slaughterhouse wastewater isolates. Regarding the virulence genes, none of the urban sewage isolates had afa, csgA, and sfa/focDE, while only sfa/focDE was not detected among the slaughterhouse wastewater isolates. Multidrug resistant isolates were significantly more prevalent in slaughterhouse wastewater (52%) than in urban sewage (23.1%). The ability to produce biofilms, which was evaluated by the microtiter plate method, was observed for 50% of the urban sewage E. coli and 68% of the slaughterhouse wastewater E. coli. The abundance of virulent antibiotic-resistant E. coli strains with the ability to form biofilms in urban and slaughterhouse wastewater makes it necessary to pay more attention to wastewater treatment processes. [ABSTRACT FROM AUTHOR]

Multidrug-resistant (MDR) Escherichia coli poses a significant threat to public health, contributing to elevated rates of morbidity, mortality, and economic burden. This study focused on investigating the antibiotic resistance profiles, resistance and virulence gene distributions, biofilm formation capabilities, and sequence types of E. coli strains resistant to six or more antibiotic classes. Among 918 strains isolated from 33 wastewater treatment plants (WWTPs), 53.6% (492/918) demonstrated resistance, 32.5% (298/918) were MDR, and over 8% (74/918) were resistant to six or more antibiotic classes, exhibiting complete resistance to ampicillin and over 90% to sulfisoxazole, nalidixic acid, and tetracycline. Key resistance genes identified included sul2, blaTEM, tetA, strA, strB, and fimH as the predominant virulence genes linked to cell adhesion but limited biofilm formation; 69% showed no biofilm formation, and approximately 3% were strong producers. Antibiotic residue analysis detected ciprofloxacin, sulfamethoxazole, and trimethoprim in all 33 WWTPs. Multilocus sequence typing analysis identified 29 genotypes, predominantly ST131, ST1193, ST38, and ST69, as high-risk clones of extraintestinal pathogenic E. coli. This study provided a comprehensive analysis of antibiotic resistance in MDR E. coli isolated from WWTPs, emphasizing the need for ongoing surveillance and research to effectively manage antibiotic resistance. [ABSTRACT FROM AUTHOR]

Simple Summary: The current study aimed to investigate the virulence and resistance genes associated with Klebsiella pneumoniae isolated from the farms of four animal species in selected areas of Xinjiang, China, encompassing cows, sheep, chickens, and pigs. The sensitivity to antibiotics was evaluated. Genetic diversity was assessed using wzi allele typing and multilocus sequence typing. The findings shed light on the antibiotic resistance patterns and genetic diversity among K. pneumoniae isolates originating from different animal sources, highlighting the potential for inter-species transmission of this bacterium. Klebsiella pneumoniae (K. pneumoniae) is recognized as a zoonotic pathogen with an increasing threat to livestock and poultry. However, research on K. pneumoniae of animal origin remains limited. To address the gap, a comprehensive investigation was carried out by collecting a total of 311 samples from the farms of four animal species (dairy cow, chicken, sheep, and pig) in selected areas of Xinjiang, China. Isolates were identified by khe gene amplification and 16S rRNA gene sequencing. Genotyping of K. pneumonia isolates was performed using wzi typing and multilocus sequence typing (MLST). PCR was employed to identify virulence and resistance genes. An antibiotic susceptibility test was conducted using the Kirby–Bauer method. The findings revealed an isolation of 62 K. pneumoniae strains, with an average isolation rate of 19.94%, with the highest proportion originating from cattle sources (33.33%). Over 85.00% of these isolates harbored six virulence genes (wabG, uge, fimH, markD, entB, and ureA); while more than 75.00% of isolates possessed four resistance genes (blaTEM, blaSHV, oqxA, and gyrA). All isolates exhibited complete resistance to ampicillin and demonstrated substantial resistance to sulfisoxazole, amoxicillin/clavulanic acid, and enrofloxacin, with an antibiotic resistance rate of more than 50%. Furthermore, 48.39% (30/62) of isolates were classified as multidrug-resistant (MDR) strains, with a significantly higher isolation rate observed in the swine farms (66.67%) compared to other farms. Genetic characterization revealed the classification of the 62 isolates into 30 distinct wzi allele types or 35 different sequence types (STs). Notably, we identified K. pneumoniae strains of dairy and swine origin belonging to the same ST42 and wzi33-KL64 types, as well as strains of dairy and chicken origin belonging to the same wzi31-KL31-K31 type. These findings emphasize the widespread occurrence of drug-resistant K. pneumoniae across diverse animal sources in Xinjiang, underscoring the high prevalence of multidrug resistance. Additionally, our results suggest the potential for animal-to-animal transmission of K. pneumoniae and there was a correlation between virulence genes and antibiotic resistance genes. Moreover, the current study provides valuable data on the prevalence, antibiotic resistance, and genetic diversity of K. pneumoniae originating from diverse animal sources in Xinjiang, China. [ABSTRACT FROM AUTHOR]

Bacterial infection poses a significant threat to the health and survival of turtles, both in their natural habitats and in captivity. Notably, the presence and interactions of bacteria, such as those belonging to the Aeromonas genus, have been identified for their adverse impact on turtle health and ecosystem dynamics. This study investigated the prevalence, antibiotic susceptibility, virulence factors, and antimicrobial resistance genes of Aeromonas hydrophila in Trachemys scripta elegans (Red-eared Slider) and Chelonia mydas (Green Turtle). The study also examined the innate immunity of T. scripta elegans and C. mydas. Buccal and cloacal swabs from 39 turtles were collected and examined alongside 45 water samples to try to isolate and amplify A. hydrophila. Polymerase chain reaction (PCR) amplifications were then sequenced to identify the presence of specific genes associated with virulence, and antimicrobial resistance. The findings revealed that A. hydrophila had a prevalence rate of 35.9% in all examined turtle species, with 14 water samples accounting for 31% of the total samples. Buccal samples from Green and Red-eared turtles showed a higher rate of A. hydrophila isolates than cloacal samples. Sensitivity to norfloxacin (85.71%), gentamycin (71.42%), and ceftriaxone (64.28%) was observed, whereas resistance to amoxicillin (64.28%), ampicillin (57.14%), and rifamycin (57.14%) was apparent. Antibiotic resistance genes, including blaTEM and blaSHV, in A. hydrophila isolates, were present in 42.86% and 28.5% of the samples, respectively. PCR analysis confirmed the genetic identity of A. hydrophila, and five virulence genes were detected: AHCYTOEN (100%), followed by aerolysin (92.8%), alt and act (85.7%), and lipase (78.5%). Genetic sequences of the isolates had a 99% identity with the reference strains in GenBank. Furthermore, the study found the innate immunity of turtles to be robust due to serum lysozyme activity, bactericidal killing assay (BKA), total leukocyte count, heterophil/lymphocyte ratio, and packed cell volume (PCV%), all of which help to protect them from infections. Overall, this research highlights the need for continued monitoring and management of antibiotic resistance in aquatic ecosystems. [ABSTRACT FROM AUTHOR]

【Objective】The study aimed to isolate Escherichia coli without virulence genes and antibiotic resistance from pig intestines and to analyze their growth performance and genetic evolution, with a view to laying a foundation for the development of microecologics or oral vaccines for pigs. 【Method】Fecal samples of 11 and 15 healthy adult pigs were collected from two pig farms in Guangdong Province, respectively. First, E. coli strains were isolated by McConkey plate and identified by 16S rRNA gene sequencing. Then, PCR was used to verify whether the selected E. coli contained 13 common virulence genes of pig pathogenic E. coli. Antimicrobial susceptibility tests were performed with 7 classes (14 kinds) of antimicrobial agents. Finally, the growth curves of selected E. coli strains were analyzed, and a phylogenetic tree was constructed based on the extracted genome sequences.【Result】Among 260 suspected E. coli strains, 206 strains were identified as E. coli by 16S rRNA gene sequencing analysis. 107 strains of E. coli did not contain 13 common virulence genes, of which 25 strains were sensitive to 7 classes (14 kinds) of antibiotics. Among the 25 E. coli strains, 23 non-pathogenic and non-antibiotic resistant E. coli strains (numbered as 2-9, 3-2, 3-4, 5-1, 6-1, 6-2, 6-9, 6-10, 8-2, 8-9, 10-5, B-4, B-6, B-7, B-10, D-10, E-2, E-10, J-1, J-4, K-6, L-6 and O-9) showed similar growth performance to E. coli Nissle 1917 and MG1655, and 2 E. coli strains (numbered as 6-4 and 5-2) showed higher growth rates than those of E. coli Nissle 1917 and MG1655. Among them, 10 strains (numbered as E-10, E-2, 6-4, 6-9, 8-2, O-9, 3-2, 6-2, J-1, K-6) had good acid tolerance and may be suitable for long-term colonization in the gastrointestinal tract of pigs. The phylogenetic tree analysis based on SNP core genome showed that 9 strains (numbered as 3-2, 6-10, 10-5, 6-1, 8-9, 5-2, L-6, O-9 and K-6) of the 25 strains were in the same group at the beginning of the phylogenetic tree, which were most likely to belong to the ancestral group of pig native E. coli. 【Conclusion】 Based on the comprehensive analysis of antibiotic sensitivity, growth performance, genome-wide evolutionary tree and gastric environment tolerance, one strain (numbered as O-9) was dominant in these four aspects. The growth kinetics of this strain is similar to or even better than that of E. coli Nissle 1977 and MG1655, which means that O-9 strain of E. coli isolates was likely to be high-quality pig native E. coli strain, with great research potential [ABSTRACT FROM AUTHOR]

Background: Companion animals, including dogs and cats, are frequently identified as sources of Pasteurella multocida, a bacterium that can be transmitted to humans and cause infections. Objectives: This survey defines the prevalence, antibiotic sensitivity, capsular types, lipopolysaccharide (LPS) types and virulence factors of P. multocida isolated from cats. Methods: A total of 100 specimens from various cat breeds were collected. P. multocida was characterized using both biochemical tests and PCR. Genotypes of isolates were determined using capsular and LPS typing methods. Additionally, virulotyping was performed by detecting the presence of 12 virulence‐associated genes. Disk diffusion was used to determine the antibiotic sensitivity of the isolates. Results: The prevalence of P. multocida in cats was 29%. Among the isolates, the majority were capsular type A (96.5%) and type D (3.4%), with a predominant presence of type A. Twenty‐six of the isolates (89.66%) belonged to LPS genotype L6, whereas three isolates (10.3%) belonged to genotype L3. Among the 12 virulence genes examined, sodC, oma87, ptfA, nanB and ompH showed remarkable prevalence (100%). The toxA gene was detected in four isolates (13.8%). Variations were observed in other virulence genes. The nanH gene was present in 93.1% of the isolates, whereas the pfhA gene was detected in 58.6% of the isolates. The exbD‐tonB, hgbB, sodA and hgbA genes showed prevalence rates of 96.5%, 96.5%, 96.5% and 82.8%, respectively. Additionally, particular capsule and LPS types were associated with specific virulence genes. Specifically, the toxA and pfhA genes were found to be more prevalent in isolates with capsular type A and LPS genotype L6. Most isolates were resistant to ampicillin, clindamycin, lincomycin, streptomycin and penicillin. Conclusions: According to this epidemiological and molecular data, P. multocida from cats possess several virulence‐associated genes and are resistant to antimicrobial medicines commonly used in humans and animals. Thus, it is crucial to consider the public health concerns of P. multocida in humans. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao is the property of Chinese Journal of Preventive Veterinary Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Escherichia coli (E. coli) is closely associated with the occurrence of puerperal metritis in dairy cows. E. coli carries some the virulence and multi-drug resistant genes, which pose a serious threat to the health of postpartum cows. In this study, E. coli was isolated and identified from the uterine contents of postpartum cows with puerperal metritis in the Ningxia region of China, and its phylogenetic subgroups were determined. Meanwhile, virulence and drug resistance genes carried by E. coli and drug sensitivity were detected, and the characteristics of virulence and drug resistance genes distribution in E. coli phylogroups were further analyzed. The results showed that the isolation rate of E. coli in puerperal metritis samples was 95.2%. E. coli was mainly divided into phylogroups B2 and D, followed by groups A and B1, and was more connected to O157:H7, O169:H4, and ECC-1470 type strains. The virulence genes were mainly dominated by ompF (100%), traT (100%), fimH (97%), papC (96%), csgA (95%), Ang43 (93.9%), and ompC (93%), and the resistance genes were dominated by TEM (99%), tetA (71.7%), aac(3)II (66.7%), and cmlA (53.5%). Additionally, it was observed that the virulence and resistance gene phenotypes could be divided into two subgroups, with subgroup B2 and D having the highest distributions. Drug sensitivity tests also revealed that the E. coli was most sensitive to the fluoroquinolones enrofloxacin, followed by macrolides, aminoglycosides, tetracyclines, β-lactams, peptides and sulfonamides, and least sensitive to lincosamides. These results imply that pathogenic E. coli, which induces puerperal metritis of dairy cows in the Ningxia region of China, primarily belongs to the group B2 and D, contains multiple virulence and drug resistance genes, Moreover, E. coli has evolved resistance to several drugs including penicillin, lincomycin, cotrimoxazole, and streptomycin. It will offer specific guidelines reference for the prevention and treatment of puerperal metritis in dairy cows with E. coli infections in the Ningxia region of China. [ABSTRACT FROM AUTHOR]

The present study involves the precise identification and safety evaluation of Enterococcus casseliflavus KB1733, previously identified using 16S rRNA analysis, through whole-genome sequencing, phenotypic analysis, and preclinical toxicity studies. Analyses based on the genome sequencing data confirm the identity of KB1733 as E. casseliflavus and show that the genes related to vancomycin resistance are only present on the chromosome, while no virulence factor genes are present on the chromosome or plasmid. Phenotypic analyses of antibiotic resistance and hemolytic activity also indicated no safety concerns. A bacterial reverse mutation test showed there was no increase in revertant colonies of heat-killed KB1733. An acute toxicity test employing heat-killed KB1733 at a dose of 2000 mg/kg body weight in rats resulted in no deaths and no weight gain or other abnormalities in the general condition of the animals, with renal depression foci and renal cysts only occurring at the same frequency as in the control. Taking the background data into consideration, the effects on the kidneys observed in the current study were not caused by KB1733. Our findings suggest that KB1733 is non-pathogenic to humans/animals, although further studies involving repeated oral toxicity tests and/or clinical tests are required. [ABSTRACT FROM AUTHOR]

Trueperella pyogenes can cause various infections in the organs and tissues of different livestock (including pigs, cows, goats, and sheep), including mastitis, endometritis, pneumonia, or abscesses. Moreover, diseases induced by T. pyogenes cause significant economic losses in animal husbandry. In recent large-scale investigations, T. pyogenes has been identified as one of the main pathogens causing endometritis in lactating cows. However, the main treatment for the above-mentioned diseases is still currently antibiotic therapy. Understanding the impact of endometritis associated with T. pyogenes on the fertility of cows can help optimize antibiotic treatment for uterine diseases, thereby strategically concentrating the use of antimicrobials on the most severe cases. Therefore, it is particularly important to continuously monitor the prevalence of T. pyogenes and test its drug resistance. This study compared the uterine microbiota of healthy cows and endometritis cows in different cattle farms, investigated the prevalence of T. pyogenes, evaluated the genetic characteristics and population structure of isolated strains, and determined the virulence genes and drug resistance characteristics of T. pyogenes. An amount of 186 dairy cows were involved in this study and 23 T. pyogenes strains were isolated and identified from the uterine lavage fluid of dairy cows with or without endometritis. [ABSTRACT FROM AUTHOR]

Objective To study the drug resistance phenotype, drug resistance gene, and virulence gene of Helicobacter pylori (Hp) in patients with failed eradication therapy, and to provide reference for remedial therapy. Methods Hp was isolated and cultured from gastric mucosa of patients with failed eradication therapy. Drug sensitivity test was used to detect the resistance of isolated strains to Metronidazole (MTZ), Clarithromycin (CLA), Levofloxacin (LEV), Amoxicillin (AMX), Tetracycline (TET), and Furazolidone (FZ). CLA and LEV resistance genes 23S rRNA, gyrA and virulence genes cagA, vacA, iceA, oipA were amplified by PCR, to further explore the relationship between drug resistance phenotype and drug resistance and virulence genes. Results In total 40 strains of Hp were isolated from 58 patients (69.0%). The resistance rates to MTZ, CLA, LEV, AMX, TET, and FZ were 100.0%, 82.5%, 72.5%, 0%, 0%, and 0%, respectively. The most common mutations of 23S rRNA and gyrA genes were T2182C (78.6%) and N87K (32.0%), respectively, and new C2165T and A2219G mutations were found in CLA resistant strains. In addition, the positive rates of virulence genes cagA, vac A si, vacA s2, vacA ml, vacA m2, tceAl, tceA2, and oipA were 97.1%, 100.0%, 0%, 42.9%, 57.1%, 62.9%, 11.4%, and 68.6%, respectively, and the difference was statistically significant. Conclusion The resistance of Hp isolates to MTZ, CLA and LEV in patients with failed eradication therapy in this area is serious, and AMX, TET and FZ are recommended for remedial therapy. The relationship between CLA resistance and C2165T and A2219G mutation and LEV resistance and vacA m2 warrants further investigation. [ABSTRACT FROM AUTHOR]

Background: It has been determined that Escherichia coli is the prevailing uropathogen (50-90%) in both complicated and uncomplicated urinary tract infections. Uropathogenic Escherichia coli (UPEC) possesses an extensive array of virulence factors, which confers the potential to induce urinary tract infections and is correlated with the development of antibiotic resistance. The purpose of this research was to detect the distribution of virulence genes and their association with antibiotic resistance rates at the molecular level. Materials & Methods: This study was conducted in the microbiology laboratory at Al-Hussein Medical city teaching hospital, Laboratory of microbiology in Karbala province, Iraq. Urine samples were collected b/w October 2023 to January 2024, and transported for further analysis using standard microbiological and biochemical techniques. The sampling technique used in the study can be described as convenience sampling. The identification of E.coli isolates was confirmed using 16rRNA, resistance and sensitivity to various antibiotics were determined with Vitek2 system. Identification of genes (fimH, KpsmII, Pap, ompT) by uniplex PCR. Results: E.Coli was shown to be the most often isolated bacterium for various urine samples, with a percentage of 16(35.5%). The most virulent gene observed were Fim H, KpsMII, pap and OmpT were16 (100%), 13 (81.3%), 9(56.3%) and 8(50%) respectively. Thirteen type of antibiotics were identified sensitive to E.coli isolate the most common resistant rate was Ticarcillin, Pipraciilin had high resistance rate 14(87.5), Ciprofloxacin 9(56.3%), Minocycline, Aztreonam, Trimethoprim had rate of 8(50). Sensitivity rate of Meropenem, Imipenem, Gentamicin was 14(87.5), Cefepime 13(81.25), Amikacin 12(75.0). Conclusion: The study concluded that there is distribution of virulence gene in E.coli isolates, specially fimH, pap correlated with resistance to antibiotics specially with MDR and XDR patterns. [ABSTRACT FROM AUTHOR]

Objective: To investigate the frequency of exoU and exoY genes in patients with Pseudomonas aeruginosa infection. Methods: In this study, 100 clinical isolates of Pseudomonas aeruginosa were collected from patients hospitalized in educational-therapeutic hospitals and were identified using standard microbiological tests. Then, the antibiotic resistance pattern of the isolates was determined by the disk agar diffusion method. The bacterial DNAs were extracted by the alkaline lysis method. Finally, the presence of exoU and exoY genes was evaluated by the PCR test. Results: In this study, 47%, 72%, 29%, 39%, 40%, and 44% of the isolates were non-susceptible to piperacillin, aztreonam, ceftazidime, imipenem, tobramycin, and ciprofloxacin, respectively. In addition, 95% and 93% of the clinical isolates carried the exoU and exoY genes. Blood and fecal isolates had both virulence genes, while only one wound isolate had neither genes. Meanwhile, all urinary isolates contained the exoY gene and only one isolate lacked the exoU gene. Also, 88 isolates simultaneously had both exoU and exoY genes. Conclusions: High prevalence of exoU and exoY genes in this region indicates a significant role of type III secretion system in pathogenesis of Pseudomonas aeruginosa. The type III secretion system may be a suitable target to reduce the pathogenicity of this bacterium. [ABSTRACT FROM AUTHOR]

This systematic review and meta-analysis investigates the prevalence of Vibrio parahaemolyticus, its virulence factors, antimicrobial resistance (AMR), and its resistance determinants in shrimp. This study was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines, to identify and select relevant peer-reviewed articles published between January 2020 and December 2022. The search strategy involved multiple online databases, including Google Scholar, PubMed, ScienceDirect, and Scopus. The inclusion criteria focused on studies that examined V. parahaemolyticus prevalence, virulence factors, and AMR in shrimp from farms to retail outlets. A total of 32 studies were analyzed, revealing a pooled estimate prevalence of V. parahaemolyticus in shrimp at 46.0%, with significant heterogeneity observed. Subgroup analysis highlighted varying prevalence rates across continents, emphasizing the need for further investigation. Virulence factor analysis identified thermostable direct hemolysin (tdh) and tdh-related hemolysin (trh) as the most common. Phenotypic AMR analysis indicated notable resistance to glycopeptides, nitrofurans, and beta-lactams. However, the correlation between antimicrobial usage in shrimp farming and observed resistance patterns was inconclusive. Funnel plots suggested potential publication bias, indicating a need for cautious interpretation of findings. This study underscores the urgency of coordinated efforts to address AMR in V. parahaemolyticus to safeguard public health and to ensure sustainable aquaculture practices. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae (Strep. agalactiae) is bovine mastitis pathogen and has thus became a matter of concern to dairy farms worldwide in terms of economic loss. The aims of this study were to (a) determine virulence genes, and (b) characterize the antimicrobial resistance (AMR) profiles and AMR genes and (c) figure out the relationship between AMR phenotypes and genotypes of Strep. agalactiae isolated from dairy cows in north China. A total of 20 virulence genes and 23 AMR genes of 140 isolates collected from 12 farms in six provinces were studied. The antimicrobial susceptibility of 10 veterinary commonly used antimicrobials were tested using the broth microdilution method. Results showed that all the isolates harbored the virulence genes lacIV, gapC, and dltA. The isolates that harbored the genes lacIII, fbsA, hylB, and cfb exhibited the high prevalence (99.29%), followed by isolates that harbored lacI (98.57%), bibA (97.86%), cylE (97.14%), lacII (92.14%), cspA (52.14%), pavA (25%), bca (2.14%), and scpB (0.71%). The fbsB, lmb, spbI, bac, and rib genes were not detected. The virulence patterns of B (fbsA_cfb_cylE_ hylB_bibA_cspA_ gapC_dltA_lacIII/IV) and C (fbsA_cfb_ bibA _ gapC_ dltA_lacIV) were dominant, accounting for 97.86% of the isolates. The following AMR genes were prevalent: pbp1A (97.14%), tet(M) (95.00%), lnu (A) (80.71%), erm (B) (75.00%), tet(O) (72.14%), blaZ (49.29%), tet(S) (29.29%), blaTEM (25.71%), erm (A) (17.14%), erm (C) (13.57%), tet (L) (10.71%), linB (2.86%), and erm (TR) (2.86%). The pbp2b, mecA1, mecC, lnu (D), erm (F/G/Q), and mef (A) genes were not detected. Eighty percent of the isolates harbored AMR genes and were highly resistant to tetracycline, followed by macrolides (10.71%), lincosamides (9.29%) and β-lactams (4.29%). In conclusion, isolates only exhibited well correlation between tetracyclines resistance phenotype and genotype, and almost all isolates harbored intact combination of virulence genes.

Vibrio rotiferianus is a pathogenic bacterium that is widely distributed in the ocean and can cause diseases in a variety of marine animals, including fish, shrimp and shellfish. In this study, we isolated and identified V. rotiferianus strain TO-01 from diseased Trachinotus ovatus. Infected fish exhibited a range of pathological symptoms, including lethargy in swimming, blackened body coloration, skin ulceration and erratic side swimming. Tissue section examination revealed pronounced damage to the liver, spleen and gills of the infected fish. An analysis of immune factor expression revealed a significant increase in the expression of inflammatory factors, including interleukin (IL)-1, IL-2, IL-6 receptor and IL-21, in the spleen and kidney of fish infected with TO-01 at 6 h post-infection. To gain a more in-depth comprehension of the pathogenicity and drug resistance of V. rotiferianus, we conducted whole genome sequencing. The total genome length of V. rotiferianus was 4734,075 bp, and 74 virulence factors were predicted within the genome. Most of these virulence genes were associated with bacterial adhesion and flagellar motility. A gene associated with group sensing, cqsA, and a gene associated with hemolysin, tlh, were also identified. The prediction of drug resistance genes was in agreement with the results of drug sensitivity assays, both of which indicated that TO-01 has significant resistance to β-lactams. The findings of this study significantly enhance our understanding of the pathogenic mechanism of V. rotiferianus and give crucial insights into disease prevention and control in the management of T. ovatus.

Background: Escherichia coli is the most common gram-negative pathogen to cause neonatal infections. Contemporary virulence characterization and antimicrobial resistance (AMR) data of neonatal E. coli isolates in China are limited. Methods: A total of 159 E. coli strains isolated from neonates were collected and classified into invasive and non-invasive infection groups, according to their site of origin. The presence of virulence genes was determined using polymerase chain reaction (PCR). All the strains were subjected to antimicrobial susceptibility testing using the broth dilution method. Results: The top three virulence genes with the highest detection rates were fimH (90.6 %), iutA (88.7 %), and kspMT II (88.1 %). The prevalences of fyuA (p = 0.023), kpsMT K1 (p = 0.019), ibeA (p

ObjectiveTo investigate the infection status， pathogenic types， antimicrobial resistance and molecular typing of diarrheagenic Escherichia coli （DEC） isolated from patients with foodborne diarrhea in Xi’an City in 2021.MethodsFecal samples from patients with diarrhea were collected from 5 sentinel hospitals for DEC isolation and the pathogenic types of DEC were confirmed via PCR to detect virulence genes. Antimicrobial susceptibility testing and molecular typing were performed using the broth microdilution method and pulsed field gel electrophoresis （PFGE）. The results were statistically analyzed using SPSS software 19.0.ResultsAmong the 389 fecal samples collected from patients with diarrhea， 40 were DEC-positive， with a positivity rate of 10.28%. A total of 41 DEC strains were identified in this study. Enterotoxigenic E. coli （ETEC） and enteroaggregative E. coli （EAEC） were predominant， accounting for 41.46% （17/41） and 39.02% （16/41） of cases， respectively. The gene types estIa/estIb （70.59%， 12/17） and astA/pic （87.50 %， 14/16） were dominant in ETEC and EAEC， respectively. Forty strains （97.56%， 40/41） were resistant to at least one antibiotic， and the resistance rates to ampicillin， tetracycline， cefotaxime， and nalidixic acid were greater than 50%. The multidrug resistance rate was 56.10% （23/41）. Cluster analysis revealed 40 PFGE patterns from the 41 DEC strains. The similarity of these patterns was 62.0%-100.0%.ConclusionThe detection rate of DEC in patients with foodborne diarrhea in Xi’an City in 2021 was relatively high， with the major pathotypes being ETEC and EAEC. The PFGE patterns of the strains were diverse. Finally， the antimicrobial and multidrug resistance of the strains was relatively severe.

This study investigated virulence - associated resistance genes and phylogenetic analysis of Escherichia coli (E. coli) isolated from various meat types in Ghana. The prevalence of virulence genes detected among E. coli isolates were 71.4%, 24.5%, 20.4%, 18.4%, and 4.1% for ibeA, iutA, KpsMTII, papGIII, and sfa, respectively. The E. coli strains belonged to four major phylogenetic groups; group A (14.3%), group C (34.7%), group D (30.8%), and group F (6.1%). E. coli resistance to relevant antibiotics ranged between 12.5% and 85.7%. The prevalent resistance gene was tet A (16.3%), tet B (4.08%), and floR (4.08%). One E. coli isolate from local chicken harboured blaCTX-M and another from local guinea fowl meat carried blaTEM. Virulence and resistance genes occurred (P

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Background: With the increase in human population, the consumption of livestock products such as sheep meat has also increased. Sheep are the reservoir and shedder of Escherichia coli that can be transmitted to humans. Aims: Characterization of fecal E. coli isolated from sheep in slaughterhouse. Methods: Stool specimens were collected from 30 apparently healthy sheep from different flocks in Shiraz industrial slaughterhouse. The resistance of E. coli isolates against 10 antibiotics was determined by disk diffusion method. The presence of three major extended spectrum beta-lactamase (ESBL) genes and five tetracycline resistance genes as well as seven virulence genes were investigated by polymerase chain reaction (PCR) technique. Using the microtiter plate method, the biofilm formation ability of E. coli isolates was investigated. Results: The highest frequency of resistance was to amoxicillin (100%) followed by tetracycline (25%). All E. coli isolates were susceptible to gentamicin and nitrofurantoin, and only one isolate was resistant to the tested third-generation cephalosporins. Multidrug resistance phenotype was observed in 16.7% of the isolates. blaTEM (25%) was the most prevalent ESBL gene and tetA (62.5%) was the most prevalent tetracycline resistance gene in the isolates. crl, csgA, fimH, and bcsA genes were present in all isolates, and the prevalence of papC and afa genes was 95.8% and 83.3%, respectively. In total, 62.5% of the isolates were biofilm producers. Conclusion: According to the concept of One Health, the presence of virulent antibiotic-resistant biofilm producing strains of E. coli in sheep is a risk to public health. [ABSTRACT FROM AUTHOR]

This work aimed to identify the chemical compounds of Cinnamomum burmannii leaf essential oil (CBLEO) and to unravel the antibacterial mechanism of CBLEO at the molecular level for developing antimicrobials. CBLEO had 37 volatile compounds with abundant borneol (28.40%) and showed good potential to control foodborne pathogens, of which Staphylococcus aureus had the greatest inhibition zone diameter (28.72 mm) with the lowest values of minimum inhibitory concentration (1.0 μg/mL) and bactericidal concentration (2.0 μg/mL). To unravel the antibacterial action of CBLEO on S. aureus, a dynamic exploration of antibacterial growth, material leakage, ROS formation, protein oxidation, cell morphology, and interaction with genome DNA was conducted on S. aureus exposed to CBLEO at different doses (1/2–2×MIC) and times (0–24 h), indicating that CBLEO acts as an inducer for ROS production and the oxidative stress of S. aureus. To highlight the antibacterial action of CBLEO on S. aureus at the molecular level, we performed a comparative association of ROS accumulation with some key virulence-related gene (sigB/agrA/sarA/icaA/cidA/rsbU) transcription, protease production, and biofilm formation in S. aureus subjected to CBLEO at different levels and times, revealing that CBLEO-induced oxidative stress caused transcript suppression of virulence regulators (RsbU and SigB) and its targeted genes, causing a protease level increase destined for the biofilm formation and growth inhibition of S. aureus, which may be a key bactericidal action. Our findings provide valuable information for studying the antibacterial mechanism of essential oil against pathogens. [ABSTRACT FROM AUTHOR]

To identify the cause of mass mortality of adult Macrobrachium rosenbergii in a farm in Gaoyou City, Jiangsu Province, China, a dominant strain named DKQ-1 was isolated from the hepatopancreas of dying M. rosenbergii and identified as Aeromonas dhakensis by purification culture, biochemical characterization, and 16S rRNA and gyrB gene sequence analysis. The results of the challenge test revealed that the strain was highly pathogenic and the 50% lethal dose (LD50) in 72 h to M. rosenbergii was 1.54×105 CFU/mL. The amplification results of virulence genes show that strain DKQ-1 carried 9 virulence genes, including ascV, aexT, aer, act, lip, ompAI, gcaT, acg, and exu, supporting the strong virulence of strain DKQ-1 to M. rosenbergii. Histopathological observation of the hepatopancreas, gills, and intestines indicated that DKQ-1 injection into M. rosenbergii could cause serious tissue damage, which further supported the strong virulence of this strain. In addition, a drug susceptibility test revealed that strain DKQ-1 was sensitive to 16 kinds of antibiotics, resistant to 9 kinds of antibiotics, and had intermediate resistance to spectinomycin and kanamycin. This study is the first report of A. dhakensis isolated from M. rosenbergii and provided a reference for the pathogen identification of bacterial diseases in M. rosenbergii, and for the prevention and treatment caused by A. dhakensis. [ABSTRACT FROM AUTHOR]

Background: Proteus mirabilis has been identified as an important zoonotic pathogen, causing several illnesses such as diarrhoea, keratitis and urinary tract infections. Objective: This study assessed the prevalence of P. mirabilis in broiler chickens, its antibiotic resistance (AR) patterns, ESBL‐producing P. mirabilis and the presence of virulence genes. Methods: A total of 26 isolates were confirmed as P. mirabilis from 480 pooled broiler chicken faecal samples by polymerase chain reaction (PCR). The disk diffusion method was used to evaluate the antibacterial susceptibility test, while nine virulence genes and 26 AR genes were also screened by PCR. Results: All 26 P. mirabilis isolates harboured the ireA (siderophore receptors), ptA, and zapA (proteases), ucaA, pmfA, atfA, and mrpA (fimbriae), hlyA and hpmA (haemolysins) virulence genes. The P. mirabilis isolates were resistant to ciprofloxacin (62%) and levofloxacin (54%), while 8 (30.7%) of the isolates were classified as multidrug resistant (MDR). PCR analysis identified the blaCTX‐M gene (62%), blaTEM (58%) and blaCTX‐M‐2 (38%). Further screening for AMR genes identified mcr‐1, cat1, cat2, qnrA, qnrD and mecA, 12%, 19%, 12%, 54%, 27% and 8%, respectively for P. mirabilis isolates. The prevalence of the integron integrase intI1 and intI2 genes was 43% and 4%, respectively. Conclusions: The rise of ciprofloxacin and levofloxacin resistance, as well as MDR strains, is a public health threat that points to a challenge in the treatment of infections caused by these zoonotic bacteria. Furthermore, because ESBL‐producing P. mirabilis has the potential to spread to humans, the presence of blaCTX‐M‐producing P. mirabilis in broilers should be kept under control. This is the first study undertaken to isolate P. mirabilis from chicken faecal samples and investigate its antibiotic resistance status as well as virulence profiles in South Africa. [ABSTRACT FROM AUTHOR]

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Members of the Bacillus cereus group are spore-forming Gram-positive bacilli that are commonly associated with diarrheal or emetic food poisoning. They are widespread in nature and frequently present in both raw and processed food products. Here, we genetically characterized 24 B. cereus group isolates from foodstuffs. Whole-genome sequencing (WGS) revealed that most of the isolates were closely related to B. cereus sensu stricto (12 isolates), followed by B. pacificus (5 isolates), B. paranthracis (5 isolates), B. tropicus (1 isolate), and "B. bingmayongensis" (1 isolate). The most detected virulence genes were BAS_RS06430, followed by bacillibactin biosynthesis genes (dhbA, dhbB, dhbC, dhbE, and dhbF), genes encoding the three-component non-hemolytic enterotoxin (nheA, nheB, and nheC), a gene encoding an iron-regulated leucine-rich surface protein (ilsA), and a gene encoding a metalloprotease (inhA). Various biofilm-associated genes were found, with high prevalences of tasA and sipW genes (matrix protein-encoding genes); purA, purC, and purL genes (eDNA synthesis genes); lytR and ugd genes (matrix polysaccharide synthesis genes); and abrB, codY, nprR, plcR, sinR, and spo0A genes (biofilm transcription regulator genes). Genes related to fosfomycin and beta-lactam resistance were identified in most of the isolates. We therefore demonstrated that WGS analysis represents a useful tool for rapidly identifying and characterizing B. cereus group strains. Determining the genetic epidemiology, the presence of virulence and antimicrobial resistance genes, and the pathogenic potential of each strain is crucial for improving the risk assessment of foodborne B. cereus group strains. [ABSTRACT FROM AUTHOR]

Background: In the present study, we examine the prevalence of phylogenetic groups, O-serogroups, adhesin genes, antimicrobial resistance, the level of gene expression associated with biofilm formation, and the presence of extended-spectrum beta-lactamase (ESBL) in UPEC strains isolated from both pediatric and adult patients. Methods: In this cross-sectional study, 156 UPEC isolates were collected from UTI patients. ESBL-producing isolates were detected using the double-disc synergy (DDS) method, and biofilm formation was assessed through a microplate assay. The presence of O-serogroups, adhesion factors and resistance genes, including ESBLs and PMQR genes, was detected by PCR, and isolates were categorized into phylogenetic groups using multiplex PCR. Additionally, the quantitative real-time PCR method was also used to determine the expression level of genes related to biofilm. Results: During the study period, 50.6% (79/156) of the samples were obtained from children, and 49.4% (77/156) were from adults. The highest rate of resistance was to NA (91.7%), while FM (10.9%) had the lowest rate of antibiotic resistance. In addition, 67.9% (106/156) of UPEC isolates were ESBL producers. Most of UPEC isolates belonged to phylogenetic group B2 (37.1%). This study revealed that blaCTX-M and qnrS are widely distributed among UPEC isolates. The mean expression levels of fimA genes were significantly higher in non-biofilm producers than in biofilm producers (p < 0.01). Conclusions: The high antibiotic resistance rates in this study highlight the significance of local resistance monitoring and investigating underlying mechanisms. Our findings indicate the dominance of phylogroup B2 and group D as the prevailing phylogenetic groups. Consequently, it is imperative to investigate the epidemiological aspects and characterize UPEC isolates across diverse regions and time frames. [ABSTRACT FROM AUTHOR]

Simple Summary: Wildlife can host zoonotic pathogens and transfer them to humans via food of animal origin. Moreover, European Union legislation regulates the hygiene of food of animal origin. In the present study, we investigated major zoonotic bacterial and viral pathogens (Salmonella spp., Yersinia enterocolitica, Listeria monocytogenes, Shiga-toxin-producing Escherichia coli (STEC), and hepatitis E virus (HEV)) in samples of wildlife and game meat from northwest Italy. Only a few samples were found to be contaminated with zoonotic bacteria, but they carried pathogenicity and antibiotic-resistance genes. HEV was not detected in any of the samples. Notwithstanding the low frequency of zoonotic pathogens, hygiene rules during the manipulation of game meat are essential to ensure consumer food safety. Wildlife can represent a reservoir of zoonotic pathogens and a public health problem. In the present study, we investigated the spread of zoonotic pathogens (Salmonella spp., Yersinia enterocolitica, Listeria monocytogenes, Shiga-toxin-producing Escherichia coli (STEC), and hepatitis E virus (HEV)) considering the presence of virulence and antibiotic resistance genes in game meat from animals hunted in northwest Italy. During two hunting seasons (2020 to 2022), samples of liver and/or muscle tissue were collected from chamois (n = 48), roe deer (n = 26), deer (n = 39), and wild boar (n = 35). Conventional microbiology and biomolecular methods were used for the detection, isolation, and characterization of the investigated pathogens. Two L. monocytogenes serotype IIa strains were isolated from wild boar liver; both presented fosfomycin resistance gene and a total of 22 virulence genes were detected and specified in the text. Eight Y. enterocolitica biotype 1A strains were isolated from chamois (2), wild boar (5), and deer (1) liver samples; all showed streptogramin and beta-lactam resistance genes; the virulence genes found were myfA (8/8 strains), ymoA (8/8), invA (8/8), ystB (8/8), and ail (4/8). Our data underscore the potential role of wildlife as a carrier of zoonotic and antibiotic-resistant pathogens in northwest Italy and a food safety risk for game meat consumers. [ABSTRACT FROM AUTHOR]

Introduction: Aeromonas hydrophila and methicillin-resistant Staphylococcus aureus (MRSA) are potent bacterial pathogens posing major hazards to human health via consuming fish harboring these pathogens or by cross-contamination beyond the contaminated environment. The aim of this study was to determine risk variables associated with the presence of certain pathogenic bacteria from Mugil cephalus fish in retail markets in Egypt. The virulence genes of A. hydrophila and S. aureus were also studied. Furthermore, the antibiotic sensitivity and multidrug resistance of the microorganisms were evaluated. Methods: In a cross-sectional investigation, 370 samples were collected from mullet skin and muscle samples, washing water, fish handlers, knives, and chopping boards. Furthermore, fish handlers' public health implications were assessed via their response to a descriptive questionnaire. Results: S. aureus and Aeromonas species dominated the investigated samples with percentages of 26.76% and 30.81%, respectively. Furthermore, A. hydrophila and MRSA were the predominant recovered bacterial pathogens among washing water and knives (53.85% and 46.66%, respectively). The virulence markers aerA and hlyA were found in 90.7% and 46.5% of A. hydrophila isolates, respectively. Moreover, the virulence genes nuc and mec were prevalent in 80% and 60% of S. aureus isolates, respectively. Antimicrobial susceptibility results revealed that all A. hydrophila isolates were resistant to amoxicillin and all MRSA isolates were resistant to amoxicillin and ampicillin. Remarkably, multiple drug resistance (MDR) patterns were detected in high proportions in A. hydrophila (88.37%) and MRSA (100%) isolates. The prevalence of Aeromonas spp. and S. aureus had a positive significant correlation with the frequency of handwashing and use of sanitizer in cleaning of instruments. MRSA showed the highest significant prevalence rate in the oldest age category. Conclusion: The pathogenic bacteria recovered in this study were virulent and had a significant correlation with risk factors associated with improper fish handling. Furthermore, a high frequency of MDR was detected in these pathogenic bacteria, posing a significant risk to food safety and public health. [ABSTRACT FROM AUTHOR]

Copyright of China Tropical Medicine is the property of China Tropical Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Aims This study aimed to characterize the first complete genome of Corynebacterium parakroppenstedtii and clarify the evolutionary relationship in the Corynebacterium kroppenstedtii complex (CKC) by using comparative genomics analysis. Methods and results The genome of isolate yu01 from a breast specimen was sequenced, and 35 CKC genomes were collected. Analysis of 16S rRNA, rpoB , and fusA suggested ambiguous identification, whereas ANI analysis assigned isolate yu01 as Coryne. parakroppenstedtii. The fourth genospecies " Corynebacterium aliikroppenstedtii " was identified in CKC. Comparative genomics analysis suggested that the genomic arrangement in CKC was highly conserved. A total of 43 potential virulence genes and 79 species-specific genes were detected. Most genome-based phylogenetic analysis were incapable of resolving the interspecific evolutionary relationships among CKCs. A total of 20 core genes were found to be distinguishable in CKC. Conclusions This study suggested the limited divergence and unavailability of normal single gene-based identification in CKC and questioned the precise species of strains associated with mastitis, identified as Coryne. kroppenstedtii in previous studies. The 20 genes showed potential to enhance the methods for the identification and epidemiological investigation of CKC. [ABSTRACT FROM AUTHOR]

Aims We aimed to investigate the prevalence, pathology, and characterization of Streptococcus dysgalactiae subsp. equisimilis (SDSE) in slaughtered pigs of India. Methods and results We collected 1254 morbid tissues (lungs—627 and spleen—627) and 627 heart-blood from 627 slaughtered pigs. The bacterial isolation, antibiogram, virulence gene profiling, and mouse pathogenicity testing were performed for the detection and characterization of SDSE. A total of 177 isolates (heart-blood—160 and tissues—17) were recovered from 627 slaughtered pigs with higher isolation rate in heart-blood (25.51%). The prevalence of SDSE was 11% in morbid tissues by polymerase chain reaction. Majority of isolates showed higher detection of streptolysin O, followed by streptokinase and extracellular phospholipase A virulence genes with higher degree of resistance to azithromycin, clindamycin, erythromycin, and penicillin antibiotics. Mouse pathogenicity testing confirmed virulence based on histopathological lesions and re-isolation of SDSE. Conclusions Our findings highlight the high prevalence of SDSE in slaughtered pigs. The presence of virulence genes and mouse pathogenicity testing confirm their pathogenic potential. [ABSTRACT FROM AUTHOR]