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Characterisation of Yersinia enterocolitica strains isolated from wildlife in the northwestern Italian Alps
2022
Carella, Emanuele | Romano, Angelo | Domenis, Lorenzo | Robetto, Serena | Spedicato, Raffaella | Guidetti, Cristina | Pitti, Monica | Orusa, Riccardo
Yersiniosis is a zoonosis causing gastroenteritis, diarrhoea, and occasionally reactive arthritis and septicaemia. Cases are often linked to meat consumption and the most common aetiological agent is the Gram-negative bacilliform Yersinia enterocolitica bacterium. The occurrence of Yersinia spp. among wild animals has mostly been studied in wild boar, but it has seldom been in other species. A total of 1,868 faecal samples from animals found dead or hunted were collected between 2015 and 2018 in the Valle d’Aosta region of the northwestern Italian Alps. Alpine ibex faecal samples were collected during a health monitoring program in 2018. Bacteria were isolated via PCR and confirmed as Y. enterocolitica biochemically. Strain antimicrobial susceptibility was tested by Kirby–Bauer disc diffusion, and the presence of virulence factors and antimicrobial resistance genes was investigated using whole-genome sequencing. Yersinia enterocolitica strains of biotype 1A were detected in six faecal samples from red deer (0.93%), roe deer (0.49%) and red foxes (0.7%). Strains found in beech martens (3.57%) and Alpine ibex (2.77%) belonged to biotypes 1B and 5, respectively and harboured the pYPTS01 plasmid that had only been detected in Y. pseudotuberculosis PB1/+. All the isolates were resistant to ampicillin and erythromycin. The biovar 1A strains exhibited different virulence factors and behaved like non-pathogenic commensals. The strain from an Alpine ibex also harboured the self-transmissible pYE854 plasmid that can mobilise itself and the pYPTS01 plasmid to other strains. The beech marten could be considered a sentinel animal for Y. enterocolitica. Phenotypic resistance may account for the ability of all the strains to resist β-lactams.
Show more [+] Less [-]First report of the presence of enterotoxin gene in coagulase-negative staphylococci recovered from meat of snails (Achatina achatina)
2022
Okafor, Arthur C. | Ogbo, Frank C. | Akharaiyi, Fred C. | Oladeinde, Henry B.
It has been suggested that coagulase-negative staphylococci can serve as reservoirs of virulence genes for other bacteria. This study assessed the presence of such genes in selected isolates recovered from meat of the giant African snail (Achatina achatina). Virulence genes were detected using a polymerase chain reaction targeting specific primers. Two representative isolates were identified using 16S rRNA gene sequencing. The results showed that the staphylococcal enterotoxin A gene (sea) was present in five out of the eight isolates studied. The isolates expressed resistance mainly to three antibiotics: chloramphenicol, norfloxacin and cloxacillin in descending order of incidence. Most importantly, the Staphylococcus sciuri isolate NEDU 181, in addition to being resistant to the three aforementioned antibiotics, also harboured the sea gene. Our findings demonstrate, for the first time, the presence of toxigenic and antibiotic-resistant coagulase-negative Staphylococcus spp. in commercially-available fresh snail meat. With staphylococcal enterotoxin A known to survive cooking temperature, this presents a food safety concern.
Show more [+] Less [-]Genetic and pathogenic characterisation of a virulent Akabane virus isolated from goats in Yunnan, China
2022
Gao, Hua-Feng | Wang, Jin-Ping | Yang, Zhen-Xing | Xie, Jia-Rui | He, Yu-Wen | Hong, Qiong-Hua | Xin, Ai-Guo
Akabane virus (AKAV) has been detected in a variety of host species in China, but there are only limited records of its occurrence in goats. However, more attention needs to be paid to understanding the diversity of viruses in this species. The aim of the study was to explore the genotype characteristics and variation trend of AKAV and their relationship with virulence in Yunnan, China. Blood samples were collected from goats during routine surveillance of goat diseases in Yunnan province in 2019. The AKAV CX-01 strain was isolated using BHK-21 cells. To understand pathogenicity, the virus was intraperitoneally (IP) and intracerebrally (IC) inoculated into suckling mice and tissue samples were subsequently analysed histopathologically and immunohistochemically. Akabane virus CX-01 strain induced encephalitis and impairment of the central nervous system with fatal consequences. Phylogenetic analysis based on the ORF sequences of the small segments indicated that the AKAV isolate used was most closely related to the GD18134/2018 Chinese midge and bovine NM BS/1strains, while phylogenetic analysis based on the medium segments showed a close relationship between CX-01 and the Chinese GLXCH01 strain. The CX-01 isolate was related to AKAV genogroup Ia and probably originated from a recombination of different strains.
Show more [+] Less [-]Prevalence and Characterization of Some Pathogenic Bacteria in Fermented Milk Products and Mish Cheese in Dakahalia Governorate, Egypt
2022
Rana Abd Elkader Mahmoud Abd El latif | Mohammed El Sherbini El-sayed Ali | Adel Abdelkhalek
Bacterial contamination of fermented dairy products has serious implications for both safeties of the final products, and the transmission of foodborne pathogens to the consumers. In this regard, 175 samples of fermented dairy products including 50 each of plain yoghurt, fruit yoghurt, laban rayeb, and 25 from mish cheese were randomly collected from different supermarkets and retail shops in Dakahlia Governorate, Egypt. Samples were examined bacteriologically for the prevalence of Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), particularly, E. coli O157. The recovered isolates were examined for the detection of toxin, and virulence-associated genes using PCR. The obtained results showed that the average counts (CFU/g) of total bacterial, psychrotrophic, coliform, and S. aureus were 1.72x105, 9.9 x103, 5.08x102, and 3.07 x102 for the plain yoghurt, 7.1 x 104, 2.3 x 103, 9.5 x10, and 3.3 x10 for fruit yoghurt, 6.07x104, 6.1 x 103, 9.8 x10, and 1.35x102 for laban rayeb, and 1.2 x 106, 2.5 x104, 8.3 x10, and 7.2 x103 for Mish cheese, respectively. E. coli and E. coli O157 were detected in 13 out of 175 (7.43%) and 4 out of 175 (2.3%) samples, respectively. Using PCR for 9 selected E. coli isolates showed that 7 out of 9 E. coli isolates were positive for the stx1 gene, 5 out of 9 were positive for stx2, and 3 out of 9 were positive for eaeA, and 4 out of 9 were positive for hylA. S. aureus showed that 55.6% of the recovered isolates were coagulase positive. Ten randomly selected S. aureus isolates tested positive for nuc (thermonuclease genes), while mecA (methicillin-resistant S. aureus “MRSA”) gene was detected in 20% of the examined isolates. Therefore, strict hygienic measures should be adopted during all steps of the manufacture of such dairy products.
Show more [+] Less [-]Effect of aromatic oils on the expression of some virulence-associated and antimicrobial resistance genes of Escherichia coli isolated from broilers
2022
Walid Hamdy Hassan (Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt) | Hala Sayed Hassan Salam (Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt) | Wafaa Mohamed Hassan (Microbiology-Reference Laboratory of Quality Control on Poultry Production, Animal Health Research Institute AHRI, Giza Governorate, Egypt) | Salama Abohamra Sayed Shany (Department of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt) | Ghada Sayed Ibrahim Osman (Department of Bacteriology, Animal Health Research Institute, Beni-Suef Laboratory, Agricultural Research Center, Beni-Suef, Egypt.)
Objectives: This study aimed to prove the effects of Escherichia coli isolates isolated from dis¬eased broilers to form biofilms, describe their antimicrobial sensetivity, and determine the effect of allicin and cinnamon essential oils on the expression of some genes (fimH, int1, and luxS) through quantitative polymerase chain reaction (q-PCR). Materials and Methods: 140 samples were obtained from diseased broilers in Beni-Suef Governorate, Egypt. These samples were examined by conventional bacteriology methods to detect the causative agent. The antimicrobial susceptibility of the isolated bacteria was assessed using the disc diffusion method, The ability of yeast extract-casamino acids Congo Red Agar to generate phenotypic biofilms was next tested. The presence of resistance and virulence genes in some multidrug resistant isolates was genotypically investigated. The antibacterial effects of allicin and cinnamon oil were evaluated against the growth of multidrug-resistant E. coli. Finally, q-PCR was utilized to assess changes in some genes' expression. Results: Escherichia coli was isolated from 61 samples (43.6%). An antimicrobial susceptibility test revealed that multidrug-resistance (MDR) (could resist more than three antimicrobial classes) E. coli prevalence was 100%. 40.8% of isolates phenotypically produce biofilms. The detection of resistance and virulence genes by PCR showed that all tested isolates carry aadB, fimH, int1, qnrS, and luxS genes, while only 40% harbor iss genes. q-PCR showed that after treatment with allicin and cinnamon oils, gene expression went down. Conclusion: This investigation highlights that E. coli showed resistance against most of the tested antimicrobials; all isolates were MDR. The study showed wide dissemination of virulence and resistance genes among E. coli. Allicin and cinnamon oils have antimicrobial activities and could be used as alternatives to synthetic antimicrobial agents. [J Adv Vet Anim Res 2022; 9(2.000): 191-202]
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