Contamination of Kazakhstan cheeses originating from Escherichia coli and its resistance to antimicrobial drugs.

Background and Aim: Escherichia coli, a commensal intestine bacterium of vertebrates, is widely distributed in the environment and indicates the microbiological quality of food products in relation to coliforms. In addition, virulent strains, particularly E. coli O157:H7, cause outbreaks of toxic infections caused by consuming dairy products. Because food safety studies regarding E. coli have not been conducted in Central Asia, this research aimed to study the characteristics of contamination, microbiological and genotypic properties, and resistance to antimicrobial agents of E. coli strains that contaminate various types of commercialized cheeses originating from Kazakhstan. Materials and Methods: In retail outlets, 207 samples of three types of cheese produced by 22 industrial and eight small enterprises in the central, eastern, southern, and northern regions of Kazakhstan were selected in 2020-2023. E. coli contamination was examined using standard microbiological, mass spectrometric, and molecular genetic methods. The discodiffuse European Committee on Antimicrobial Susceptibility Testing method was used to test the resistance of the identified E. coli isolates (65/207; 31.4%) to 20 antibacterial drugs. The Shiga toxin-producing E. coli (VT1 and VT2) and E. coli O157:H7 (eae) genes were investigated in all E. coli isolates using multiplex polymerase chain reaction. Results: An average of 31.4% samples of commercial Kazakhstani cheeses of various types were found to be contaminated with E. coli in almost all geographical regions of Kazakhstan, regardless of the productivity of the dairy enterprises. Soft cheeses produced by small farms (80% of samples) packaged at the retail site (100%) were the most contaminated with E. coli. The microbiological index (colony-forming unit/g) was unsatisfactory and unsuitable in 6.2% of such cheese samples. For the first time in Central Asia, the enteropathogenic strain E. coli O157:H7 was detected in 0.5% of cheese samples. E. coli isolates from cheese samples were resistant to 65% of antibacterial drugs and contained resistance genes to ß-lactams, sulfonamides, and quinolones groups. At the same time, 25% of the E. coli isolates were multi-resistant to three or more antimicrobial agents. Conclusion: The high level of contamination caused by multi-antibiotic resistant E. coli strains, including pathogenic pathogens, poses a risk to public health and highlights the need for further research on the monitoring and control of coliform enteropathogens in food products.

Prevalence and antimicrobial resistance of Enterobacteriaceae in the north of Kazakhstan.

Background: An increasing number of drugs are used each year in the treatment of small pets (cats and dogs), including medicines (cephalosporins and fluoroquinolones) used in human therapy. Aim: The purpose of this study was to isolate and explore the antibiotic resistance of opportunistic Enterobacteriaceae (Escherichia coli, Klebsiella, Proteus, Ci trobacter, Enterobacter) from cats and dogs, and to isolate resistance genes in the microorganisms. Methods: In 2021, 808 samples of biological material from small domestic animals were collected in veterinary clinics in Kostanay. From these, 210 microorganisms were isolated and identified. Results: A large majority of the strains sampled belonged to E. coli-149 (70.9%), Enterobacter-11 (5.2%), Klebsiella-28 (13.3%), Proteus-12 (5.7%) and 10 Citrobacter isolates (4.8%). In all isolates identified, antibiotic resistance/sensitivity was determined by disc-diffusion method to ampicillin, cefoxitin, gentamicin, levomycetin, tetracycline, ciprofloxacin, norfloxacin, ofloxacin, cefoperazone, cefpodoxime, streptomycin, kanamycin, doxycycline, gemifloxacin, nalidixic acid, furazolidone, furadonine, amoxicillin, and enrofloxacin. Conclusion: The study has demonstrated that the greatest number of Enterobacteriaceae were sensitive to the action of meropenem, which belongs to the group of beta-lactam antibiotics; resistance was demonstrated against tetracycline, doxycycline, ampicillin, amoxicillin, ofloxacin, and cefpodoxime. The most common genes encoding antimicrobial resistance were as follows: BlaTEM and OXA in 41 and 28 isolates, respectively, encoding resistance to beta-lactams; StrA and StrB in 45 and 48 isolates encoding aminoglycosides; and tetA and tetB in 43 and 28 isolates encoding tetracyclines. Obtained data demonstrate that uncontrolled and frequent use of beta-lactam and tetracycline antibacterials, in cats and dogs, results in the spread of genotypic resistance among micro-organisms of the family Enterobacteriaceae.

Identification of the causative agent of canine babesiosis in the North of Kazakhstan.

Background: Canine babesiosis is a common disease in the northern part of the Republic of Kazakhstan, in particular in the Kostanay region. In recent years, a large number of cases of the disease with a variety of clinical symptoms have been registered. Aim: The purpose of the study was to monitor the spread, characterization, and identify the Babesia species involved of Babesia species in ticks and blood of dogs in the Kostanai region. Methods: The research work began in 2017 with the study of the spread of babesiosis in dogs in the Kostanay region according to the reports of veterinary clinics. The collection of ticks from the territory and from dogs was carried out in 2017-2021. Results: As a result of the research work, the presence in the city and some areas of the Kostanay region of two species of ixodid Dermacentor reticulatus and Dermacentor marginatus, was established. Of these, one species was identified in dogs, which serves as a carrier of canine babesiosis-D. reticulatus. In all 31 DNA samples from the blood of dogs diagnosed with babesiosis, a fragment of the 18S rRNA gene was amplified. The nucleotide sequence was obtained for 30 samples (96.8%), in one sample a low luminescence intensity of a specific PCR product was observed. Two Babesia canis haplotypes were distinguished on the basis of two nucleotide substitutions (GA→AG) observed in the sequences of the 18S rRNA gene. Conclusions: In conclusion, the results of this study provide insight into the distribution of B. canis haplotypes in dogs in the Kostanay region, and canine babesiosis is caused solely by the large Babesia species B. canis.

Prevalence and resistance to antibacterial agents in Salmonella enterica strains isolated from poultry products in Northern Kazakhstan.

Background and Aim: Salmonella is one of the main causative agents of foodborne infections. The source of the pathogen, in most cases, is poultry products. The intensification of poultry farming and the constant and uncontrolled use of antimicrobials has led to an increase in the level of antibiotic resistance, especially in developing countries. This study aimed to determine the level of sensitivity to antimicrobial agents in Salmonella enterica strains isolated from poultry products in Northern Kazakhstan, as well as to determine the genetic mechanisms of resistance and the presence of integrons. Materials and Methods: In total, 398 samples of poultry products sold in Northern Kazakhstan were selected. Salmonella strains were isolated from product samples using microbiological methods. Salmonella was identified based on morphological, biochemical, and serological methods, as well as polymerase chain reaction (PCR). Sensitivity testing for antimicrobial agents was performed using the disk diffusion method. The detection of resistance genes was performed using PCR and gel electrophoresis. Results: Out of 398 samples of poultry products, a total of 46 Salmonella isolates were obtained. Most of the isolates belong to the serovar Salmonella Enteritidis (80.4%). The assessment of sensitivity to antibacterial agents showed that Salmonella was mainly resistant to nalidixic acid (63%), furadonin (60.9%), ofloxacin (45.6%), and tetracycline (39.1%). In 64.3% of cases, Salmonella was resistant to three or more groups of antibacterial agents. Resistance genes such as tetA, tetB, blaTEM, aadA, sul3, and catII, as well as integrons of two classes (teg1 and teg2), were identified. Conclusion: Poultry products contain antimicrobial-resistant strains of Salmonella, as well as genes encoding resistance mechanisms. The results emphasize the need for constant monitoring of not only pathogenic microorganisms but also their sensitivity to antimicrobial agents. The potential threat to human health requires a unified approach to the problem of antibiotic resistance from representatives of both public health and the agroindustrial complex.

Antibiotic resistance of enterobacterial pathogens isolated on the territory of the Northern Kazakhstan.

The purpose of this study was to isolate and identify strains of infectious disease pathogens and to determine their antibacterial drug resistance. From 2018-2020, 5322 samples of biological material from animals and birds, products of animal origin were taken at retail outlets, livestock enterprises in Kostanay and North Kazakhstan regions. In the study territories of 20 districts and 4 cities were covered. Phenotypic resistance analysis of the recovered isolates of Salmonella spp. showed a high level of resistance to antibiotics of the tetracyclines (82/137) and nitrofurans (81/137) groups. S. aureus isolates had increased resistance to the ß-lactams (124/131) and macrolides (96/131) groups. As a result, the largest number of genes encoding resistance was found to the ß-lactams, aminoglycosides, tetracyclines and macrolides groups. The study was carried out within the framework of the project AR05131447 "Monitoring of antibiotic resistance of pathogens of enteropathogenic zooanthroponosis diseases in the Northern Kazakhstan".

Antibiotic resistance and biofilm formation in Staphylococcus aureus isolated from dairy cows at the stage of subclinical mastitis in northern Kazakhstan.

Staphylococcus aureus is an important causative agent of subclinical bovine mastitis worldwide. The aim of this research was to study the ability of S. aureus to form biofilms. Additionally, we examined the genes involved in cell resistance and sensitivity to antibiotics. Samples were collected from December 2020 to May 2021 from Simmental and black-and-white cows. The study was carried out on a total number of 643 cows, of which 278 (23 %) were in the subclinical mastitis stage. Finally, 64 S. aureus isolates were isolated and identified. The highest level of phenotypic resistance was observed to antibiotics of the tetracycline (tetracycline - 48.4 %, doxycycline - 32.8 %) and ß -lactam (ampicillin - 45.3 %, penicillin - 45.3 %) groups. The genes encoding antibiotic resistance were characterized with the polymerase chain reaction method: blaZ in 30 isolates, mecA in 1 isolate, ermC in 15 isolates, aph (3) in 2 isolates, tetK in 19 isolates, tetM in 9 isolates. The tested S. aureus isolates had the ability to form biofilms in 76.6 % ( 49 / 64 ) of cases. Of these, 69.4 % were resistant to at least one antibiotic. The obtained results have shown that S. aureus, identified in cows with subclinical mastitis, was resistant mainly to tetracycline and ß -lactam antibiotics. In addition, S. aureus isolates expressed resistance genes to the above drugs and had the ability to form biofilm. This study will help to identify the extent of antibiotic resistance and monitor S. aureus contamination of raw milk.