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.

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.

Study of the antibiotic residues in poultry meat in some of the EU countries and selection of the best compositions of lactic acid bacteria and essential oils against Salmonella enterica.

In this study, the presence of antibiotics (ANB) residues was evaluated in poultry meat purchased from German and Lithuanian markets. In addition, the antimicrobial activity of 13 lactic acid bacteria (LAB) strains, 2 essential oils (EO) (Thymus vulgaris and Origanum vulgare L.), and their compositions were tested for the purpose of inhibiting antibiotic-resistant Salmonella spp. ANB residues were found in 3 out of the 20 analyzed poultry meat samples: sample no. 8 contained enrofloxacin (0.46 µg/kg), sample no. 14 contained both enrofloxacin and doxycycline (0.05 and 16.8 µg/kg, respectively), and sample no. 18 contained enrofloxacin (2.06 µg/kg). The maximum residue limits (MRLs) for the sum of enrofloxacin and ciprofloxacin and for doxycycline in the poultry muscle are 100 µg/kg. Finally, none of the tested poultry meat samples exceeded the suggested MRLs; however, the issue of ANB residues still requires monitoring of the poultry industry in Germany, Poland, and Lithuania, despite the currently established low ANB concentrations. These findings can be explained by the increased use of alternatives to ANB in the poultry industry. Our results showed that an effective alternative to ANB, which can help to reduce the occurrence of antibiotic-resistant salmonella, is a composition containing 1.0% of thyme EO and the following LAB strains: Lactobacillus plantrum LUHS122, Enteroccocus pseudoavium LUHS242, Lactobacillus casei LUHS210, Lactobacillus paracasei LUHS244, Lactobacillus plantarum LUHS135, Lactobacillus coryniformins LUHS71, and Lactobacillus uvarum LUHS245, which can be recommended for poultry industry as components of feed or for the treatment of surfaces, to control the contamination with Salmonella strains. However, it should be mentioned that most of the tested LAB strains were inhibited by thyme EO at the concentrations of 0.5 and 1.0%, except for LUHS122, LUHS210, and LUHS245. Finally, it can be noted that the agents responsible for the inhibitory effect on Salmonella are not the viable LAB strains but rather their metabolites, and further studies are needed to identify which metabolites are the most important.