Analyzing the Sensitivity and Antibiotic Resistance of Enterobacteriaceae Isolates Isolated from Animal Products in Almaty Region, Kazakhstan

Analyzing the Sensitivity and Antibiotic Resistance of Enterobacteriaceae Isolates Isolated from Animal Products in Almaty Region, Kazakhstan | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Analyzing the Sensitivity and Antibiotic Resistance of Enterobacteriaceae Isolates Isolated from Animal Products in Almaty Region, Kazakhstan Z. A. Latypova, A. S. Nurpeisova, M. T. Nurgalieva, A. B. Bizhanov, and 16 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4575952/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: This article presents the results of studies on the antibiotic resistance of Escherichia coli and Salmonella enterica isolates isolated from animal products of the Almaty region, Kazakhstan using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood. Results: The results showed that the Almaty region can be classified as region with low resistance rates of E. coli to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of E. coli to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin, ciprofloxacin. The use of a domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro and macronutrients for reliable and high-quality laboratory analyses. Conclusions: It is assumed that the irrational use of fluoroquinolones in animal husbandry leads to an increase in the resistance of microorganisms that cause infectious diseases common to humans and animals, since the above types of antibiotics (fluoroquinolones) are the most important drugs for the treatment of bacterial infections in medicine and veterinary medicine. The results indicate that pathogens of enteropathogenic diseases resistant to antibacterial drugs are circulating in the territory of the Almaty region of Kazakhstan and the use of a domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro and macronutrients for reliable and high-quality laboratory analyses. antibiotic resistance Salmonella enterica Escherichia сoli animal products culture media for the cultivation of microorganisms with the addition of defibrinated horse blood Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Diseases and deaths from various infections caused by contaminated food pose a constant threat to public health and a serious obstacle to socioeconomic development worldwide. The Global Antimicrobial Resistance and Use Surveillance System (GLASS), which is based on the World Health Organization (WHO) has identified widespread antibiotic resistance. According to the presented data, 70% of known bacteria have developed resistance to one or more antibiotics. The greatest amount of data on antimicrobial resistance has been obtained for the following bacterial species: Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus Streptococcus pneumoniae and Salmonella spp . [ 1 ]. Of particular for importance of such infectious diseases in medicine, veterinary medicine and agriculture determine the increased resistance of pathogens circulating in the environment to antimicrobial agents [ 2 , 3 ]. Аntimicrobial agents, including antibiotics and antiviral, antifungal and antiparasitic drugs, are medicines used to prevent and treat infections in humans, animals and plants [ 4 ]. In May of 2024, the WHO updated the Bacterial Priority Pathogens List (BPPL) 2024, which includes 15 families of antibiotic-resistant bacteria grouped into critical, high and medium categories for prioritization. Four new pathogen–antibiotic combinations were added to BPPL 2024. The updated list contains new data. А separate item in the high-priority group included third-generation cephalosporin-resistant bacteria of the order Enterobacterales, which requires increased attention and special measures. Bacteria of the genus Salmonella are classified as pathogens of a high-priority group. BPPL 2024 contains recommendations for the development of new and necessary treatment methods designed to stop the spread of antimicrobial resistance (AMR) [ 5 ]. In current clinical veterinary practice, the rational use of antibacterial drugs is important. It is known that inappropriate antibiotic prescribing in animal therapy inevitably leads to an increase in the level of tolerance to these drugs in circulating strains of pathogenic and conditionally pathogenic microorganisms. To curb the increase in inantibiotic resistance of pathogens of infectious diseases of farm animals and birds, it is necessary to take an evidence-based approach for the use of antibiotic, to reduce their unnecessary and mass use [ 3 ]. The aim of this study was to assess the sensitivity or antibiotic resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood. Material/Methods The material for study consisted of cultures of E.сoli and Salmonella (n=186) isolated from the territory of the Almaty region of the Republic of Kazakhstan. The sources of the microorganism isolates were 5 types of animal product, namely, beef (n=123), lamb (n=119), pork (n=122), and poultry (n=114), and poultry eggs (n=122), which were purchased by random sampling in large retail chains, individual food and retail stores shops, supermarkets and markets in the Almaty region. All the studies were carried out in the laboratories of the Kazakh Research Veterinary Institute LLP. The identification of microorganisms from animal products was carried out according to methods generally accepted in veterinary laboratory practice. In accordance with the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST), sensitivity assessment and interpretation of the results of studies on the sensitivity of E. coli and Salmonella to antibiotics were carried out using the disc diffusion method of Mueller-Hinton Agar (Oxoid, UK) with and without the addition of 5% defibrinated horse blood. Version 10.0, 2020. http://www.eucast.org [6,7]. During the sensitivity determination, commercial antibiotic discs of certain types were used: cefotaxime 30 mcg, ampicillin/sulbactam 20 mcg, norfloxacin 10 mcg, pefloxacin 5 mcg, ceftriaxone 30 mcg, tigecycline 15 mcg, meropenem 10 mcg, levofloxacin 5mcg, trimethoprim 5 mcg, gentamicin 10 mcg, chloramphenicol 30 mcg, amoksiklav 30 mcg. Cultures of Escherichia coli ATCC 25922 and ATCC 35218 were used for sensitivity determination. Molecular genetics testing DNA isolation Genomic DNA was isolated from daily bacterial cultures using a PureLink Genomic DNA Kit, - according to the manufacturer’s protocol (Invitrogen, Carlsbad, USA). The DNA concentration in the samples was determined on a Qubit® 2.0 fluorimeter using a QubitTM dsDNA HS Assay Kit (Life Technologies, Oregon, USA). Amplification of 16S rRNA The 16S rRNA gene was used as a genetic marker. To amplify the 16S rRNA gene, 25 µl of the following reaction mixture was prepared: 12.5 µl of Q5® Hot Start High-Fidelity 2X Master Mix (New England Biolabs Ins., USA); a pair of universal primers; 8F (5'-AGAGTTTGATCCTGGCTCAG-3') and 806R (5'-GGACTACCAGGGTATCTAAT-3') (1.2 µl in 10 µM concentration); and DNA and water matrix (up to 25 µl) [5]. The amplification mode consisted of the following cycles: 95°C for 5 min, 95°C for 30 seconds, 55°C for 40 seconds, 72°C for 50 seconds for 30 cycles; and elongation at 72°C for 10 min. The PCR products were purified using CleanSweep™ PCR Purification (Life Technologies, Carlsbad, CA). 16S rRNA sequencing. Sequencing of the 16S rRNA gene of bacteria was performed using the Big Dye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, USA) according to the manufacturer’s protocol (BigDye® Terminator v3.1 Cycle Sequencing Kit Protocol Applied Biosystems, USA), followed by fragment separation on an automatic genetic analyser 3500 DNA Genetic Analyser (Applied Biosystems, Hitachi, Tokyo Japan). The sequencing results were processed in the Seq program (Applied Biosystems). The search for homologous nucleotide sequences of 16S rRNA genes was carried out using the Basic Local Alignment Search Tool (BLAST) in the International GenBank database of the National Center for Biotechnology Information of the USA (htpp://www.ncbi.nlm. nih.gov) [8]. Phylogenetic analysis was performed using of the Molecular Evolutionary Genetics Analysis (MEGA6) software and nucleotide sequence alignment was performed using the ClustalW algorithm. Phylogenetic identification was carried out using the neighbor joining algorithm BLASTN (NJ) program. Statistical processing Statistical processing of the research results was carried out using standard methods of descriptive statistics via the Python program. Results According to the research, 111 E. coli isolates (18.3%) and 75 Salmonella isolates (12.5%) were isolated, including 22.52% (n = 25) of the E. coli isolates from poultry, 24.32% (n = 27) from lamb, 9.9% (n = 11) from beref, 34.23% (n = 38) from pork and 9% (n = 10) from poultry eggs. Salmonella isolates were isolated from poultry – (37.3%, (n = 28), lamb (14.66%, (n = 11), beef (14.66%, (n = 11), pork (14.66%, (n = 11) and poultry eggs (18.66%, (n = 14). Fourteen isolates lost their viability during storage. As shown in Fig. 1 , the isolates were highly sensitive to most antibiotics, such as cefotaxime, ampicillin/sulbactam, ceftriaxone, tigecycline, meropenem, trimethoprim, gentamicin, chloramphenicol and amoksiklav, with 100% sensitivity and 0% resistance. Moreover, 15.15% and 25.76% of the isolates were resistant to norfloxacin and pefloxacin, respectively. Levofloxacin showed a small percentage of resistance (7.58%) and high sensitivity (92.4%). Figure 2 shows the resistance and sensitivity of 106 E. coli isolates to various antibiotics. High sensitivity to most antibiotics, such as cefotaxime, ampicillin/sulbactam, ceftriaxone, tigecycline, meropenem, trimethoprim, chloramphenicol and amoksiklav was detected, with high sensitivity (more than 97%) and low resistance. Moderate levels of resistance were detected for gentamicin and pefloxacin (16.04% and 15.09%), respectively. There was a greater percentage of resistant to norfloxacin (29.25%) than to other antibiotics. As shown in Fig. 1 , for E. coli , the most effective antibiotics are trimethoprim, ceftriaxone, and tigecycline. Some antibiotics, such as cefotaxime, ampicillin/sulbactam and levofloxacin, are less effective. Figure 2 shows the most effective antibiotics for Salmonella infection: trimethoprim, gentamicin, chloramphenicol and amoksiklav. These data show that trimethoprim, ceftriaxone and tigecycline are the most effective antibiotics for both types of bacteria, while some antibiotics have limited efficacy. The average values of zones of growth suppression show how effective each antibiotic is against these E. coli and Salmonella test samples. For example, meropenem shows high efficacy against E. coli with the highest average value zones of suppression (33.78 mm) and a low standard deviation (1.97 mm), which indicates the stability of its action. For Salmonella, pefloxacin also showed high efficacy, with an average value of 25.5 mm, but the standard deviation was greater (5.74 mm), indicating greater variability of action between samples. Compared with meropenem and pefloxacin, ampicillin/sulbactam showed lower efficacy, especially against E. coli , with lower mean zones of suppression and greater standard deviations, indicating a less predictable effect. These data can help in choosing the most effective antibiotic for the treatment of infections caused by E. coli and Salmonella, taking into account the average zones of suppression and the variability of drug action. Thus, monitoring studies of animal products have allowed us to conclude that poultry, lamb and pork samples are the most contaminated with E. coli − 22.52%, 24.32% and 34.23%, respectively. The results of research products to identify Salmonella showed that these pathogens were found quite often in poultry and poultry eggs, at 37.3% and 18.66%, respectively. In total, 186 isolates were isolated, including: E. coli − 111, and Salmonella − 75, which were used for further research. Sensitivity to 12 antibacterial drugs was studied in 172 isolated isolates by the disc diffusion method. All isolates isolated from animal products showed varying degrees of sensitivity and resistance to antibiotics. The interpretation of the results in determining sensitivity was carried out using the recommendations of EUCAST (v 6.0) [29]. The results of the research are shown in Fig. 5. Table 1 Resistance of E. coli and Salmonella to various antibiotics Name of the antibiotic E. coli resistance Salmonella resistance Cefotaxime 30 mcg 2,83 0 Ampicillin/Sulbactam 20 mcg 2,83 0 Norfloxacin 10 mcg 29,25 15,15 Pefloxacin 5 mcg 15,09 25,76 Ceftriaxone 30 mcg 0 0 Tigecycline 15 mcg 0 0 Meropenem 10 mcg 0,94 0 Levofloxacin 5mcg 2,83 7,58 Trimethoprim 5 mcg 0 0 Gentamicin 10 mcg 16,04 0 Chloramphenicol 30 mcg 0 0 Amoksiklav 30 mcg 0 0 As shown in from Table 1 and Fig. 3 , cefotaxime and ampicillin/sulbactam had the same resistance in E. coli (2.83%), and no resistance was detected in Salmonella. Norfloxacin shows high-level resistance in E. coli (29.25%) and moderate resistance in Salmonella (15.15%). Pefloxacin had significant resistance in both bacteria: 15.09% for E. coli and 25.76% for Salmonella. Meropenem resistance was low shows in E. coli (0.94%), and no resistance was detected in Salmonella. Levofloxacin had the same resistance in E. coli (2.83%) cefotaxime, ampicillin/sulbactam and moderate resistance in Salmonella (7.58%). Gentamicin showed resistance in E. coli (16.04%) and zero resistance in Salmonella. The isolates were found to be resistant to at least one of the tested antibacterial drugs. Some of the studied bacteria were resistant to several antibiotics in different groups and exhibited polyresistance. Thus, 74 (43%) E. coli isolates were resistant to fluoroquinolones (first, second and third generation), β-lactam antibiotics (first, second generation) and aminoglycosides. According to the findings, the frequency of resistance to cefotaxime in E. coli was 2.83% (n = 3), that to ampicillin/sulbactam was 2.83% (n = 3), that to norfloxacin was 29.25% (n = 31), that to pefloxacin was 15.09% (n = 16), that to meropenem was 0.94% (n = 1), that to levofloxacin was 2.83% (n = 3), and that to gentamicin was 16.04% (n = 17). Among the isolates of the genus Salmonella, 32 (18%) were resistant to only the antibacterial drugs fluoroquinolones (first, second and third generation). Thus, resistance to norfloxacin was 15.15% (n = 10), resistance to pefloxacin was 25.76% (n = 17), and resistance to levofloxacin was 7.58% (n = 5). The tested isolates of the genus Salmonella showed sensitivity to the antibiotics β-lactam antibiotics and aminoglycosides. Isolates of E. coli and Salmonella resistant to ceftriaxone, tigecycline, chloramphenicol and amoksiklav were not isolated. Overall, it was experimentally determined that the highest percentage of resistance in isolated E. coli and Salmonella isolates was found for norfloxacin (29.25% and 15.15%, respectively) and pefloxacin (15.09% and 25.76%, respectively), and the lowest percentage was found for meropenem (0.94% and 0.00%, respectively). Notably, of the 172 Enterobacteriaceae isolates studied, 66 (38%) showed sensitivity to all groups of tested antibiotics. In the process of identifying antibiotic-resistant and antibiotic-sensitive Enterobacteriaceae, a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood was tested. The detection of hemolytic activity suggested that all the studied cultures had the necessary phenotypic properties. During cultivation on a dense medium of the Enterococcus faecalis strain, the formation of small, rounded, convex, shiny colonies ranging in size from 0.5 mm to 1.0 mm, without hemolysis zones was noted. The growth of Staphylococcus aureus was characterized by the presence of colonies ranging in size from 0.5 mm to 1.5 mm. Hemolysis zones were established around each colony, the size of which did not exceed 2.0 mm. During cultivation of the Streptococcus pyogenes strain on agar, the growth of gray colonies with a size of 1.0 mm and a clearly defined hemolysis zone reaching 3.0–4.0 mm was noted. It should be noted that during the incubation of microorganisms on plain agar with the addition of blood, the indicated sizes of the hemolysis zone were noted after 24 hours of incubation, and after 72 hours of growth, a significant increase in their sizes was observed. It was also found that during cultivation of Staphylococcus aureus and Streptococcus pyogenes on a medium containing the proposed component from defibrated horse blood, the enlightening zones were clearer and more extensive, and more pronounced lysis was observed. To confirm the species identity of Enterobacteriaceae isolated from meat, 2 isolates of Salmonella and E. coli were sequenced, and the results are shown in Figs. 4 and 5. Figure 4. A phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the S. enterica sample with reference sequences from the NCBI database. The S. enterica sample is listed as having the isolate 7 gene for 16S rRNA Figure 4 provides a phylogenetic tree representing the evolutionary relationships between different strains of S. enterica (7) based on similarities and differences in their genetic characteristics. Sample "7" is located at the top of the tree and branches closest to the group consisting of various subspecies of Salmonella enterica . This proximity suggests that they are closely related to these bacteria, and "7" probably refers to a specific strain or sequence within this group. The tree shows that the Salmonella enterica group forms a monophyletic clade, meaning that all strains in this group have one common ancestor that has nothing in common with any strains outside this group. Sample "7" appears to be part of this clade. From the figure, it can be concluded that sample "7" represents a unique sequence or strain of Salmonella enterica , which may be an important discovery for studies of pathogenicity, antibiotic resistance or the evolutionary biology of this species. The degree of homology with the nearest strain NR 104709.1:40–767 Salmonella enterica subsp. enterica strain LT2 was 100.00%. Figure 5. A phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the E. coli sample with reference sequences from the NCBI database. The E. coli sample used for the 16S rRNA gene is listed as isolate 11 gene Figure 5 provides a phylogenetic tree representing the evolutionary relationships between different strains of E.coli (11) based on similarities and differences in their genetic characteristics. The position on the tree "Isolate 11 gene for 16S RNA" is located on a separate branch, which makes it relatively close to the "Leclercia adecarboxylata gene for 16S RNA partial sequence strain: NBRC 102595". The proximity on the tree indicates that these two strains may have a relatively recent common ancestor compared to other represented species and strains. The scale band represents a difference of 0.5% in the nucleotide sequence of the 16S rRNA gene, which means that a line of this length represents a sequence divergence of 0.5% from the branching point. The shorter the branch leading to isolate 11, the more genetically similar it is to the sequence at the point where it branches off from Leclercia adecarboxylata. The topology of the tree indicates the nature of the evolutionary divergence. Isolate 11 was separated from the cluster, which included several strains of Escherichia and another strain of Leclercia. This suggests that Isolate 11 is genetically sufficiently distinct to form its own line within this group. The tree also demonstrated a wide range of genetic diversity among various species and strains of Escherichia. For example, has been shown that E. coli and E. fergusonii are genetically different from the Isolate 11 sample. Thus, Isolate 11 16S is 100% homologous to other bacteria of the genus Escherichia and has special genetic characteristics that distinguish it from other species and strains of Escherichia. Discussion The need to develop new ways to solve the problems of high levels of resistance of bacteria, viruses, fungi, and parasites and progressive drug resistance is a global task of modern medicine [ 9 , 10 ]. The problem of resistance to drugs, antibiotics or antimicrobials is growing rapidly worldwide. Antibiotic therapy is one of the main approaches of modern medicine to fight infections. The "golden era" of antibiotics lasted from the 1930s to the 1960s, when many antibiotics appeared. Unfortunately, this era has ended because researchers have been unable to maintain the pace of antibiotic discovery in the face of new resistant pathogens. Constant failures in the development or discovery of new antibiotics and their unreasonable use are predisposing factors associated with the emergence of antibiotic resistance in many infectious diseases of animals and humans [ 9 , 11 – 13 ]. The main causes of many infectious diseases in animals and humans are gram-negative bacteria, among which representatives of the Enterobacteriaceae family occupy a leading position. The greatest amount of data on antimicrobial resistance was obtained for the following bacterial species: Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Streptococcus pneumoniae , and Salmonella spp [ 14 , 15 ]. The obtained results, to our knowledge, represent an assessment of the burden of antibiotic sensitivity and resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan, which poses a serious global health threat that requires greater attention, funding and capacity building, research and development, as well as prioritization of specific pathogens from the broader global health community. The findings include an assessment of the burden of antibiotic sensitivity and resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan, which represents a serious global health threat and requires greater attention, funding and capacity-building, research and development, as well as prioritization of specific pathogens by the broader global health community. The results of the research presented in the article using a new domestic component of culture media for cultivating microorganisms from defibrinated horse blood showed that the Almaty region can be classified as region with low resistance rates of E. coli to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of E. coli to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin and ciprofloxacin. Today, for the first time, the WHO publishes a Bacterial Priority Pathogens List of antibiotic–resistant “priority pathogens” including 15 families of antibiotic-resistant bacteria that pose the greatest threat to human health. The list was intended to inform research and development priorities related to new antibiotics and to pay special attention to multidrug-resistant pathogens that cause severe and often fatal infections in health facilities [ 14 ]. There is much scientific evidence proving that antibiotics, of course, can save millions of lives, but at the same time they can also harm, and even future generations, if used improperly. This fact raises the question of the rationality of the use of antibacterial agents and suggests that despite all efforts, there is a lack of knowledge in the global medical community regarding the potentially dangerous aspects of antibiotic therapy. The irrational use of antibiotics, which are far from WHO recommendations and are mainly in injectable form, has been reported by Ethiopian scientists [ 16 ]. E.coli and Salmonella are among the most common bacteria in the world. Many methods have been developed to diagnose these infections, one of which is bacteriological examination. The complexity and complexity of isolating U.coli and Salmonella limits the use of this method in routine practice in practical healthcare, but it remains the only method for phenotypic determination of the sensitivity of strains to antibacterial drugs. Monitoring of the increasing resistance of these infections, correction of eradication therapy regimens considering the sensitivity of isolated strains to chemotherapeutic drugs, and testing of new drugs are indispensable bacteriological methods for practical and scientific purposes. An obligatory component of the medium should be an additive of 5–10% of the blood or serum of animals (horse, sheep) [ 17 ]. E.coli and Salmonella are among the most common bacteria in the world. Many methods have been developed to diagnose these infections, one of which is bacteriological research. The complexity of isolating E.coli and Salmonella limits the use of this method in routine practical healthcare practice, but it remains the only method for phenotypic determination of the sensitivity of strains to antibacterial drugs. The monitoring of the increasing resistance of these infections, the correction of eradication therapy regimens considering the sensitivity of isolated strains to chemotherapeutic drugs, and the testing of new drugs make this bacteriological method indispensable for practical and scientific purposes. An obligatory component of the medium should be an additive of 5–10% of the blood or serum of animals (horse, sheep) [ 18 ]. Therefore, a component for media has been proposed for conducting bacteriological studies, allowing us to obtain media enriched with micro- and macronutrients for reliable and high-quality laboratory analyses. The use of the proposed component obtained from defibrinated horse blood in the cultivation of microorganisms will contribute to the identification, differentiation and determination of the hemolytic activity of microorganisms that cause diseases in humans and animals. This work also has the goal of authenticating the growth of E. coli and Salmonella strains via phylogenetic analysis. For this purpose, we studied the 16S RNA of the bacterium. Ribosomal RNA 16S is a component of the 30S subunit of prokaryotic ribosomes and has high conservation, which makes it a good candidate for studying evolutionary relationships [ 19 ]. The studied E. coli gene, "Isolate 11", is part of a phylogenetic analysis showing its genetic relationship with other bacteria of the genus Escherichia with a difference of 0.5% in the nucleotide sequence of the 16S rRNA gene. Its location on the tree suggests that it has special genetic characteristics that distinguish it from other species and strains of Escherichia. The studied Salmonella gene, designated "7", is included in the exclusive group of Salmonella enterica , which is formed from one exceptional ancestral line. The "7" sample corresponds to a distinctive sequence or variant of Salmonella enterica , and this discovery may be crucial for investigating its virulence, drug resistance, or genetic development over time. This phylogenetic analysis is crucial for epidemiological tracking, understanding bacterial evolution, and the potential development of targeted therapies or interventions [ 20 ]. In the context of phylogenetic trees, it is important to consider that exact interpretation depends on the methods used to construct the tree, such as the specific genetic markers being analysed and the algorithms used in the analysis. Moreover, although bootstrap values provide statistical support, they do not guarantee that the tree perfectly reflects the true evolutionary history due to potential genetic recombination, horizontal gene transfer, and other factors that may complicate bacterial phylogenetics [ 21 ]. Therefore, this work analysed the sensitivity of Enterobacteriaceae isolates isolated from animal products to a wide range of antibiotics used in practice. The obtained results suggest that regions with low resistance rates of E. coli to cefotaxime, ampicillin/sulbactam, levofloxacin, and meropenem and Salmonella to levofloxacin and high resistance rates of E. coli to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin, ciprofloxacin. These facts support the rational use of antibacterial agents, and ssugges that despite all efforts, there is a lack of knowledge in the medical and veterinary communities regarding the potentially dangerous aspects of antibiotic therapy. Conclusions Pathogens of enteropathogenic diseases that are resistant to antibacterial drugs are circulating in the Almaty region of Kazakhstan. Using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro- and macronutrients for reliable and high-quality laboratory analyses. According to the data obtained, the low resistance rates of E.coli to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of E. coli to norfloxacin and ciprofloxacin, gentamicin, and Salmonella to norfloxacin and ciprofloxacin were detected in the Almaty region. Circulating pathogens of the enteropathogenic diseases E. coli and Salmonella in the Almaty region, Kazakhstan have been confirmed by phylogenetic analysis to be genuine pathogens with special genetic characteristics that distinguish them from other species and strains. Such phylogenetic analysis is crucial for epidemiological tracking, understanding bacterial evolution, and the potential development of targeted therapies or interventions. Abbreviations The Global Antimicrobial Resistance and Use Surveillance System (GLASS) World Health Organization (WHO) The Bacterial Priority Pathogens List (BPPL) Antimicrobial resistance (AMR) The European Committee on Antimicrobial Susceptibility Testing (EUCAST) The Basic Local Alignment Search Tool (BLAST) Declarations Author Contributions: Conceptualization, Z.L. and A.N. contributed to the conceptualization, supervision, editing, and project administration. Z.L., A.N., Zh.A. M.N., A.B., and M.K. contributed to the writing of the original draft, writing, review, and editing. Sh.Y., K.R. and S.Sh.; methodology, Z.L., S.Sh. and Zh.A.; validation, A.M., V.S., M.Y., M.N., M.S., A.B. and M.K.; formal analysis, Z.L., S.Sh., M.N., and A.N.; investigation, Z.L., S.Sh., Sh.E., G.K., Zh.T., O.N., K.R., N.I., A.M., V.S., M.N., M.S., Zh.A. and M.K.; resources, A.M., V.S., M.Y., M.N., M.S., B.A. All authors have read and agreed to the published version of the manuscript. Funding: This work was supported by the Ministry of Healthcare of the Republic of Kazakhstan, within the framework of the PTF IRN BR218004/0223 "Improving biosafety measures in Kazakhstan: countering dangerous and especially dangerous infections" for 2023-2025, through the program of the Applied Epidemiology Field Epidemiology Training Program (FETP), Centers for Disease Control and Prevention in the Central Asia Region (CDC/CAR). Availability of data and materials: All the data analysed or generated during this study are included in this published article and its Supporting Information files. Any additional information is available from the corresponding author upon reasonable request. Conflicts of Interest: The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. References Havelaar AH, Kirk MD, Torgerson PR, Gibb HJ, Hald T, Lake RJ, Praet N, Bellinger DC, de Silva NR, Gargouri N, Speybroeck N, Cawthorne A, Mathers C, Stein C, Angulo FJ, Devleesschauwer B. World Health Organization Global Estimates and Regional Comparisons of the Burden of Foodborne Disease in 2010. World Health Organization Foodborne Disease Burden Epidemiology Reference Group. PLoS Med. 2015;12(12):e1001923. 10.1371/journal.pmed.1001923. eCollection . Hariharan P, Bharani T, Franklyne JS, Biswas P, Solanki SS, Paul-Satyaseela M. Antibiotic susceptibility pattern of Enterobacteriaceae and non fermenter gram-negative clinical isolates of microbial resource orchid. J Nat Sci Biol Med. 2015 Jan-Jun;6(1):198–201. 10.4103/0976-9668.149121 . PMID: 25810660; PMCID: PMC4367035. English BK, Gaur AH. The use and abuse of antibiotics and the development of antibiotic resistance. Adv Exp Med Biol. 2010;659:73–82. Puvača N. Antimicrobial Resistance and Treatment in Companion, Food and Exotic Animals. Antibiot (Basel). 2022;11(10):1360. 10.3390/antibiotics11101360 . PMID: 36290017; PMCID: PMC9598238. WHO Bacterial Priority Pathogens List. 2024: bacterial pathogens of public health importance to guide research, development and strategies to prevent and control antimicrobial resistance. World Health Organization, 2024. European Committee on Antimicrobial Susceptibility Testing (EUCAST.) Disco-diffuse EUCAST method. - Version 10.0, 2020. http://www.eucast.org . Mueller JH, Hinton J. (1941). A protein-free medium for primary isolation of the gonococcus and meningococcus, Proc. Soc. Exper. Biol. Med. 48: 330. The National Center for Biotechnology Information. NCBI databases htpp://www.ncbi.nlm.nih.gov . Aslam B, Wang W, Arshad MI, Khurshid M, Muzammil S, Rasool MH, Nisar MA, Alvi RF, Aslam MA, Qamar MU, Salamat MKF, Baloch Z. Antibiotic resistance: a rundown of a global crisis. Infect Drug Resist. 2018;11:1645–58. PMID: 30349322; PMCID: PMC6188119. Dhingra S, Rahman NAA, Peile E, Rahman M, Sartelli M, Hassali MA, Islam T, Islam S, Haque M. Microbial Resistance Movements: An Overview of Global Public Health Threats Posed by Antimicrobial Resistance, and How Best to Counter. Front Public Health. 2020;8:535668. 10.3389/fpubh.2020.535668 . PMID: 33251170; PMCID: PMC7672122. Nathan C, Cars O. Antibiotic resistance-problems, progress, and prospects. N Engl J Med. 2014;371(19):1761–3. Robert C. Hospital infection: a historical review. Br Med Bull. 1944;2:272–6. 10.1093/oxfordjournals.bmb.a071061 . Smith PW, Watkins K, Hewlett A. Infection control through the ages. Am J infect control. 2012;40:35–42. 10.1016/j.ajic.2011.02.019 . Antimicrobial Resistance Collaborators. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022;399(10325):629–655. doi: 10.1016/S0140-6736(21)02724-0. Epub 2022 Jan 19. Erratum in: Lancet. 2022;400(10358):1102. 10.1016/S0140-6736(21)02653-2 . Bao L, Peng R, Ren X, Ma R, Li J, Wang Y. Analysis of some common pathogens and their drug resistance to antibiotics. Pak J Med Sci. 2013;29(1):135–9. 10.12669/pjms.291.2744 . Kebede HK, Gesesew HA, Woldehaimanot TE, Goro KK. Antimicrobial use in paediatric patients in a teaching hospital in Ethiopia. PLoS ONE. 2017;12(3):e0173290. 10.1371/journal. pone.0173290 . Kowalska-Krochmal B, Dudek-Wicher R. The Minimum Inhibitory Concentration of Antibiotics: Methods, Interpretation, Clinical Relevance. Pathogens. 2021;10(2):165. 10.3390/pathogens10020165 . Gajic I, Kabic J, Kekic D, Jovicevic M, Milenkovic M, Mitic Culafic D, Trudic A, Ranin L, Opavski N. Antimicrobial Susceptibility Testing: A Comprehensive Review of Currently Used Methods. Antibiot (Basel). 2022;11(4):427. 10.3390/antibiotics11040427 . Grinevich D, Harden L, Thakur S, Callahan B. Serovar-level identification of bacterial foodborne pathogens from full-length 16S rRNA gene sequencing. mSystems. 2024;9(3):e0075723. Baquero F, Martínez JL, Lanza F, Rodríguez-Beltrán V, Galán J, San Millán JC, Cantón A, Coque R. Evolutionary Pathways and Trajectories in Antibiotic Resistance. Clin Microbiol Rev. 2021;34(4):e0005019. Didelot X. Phylogenetic Analysis of Bacterial Pathogen Genomes. Methods Mol Biol. 2023;2674:87–99. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4575952","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":317144573,"identity":"b3d97c05-fbcb-44f0-a14a-fcfd7d09d9da","order_by":0,"name":"Z. A. Latypova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"Z.","middleName":"A.","lastName":"Latypova","suffix":""},{"id":317144574,"identity":"9626d896-8b62-4b12-94b4-1715cbddb400","order_by":1,"name":"A. S. Nurpeisova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"A.","middleName":"S.","lastName":"Nurpeisova","suffix":""},{"id":317144575,"identity":"d2e573c0-71b9-4708-bbe7-aeac9731fce6","order_by":2,"name":"M. T. Nurgalieva","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAt0lEQVRIiWNgGAWjYBACCTiLvYeZFC0JDAw8PGdI1iKRQ6QWyfbjzx78/GGTZy/59rAB457DhLVI8ySkG/YkpBXzSOclJzA8I0KLHEPCMQmehMOJPdI5xgcYDhCjhf9hm+SfhP+JPZJniNQiLZHMBnTcgcQeCR7jBKK0SM54xm4sk5ac2HMmL9kg4UA6YS0S59OfPXxjY5fY3n72sMSHA9aEtQABG4KZQJQGFC2jYBSMglEwCrABABYXNy1wJ1wcAAAAAElFTkSuQmCC","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":true,"prefix":"","firstName":"M.","middleName":"T.","lastName":"Nurgalieva","suffix":""},{"id":317144576,"identity":"f6b41c0a-6615-40a0-9928-211e6fb2b2c4","order_by":3,"name":"A. B. Bizhanov","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"A.","middleName":"B.","lastName":"Bizhanov","suffix":""},{"id":317144578,"identity":"7a011065-70c6-4938-b1bb-ec7f10242f2b","order_by":4,"name":"M. R. Yussupov","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"M.","middleName":"R.","lastName":"Yussupov","suffix":""},{"id":317144580,"identity":"30440040-0e10-416f-aa0a-942e7a8f2115","order_by":5,"name":"Zh. S. Abay","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"Zh.","middleName":"S.","lastName":"Abay","suffix":""},{"id":317144581,"identity":"69e0147e-e283-43ea-a61a-0a633f65110c","order_by":6,"name":"S. T. Sarbakanova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"T.","lastName":"Sarbakanova","suffix":""},{"id":317144582,"identity":"2a207c5c-696f-4195-99f3-25f64dc0d293","order_by":7,"name":"E. B. Shakibaev","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"E.","middleName":"B.","lastName":"Shakibaev","suffix":""},{"id":317144583,"identity":"3320a1af-3386-4bb6-bcd5-c94338ca0d50","order_by":8,"name":"R. A. Kerimbaeva","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"R.","middleName":"A.","lastName":"Kerimbaeva","suffix":""},{"id":317144584,"identity":"91fc9821-0a47-4beb-b882-789eb9027629","order_by":9,"name":"G. M. Kobdikova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"G.","middleName":"M.","lastName":"Kobdikova","suffix":""},{"id":317144585,"identity":"aff264dd-60f9-4d47-9184-8228b87a86c2","order_by":10,"name":"Zh. S. Tursynova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"Zh.","middleName":"S.","lastName":"Tursynova","suffix":""},{"id":317144587,"identity":"777d3f9c-e4b1-4815-8a60-59ce25126b26","order_by":11,"name":"O. O. Nurlybayev","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"O.","middleName":"O.","lastName":"Nurlybayev","suffix":""},{"id":317144589,"identity":"1a5a9e91-d646-44bc-abc3-a129c00988ff","order_by":12,"name":"N. K. Ibrayim","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"N.","middleName":"K.","lastName":"Ibrayim","suffix":""},{"id":317144591,"identity":"71d71ee0-93f0-4d0d-89a5-498836b5bf1d","order_by":13,"name":"A. K. Mussayeva","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"A.","middleName":"K.","lastName":"Mussayeva","suffix":""},{"id":317144593,"identity":"14596eaf-d24c-40d5-8507-f5aca4021380","order_by":14,"name":"H. S. Yoo","email":"","orcid":"","institution":"Seoul National University","correspondingAuthor":false,"prefix":"","firstName":"H.","middleName":"S.","lastName":"Yoo","suffix":""},{"id":317144594,"identity":"d40edd3c-7b95-42ab-983d-7c16a8bb442c","order_by":15,"name":"S. B. Mamanova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"B.","lastName":"Mamanova","suffix":""},{"id":317144595,"identity":"fc48d23a-4d55-4b26-a999-ce88a7fd31d3","order_by":16,"name":"M. S. Serikov","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"M.","middleName":"S.","lastName":"Serikov","suffix":""},{"id":317144596,"identity":"6d5dade2-66ef-40fc-a844-8cee97ca4764","order_by":17,"name":"V. Suchshikh","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"V.","middleName":"","lastName":"Suchshikh","suffix":""},{"id":317144597,"identity":"371d2846-10e9-42a1-8db8-fa1bb6f04f1b","order_by":18,"name":"Zh. A. Abdraimova","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"Zh.","middleName":"A.","lastName":"Abdraimova","suffix":""},{"id":317144598,"identity":"f345bea0-d6e7-4115-beb2-1b91051f5a3f","order_by":19,"name":"M. M. Kassenov","email":"","orcid":"","institution":"Kazakh Scientific Research Veterinary Institute","correspondingAuthor":false,"prefix":"","firstName":"M.","middleName":"M.","lastName":"Kassenov","suffix":""}],"badges":[],"createdAt":"2024-06-13 11:43:00","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4575952/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4575952/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":59949447,"identity":"ca1f79d3-da19-4b34-8528-0306ba90c511","added_by":"auto","created_at":"2024-07-09 17:05:25","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":94769,"visible":true,"origin":"","legend":"\u003cp\u003eResistance and sensitivity of Salmonella isolates\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/6830b16f6bfa64e016f7b9f6.jpg"},{"id":59949805,"identity":"9140c4f0-38ee-404e-a8c2-4a4526c63cdd","added_by":"auto","created_at":"2024-07-09 17:13:25","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":91735,"visible":true,"origin":"","legend":"\u003cp\u003eResistance and sensitivity of the \u003cem\u003eE. coli\u003c/em\u003e isolates\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/9f1a8944f41aa3d9f10571f1.jpg"},{"id":59949449,"identity":"a6fa51d1-80b4-484d-b47a-70a34257ec13","added_by":"auto","created_at":"2024-07-09 17:05:25","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":71096,"visible":true,"origin":"","legend":"\u003cp\u003eResistance of isolated isolates of \u003cem\u003eE. coli\u003c/em\u003e and Salmonella to various antibiotics, %\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/a4c47ff54a934fa38951562a.jpg"},{"id":59949451,"identity":"e18a1079-b9f0-4a30-8dde-cff34afb9748","added_by":"auto","created_at":"2024-07-09 17:05:26","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":102356,"visible":true,"origin":"","legend":"\u003cp\u003eA phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the \u003cem\u003eS. enterica\u003c/em\u003esample with reference sequences from the NCBI database. The \u003cem\u003eS. enterica\u003c/em\u003e sample is listed as having the isolate 7 gene for 16S rRNA\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/e83002f82eb9b68cc45a5fe8.jpg"},{"id":59949448,"identity":"f666b4f2-0462-4a87-a324-51c472fc0142","added_by":"auto","created_at":"2024-07-09 17:05:25","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":93964,"visible":true,"origin":"","legend":"\u003cp\u003eA phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the \u003cem\u003eE. coli\u003c/em\u003e sample with reference sequences from the NCBI database. The \u003cem\u003eE. coli\u003c/em\u003e sample used for the 16S rRNA gene is listed as isolate 11 gene\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/495aa89fc77d7f8fbdde5b0d.jpg"},{"id":70136446,"identity":"4c050e06-3c52-42d6-98c5-fa0aa5e9130b","added_by":"auto","created_at":"2024-11-28 17:16:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":929479,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4575952/v1/78e96850-67de-4625-956c-cd923e558676.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eAnalyzing the Sensitivity and Antibiotic Resistance of Enterobacteriaceae Isolates Isolated from Animal Products in Almaty Region, Kazakhstan\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDiseases and deaths from various infections caused by contaminated food pose a constant threat to public health and a serious obstacle to socioeconomic development worldwide.\u003c/p\u003e \u003cp\u003eThe Global Antimicrobial Resistance and Use Surveillance System (GLASS), which is based on the World Health Organization (WHO) has identified widespread antibiotic resistance. According to the presented data, 70% of known bacteria have developed resistance to one or more antibiotics. The greatest amount of data on antimicrobial resistance has been obtained for the following bacterial species: \u003cem\u003eEscherichia coli, Klebsiella pneumoniae, Staphylococcus aureus Streptococcus pneumoniae\u003c/em\u003e and \u003cem\u003eSalmonella spp\u003c/em\u003e. [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOf particular for importance of such infectious diseases in medicine, veterinary medicine and agriculture determine the increased resistance of pathogens circulating in the environment to antimicrobial agents [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eАntimicrobial agents, including antibiotics and antiviral, antifungal and antiparasitic drugs, are medicines used to prevent and treat infections in humans, animals and plants [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn May of 2024, the WHO updated the Bacterial Priority Pathogens List (BPPL) 2024, which includes 15 families of antibiotic-resistant bacteria grouped into critical, high and medium categories for prioritization. Four new pathogen\u0026ndash;antibiotic combinations were added to BPPL 2024. The updated list contains new data. А separate item in the high-priority group included third-generation cephalosporin-resistant bacteria of the order Enterobacterales, which requires increased attention and special measures.\u003c/p\u003e \u003cp\u003eBacteria of the genus Salmonella are classified as pathogens of a high-priority group. BPPL 2024 contains recommendations for the development of new and necessary treatment methods designed to stop the spread of antimicrobial resistance (AMR) [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn current clinical veterinary practice, the rational use of antibacterial drugs is important. It is known that inappropriate antibiotic prescribing in animal therapy inevitably leads to an increase in the level of tolerance to these drugs in circulating strains of pathogenic and conditionally pathogenic microorganisms.\u003c/p\u003e \u003cp\u003eTo curb the increase in inantibiotic resistance of pathogens of infectious diseases of farm animals and birds, it is necessary to take an evidence-based approach for the use of antibiotic, to reduce their unnecessary and mass use [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe aim of this study was to assess the sensitivity or antibiotic resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood.\u003c/p\u003e"},{"header":"Material/Methods","content":"\u003cp\u003eThe material for study consisted of cultures of \u003cem\u003eE.сoli\u003c/em\u003e and Salmonella\u003cem\u003e\u0026nbsp;\u003c/em\u003e(n=186) isolated from the territory of the Almaty region of the Republic of Kazakhstan.\u003c/p\u003e\n\u003cp\u003eThe sources of the microorganism isolates were 5 types of animal product, namely, beef (n=123), lamb (n=119), pork (n=122), and poultry (n=114), and poultry eggs (n=122), which were purchased by random sampling in large retail chains, individual food and retail stores shops, supermarkets and markets in the Almaty region.\u003c/p\u003e\n\u003cp\u003eAll the studies were carried out in the laboratories of the Kazakh Research Veterinary Institute LLP.\u003c/p\u003e\n\u003cp\u003eThe identification of microorganisms from animal products was carried out according to methods generally accepted in veterinary laboratory practice.\u003c/p\u003e\n\u003cp\u003eIn accordance with the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST), sensitivity assessment and interpretation of the results of studies on the sensitivity of \u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003eand Salmonella to antibiotics were carried out using the disc diffusion method of Mueller-Hinton Agar (Oxoid, UK) with and without the addition of 5% defibrinated horse blood. Version 10.0, 2020. http://www.eucast.org [6,7].\u003c/p\u003e\n\u003cp\u003eDuring the sensitivity determination, commercial antibiotic discs of certain types were used: cefotaxime 30 mcg, ampicillin/sulbactam 20 mcg, norfloxacin 10 mcg, pefloxacin 5 mcg, ceftriaxone 30 mcg, tigecycline 15 mcg, meropenem 10 mcg, levofloxacin 5mcg, trimethoprim 5 mcg, gentamicin 10 mcg, chloramphenicol 30 mcg, amoksiklav 30 mcg.\u003c/p\u003e\n\u003cp\u003eCultures of \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922 and ATCC 35218 were used for sensitivity determination.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMolecular genetics testing\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eDNA isolation\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eGenomic DNA was isolated from daily bacterial cultures using a PureLink Genomic DNA Kit, - according to the manufacturer\u0026rsquo;s protocol (Invitrogen, Carlsbad, USA). The DNA concentration in the samples was determined on a Qubit\u0026reg; 2.0 fluorimeter using a QubitTM dsDNA HS Assay Kit (Life Technologies, Oregon, USA).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAmplification of 16S rRNA\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe 16S rRNA gene was used as a genetic marker. To amplify the 16S rRNA\u0026nbsp;gene, 25 \u0026micro;l of\u0026nbsp;the following reaction mixture was prepared: 12.5 \u0026micro;l of Q5\u0026reg; Hot Start High-Fidelity 2X Master Mix (New England Biolabs Ins., USA); a pair of universal primers; 8F (5\u0026apos;-AGAGTTTGATCCTGGCTCAG-3\u0026apos;) and 806R (5\u0026apos;-GGACTACCAGGGTATCTAAT-3\u0026apos;) (1.2 \u0026micro;l in 10 \u0026micro;M concentration); and DNA and water matrix (up to 25 \u0026micro;l) [5]. The amplification mode consisted of the following cycles: 95\u0026deg;C \u0026nbsp;for 5 min, 95\u0026deg;C for 30 seconds, 55\u0026deg;C for 40 seconds, 72\u0026deg;C for 50 seconds for 30 cycles; and elongation at 72\u0026deg;C for 10 min.\u003c/p\u003e\n\u003cp\u003eThe PCR products were purified using CleanSweep\u0026trade; PCR Purification (Life Technologies, Carlsbad, CA).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e16S rRNA sequencing.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eSequencing of the 16S rRNA gene of bacteria was performed using the Big Dye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, USA) according to the manufacturer\u0026rsquo;s protocol (BigDye\u0026reg; Terminator v3.1 Cycle Sequencing Kit Protocol Applied Biosystems, USA), followed by fragment separation on an automatic genetic analyser 3500 DNA Genetic Analyser (Applied Biosystems, Hitachi, Tokyo Japan). The sequencing results were processed in the Seq program (Applied Biosystems).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe search for homologous nucleotide sequences of 16S rRNA genes was carried out using the Basic Local Alignment Search Tool (BLAST) in the International GenBank database of the National Center for Biotechnology Information of the USA (htpp://www.ncbi.nlm. nih.gov) [8]. Phylogenetic analysis was performed using of the Molecular Evolutionary Genetics Analysis (MEGA6) software and nucleotide sequence alignment was performed using the ClustalW algorithm. Phylogenetic identification was carried out using the neighbor joining algorithm BLASTN (NJ) program.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eStatistical processing\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eStatistical processing of the research results was carried out using standard methods of descriptive statistics via the Python program.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eAccording to the research, 111 \u003cem\u003eE. coli\u003c/em\u003e isolates (18.3%) and 75 Salmonella isolates (12.5%) were isolated, including 22.52% (n\u0026thinsp;=\u0026thinsp;25) of the \u003cem\u003eE. coli\u003c/em\u003e isolates from poultry, 24.32% (n\u0026thinsp;=\u0026thinsp;27) from lamb, 9.9% (n\u0026thinsp;=\u0026thinsp;11) from beref, 34.23% (n\u0026thinsp;=\u0026thinsp;38) from pork and 9% (n\u0026thinsp;=\u0026thinsp;10) from poultry eggs.\u003c/p\u003e \u003cp\u003eSalmonella isolates were isolated from poultry \u0026ndash; (37.3%, (n\u0026thinsp;=\u0026thinsp;28), lamb (14.66%, (n\u0026thinsp;=\u0026thinsp;11), beef (14.66%, (n\u0026thinsp;=\u0026thinsp;11), pork (14.66%, (n\u0026thinsp;=\u0026thinsp;11) and poultry eggs (18.66%, (n\u0026thinsp;=\u0026thinsp;14). Fourteen isolates lost their viability during storage.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the isolates were highly sensitive to most antibiotics, such as cefotaxime, ampicillin/sulbactam, ceftriaxone, tigecycline, meropenem, trimethoprim, gentamicin, chloramphenicol and amoksiklav, with 100% sensitivity and 0% resistance.\u003c/p\u003e \u003cp\u003eMoreover, 15.15% and 25.76% of the isolates were resistant to norfloxacin and pefloxacin, respectively. Levofloxacin showed a small percentage of resistance (7.58%) and high sensitivity (92.4%).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the resistance and sensitivity of 106 \u003cem\u003eE. coli\u003c/em\u003e isolates to various antibiotics. High sensitivity to most antibiotics, such as cefotaxime, ampicillin/sulbactam, ceftriaxone, tigecycline, meropenem, trimethoprim, chloramphenicol and amoksiklav was detected, with high sensitivity (more than 97%) and low resistance.\u003c/p\u003e \u003cp\u003eModerate levels of resistance were detected for gentamicin and pefloxacin (16.04% and 15.09%), respectively. There was a greater percentage of resistant to norfloxacin (29.25%) than to other antibiotics.\u003c/p\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, for \u003cem\u003eE. coli\u003c/em\u003e, the most effective antibiotics are trimethoprim, ceftriaxone, and tigecycline. Some antibiotics, such as cefotaxime, ampicillin/sulbactam and levofloxacin, are less effective.\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the most effective antibiotics for Salmonella infection: trimethoprim, gentamicin, chloramphenicol and amoksiklav.\u003c/p\u003e \u003cp\u003eThese data show that trimethoprim, ceftriaxone and tigecycline are the most effective antibiotics for both types of bacteria, while some antibiotics have limited efficacy.\u003c/p\u003e \u003cp\u003eThe average values of zones of growth suppression show how effective each antibiotic is against these \u003cem\u003eE. coli\u003c/em\u003e and Salmonella test samples. For example, meropenem shows high efficacy against \u003cem\u003eE. coli\u003c/em\u003e with the highest average value zones of suppression (33.78 mm) and a low standard deviation (1.97 mm), which indicates the stability of its action.\u003c/p\u003e \u003cp\u003eFor Salmonella, pefloxacin also showed high efficacy, with an average value of 25.5 mm, but the standard deviation was greater (5.74 mm), indicating greater variability of action between samples.\u003c/p\u003e \u003cp\u003eCompared with meropenem and pefloxacin, ampicillin/sulbactam showed lower efficacy, especially against \u003cem\u003eE. coli\u003c/em\u003e, with lower mean zones of suppression and greater standard deviations, indicating a less predictable effect.\u003c/p\u003e \u003cp\u003eThese data can help in choosing the most effective antibiotic for the treatment of infections caused by \u003cem\u003eE. coli\u003c/em\u003e and Salmonella, taking into account the average zones of suppression and the variability of drug action.\u003c/p\u003e \u003cp\u003eThus, monitoring studies of animal products have allowed us to conclude that poultry, lamb and pork samples are the most contaminated with \u003cem\u003eE. coli\u003c/em\u003e \u0026minus;\u0026thinsp;22.52%, 24.32% and 34.23%, respectively.\u003c/p\u003e \u003cp\u003eThe results of research products to identify Salmonella showed that these pathogens were found quite often in poultry and poultry eggs, at 37.3% and 18.66%, respectively.\u003c/p\u003e \u003cp\u003eIn total, 186 isolates were isolated, including: \u003cem\u003eE. coli\u003c/em\u003e \u0026minus;\u0026thinsp;111, and Salmonella \u0026minus;\u0026thinsp;75, which were used for further research.\u003c/p\u003e \u003cp\u003eSensitivity to 12 antibacterial drugs was studied in 172 isolated isolates by the disc diffusion method.\u003c/p\u003e \u003cp\u003eAll isolates isolated from animal products showed varying degrees of sensitivity and resistance to antibiotics. The interpretation of the results in determining sensitivity was carried out using the recommendations of EUCAST (v 6.0) [29]. The results of the research are shown in Fig.\u0026nbsp;5.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResistance of \u003cem\u003eE. coli\u003c/em\u003e and Salmonella to various antibiotics\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eName of the antibiotic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e resistance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSalmonella resistance\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefotaxime 30 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2,83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmpicillin/Sulbactam 20 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2,83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNorfloxacin 10 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e29,25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15,15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePefloxacin 5 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15,09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25,76\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCeftriaxone 30 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTigecycline 15 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMeropenem 10 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0,94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLevofloxacin 5mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2,83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e7,58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrimethoprim 5 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGentamicin 10 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16,04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChloramphenicol 30 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmoksiklav 30 mcg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAs shown in from Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, cefotaxime and ampicillin/sulbactam had the same resistance in \u003cem\u003eE. coli\u003c/em\u003e (2.83%), and no resistance was detected in Salmonella. Norfloxacin shows high-level resistance in \u003cem\u003eE. coli\u003c/em\u003e (29.25%) and moderate resistance in Salmonella (15.15%). Pefloxacin had significant resistance in both bacteria: 15.09% for \u003cem\u003eE. coli\u003c/em\u003e and 25.76% for Salmonella. Meropenem resistance was low shows in \u003cem\u003eE. coli\u003c/em\u003e (0.94%), and no resistance was detected in Salmonella. Levofloxacin had the same resistance in \u003cem\u003eE. coli\u003c/em\u003e (2.83%) cefotaxime, ampicillin/sulbactam and moderate resistance in Salmonella (7.58%). Gentamicin showed resistance in \u003cem\u003eE. coli\u003c/em\u003e (16.04%) and zero resistance in Salmonella.\u003c/p\u003e \u003cp\u003eThe isolates were found to be resistant to at least one of the tested antibacterial drugs. Some of the studied bacteria were resistant to several antibiotics in different groups and exhibited polyresistance.\u003c/p\u003e \u003cp\u003eThus, 74 (43%) \u003cem\u003eE. coli\u003c/em\u003e isolates were resistant to fluoroquinolones (first, second and third generation), β-lactam antibiotics (first, second generation) and aminoglycosides.\u003c/p\u003e \u003cp\u003eAccording to the findings, the frequency of resistance to cefotaxime in \u003cem\u003eE. coli\u003c/em\u003e was 2.83% (n\u0026thinsp;=\u0026thinsp;3), that to ampicillin/sulbactam was 2.83% (n\u0026thinsp;=\u0026thinsp;3), that to norfloxacin was 29.25% (n\u0026thinsp;=\u0026thinsp;31), that to pefloxacin was 15.09% (n\u0026thinsp;=\u0026thinsp;16), that to meropenem was 0.94% (n\u0026thinsp;=\u0026thinsp;1), that to levofloxacin was 2.83% (n\u0026thinsp;=\u0026thinsp;3), and that to gentamicin was 16.04% (n\u0026thinsp;=\u0026thinsp;17).\u003c/p\u003e \u003cp\u003eAmong the isolates of the genus Salmonella, 32 (18%) were resistant to only the antibacterial drugs fluoroquinolones (first, second and third generation). Thus, resistance to norfloxacin was 15.15% (n\u0026thinsp;=\u0026thinsp;10), resistance to pefloxacin was 25.76% (n\u0026thinsp;=\u0026thinsp;17), and resistance to levofloxacin was 7.58% (n\u0026thinsp;=\u0026thinsp;5). The tested isolates of the genus Salmonella showed sensitivity to the antibiotics β-lactam antibiotics and aminoglycosides. Isolates of \u003cem\u003eE. coli\u003c/em\u003e and Salmonella resistant to ceftriaxone, tigecycline, chloramphenicol and amoksiklav were not isolated.\u003c/p\u003e \u003cp\u003eOverall, it was experimentally determined that the highest percentage of resistance in isolated \u003cem\u003eE. coli\u003c/em\u003e and Salmonella isolates was found for norfloxacin (29.25% and 15.15%, respectively) and pefloxacin (15.09% and 25.76%, respectively), and the lowest percentage was found for meropenem (0.94% and 0.00%, respectively).\u003c/p\u003e \u003cp\u003eNotably, of the 172 Enterobacteriaceae isolates studied, 66 (38%) showed sensitivity to all groups of tested antibiotics.\u003c/p\u003e \u003cp\u003eIn the process of identifying antibiotic-resistant and antibiotic-sensitive Enterobacteriaceae, a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood was tested. The detection of hemolytic activity suggested that all the studied cultures had the necessary phenotypic properties.\u003c/p\u003e \u003cp\u003eDuring cultivation on a dense medium of the \u003cem\u003eEnterococcus faecalis\u003c/em\u003e strain, the formation of small, rounded, convex, shiny colonies ranging in size from 0.5 mm to 1.0 mm, without hemolysis zones was noted.\u003c/p\u003e \u003cp\u003eThe growth of \u003cem\u003eStaphylococcus aureus\u003c/em\u003e was characterized by the presence of colonies ranging in size from 0.5 mm to 1.5 mm. Hemolysis zones were established around each colony, the size of which did not exceed 2.0 mm.\u003c/p\u003e \u003cp\u003eDuring cultivation of the \u003cem\u003eStreptococcus pyogenes\u003c/em\u003e strain on agar, the growth of gray colonies with a size of 1.0 mm and a clearly defined hemolysis zone reaching 3.0\u0026ndash;4.0 mm was noted.\u003c/p\u003e \u003cp\u003eIt should be noted that during the incubation of microorganisms on plain agar with the addition of blood, the indicated sizes of the hemolysis zone were noted after 24 hours of incubation, and after 72 hours of growth, a significant increase in their sizes was observed.\u003c/p\u003e \u003cp\u003eIt was also found that during cultivation of \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and \u003cem\u003eStreptococcus pyogenes\u003c/em\u003e on a medium containing the proposed component from defibrated horse blood, the enlightening zones were clearer and more extensive, and more pronounced lysis was observed.\u003c/p\u003e \u003cp\u003eTo confirm the species identity of \u003cem\u003eEnterobacteriaceae\u003c/em\u003e isolated from meat, 2 isolates of Salmonella and \u003cem\u003eE. coli\u003c/em\u003e were sequenced, and the results are shown in Figs.\u0026nbsp;4 and 5.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure 4. A phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the \u003cem\u003eS. enterica\u003c/em\u003e sample with reference sequences from the NCBI database. The \u003cem\u003eS. enterica\u003c/em\u003e sample is listed as having the isolate 7 gene for 16S rRNA\u003c/p\u003e \u003cp\u003eFigure 4 provides a phylogenetic tree representing the evolutionary relationships between different strains \u003cem\u003eof S. enterica\u003c/em\u003e (7) based on similarities and differences in their genetic characteristics.\u003c/p\u003e \u003cp\u003eSample \"7\" is located at the top of the tree and branches closest to the group consisting of various subspecies of \u003cem\u003eSalmonella enterica\u003c/em\u003e. This proximity suggests that they are closely related to these bacteria, and \"7\" probably refers to a specific strain or sequence within this group. The tree shows that the \u003cem\u003eSalmonella enterica\u003c/em\u003e group forms a monophyletic clade, meaning that all strains in this group have one common ancestor that has nothing in common with any strains outside this group. Sample \"7\" appears to be part of this clade.\u003c/p\u003e \u003cp\u003eFrom the figure, it can be concluded that sample \"7\" represents a unique sequence or strain of \u003cem\u003eSalmonella enterica\u003c/em\u003e, which may be an important discovery for studies of pathogenicity, antibiotic resistance or the evolutionary biology of this species. The degree of homology with the nearest strain NR 104709.1:40\u0026ndash;767 \u003cem\u003eSalmonella enterica subsp. enterica strain\u003c/em\u003e LT2 was 100.00%.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure 5. A phylogenetic tree was constructed by comparing the nucleotide sequence of the 16S rRNA gene of the \u003cem\u003eE. coli\u003c/em\u003e sample with reference sequences from the NCBI database. The \u003cem\u003eE. coli\u003c/em\u003e sample used for the 16S rRNA gene is listed as isolate 11 gene\u003c/p\u003e \u003cp\u003eFigure 5 provides a phylogenetic tree representing the evolutionary relationships between different strains of \u003cem\u003eE.coli\u003c/em\u003e (11) based on similarities and differences in their genetic characteristics.\u003c/p\u003e \u003cp\u003eThe position on the tree \"Isolate 11 gene for 16S RNA\" is located on a separate branch, which makes it relatively close to the \"Leclercia adecarboxylata gene for 16S RNA partial sequence strain: NBRC 102595\".\u003c/p\u003e \u003cp\u003eThe proximity on the tree indicates that these two strains may have a relatively recent common ancestor compared to other represented species and strains. The scale band represents a difference of 0.5% in the nucleotide sequence of the 16S rRNA gene, which means that a line of this length represents a sequence divergence of 0.5% from the branching point.\u003c/p\u003e \u003cp\u003eThe shorter the branch leading to isolate 11, the more genetically similar it is to the sequence at the point where it branches off from Leclercia adecarboxylata. The topology of the tree indicates the nature of the evolutionary divergence. Isolate 11 was separated from the cluster, which included several strains of Escherichia and another strain of Leclercia. This suggests that Isolate 11 is genetically sufficiently distinct to form its own line within this group.\u003c/p\u003e \u003cp\u003eThe tree also demonstrated a wide range of genetic diversity among various species and strains of Escherichia. For example, has been shown that \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eE. fergusonii\u003c/em\u003e are genetically different from the Isolate 11 sample.\u003c/p\u003e \u003cp\u003eThus, Isolate 11 16S is 100% homologous to other bacteria of the genus Escherichia and has special genetic characteristics that distinguish it from other species and strains of Escherichia.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe need to develop new ways to solve the problems of high levels of resistance of bacteria, viruses, fungi, and parasites and progressive drug resistance is a global task of modern medicine [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe problem of resistance to drugs, antibiotics or antimicrobials is growing rapidly worldwide. Antibiotic therapy is one of the main approaches of modern medicine to fight infections. The \"golden era\" of antibiotics lasted from the 1930s to the 1960s, when many antibiotics appeared. Unfortunately, this era has ended because researchers have been unable to maintain the pace of antibiotic discovery in the face of new resistant pathogens. Constant failures in the development or discovery of new antibiotics and their unreasonable use are predisposing factors associated with the emergence of antibiotic resistance in many infectious diseases of animals and humans [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan additionalcitationids=\"CR12\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe main causes of many infectious diseases in animals and humans are gram-negative bacteria, among which representatives of the Enterobacteriaceae family occupy a leading position. The greatest amount of data on antimicrobial resistance was obtained for the following bacterial species: \u003cem\u003eEscherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Streptococcus pneumoniae\u003c/em\u003e, and \u003cem\u003eSalmonella spp\u003c/em\u003e [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe obtained results, to our knowledge, represent an assessment of the burden of antibiotic sensitivity and resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan, which poses a serious global health threat that requires greater attention, funding and capacity building, research and development, as well as prioritization of specific pathogens from the broader global health community.\u003c/p\u003e \u003cp\u003eThe findings include an assessment of the burden of antibiotic sensitivity and resistance of Enterobacteriaceae isolates isolated from animal products of the Almaty region of Kazakhstan, which represents a serious global health threat and requires greater attention, funding and capacity-building, research and development, as well as prioritization of specific pathogens by the broader global health community.\u003c/p\u003e \u003cp\u003eThe results of the research presented in the article using a new domestic component of culture media for cultivating microorganisms from defibrinated horse blood showed that the Almaty region can be classified as region with low resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin and ciprofloxacin.\u003c/p\u003e \u003cp\u003eToday, for the first time, the WHO publishes a Bacterial Priority Pathogens List of antibiotic\u0026ndash;resistant \u0026ldquo;priority pathogens\u0026rdquo; including 15 families of antibiotic-resistant bacteria that pose the greatest threat to human health. The list was intended to inform research and development priorities related to new antibiotics and to pay special attention to multidrug-resistant pathogens that cause severe and often fatal infections in health facilities [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThere is much scientific evidence proving that antibiotics, of course, can save millions of lives, but at the same time they can also harm, and even future generations, if used improperly. This fact raises the question of the rationality of the use of antibacterial agents and suggests that despite all efforts, there is a lack of knowledge in the global medical community regarding the potentially dangerous aspects of antibiotic therapy. The irrational use of antibiotics, which are far from WHO recommendations and are mainly in injectable form, has been reported by Ethiopian scientists [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eE.coli\u003c/em\u003e and Salmonella are among the most common bacteria in the world. Many methods have been developed to diagnose these infections, one of which is bacteriological examination. The complexity and complexity of isolating U.coli and Salmonella limits the use of this method in routine practice in practical healthcare, but it remains the only method for phenotypic determination of the sensitivity of strains to antibacterial drugs. Monitoring of the increasing resistance of these infections, correction of eradication therapy regimens considering the sensitivity of isolated strains to chemotherapeutic drugs, and testing of new drugs are indispensable bacteriological methods for practical and scientific purposes. An obligatory component of the medium should be an additive of 5\u0026ndash;10% of the blood or serum of animals (horse, sheep) [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eE.coli\u003c/em\u003e and Salmonella are among the most common bacteria in the world. Many methods have been developed to diagnose these infections, one of which is bacteriological research. The complexity of isolating \u003cem\u003eE.coli\u003c/em\u003e and Salmonella limits the use of this method in routine practical healthcare practice, but it remains the only method for phenotypic determination of the sensitivity of strains to antibacterial drugs. The monitoring of the increasing resistance of these infections, the correction of eradication therapy regimens considering the sensitivity of isolated strains to chemotherapeutic drugs, and the testing of new drugs make this bacteriological method indispensable for practical and scientific purposes. An obligatory component of the medium should be an additive of 5\u0026ndash;10% of the blood or serum of animals (horse, sheep) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTherefore, a component for media has been proposed for conducting bacteriological studies, allowing us to obtain media enriched with micro- and macronutrients for reliable and high-quality laboratory analyses.\u003c/p\u003e \u003cp\u003eThe use of the proposed component obtained from defibrinated horse blood in the cultivation of microorganisms will contribute to the identification, differentiation and determination of the hemolytic activity of microorganisms that cause diseases in humans and animals.\u003c/p\u003e \u003cp\u003eThis work also has the goal of authenticating the growth of \u003cem\u003eE. coli\u003c/em\u003e and Salmonella strains via phylogenetic analysis. For this purpose, we studied the 16S RNA of the bacterium. Ribosomal RNA 16S is a component of the 30S subunit of prokaryotic ribosomes and has high conservation, which makes it a good candidate for studying evolutionary relationships [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe studied \u003cem\u003eE. coli\u003c/em\u003e gene, \"Isolate 11\", is part of a phylogenetic analysis showing its genetic relationship with other bacteria of the genus Escherichia with a difference of 0.5% in the nucleotide sequence of the 16S rRNA gene. Its location on the tree suggests that it has special genetic characteristics that distinguish it from other species and strains of Escherichia.\u003c/p\u003e \u003cp\u003eThe studied Salmonella gene, designated \"7\", is included in the exclusive group of \u003cem\u003eSalmonella enterica\u003c/em\u003e, which is formed from one exceptional ancestral line. The \"7\" sample corresponds to a distinctive sequence or variant of \u003cem\u003eSalmonella enterica\u003c/em\u003e, and this discovery may be crucial for investigating its virulence, drug resistance, or genetic development over time.\u003c/p\u003e \u003cp\u003eThis phylogenetic analysis is crucial for epidemiological tracking, understanding bacterial evolution, and the potential development of targeted therapies or interventions [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn the context of phylogenetic trees, it is important to consider that exact interpretation depends on the methods used to construct the tree, such as the specific genetic markers being analysed and the algorithms used in the analysis. Moreover, although bootstrap values provide statistical support, they do not guarantee that the tree perfectly reflects the true evolutionary history due to potential genetic recombination, horizontal gene transfer, and other factors that may complicate bacterial phylogenetics [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTherefore, this work analysed the sensitivity of Enterobacteriaceae isolates isolated from animal products to a wide range of antibiotics used in practice.\u003c/p\u003e \u003cp\u003eThe obtained results suggest that regions with low resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to cefotaxime, ampicillin/sulbactam, levofloxacin, and meropenem and Salmonella to levofloxacin and high resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin, ciprofloxacin.\u003c/p\u003e \u003cp\u003eThese facts support the rational use of antibacterial agents, and ssugges that despite all efforts, there is a lack of knowledge in the medical and veterinary communities regarding the potentially dangerous aspects of antibiotic therapy.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003ePathogens of enteropathogenic diseases that are resistant to antibacterial drugs are circulating in the Almaty region of Kazakhstan. Using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro- and macronutrients for reliable and high-quality laboratory analyses.\u003c/p\u003e \u003cp\u003eAccording to the data obtained, the low resistance rates of \u003cem\u003eE.coli\u003c/em\u003e to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to norfloxacin and ciprofloxacin, gentamicin, and Salmonella to norfloxacin and ciprofloxacin were detected in the Almaty region.\u003c/p\u003e \u003cp\u003eCirculating pathogens of the enteropathogenic diseases \u003cem\u003eE. coli\u003c/em\u003e and Salmonella in the Almaty region, Kazakhstan have been confirmed by phylogenetic analysis to be genuine pathogens with special genetic characteristics that distinguish them from other species and strains. Such phylogenetic analysis is crucial for epidemiological tracking, understanding bacterial evolution, and the potential development of targeted therapies or interventions.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eThe Global Antimicrobial Resistance and Use Surveillance System (GLASS)\u003c/p\u003e\n\u003cp\u003eWorld Health Organization (WHO)\u003c/p\u003e\n\u003cp\u003eThe Bacterial Priority Pathogens List (BPPL)\u003c/p\u003e\n\u003cp\u003eAntimicrobial resistance (AMR)\u003c/p\u003e\n\u003cp\u003eThe European Committee on Antimicrobial Susceptibility Testing (EUCAST)\u003c/p\u003e\n\u003cp\u003eThe Basic Local Alignment Search Tool (BLAST)\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e Conceptualization, Z.L. and A.N. contributed to the conceptualization, supervision, editing, and project administration. Z.L., A.N., Zh.A. M.N., A.B., and M.K. contributed to the writing of the original draft, writing, review, and editing. Sh.Y., K.R. and S.Sh.; methodology, Z.L., S.Sh. and Zh.A.; validation, A.M., V.S., M.Y., M.N., M.S., A.B. and M.K.; formal analysis, Z.L., S.Sh., M.N., and A.N.; investigation, Z.L., S.Sh., Sh.E., G.K., Zh.T., O.N., K.R., N.I., A.M., V.S., M.N., M.S., Zh.A. and M.K.; resources, A.M., V.S., M.Y., M.N., M.S., B.A. All authors have read and agreed to the published version of the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Ministry of Healthcare of the Republic of Kazakhstan, within the framework of the PTF IRN BR218004/0223 \u0026quot;Improving biosafety measures in Kazakhstan: countering dangerous and especially dangerous infections\u0026quot; for 2023-2025, through the program of the Applied Epidemiology Field Epidemiology Training Program (FETP), Centers for Disease Control and Prevention in the Central Asia Region (CDC/CAR).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the data analysed or generated during this study are included in this published article and its Supporting Information files. Any additional information is available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u003c/strong\u003e The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eHavelaar AH, Kirk MD, Torgerson PR, Gibb HJ, Hald T, Lake RJ, Praet N, Bellinger DC, de Silva NR, Gargouri N, Speybroeck N, Cawthorne A, Mathers C, Stein C, Angulo FJ, Devleesschauwer B. World Health Organization Global Estimates and Regional Comparisons of the Burden of Foodborne Disease in 2010. World Health Organization Foodborne Disease Burden Epidemiology Reference Group. PLoS Med. 2015;12(12):e1001923. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pmed.1001923. eCollection\u003c/span\u003e\u003cspan address=\"10.1371/journal.pmed.1001923. eCollection\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHariharan P, Bharani T, Franklyne JS, Biswas P, Solanki SS, Paul-Satyaseela M. Antibiotic susceptibility pattern of Enterobacteriaceae and non fermenter gram-negative clinical isolates of microbial resource orchid. J Nat Sci Biol Med. 2015 Jan-Jun;6(1):198\u0026ndash;201. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4103/0976-9668.149121\u003c/span\u003e\u003cspan address=\"10.4103/0976-9668.149121\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 25810660; PMCID: PMC4367035.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEnglish BK, Gaur AH. The use and abuse of antibiotics and the development of antibiotic resistance. Adv Exp Med Biol. 2010;659:73\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePuvača N. Antimicrobial Resistance and Treatment in Companion, Food and Exotic Animals. Antibiot (Basel). 2022;11(10):1360. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/antibiotics11101360\u003c/span\u003e\u003cspan address=\"10.3390/antibiotics11101360\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 36290017; PMCID: PMC9598238.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO Bacterial Priority Pathogens List. 2024: bacterial pathogens of public health importance to guide research, development and strategies to prevent and control antimicrobial resistance. World Health Organization, 2024.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEuropean Committee on Antimicrobial Susceptibility Testing (EUCAST.) Disco-diffuse EUCAST method. - Version 10.0, 2020. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.eucast.org\u003c/span\u003e\u003cspan address=\"http://www.eucast.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMueller JH, Hinton J. (1941). A protein-free medium for primary isolation of the gonococcus and meningococcus, Proc. Soc. Exper. Biol. Med. 48: 330.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThe National Center for Biotechnology Information. NCBI databases \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehtpp://www.ncbi.nlm.nih.gov\u003c/span\u003e\u003cspan address=\"http://htpp://www.ncbi.nlm.nih.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAslam B, Wang W, Arshad MI, Khurshid M, Muzammil S, Rasool MH, Nisar MA, Alvi RF, Aslam MA, Qamar MU, Salamat MKF, Baloch Z. Antibiotic resistance: a rundown of a global crisis. Infect Drug Resist. 2018;11:1645\u0026ndash;58. PMID: 30349322; PMCID: PMC6188119.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhingra S, Rahman NAA, Peile E, Rahman M, Sartelli M, Hassali MA, Islam T, Islam S, Haque M. Microbial Resistance Movements: An Overview of Global Public Health Threats Posed by Antimicrobial Resistance, and How Best to Counter. Front Public Health. 2020;8:535668. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fpubh.2020.535668\u003c/span\u003e\u003cspan address=\"10.3389/fpubh.2020.535668\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 33251170; PMCID: PMC7672122.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNathan C, Cars O. Antibiotic resistance-problems, progress, and prospects. N Engl J Med. 2014;371(19):1761\u0026ndash;3.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRobert C. Hospital infection: a historical review. Br Med Bull. 1944;2:272\u0026ndash;6. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/oxfordjournals.bmb.a071061\u003c/span\u003e\u003cspan address=\"10.1093/oxfordjournals.bmb.a071061\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSmith PW, Watkins K, Hewlett A. Infection control through the ages. Am J infect control. 2012;40:35\u0026ndash;42. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ajic.2011.02.019\u003c/span\u003e\u003cspan address=\"10.1016/j.ajic.2011.02.019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAntimicrobial Resistance Collaborators. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022;399(10325):629\u0026ndash;655. doi: 10.1016/S0140-6736(21)02724-0. Epub 2022 Jan 19. Erratum in: Lancet. 2022;400(10358):1102. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/S0140-6736(21)02653-2\u003c/span\u003e\u003cspan address=\"10.1016/S0140-6736(21)02653-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBao L, Peng R, Ren X, Ma R, Li J, Wang Y. Analysis of some common pathogens and their drug resistance to antibiotics. Pak J Med Sci. 2013;29(1):135\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.12669/pjms.291.2744\u003c/span\u003e\u003cspan address=\"10.12669/pjms.291.2744\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKebede HK, Gesesew HA, Woldehaimanot TE, Goro KK. Antimicrobial use in paediatric patients in a teaching hospital in Ethiopia. PLoS ONE. 2017;12(3):e0173290. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal. pone.0173290\u003c/span\u003e\u003cspan address=\"10.1371/journal. pone.0173290\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKowalska-Krochmal B, Dudek-Wicher R. The Minimum Inhibitory Concentration of Antibiotics: Methods, Interpretation, Clinical Relevance. Pathogens. 2021;10(2):165. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/pathogens10020165\u003c/span\u003e\u003cspan address=\"10.3390/pathogens10020165\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGajic I, Kabic J, Kekic D, Jovicevic M, Milenkovic M, Mitic Culafic D, Trudic A, Ranin L, Opavski N. Antimicrobial Susceptibility Testing: A Comprehensive Review of Currently Used Methods. Antibiot (Basel). 2022;11(4):427. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/antibiotics11040427\u003c/span\u003e\u003cspan address=\"10.3390/antibiotics11040427\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrinevich D, Harden L, Thakur S, Callahan B. Serovar-level identification of bacterial foodborne pathogens from full-length 16S rRNA gene sequencing. mSystems. 2024;9(3):e0075723.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaquero F, Mart\u0026iacute;nez JL, Lanza F, Rodr\u0026iacute;guez-Beltr\u0026aacute;n V, Gal\u0026aacute;n J, San Mill\u0026aacute;n JC, Cant\u0026oacute;n A, Coque R. Evolutionary Pathways and Trajectories in Antibiotic Resistance. Clin Microbiol Rev. 2021;34(4):e0005019.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDidelot X. Phylogenetic Analysis of Bacterial Pathogen Genomes. Methods Mol Biol. 2023;2674:87\u0026ndash;99.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"antibiotic resistance, Salmonella enterica, Escherichia сoli, animal products, culture media for the cultivation of microorganisms with the addition of defibrinated horse blood","lastPublishedDoi":"10.21203/rs.3.rs-4575952/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4575952/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eThis article presents the results of studies on the antibiotic resistance of \u003cem\u003eEscherichia coli\u003c/em\u003e and \u003cem\u003eSalmonella enterica\u003c/em\u003e isolates isolated from animal products of the Almaty region, Kazakhstan using a new domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eThe results showed that the Almaty region can be classified as region with low resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to cefotaxime, ampicillin/sulbactam, levofloxacin, meropenem and Salmonella to levofloxacin and high resistance rates of \u003cem\u003eE. coli\u003c/em\u003e to norfloxacin, ciprofloxacin, gentamicin, and Salmonella to norfloxacin, ciprofloxacin. The use of a domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro and macronutrients for reliable and high-quality laboratory analyses.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e It is assumed that the irrational use of fluoroquinolones in animal husbandry leads to an increase in the resistance of microorganisms that cause infectious diseases common to humans and animals, since the above types of antibiotics (fluoroquinolones) are the most important drugs for the treatment of bacterial infections in medicine and veterinary medicine.\u003c/p\u003e\n\u003cp\u003eThe results indicate that pathogens of enteropathogenic diseases resistant to antibacterial drugs are circulating in the territory of the Almaty region of Kazakhstan and the use of a domestic component of culture media for the cultivation of microorganisms from defibrinated horse blood makes it possible to obtain media enriched with micro and macronutrients for reliable and high-quality laboratory analyses.\u003c/p\u003e","manuscriptTitle":"Analyzing the Sensitivity and Antibiotic Resistance of Enterobacteriaceae Isolates Isolated from Animal Products in Almaty Region, Kazakhstan","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-09 17:05:21","doi":"10.21203/rs.3.rs-4575952/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c362c837-d180-4a8b-977c-9b385e6199f2","owner":[],"postedDate":"July 9th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-11-28T17:08:40+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-09 17:05:21","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4575952","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4575952","identity":"rs-4575952","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}