Background and Aim: Brucellosis remains one of the most common zoonoses. The current anti-brucellosis measures are largely deemed ineffective due to a lack of specificity of conventional serological tests. This study evaluated the use of Brucella outer membrane protein (Omp)19 for serodiagnostic testing. Materials and Methods: The antigenicity of recombinant Brucella Omp19, Omp25, and Omp31 was examined in serum samples from mice and rabbits immunized with Omp19 or Brucella abortus 19 whole cell (WC) and 12 and 152 cows experimentally or naturally infected with brucellosis, respectively. Serum samples were collected from 151 cows that were vaccinated with B. abortus 19 and 12 unvaccinated heifers that were maintained on a brucellosis-free farm. Results: Immunization with Omp19 resulted in antibody production in mice after a single injection without the use of adjuvant. Serum antibodies obtained from rabbits immunized with inactivated B. abortus strain 19 WC targeted Omps by enzyme-linked immunosorbent assay (ELISA) and Western blot. Antibodies targeting Omp19 were identified in all B. abortus strain 544 experimentally infected cows at day 14 post-inoculation (p.i.); Omp25 was detected by ELISA at day 28 p.i., while an ELISA targeting Omp31 was negative for 25% of cows at this time point. Omp19 and Omp25 were readily detected by sera from cows from a new epizootic focus. Antibodies recognizing Omps were also detected in >50% of the animals maintained in a brucellosis-free herd at 10 months after vaccination. Conclusion: Brucella Omp19 in combination with Omp25 and Omp31 may be utilized as target antigens in an ELISA designed for serological testing of unvaccinated cattle.

Background and Aim: Staphylococcus aureus is the most frequent and ubiquitous cause of mastitis in cows. In recent decades, antibiotic resistance has rapidly spread among infectious disease pathogens in Kazakhstan and globally. This study examined the phenotypic and genotypic resistance of S. aureus strains obtained from cattle milk to antibiotics. Materials and Methods: In 2021 and 2022, 675 cow milk samples were collected from 16 dairy farms in Northern Kazakhstan. Staphylococcus aureus was identified using culture and biochemical methods. The nature of antibiotic resistance was determined by the disk diffusion (DD) method. The distribution of antibiotic resistance genes was determined by polymerase chain reaction. Results: Among the obtained S. aureus isolates, high levels of resistance to β-lactam antibiotics (100%), tetracyclines (95.4%), fluoroquinolones (95.4%), and macrolides (60.92%) were observed. Meanwhile, the lowest levels of resistance were identified for sulfonamides (21.84%) and aminoglycosides (27.59%). All the obtained isolates were positive for the nuc gene encoding thermonuclease. The blaZ, ermC, and tetK genes were detected in 45.9%, 77%, and 83.9% of the studied S. aureus isolates, respectively. Conclusion: The results indicate a high prevalence of antibiotic resistance in S. aureus isolated from cows with clinical and subclinical forms of mastitis in Northern Kazakhstan. In addition, the prevalence of resistance was higher when evaluated by the DD method than when detecting the specific antibiotic resistance genes blaZ, tetK, and ermC, indicating the need for deeper analysis of the phenotypic and genetic determinants of antibiotic resistance. Keywords: antibiotic resistance genes, cow milk, polymerase chain reaction, Staphylococcus aureus strains.

Background and Aim: An accurate diagnosis of Brucella-infected animals is one of the critical measures in eradication programs. Conventional serological tests based on whole-cell (WC) antigens and detecting antibodies against pathogen-associated lipopolysaccharide might give false-positive results due to the cross-reactivity with other closely related bacteria. This study evaluated the serological potential of Brucella spp. chimeric outer membrane proteins (Omps) as antigens in an indirect enzyme-linked immunosorbent assay (i-ELISA). Materials and Methods: The chimeric gene constructs of the most immunodominant regions of Brucella Omps 25+31, 25+19, and 19+31 were cloned into the pET28a expression vectors and transformed into Escherichia coli BL21 (DE3). The serological potential of chimeric proteins compared with single recombinant Omps (rOmps)19, 25, and/or 31 were studied on blood serum samples of (i) a rabbit immunized with killed Brucella abortus 19WC, (ii) mice immunized with single rOmps, (iii) cows seropositive for brucellosis by rose Bengal test, and (iv) cattle naturally and/or experimentally infected with brucellosis. Results: E. coli BL21 actively produced Brucella chimeric rOmps, the concentration of which reached a maximum level at 6 h after isopropyl-β-D-1-thiogalactopyranoside stimulation. Target proteins were antigenic and expressed in an active state, as recognized by rabbit anti-B. abortus antibodies in an i-ELISA and western blotting. Murine antibodies against the single rOmps reacted with chimeric antigens, and conversely, antichimeric antibodies found their epitopes in single proteins. Brucella chimeric rOmps showed higher antigenicity in blood sera of seropositive cattle kept in the hotbed of the infection and/or experimentally challenged with brucellosis than single proteins. Conclusion: Brucella chimeric recombinant outer membrane proteins could be a potential antigen candidate for developing an ELISA test for accurate diagnosis of bovine brucellosis.

Brucellosis remains an endemic zoonotic disease in many developing countries, causing great harm to public health and devastating losses to livestock. One of the main reasons for the low effectiveness of anti-brucellosis measures is the lack of reliable methods for diagnosing infected animals throughout their lifespan. Classical serological tests, such as the tube agglutination test, rose Bengal plate test, and complement fixation test, as well as commercial enzyme-linked immunosorbent assay kits, are based on the detection of antibodies to the cell wall polysaccharide antigens of Brucella spp. smooth strains. As a result, they do not exclude cross-reactions with related bacteria and fail to differentiate between infected and vaccinated animals. Over the past decades, many attempts have been made to identify immunoreactive and pathogen-specific protein antigens. To date, several studies have investigated Brucella spp. recombinant proteins, including cell wall proteins, as the best antigens for diagnosing brucellosis in animals and humans. However, the available results on the specificity and sensitivity of serological tests based on cell wall proteins are ambiguous and sometimes contradictory. This review aims to provide an overview of the current state of knowledge of the diagnostic value of outer membrane and/or periplasmic proteins of Brucella spp. The goal is to identify future developments that may lead to reliable antigens for serological tests. Keywords: Brucella, diagnostics, enzyme-linked immunosorbent assay, outer membrane protein, periplasmic protein.

Aim of the present study was to provide presence of opisthorchiid metacercariae in cyprinid fish Leuciscus idus in Nura-Sarysu river, Kazakhstan. Infection rate of the ides by the metacercariae was 42%. The metacercariae, similar morphologically to those of the liver flukes, were found: elliptical in shape, 0.19-0.25×0.15-0.22 mm, oral and ventral suckers nearly equal size, and excretory bladder O-shape with black content, occupying posterior part of the body. The metacercariae were divided into 2 groups with differences in size and thickness of cyst wall. Adult flukes were recovered from the Syrian hamsters infected with the opisthorch metacercariae and identified with morphological characters to Opisthorchis felineus and Metorchis bilis. DNA sequences of ITS1, ITS2, and cox1 supported the taxonomic assignment.

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