Bacteriological Methods Research Articles

The State of Colon Microbiocenosis and Antioxidant Properties of Rat Colonocytes in Environmental Dysbiosis and Use of Probiotic Bifidumbacterin® and Symbiotic Acipol®

INTRODUCTION: The microbial community of the colon is a single microecological system which quickly responds to the exposure to external and internal factors with quantitative and qualitative changes with the result of development of the condition termed dysbiosis. The changes in the composition of colon microbiocenosis can lead to a number of diseases and to the development of immunopathological and allergic conditions.
 AIM: To evaluate the state of the colon microbiocenosis and antioxidant properties of rats’ colonocytes against a background environmental dysbiosis and the use of probiotic Bifidumbacterin® and symbiotic Acipol®.
 MATERIALS AND METHODS: In the experiment, intoxication in rats was modeled with the fungicide Thiram® (dimethylcarbamothioylsulfanyl N,N-dimethylcarbamodithioate, 1.6 mg per animal of 200 g mass) for 28 days, which led to the development of environmental dysbiosis. The detected changes were subsequently corrected within 21 days with the probiotic Bifidumbacterin® (0.14 ml per animal of 200 g mass) and the symbiotic Acipol® (0.08 ml per animal of 200 g mass). The composition of the microflora of colon mucosa of rats was studied using a bacteriological method. The activity of antioxidant protection enzymes of colonocytes was studied by the content of superoxide dismutase and catalase. The condition of lipid peroxidation processes was assessed by the content of diene conjugates and malondialdehyde.
 RESULTS: In introduction of Thiram®, reduction of the amount of obligate and opportunistic representatives of colon microbiocenosis was noted, as well as intensification of lipid peroxidation processes in the conditions of weakening of antioxidant protection in colonocytes. Application of Bifidumbacterin® and Acipol® resulted in increase in the amount of obligate bacteria and decrease in the content of opportunists of the colon microbiota with the strongest effect of Acipol® on the values of the determined parameters. When studying changes in biochemical parameters in animal colonocytes, both drugs were found to contribute to the normalization of the content of malondialdehyde in the colon tissue, besides, Acipol® led to a decrease in the concentration of diene conjugates in colonocytes.
 CONCLUSION: Introduction of Thiram® in all the specified periods affected the qualitative and quantitative composition of the microbiota and biochemical parameters in the colonocytes of the animals. The use of Bifidumbacterin® and Acipol® preparations resulted in the restoration of the qualitative and quantitative composition of colonic microorganisms, and also had a positive effect on the content of lipid peroxidation products in colonocytes, but more efficient was found to be Acipol® symbiotic.

PATHOGENETIC SIGNIFICANCE OF THE CHARACTER OF WOMAN’S UROGENITAL INFECTION IN THE DEVELOPMENT OF ADENOMYOSIS

Adenomyosis is one of the important problems of modern gynecology, which leads to impaired reproductive function, reduced work capacity and quality of life of women. The prevalence of endometriosis in the female population ranges from 20-55% in patients of reproductive age. An important role in the occurrence and development of adenomyosis is played by genetic predisposition, hormonal changes and the influence of immunological factors. Of great interest is a new concept regarding the significant role of bacterial contamination of the genital tract in patients with adenomyosis. The study of the influence of microorganisms on the mechanisms of activation of the inflammatory reaction associated with the progression of endometrioid lesions is relevant and requires the search for new approaches to diagnosis and treatment of the disease.
 The purpose of the work is to study the influence of the woman’s urogenital infection in the development of adenomyosis.
 Materials and methods. 85 patients with 1st and 2nd degree adenomyosis and 30 healthy fertile women were examined. The average age of the examined was 27.3±1.6 years. A comprehensive clinical examination, ultrasound, colposcopic, hysteroscopic studies were conducted. Microbial flora analysis included bacterioscopic, bacteriological research methods with determination of sensitivity to antibiotics, and PCR method. The concentration of cytokines in the culture medium (supernatant) was determined by the enzyme immunoassay. Statistical data processing was performed using the Statsoft Statistica v6.0 and Microsoft Excel 97 software packages.
 Results. In the structure of gynecological diseases, 95.6% of patients with adenomyosis had inflammatory diseases of female genital organs, chronic herpes and cytomegalovirus infection was found in almost 93%. In women with adenomyosis, the II-III stage of vaginal cleanliness prevails with an “intermediate” type of biocenosis, which is characterized by a reduced content of lactobacilli, the presence of various types of morphotypes of gram-positive and gram-negative rods and cocci, and dysbiosis (49.4%), when a mixed bacterial flora prevails microflora, a high percentage of the combination of adenomyosis with chronic inflammatory diseases of the pelvic organs justifies the need to use antimicrobial therapy in this category of patients. When studying the cytokine profile of the cervical contents and endometrium, an increase in the concentration of the main proinflammatory cytokines IL-1β, IL-6, TNFα and a factor with chemotoxic properties IL-8 was found. The obtained data indicate that the development of adenomyosis is facilitated by a previous immuneinflammatory reaction, which is accompanied by a violation of the local cytokine balance.
 Conclusions. Microbiological examination of patients with adenomyosis revealed a violation of the microbial landscape of the reproductive tract in most of the examined. The obtained data indicate a high percentage of the combination of adenomyosis with chronic inflammatory diseases of the pelvic reproductive organs. Previous adenomyosis is an immunoinflammatory reaction, accompanied by a violation of the local cytokine balance. Increased cytokine activity and the presence of infectious agents are associated with the spread of adenomyosis.

Molecular Investigation of gyrA Mutations in Clinical Isolates of Methicillin-Resistant Staphylococcus aureus Derived from Diverse Sources

Background: Fluoroquinolones are the most effective antibiotics against Staphylococcus aureus isolates. In hospitals, excessive use of antibiotics has led to the emergence of highly resistant strains of S. aureus isolates. Objective: The aim of this study was to detect the mutations that occur in the gyrA gene encoding for DNA gyrase, which is one of the targets for fluoroquinolone resistance. Methods: Fifty clinical isolates were diagnosed as S. aureus according to molecular and bacteriological methods. The susceptibility tests were performed on all bacterial isolates by the disc diffusion method using methicillin and six fluoroquinolone antibiotics. Results: Out of fifty isolates, twelve were resistant to methicillin and all six antibiotics (nalidixic acid, lomefloxacin, ciprofloxacin, norfloxacin, ofloxacin, and levofloxacin). From the fifty isolates, 12 were resistant, 3 were intermediate, and 38 were sensitive to three or more tested antibiotics. The resistance of S. aureus isolates was also confirmed by the minimum inhibitory concentration test. The main sources of isolates were burns (10%), nose (16) wounds (8%), operation room (10%), ear (20%), urine (8%), skin (6%), and throat (22%). Twelve resistant isolates were used to examine the mutations in the gyrA gene. A direct sequence analysis found eight mutations in the gyrA gene; these mutations included 2 (25% missense mutations), 1 (12.5%) deletion mutation, and 5 (62.5%) silent mutations at various sites. Conclusion: gyrA mutations resulting from the excessive use of antibiotics may be one of the mechanisms leading to fluoroquinolone resistance.

Involvement of endoplasmic reticulum and histone proteins in immunomodulation by TLR4-interacting SPA4 peptide against Escherichia coli.

Pulmonary host defense is critical for the control of lung infection and inflammation. An increased expression and activity of Toll-like receptor 4 (TLR4) induce phagocytic uptake/clearance and inflammation against Gram-negative bacteria. In this study, we addressed the mechanistic aspect of the immunomodulatory activity of the TLR4-interacting SPA4 peptide (amino acid sequence GDFRYSDGTPVNYTNWYRGE) against Escherichia coli. Binding of the SPA4 peptide to bacteria and direct anti-bacterial effects were investigated using flow cytometric, microscopic, and bacteriological methods. The bacterial uptake and inflammatory cytokine response were studied in dendritic cells expressing endogenous basal level of TLR4 or overexpressing TLR4. The subcellular distribution and co-localization of TLR4 and bacteria were investigated by immunocytochemistry. Furthermore, we studied the cellular expression and co-localization of endoplasmic reticulum (ER) molecules (calnexin and ER membrane protein complex subunit 1; EMC1) with lysosomal-associated membrane protein 1 (LAMP1) in cells infected with E. coli and treated with the SPA4 peptide. Simultaneously, the expression of histone H2A protein was quantitated by immunoblotting. Our results demonstrate no binding or direct killing of the bacteria by SPA4 peptide. Instead, it induces the uptake and localization of E. coli in the phagolysosomes for lysis and simultaneously suppresses the secreted levels of TNF-α. Overexpression of TLR4 further augments the pro-phagocytic and anti-inflammatory activity of SPA4 peptide. A time-dependent change in subcellular distribution of TLR4 and an increased co-localization of TLR4 with E. coli in SPA4 peptide-treated cells suggest an enhanced recognition and internalization of bacteria in conjugation with TLR4. Furthermore, an increased co-localization of calnexin and EMC1 with LAMP1 indicates the involvement of ER in pro-phagocytic activity of SPA4 peptide. Simultaneous reduction in secreted amounts of TNF-α coincides with suppressed histone H2A protein expression in the SPA4 peptide-treated cells. These results provide initial insights into the plausible role of ER and histones in the TLR4-immunomodulatory activity of SPA4 peptide against Gram-negative bacteria.

Prevalence of Camel Mastitis and Its Associated Risk Factors in and around Garowe District, Puntland, Somalia

Mastitis is one of the main issues that cause economic inefficiencies in dairy farms. It has numerous infectious and non-infectious origins and is divided into clinical and subclinical forms. A cross sectional study of camel mastitis was conducted on 88 lactating camels from Garowe District Puntland between May to August 2023 to estimate the prevalence and causes of mastitis, as well the risk factors involved on disease. Prevalence of mastitis was assessed by using California mastitis test (CMT) and bacterial cultured. The overall prevalence of mastitis at animal level was camels examined for mastitis was found a prevalence of 39.7% (35/88) in the current study. The overall quarter level prevalence was 47.7% (168/352). Among these 10(11.4%) and 25(28.4%) were found to be positive with clinical and sub-clinical mastitis respectively, based on clinical examination and mastitis indicator paper (Table 2). Regarding Result analysis showed that there was a statistically significant association (P<0.05) among four the risk factors (tick infected, wound stage of lactation and poor hygienic). Tick infestation and udder lesions were significantly associated with camel mastitis (P < 0.05). Bacteriological examination of CMT along with bacteriological culture methods were used. The study cultured a positive camel milk samples revealed that Staphylococcus spp was the major causative agents for both clinical and sub clinical camel mastitis (45.7%) followed by Streptococcus agalactiae (25.7%), E. coli (14.3%), Klebsiella spp. (8.6%) and Micrococcus (5.7%) respectively. The study demonstrated that camel mastitis is a problem which warrants appropriate control measures in order to improve the health of camels and quality of camel milk production in the study area. The isolation of genera of pathogenic bacteria from the camel milk samples suggests the need for strict hygienic measures during the production and handling of camel milk to reduce public health hazards. Furthermore, public education should be given to improve their awareness about the importance of proper herd health management and hygienic milking practices. In order to minimize the adverse effect of mastitis on the yield, quality of milk and zoonotic impact of the pathogen. The prevalence of camel mastitis in the study area was found to be significantly high. Therefore, implementation of integrated approaches has great importance in the study sites for the prevention and control of mastitis hence minimizing economic loss and prevents significant public health risks.

Characterization and antimicrobial susceptibility of Escherichia coli, Salmonella enterica isolated from Uvurkhangai province in Mongolia

The incidence of infectious diseases caused by the Enterobacteriaceae family of Uvurkhangai province in Mongolia is not decreasing and is recurring despite the implementation of preventive measures in the province. Nowadays, when the resistance of microorganisms negatively affects the health of humans and animals, food safety, the environment, and the economy, the lack of effective antibiotics for treatment is the reason for the research work on this topic. Based on the outbreak and surveillance of infectious diseases for the past 5 years, the pathological materials were received in the veterinary laboratory, two strains of Escherichia coli and also two strains Salmonella enterica subsp. abortus equi were isolated, and identified by conventional bacteriological and molecular methods. The antimicrobial susceptibility of the pathogen was determined using the disc diffusion method. The strains of E. coli were resistant to several antimicrobials, including vancomycin and neomycin, intermediate to colistin. The Salmonella enterica subsp. abortus equi showed resistance to ampicillin, vancomycin, gentamicin, and neomycin. Due to the presence of antimicrobial resistance, the results suggest a risk to public health as these pathogens. However, the four isolated bacteria in the family of Enterobacteriaceae showed susceptibility to groups of tetracycline and streptomycin. As a result of this study, it was determined that treatment with tetracycline and streptomycin is effective against bacterial infections caused by the Enterobacteriaceae group.

RELATIONSHIP BETWEEN THE ACNE CLINICAL COURSE AND SEVERITY AND THE STATE OF MICROBIOTA OF THE SKIN AND LARGE INTESTINE

A high incidence of acne, tendency to its chronic course with the formation of resistance to treatment and frequent development of deep forms determine an important medical and social significance of the problem.
 Aim. To study and evaluate the degree of changes in the microbiocenosis of the skin and large intestine in patients with acne, depending on their clinical course.
 Materials and Methods. We examined 85 patients with acne, aged 18 to 45 years, 49 (57.65±5.36%) women, 36 (42.35±5.36%) men and 35 apparently healthy persons who made up the control group. The composition of microbiota of the skin and large intestine was examined using a bacteriological method.
 Results and Discussion. It was found that there is a significantly lower content of bifidobacteria and lactobacilli, bacterioids and total E. coli count in the large intestine in patients with acne associated with a higher amount of E. coli with enzyme properties, conditionally pathogenic enterobacteria, enterococci and fungi of the genus Candida. When using Friedman's non-parametric analysis of variance, it was found that there is a statistically significant difference between the frequency of detection of normocenosis and the stage of the large intestinal dysbiosis and the development of clinical forms of acne. At that, normocenosis was detected more often in patients with comedonal acne, while papular and nodular acne occurred more often in patients with manifestations of different stages of dysbiosis. An increase in seeding with associations of the alpha-hemolytic streptococcus + Staphylococcus haemolyticus + Micrococcus, beta-hemolytic streptococcus + Staphylococcus aureus + E.Coli + Candida albicans and a significant increase in the growth of alpha-hemolytic streptococcus, Staphylococcus haemoliticus and Staphylococcus aureus were found in the microbiocenosis of the skin in patients with acne. At that, all of the above microorganisms were not cultured in the patients of the control group.
 Conclusion. The qualitative and quantitative changes in the parameters of microbiocenosis of the skin and large intestine, which depend on the clinical course of dermatosis, were found in the examined patients with acne. This indicates the need for additional examination of such patients with the aim of prescribing a pathogenetically substantiated therapy.

Biological Properties and Antibiotic Resistance of <i>Klebsiella pneumoniae</i> and Its Role in the Etiological Structure of Community-Acquired Pneumonia Pathogens

Background. In a novel coronavirus pandemic, the most common complications of viral pneumonia are secondary infections of bacterial and fungal etiology. At the same time, the spread of multidrug-resistant bacteria remains a global threat to public health. One of such microorganisms of the Enterobacteriaceae family is Klebsiella pneumoniae, which belongs to dangerous resistant pathogens of the ESKAPE group.The aim of the study was the comparative analysis of the biological properties of classical and hypermucoid strains of K.pneumoniae isolated from patients with community-acquired pneumonia (CAP): characterization of their sensitivity to antibacterial drugs, Klebsiella bacteriophage, and a disinfectant (polyhexamethylene guanidine hydrochloride), as well as assessment of the strains’ virulence in the model of experimental infection in white mice.Material and methods. 56 strains of Klebsiella isolated in diagnostic quantities from patients’ sputum samples were studied. Species identification of cultures was carried out using bacteriological and mass spectrometric methods. The sensitivity of bacteria to antibiotics, bacteriophage, and the disinfectant was determined in accordance with regulatory documents.Results. In the course of the study, 243 gram-negative cultures were isolated, of which 30% were bacteria of the genus Klebsiella spp. An analysis of their species composition showed that K.pneumoniae occupied a dominant place in the structure. Based on colony morphology, Burri-Gins smear staining, and a positive string test, 14 strains with a hypermucoid phenotype were identified. These strains differed from the classic K.pneumoniae strain by the presence of a thicker capsule in smears, virulence in white mice (DCL≤103 mc), and increased resistance to commercial Klebsiella bacteriophage. At the same time, they were characterized by a wider spectrum of sensitivity to antibiotics. There were no significant differences in sensitivity to the disinfectant in strains of both morphotypes.Conclusion. The results obtained demonstrated the important role of K.pneumoniae in the etiological structure of CAP pathogens.

Comparative Analysis of the Pathogen Structure in Patients with Community-Acquired and Nosocomial Pneumonia in Medical Organizations of the Rostov, Tyumen Regions and Khabarovsk Territory at the Current Stage of a New Coronavirus Infection Pandemic

The aim of the study was to conduct a comparative analysis of the spectrum and antibiotic resistance of secondary pneumonia pathogens isolated in the territories of the Rostov, Tyumen Regions and Khabarovsk Territory against the background of a new coronavirus infection pandemic.Materials and methods. We investigated sputum samples from coronavirus-positive and coronavirus-negative patients with community-acquired pneumonia from medical organizations using bacteriological method, PCR mass spectrometry.Results and discussion. The study of the etiological structure of secondary pneumonia agents isolated from patients in medical organizations of the Southern, Ural and Far Eastern Federal Districts has revealed that the dominant cultures in SARS‑CoV‑2 “+” and SARS‑CoV‑2 “–” patients were yeast and yeast-like fungi. It has been found that under diversity of isolated fungi, Candida albicans species prevailed. The bacterial microflora is represented by a variety of gram-positive and gram-negative bacteria, of which Staphylococcus aureus and Klebsiella pneumoniaе were most often present in sputum. It has also been established that even before hospitalization of patients, community-acquired pneumonia could be caused by microorganisms of the ESKAPE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniaе, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.), which are usually considered as polyantibiotic-resistant pathogens of nosocomial infections. Moreover, in coronavirus-positive patients with secondary community-acquired pneumonia, those pathogens were isolated 2–3 times more frequently than in coronavirus-negative ones. Assessment of sensitivity/resistance of isolated strains to antibacterial drugs has revealed a general trend: the majority of the strains, regardless of the type, were characterized by a narrow spectrum of sensitivity, having 3 or more markers of antibiotic resistance. This confirms the necessity and expediency of microbiological support of the patient during the entire infectious process. The most adequate drugs of choice, providing activity against 60–70 % of strains of the Enterobacteriaceae family, are amikacin and cefoperazone/sulbactam.

Prevalence of subclinical mastitis, associated risk factors and antimicrobial susceptibility pattern of bacteria isolated from milk of dairy cattle in Kajiado Central sub-county, Kenya.

Literature is scarce on the occurrence of bovine mastitis and antimicrobial resistance among dairy animals kept by pastoralists in the Kenya. A cross-sectional study was carried out to investigate the prevalence and risk factors of subclinical mastitis (SCM) and evaluate the antibiotic sensitivity of bacteria isolated from dairy cattle kept by farmers in Kajiado Central sub-county, Kenya. A total of 202 lactating cows from 40 farms were sampled. Milk from the cows was screened for SCM using the California mastitis test, and the bacteria present in the milk samples were determined using standard bacteriological methods. The sensitivity of the isolated coagulase-negative staphylococci (CNS) and Staphylococcus aureus against antibiotics was tested using the Kirby-Bauer disk diffusion method. The prevalence of SCM at quarter- and cow-level was 31.7% and 53%, respectively. The prevalence of SCM was significantly higher (p<0.05) in exotic breeds of cattle and those kept under an extensive system of production. A total of 19 bacterial species were isolated with the majority being CNS (40.1%), S. aureus (15.8%) and Micrococcus spp. (10.4%). S. aureus isolates showed varied resistance to the tested antibiotics with the highest resistance being against ceftazidime (75%), amoxycillin (50%) and streptomycin (46.9%). Several S. aureus isolates were resistant to oxacillin (34.4%) and cefoxitin (12.5%). CNSs were more resistant against ceftazidime (79.1%), amoxycillin (34.6%) and oxacillin (32.1%). Majority (92%-100%) of the Staphylococcus spp. were highly sensitive to ciprofloxacin a fluoroquinolone and augmentin. The high prevalence of SCM and bacteria resistant to antibiotics shows a need for animal health professionals and farmers to develop strategies for the management of mastitis and antibiotic resistance in dairy cows in the study area.

Assessment of the Effectiveness of Using Magnoimmunosorbents for the Selective Concentration of Anthrax Agent Spores

The aim of the study was to assess the effectiveness of the developed anthrax magnoimmunosorbents (MIS) for the selective concentration of Bacillus anthracis spores and to increase the sensitivity of anthrax agent detection techniques, including when testing soil samples.Materials and methods. We used 10 vaccine strains of B. anthracis and 30 strains of closely related bacilli of the genus Bacillus (B. cereus – 15, B. thuringiensis – 10, B. megaterium – 5) with typical species properties. The work was performed on three experimental batches of magnoimmunosorbents. DNA extraction and PCR setting was carried out in compliance with the instructions for reagent panel for B. anthracis DNA detection “ApliSens Bacillus anthracis-FRT”.Results and discussion. It is shown that when using MIS, the sensitivity of the cultural method is increased by at least 7 times (taking into account the possibility of sorption of 1–10 or more spores on a sorbent particle). The sensitivity of the PCR method is improved by 10 times and amounts to 50 B. anthracis spores per 1 ml for the samples concentrated with the help of MIS. The sensitivity of the bacteriological method using MIS increases by a factor of 7.5 when testing the artificially contaminated with B. anthracis soil samples. Hence, application of the developed MIS makes it possible to significantly enhance the sensitivity of anthrax agent detection methods and can be considered as an effective means of sample preparation for the investigation of environmental objects (soil).

Some aspects of nasopharyngeal microflora in children with psoriasis

Objective — to study the state of nasopharyngeal microflora in children with psoriasis depending on the clinical features of dermatosis and determine antibiotic susceptibility of isolated microorganisms. Materials and methods. The isolation of nasopharyngeal microorganisms and determination of their antibiotic susceptibility were performed in 57 children with psoriasis aged 4 to 17 using the bacteriological method (culture). Microbial growth was assessed according to four degrees of growth. Susceptibility to antibacterial drugs was evaluated based on the following criteria: sensitive, moderately resistant, and resistant. Study materials were statistically processed using the StatTech v. 3.1 software. Results and discussion. Most often, Staphylococcus aureus (S. aureus) associations were cultured from the nasopharynx (56.15 %) of children with psoriasis — in 12.28 % of cases, Enterococcus faecium (Ent. faecium) — in 21.05 %. Statistically significant differences in children with psoriasis were found between the colony growth rates of Ent. faecium and S. aureus (p &lt; 0.001): massive microbial growth of Ent. faecium prevailed (64.91 %), and S. aureus was not isolated at all in the majority of children (29.82 %). Ent. faecium growth rates were statistically significantly different (p = 0.016) depending on psoriasis severity according to PASI: massive growth of microorganisms at PASI &gt; 10 was found in 87.5 % of cases, and at PASI ≤ 10 — in 54.2 % of cases. The proportion of Ent. faecium resistant to antimicrobial drugs (43.18 ± 7.47) % was found to be more than two times higher than that of S. aureus (20.00 ± 6.32) %. The susceptibility testing for each microorganism individually showed that enterococci were 28 to 100 % susceptible to antibacterial drugs, while staphylococci had 42 to 100 % susceptibility. Statistically significant differences were found between the percentage of sensitivity of Ent. faecium to antibacterial drugs between the group of children with the first diagnosis of psoriasis and the group of children with its recurrences (p = 0.005), as well as by the duration of the disease (p &lt; 0.001). Conclusions. The results of the study of children with psoriasis showed that there were bacterial associations isolated from the nasopharynx in the most significant number of cases with the predominance of Ent. faecium, which had massive growth in almost 65 % of cases and affected psoriasis severity according to PASI (OR 5.923; 95 % CI 1.387—25.300). The proportion of Ent. faecium resistant to antibiotics was higher than of S. aureus (p &lt; 0.05), but with increasing duration of psoriasis, Ent. faecium susceptibility to antimicrobials increased (r = 0.439; p = 0.004). In general, S. aureus isolated from the nasopharynx of children with psoriasis had more pronounced antibiotic susceptibility than Ent. faecium.

Epizootiological and Epidemiological Situation on Leptospirosis in the Russian Federation over the Period of 2013–2022 and the Forecast for 2023

The aim of the work was to analyze the epizootic and epidemiological situation on leptospirosis in the territory of the Russian Federation in 2013–2022 and to forecast its development in 2023. The long-term dynamics of leptospirosis incidence in the Russian Federation tends to decrease. Cases were registered in 58 constituent entities of the Russian Federation, in all federal districts. The highest incidence rates were noted in the North-Western Federal District. When studying the material from small mammals using bacteriological, immunological and molecular-biological methods, Leptospira circulation was detected in 52 entities of the Russian Federation, in all federal districts. In 2023, sporadic cases of infection are to be expected in the territories of the North-Western Federal District, the Central Federal District, and the Southern Federal District; imported cases of infection from countries with subequatorial and equatorial climates are not excluded.

Review on the Epidemiology and Public Health Importance of Camel Tuberculosis

Camel (Camelus dromedarius), Camels are an important livestock in Ethiopia and are raised in Somali, Afar, and Oromia (Kereyu, Borena, and Guji). They are used for various purposes such as transport, drafting power, ploughing land, festivals, and rivalry as in dashing. The pastoral community utilizes their products such as milk and meat. However, they are also, susceptible to various diseases which can affect their health. One of the major diseases that affect camels is tuberculosis. Camel tuberculosis is a persistent disease that is characterized by the improvement of granulomas inside the breathing tract and associated lymph nodes. The mycobacteria are discharged from those granulomas and might contaminate different prone animals Camel tuberculosis is a chronic disease that is characterized by the development of granulomas in the respiratory tract and related lymph nodes. The mycobacteria are discharged from these granulomas and can contaminate other susceptible animals. Camel tuberculosis is a chronic disease that affects the respiratory system and the lymph nodes of the infected animals. It can also pose a public health risk in areas where people consume raw camel milk or have close contact with camels. The disease is caused by different types of mycobacteria, such as Mycobacterium caprae, Mycobacterium tuberculosis, and Mycobacterium bovis. The diagnosis of camel tuberculosis can be based on clinical signs, history, tuberculin skin tests, post-mortem examinations, bacteriological and molecular methods. Public awareness and control measures are needed to reduce the transmission of the disease and its economic impact.

DEVELOPMENT OF THE TECHNOLOGY OF DEEP CULTIVATION OF A TRANSFORMED B. SUBTILIS STRAIN

Deep cultivation of recombinant cultures in bioreactors makes it possible to accumulate a large number of specific antigens used in the production of immunobiological preparations and diagnostic test systems in a short period of time, taking into account stimulating and limiting growth factors. The main purpose of the research was to determine the influence of the factors of seed concentration, pH level and the level of dissolved oxygen on deep cultivation, with the control of accumulation kinetics by turbidimetry. During the research, it was found that the most optimal inoculation concentration is 1 million cells/cm3 with a maximum final concentration of B. subtilis 4x106 cells/cm3, the most optimal pH level is a dynamic mode from 8.0 to 6.0 with a maximum final concentration of 5x106 cells/cm3, the most optimal level of dissolved oxygen is stationary mode with a minimum oxygen level of 25%, with a maximum final concentration of 4x106 cells/cm3.Bacteriological, technological and biochemical methods were used in the work to develop the modes of deep cultivation of the transformed B. subtilis strain with control of pH parameters and the level of dissolved oxygen.The scientific significance of the research is based on the development of deep cultivation, with an assessment of external growth factors and their influence on the kinetics of accumulation of the transformed B. subtilis strain, which will provide effective conditions for the growth and production of microorganisms. The practical significance of the research is based on the possibility of optimizing the biotechnological stages of the cultivation of the transformed B. subtilis strain, which will positively affect the efficiency of submerged cultivation as the main method for the accumulation of specific antigens.

Isolation of a rare pathogen &lt;i&gt;Chryseobacterium indologenes&lt;/i&gt; in patients with pneumonia

Introduction. Chryseobacterium indologenes bacteria are rarely detected, poorly studied opportunistic bacteria of the nonfermenting gram-negative bacilli group that are characterized by high intrinsic and acquired multidrug resistance and involvement in healthcare associated infections.Aim. To analyze detection rate of C. indologenes strains from various biomaterials obtained from patients with pneumonia, as well as levels of pathogen drug resistance to antimicrobial drugs.Materials and methods. The research included 1740 clinical samples obtained from patients with pneumonia hospitalized in in-patient facilities of the Khabarovsk city during the COVID-19 pandemic. Clinical samples included 1305 respiratory samples (744 sputum samples and 561 nasopharyngeal swabs), 435 samples of autopsy material (lung tissue) obtained from patients with a fatal outcome of the disease. The study was performed via classical bacteriological method. Pathogens were identified with Vitek 2 bacteriological analyzer and Vitek MS mass spectrometer. Sensitivity to antimicrobial drugs was determined by disk-diffusion method and in a bacteriological analyzer.Results. A comparative analysis of detection of the entire nonfermenting gram-negative bacteria (NFGNB) group and C. indologenes from various clinical samples was carried out. Detection rate of NFGNB equaled 17.3%. NFGNB were isolated from autopsy material most often ‒ in 31.7% of cases, less often ‒ from sputum and nasopharyngeal smears (13.8% and 10.7%, respectively). C. indologenes was detected in 0.5% of the cases (8 isolates out of 1740 samples). Isolation of this pathogen was most frequent in nasopharyngeal smears – in 1.1% (6 cases out of 561 samples), least frequent ‒ from sputum, which totaled 0.3% (2 isolates from 744 samples). The detection rate of the pathogen in respiratory samples equaled 0.6% (8 isolates out of 1305 samples). This pathogen was not found in the autopsy material. Carbapenem-resistant variants of C. indologenes were isolated in 75% (6 cases out of 8) of the cases.Conclusion. Further intensifying of diagnostic methods will result in more frequent detection of these unusual pathogens, clinical significance of which needs more justification.

Asymptomatic gastrointestinal carriage of multidrug-resistant carbapenemase-producing Enterobacteriaceae among children under five years in a Kenyan hospital

ObjectivesAsymptomatic gastrointestinal carriage of carbapenem-resistant Enterobacteriaceae (CRE) is a threat to global health in developing countries with inadequate safe drinking water, poor hygiene, and weak antimicrobial stewardship; however, epidemiological data to guide CRE infection prevention and control is limited in these settings. We assessed asymptomatic CRE and carbapenem-producing Enterobacteriaceae (CPE) fecal carriage rates and associated risk factors among hospitalized children aged under 5 years. MethodsWe adopted a cross-sectional study at Mama Lucy Kibaki Hospital in Nairobi-City County, Kenya, between June and September 2022. We collected demographic and clinical characteristics using a structured questionnaire and clinical reports and analyzed stool/rectal swab samples by standard and automated bacteriological methods. ResultsAsymptomatic CRE and CPE fecal carriage rate was 2.25% (6/267), with six isolates recovered, predominated by Escherichia coli (33.33%) and Enterobacter cloacae subsp dissolvens (33.33%). Third-generation cephalosporin and ciprofloxacin resistance were highest in Citrobacter farmer and E. cloacae subsp cloacae. All CRE and CPE were multidrug-resistant, and except E. cloacae subsp cloacae, were 100% colistin-resistant. ConclusionsAsymptomatic gastrointestinal carriage of multidrug-resistant-CRE among hospitalized children under 5 years, presents a substantial public health threat. This calls for continuous surveillance including molecular characterization of isolates, to inform infection prevention and antimicrobial stewardship adherence in line with local and global plans on AMR.

Molecular and biological Study of Colibacillosis Susceptibility in broiler

This study was aimed to Detection of virulence genes(rfbE) It was carried in College of Veterinary Medicine at Baghdad university, During a time span of November (2021) to March (2022). (100) samples were collected from diarrheal chicken . 100 chicks were used in the present study, One day old chicks were weighed (Average of chick weight was 160 gm±2) then transferred to the poultry house. Chicks in wards that were divided by wood barriers. Diagnosis of bacterial colonies were first done by the bacteriological methods. From (100) fecal samples of diarrheal cases of chicken , 90(90%) isolates were Escherichia coli.In the present study using vitek 2 compact system and Api 20 Escherichia coli. for the identification isolated bacteria. PCR technique was used to Detection of virulence genes(rfbE) ,The result for rfbE gene in this study was showed that this gene was detected in(60) isolates of (90) samples of chicken observed carried the rfbE gene which represent (67%).

&lt;i&gt;Helicobacter pylori&lt;/i&gt; Persistence in Patients with Peptic Ulcer Disease and Chronic Gastritis in the Treatment with Antibiotics and Acid-Suppressive Agents

Background. Chronic gastritides still have no division into forms according to the data of the genotyping of Helicobacter pylori isolates. Monitoring the results of anti-Helicobacter pylori therapy cannot be limited to eradication, by taking into account the data on Helicobacter pylori detection in healthy individuals. Is increased acid production associated with the presence of virulent or nonvirulent strains?Objective. To search for the predominance of cagA or any allelic combination of vacA in the isolates from patients with mild and severe chronic gastritis and from those with peptic ulcer disease. To track the time course of changes in the disappearance and repeated gastric colonization with different Helicobacter pylori strains in patients with peptic ulcer disease immediately after anti-Helicobacter pylori therapy and 1.5–2 months later. To confirm or refute the fact that the increased acid production is associated with the presence of virulent Helicobacter pylori strains rather than with the persistence of nonvirulent strains.Methods. The Helicobacter pylori isolates from the gastric biopsy specimens taken from patients with chronic gastritis and peptic ulcer disease were genotyped for cagA and allelic combinations of vacA. The strains from patients who had received anti-Helicobacter pylori therapy were genotyped using the variable number tandem repeat (VNTR) method. Patients with an exacerbation of peptic ulcer disease and those wth an exacerbation of chronic pancreatitis with concomitant gastritis were compared from the level of acid production and from the belonging of Helicobacter pylori to virulent strains.Results. Patients with mild chronic gastritis were observed to have an abundance of nonvirulent allelic combination of vacAs2m2, which differed significantly from its frequency in both the patients with severe chronic gastritis and those with peptic ulcer disease. In the latter patients, the differences between the background bacteriological pattern and the pattern observed after 1.5–2 months had a high degree of significance; in these follow-up periods, in contrast to the control immediately after therapy, the number of eradications increased and the number of cagA-containing strains decreased due to the change of strains. Acid production was significantly higher in the patients with peptic ulcer disease than in those with chronic pancreatits and concomitant gastritis; the former patients show a statistically significant predominance of virulent strains; the latter patients had nonvirulent strains (at the same rate of Helicobacter pylori detection).Conclusion. Filling the blank spots in the study of helicobacteriosis remains relevant due to the progress of genetic methods in bacteriology and to the introduction of novel approaches to assessing the results of anti-Helicobacter pylori therapy.

SCREENING FOR OXACILLIN RESISTANT SALMONELLA SPECIES ISOLATED FROM PALMS OF SOME PRIMARY SCHOOL PUPILS WITHIN KADUNA SOUTH LGA, NIGERIA

Salmonella species is the causative agent of typhoid fever which is a disease characterized by high mortality and morbidity worldwide. This study was carried out to screen oxacillin resistant- Salmonella species isolated from palms of some primary school pupils and food vendors within Kaduna South LGA, Nigeria. Three hundred (300) swab samples were collected from pupils and food vendors of the three selected primary schools as the sampling location. Salmonella enterica was isolated and identified using standard bacteriological methods. Isolates were susceptible to ciprofloxacin 28 (70.0%), Augmentin® 25 (65.0%) amoxicillin 25 (62.5%) and vancomycin 22 (55.0%) but showed resistance to oxacillin 9 (22.5%), ampicillin 12 (30.0%), chloramphenicol 15 (37.5%) and tetracycline 16 (40.0%). Antibiotics that exhibited intermediate susceptibility were methicillin 8 (20.0%) and gentamicin 11 (27.5%). Ciprofloxacin was the most effective antimicrobial agent against Salmonella enterica. The presence of oxacillin-resistant Salmonella enterica was detected via Polymerase Chain Reaction (PCR) and agarose gel electrophoresis. The study recommends that regular monitoring of antibiotic susceptibility pattern and good hygiene practices such as hand washing with soap and water and the use of alcoholic based hand sanitizers should be encouraged.