Conjunctivities of newborns

The article is focused on the peculiarities of the clinical course of separate forms of neonatal conjunctivitis, depending on the etiological factor. It was found that more often the disease refers to nosocomial eye infection and bacterial nature. We performed the bacteriological analysis of the contents of the conjunctival cavity of 50 newborn patients being treated in the neonatal department. Our analysis indicated the leading role of gram-positive bacteria Staph. epidermidis (59.7%) and Staph. aureus (21.7%) in the development of the inflammatory process. The share of other types of pathogens, including gram-negative minor and various pathogens, is from 0.54% to 3.2%. The cause of nosocomial infection is considered to be the pathogen that circulates in the department and acquires the features of a hospital strain. These are consistent with the results of similar studies conducted by both domestic and foreign clinicians, which are also presented in the article. Particular attention is paid to the causative agents of intrauterine infections that are dangerous for the anterior section: gonococcus, chlamydia, herpes simplex, etc. These agents often cause serious diseases in newborns (gonoblenorrhea, ophthalmic chlamydiosis, and ophthalmic herpes), in which the cornea and vision are often affected. The article highlights the measures of primary prevention of intrapartum infection of the ocular surface in newborns, adopted in Russia. The paper presents modern approaches to selecting drugs for local antibacterial therapy of neonatal conjunctivitis, considering age restrictions for their use. Methods of laboratory diagnostics and their validity for the etiology of conjunctivitis have been described in detail. For example, the bacteriological method (inoculations in various culture media) is considered a reference (specificity 100%). The culture medium can be used to isolate bacteria, chlamydia, and mycoplasma, which allows getting clear results even with a minimal amount of microflora

The Prevalence of Uropathogenic Escherichia coli Strains among Outpatients with Urinary Tract Infection in Zakho Hospitals-Zakho City, Duhok Province/ Iraq

This study involved the prevalence of uropathogenic Escherichia coli (UPEC) among outpatients of UTI attending three major hospitals in Zakho city. Four hundred urine samples were collected from patients of UTI of both sexes and different ages (≤ 1 year to over 50 years), during the period from July 2018 until January 2019. All urine samples were analyzed by conventional bacteriological method for the presence of Escherichia coli (E. coli), while molecular method was used for the presence of species-specific uidA gene in the isolated E. coli. Out of 400 samples, 141 (35.25%) were infected with UPEC from enrolled patients. The rate was higher in females than males (90.78% vs 9.22%), respectively. In both sexes, the age group 41-50 years in both sexes showed the highest rate (46.67%) of infection, and statistically this rate of infection was significant (p< 0.013) among both sexes and various age groups. Furthermore, in all ages, married patients showed slightly higher prevalence than un-married one (38% vs 32.5%), but this difference was statistically non-significant (p>0.05%). The rate of UTI was higher among urban inhabitants (40.56%) than others. During the months of the year, the peak (90.48%) in both sexes was during December while the lowest rates (13.64%) was during January.

Molecular Characterization of Extended Spectrum Beta – Lactamase Producing Escherichia Coli Isolated from Pregnant Women with Urinary Tract Infections Attending Ante–Natal Clinics in Ilorin Metropolis

Background: The emergence of multidrug- resistance Enterobacteriaceae especially in E. coli bacteria associated with Urinary Tract Infections (UTIs) in pregnancy is a serious menace globally posing health challenges and confounding successful empirical treatment as well as increasing pregnancy – related complications.Objectives: The aim of this study is to determine the phenotypic and genotypic characteristics of Extended Spectrum Beta – Lactamases (ESBLs) producing E. coli (ESBLs – EC) isolates in pregnant women attending ante – natal clinics within Ilorin - Kwara State, Nigeria.Materials and methods: A total of 53 non - repeated E. coli isolates from urine samples of pregnant women were presumptively identified using standard bacteriological method and confirmed by commercially available Microgen® Identification Kits. Phenotypic detection of ESBLs was determined using antibiotics susceptibility test and double disc synergy Method for screening and confirmation respectively. Polymerase Chain Reaction (PCR) was further used for the genotypic detection of ESBLs genes.Results: A total 88.67% (47/53) of E. coli exhibited resistance to the cephalosporins of which aztreonam was the highest (75.47%) and the least was cefpodoxime (35.84%) while 85.10% were confirmed positive for ESBL production. The genotypic detection showed the most occurring genotype was blaTEM (50%) blaOXA (27.7%), blaGES (22.5%), blaSHV (15%), blaCTXM and blaVEB (7.5%) while sixty – four (64%) of isolates co – harbored two or more gene. BlaTEM and blaOXA were dominant.Conclusion: This study showed high resistance of E. coli to the third generation cephalosporins harboring different ESBL genes which increases UTIs complexity and limit therapeutic options in pregnancy. Therefore, continuous monitoring of resistance in E. coli, effective appraisal of antibiotic control policies and rational use of antibiotics is therefore encouraged.

Promising Diagnostic Markers of Colon Cancer

Background. The incidence of colon cancer over the past decade has been growing markedly in the Russian Federation, with about 50 % cases detected at stagesIII–IV of the disease, when a clear clinical picture of the disease appears. In this regard, the search for new methods for early diagnosis of RTK is undoubtedly relevant.Objective. To determine the standard composition of the aerobic parietal colon microbiota and the level of cytokines (chemokines and growth factors) in patients with cancer of the left half of the colon and to assess the possibility of using these data in the diagnosis of the tumor process.Materials and methods. Blood tests were performed on the day of the study using two test systems (BioLegend): multiplex kit for determining growth factors, chemokine multiplex kit. The composition of the intestinal microbiota was determined in colon biopsy specimens by the bacteriological method using the standard test systems StaphyTest, StreptoTest, and EnteroTest.Results. There is an increase in the number of Clostridiumspp. and a decrease in Bifidobacteriumspp., E. coli in the colon during the transformation of a healthy person’s mucosa into a malignant tumor (p < 0.05); a clear tendency was revealed for both an increase (EGF, HGF, M-CSF, PDGF-AA, PDGF-BB, IP-10) and a decrease (MCP-1, RANTES) of the level of chemokines and growth factors under colon cancer conditions. In addition to general quantitative changes in the intestinal microbiota, the level of the investigated substances, a statistically significant dependence was established on the sex, age of the patient, as well as the degree of differentiation and form of tumor growth.Conclusion. It was established that changes in the quantitative composition of the intestinal microbiota, the level of some biologically active substances that occur precisely in the conditions of colon cancer, can be interconnected and interdependent, and also serve as an additional diagnostic marker in the detection of a malignant tumor.

The Biochemical Properties Peculiarities of Representatives of Periodontal Pocket Microbiota during Generalized Periodontitis

Chronic inflammatory-dystrophic lesions of periodontal tissues are widespread in Ukraine and the world diseases with complex etiology and pathogenesis. The purpose of the study: to establish the biochemical and cultural properties of microorganisms in periodontal pockets in patients with acute generalized periodontitis using a bacteriological method. Materials and methods: the contents of the periodontal pocket were taken from 10 patients with subsequent seeding on nutrient and differential diagnostic media in the bacteriological laboratory. Cultural, as well as the biochemical, and enzymatic properties of bacterial isolates were identified. Results: Pathogenic and opportunistic microorganisms (aerobes and facultative anaerobes), which had a wide range of biochemical properties (additional factors of colonization and pathogenic action) were identified. Purulent microflora was detected in 70.00% of cases. Isolates of Streptococcus, Staphylococcus and P. aeruginosa were predominantly β-hemolytic (except for E. faecalis and S. pneumoniae). The identified bacteria of the intestinal group and the Klebsiella family were lactose- and glucose-positive, K. oxytoca was the most enzymatically active microorganism. Conditionally pathogenic microorganisms of periodontal tissues and oral cavity, as well as the human body as a whole, belong to the facultative microbiota. It is shown that the level of inflammatory process varies with the number of associates of opportunistic pathogens isolated from the inflammatory process. In titers of 102–104 CFU / ml, representatives of the facultative microbiota, as a rule, do not show their pathogenic qualities. At the same time, biochemical properties are essential in the potential ability of opportunistic pathogens to acquire signs of pathogenicity. The titer of bacteria, their hemolytic and proteolytic properties significantly affect the level of virulence of microorganisms. The ability to hemolysis of erythrocytes, found in most isolates, allows to attribute the isolated strains to potentially pathogenic. It is established that the persistence of opportunistic pathogens with a high level of virulence determines the course and intensity of the inflammatory process of periodontal tissues. Conclusions: The study of the role of pathogenic and opportunistic microorganisms in the development and maintenance of long-term inflammation of periodontal tissues is a topical issue in medical science and is necessary for the verification of treatment regimens and compliance with infection control

SPECTRUM OF PATHOGENS CAUSING PURULENT INFLAMMATORY DISEASES IN SURGICAL PATIENTS

This article for the first time describes the frequency rate of detecting various surgical infections depending on the number of pathogens and the spectrum of the microorganisms found in surgical patients of the Poltava Regional Clinic Hospital, Ukraine. The objectives include the identification of the etiological spectrum of pathogens causing purulent inflammatory diseases in inpatients of the surgical unit. Materials and methods. On the basis of the bacteriological laboratory, we studied 121 biological samples taken from the patients of the surgical unit. Microflora isolation was performed on nutrient media by bacteriological method. Microorganisms isolated from various biosubstrates were identified using API biochemical test systems (BioMerieux, France). Based on the findings obtained, the occurrence rate of certain pathogens and their associations was calculated (%). Results and discussion. The study demonstrated the prevalence of mono-infections, which make up 67%, caused by Staphylococci, mainly S. Aureus in the surgical patients. In general, gram-negative bacteria are prevalent, among which Klebsiella and Acinetobacter are found out as the most prevalent pathogens; among Gram-positive microorganisms, Enterococci were identified more often in association with other bacteria. Conclusions. The study has shown among the causative agent resulting in diseases requiring surgical treatment, gram-negative bacteria predominate, among which Klebsiella and Acinetobacter are the most often detected. Among gram-positive cocci, Enterococci are typically detected in the association with other bacteria; most often, this pathogen is isolated in monoculture, mainly represented by staphylococci.

Modern methods for species identification of thermophilic bacteria of Campylobacter species

Emergent thermophilic Campylobacter hepaticus is the causative agent of Spotty Liver Disease (SLD) in laying hens. C. hepaticus is difficult to cultivate because commercial media for the isolation and cultivation of Campylobacter contain cefoperazone, which inhibits many isolates of the C. hepaticus species. Campylobacter was isolated using modified Preston broth, incubated at 37 °C under microaerophilic conditions for 7 days and then subcultured onto selective Preston agar, erythritol agar with Oxoid selective additives and 5–7% defibrinated horse blood. Commercial test systems (API Campy) were used for identification. The use of the classical bacteriological diagnostic method, which is considered the 'gold' standard, is limited due to the difficulties of cultivation. The identification of new Campylobacter species requires revision of phenotypic identification algorithms. Specific primers for the identification of new Campylobacter species also need to be developed. In our studies, using the KAM-BAC kit, we detected Campylobacter jejuni DNA in clinically healthy birds. Consequently, the carriage of Campylobacter is massive. 30 samples of test material were examined using the molecular-biological method, and 60 samples using the bacteriological method. Analyzing the results of Campylobacter detection, it should be noted that thermophilic Campylobacteria were isolated from 60 clinical samples by the bacteriological method in 5,0% (3 Campylobacter cultures), and from 30 samples by the molecular-biological method in 27,0% (8 positive samples). Based on the analysis of the study results, it is necessary to conduct an in-depth study of the natural sources of Campylobacter hepaticus distribution, virulence factors, pathogenesis and mechanisms of infections caused by these emergent pathogens. The most promising research in the study of the causative agents of Campylobacteriosis in birds will be based on the application of innovative genomic technologies based on multiplex polymerase chain reactions and genome sequencing of Campylobacter hepaticus.

DEVELOPMENT OF QUICK SCHEME IDENTIFICATION OF LISTERIA USING PHAGE BIOPREPARATION L.M 4 УЛГАУ

The causative agent Listeria monocytogenes causes listeriosis, a severe foodborne illness associated with high mortality. Rapid and sensitive methods are required to detect and identify this pathogen in the food production process. The article presents the results of research on the development of technological parameters for the production of biopreparation based on the bacteriophage L. m 4 UlSAU for quick identification of bacteria of the genus Listeria with its help. It was established that for the production of L. m 4 phage of UlSAU with maximum titers, the optimal parameters are: multiplicity of MOI 1 infection, cultivation temperature-28°C, incubation time of the phage/ culture system – 6 hours. Based on these results, we have offered technological scheme for the production of phage biopreparation L. m 4 UlSAU, which includes the following steps: confirmation of biological properties of indicator phage and increasing its titer (if necessary), verification and confirmation of biological properties of indicator culture, production of phage biopreparation and control of its indicators. Accelerated scheme for Listeria identification was offered with the help of prepared biological product based on L. m 4 bacteriophage of UlSAU. The scheme was tested on samples of chicken meat and minced meat semi finished products artificially contaminated with Listeria monocytogenes bacteria in concentrations 101-105 CFU/ml. It was established that the proposed scheme of listeria identification allows to reduce the duration of studies by 84 hours in comparison with the traditional bacteriological method and to detect listeria at a concentration of 100 CFU/ml (g) in 82 hours. The phagoidentification time at the pure culture typing stage is 18 hours.

Indirect hemagglutination reaction in ram infectious epididymitis for indication of Brucella ovis antigen in biomaterial

The reaction of indirect hemagglutination with an antibody erythrocyte diagnosticum is one of the promising methods for identifying the causative agent of infectious diseases or antigen. This preparation has not yet been developed for the diagnosis of infectious epididymitis of rams. The results of the preparation of an antibody erythrocyte diagnosticum in the reaction of indirect hemagglutination for the detection of the causative agent of infectious epididymitis in various biological material are presented in the article. As a result of scientific research, authors developed the method for obtaining an original Brucella ovis antibody diagnosticum for the reaction of indirect hemagglutination by sensitizing sheep erythrocytes with hyperimmune Brucella ovis serum using alizarin blue indicator of the epididymitic pathogen or antigen in biomaterial as the conjugate. Studies have shown the specificity and higher sensitivity of the indirect hemagglutination reaction with the antibody erythrocyte diagnosticum, compared with the antibody neutralization reaction and the bacteriological method, and its suitability for the indication of Brucella ovis antigen in biomaterial and environmental objects. It was also found that the reaction of indirect hemagglutination with antibody diagnosticum is much faster than the reaction of neutralization of antibodies and the bacteriological method. Research to improve the diagnosis of infectious ram epididymitis caused by Brucella ovis has been completed with the development of the antibody erythrocyte diagnosticum for the indirect hemagglutination reaction.The high specificity and activity of this preparationwas established by the authors. As a result of the studies carried out to test the diagnostic value of the antibody diagnosticum, a higher sensitivity of the indirect hemagglutination reaction using the new antibody diagnosticum, compared with the neutralization reaction of antibodies and the bacteriological method, and the suitability of using the diagnosticum for the indication of the Brucella ovis antigen in biomaterial, was established, which meets the requirements for express methods for detecting antigens in pathological material.

Electro-Optical Analysis of Cell Viability of Tularemia Microbe Vaccine Strain

Objective: to study the possibility of applying electro-optical analysis for the assessment of cell viability of tularemia microbe vaccine strain at different stages of experimental live tularemia vaccine production.Materials and methods. The research object was a cell culture of Francisella tularensis 15 NIIEG. Investigations were carried out at all stages of experimental live tularemia vaccine (ELTV) manufacturing according to an advanced technology: cultivation, concentrating, diafiltration, mixing with drying media, stabilization, and storage (two-year period of observation). Electro-optical analysis by the parameter “polarizability anisotropy” of bacterial cell was conducted with the help of EloTrace (EloSystems, Germany). Total concentration of cells was evaluated using density metering at 590 nm and spectrometry – at 650 nm. Viability was assessed through inoculation of plates with FT-agar.Results and discussion. The experiment has demonstrated that the change in polarizability anisotropy of the cell at the frequencies of 900 kHz and 2100 kHz, reflecting the state of cytoplasm and cytoplasmic membrane, respectively, is the earliest response to changes in vital indicators of bacterial culture in the process of cultivation. Thereby, the decrease in viability of F. tularensis cells occurrs well before the decrease in cell concentration. We have shown the preservation of viability of F. tularensis 15 NIIEG cells at all stages of experimental live tularemia vaccine production. Electro-optical analysis allows for registering the changes in vital parameters of microorganism cells in real-time mode, while the assessment of viability applying bacteriological method takes up to 5 days. Different stages of tularemia vaccine manufacturing have impact on the vital indicators of F. tularensis cells, and electro-optical analysis is a prospect method of control of such parameter as “Specific activity (the number of live microbial cells)”.