Resident Microflora Research Articles

Роль пищевых волокон в коррекции нарушений микробиоценоза кишечника на фоне антибактериальной терапии

Antibiotic-associated gut microbiocenosis disorders develop on average in every third patient in the form of both mild disorders and severe life-threatening conditions. The negative impact of antibacterial agents on the gut microbiota causes a decrease in the number and species diversity of microorganisms that produce butyric acid. Low concentrations of butyric acid can often induce inflammatory and atrophic processes of the gut mucosa, the water-electrolyte balance regulation disorders and, as a result, the gut motility and functions disorders. Long-term gut microbiocenosis disorders themselves can cause the development of the disease and in future, they become factors of its progression, thereby launching a whole cascade of new pathological processes. The article analyzes a representative clinical data published in authoritative international magazines from 1990 to 2020, indicating the effectiveness of the dietary fiber (DF) management for the correction of gut microbiocenosis disorders caused by antibacterial drugs intake. Partially hydrolyzed guar gum, registered in our country under the trade name “OptiFiber”, due to its effects such as the gut regulation, abdominal pain reduction, flatus and bloating reduction, increase the resident gut microflora amount and its metabolic activity, helps to restore and maintain of the gut eubiosis. Conclusion. “OptiFiber” can be administered for a long-term intake in order to increase the effectiveness of antibiotic therapy for various infectious diseases, as well as for the correction and prevention of gut microbiocenosis disorders associated with antibacterial drugs intake

PSXVI-14 Study of the antagonistic properties of C. utilis in relation to the resident microflora of the animal organism

Abstract C. utilis yeast is widely used in industry for the production of fodder protein, amino acids, vitamins, and enzymes. We have studied the antimicrobial activity of C. utilis CY 302 against such collection strains as E. coli 3912/41, S. flexneri 1a 8516, S. enterica serovar Typhimurium 79, S. epidermidis ATCC 14990, S. aureus ATCC 25923, Proteus vulgaris HX 19 222, L. casei subsp. Rhamnosus ATCC 7469, B. breve ATCC 15701, C. albicans ATCC 10231 by using the agar blocks method and by measuring the inhibition growth zone (IGZ, mm) accompanied by the target cells death zone. The yeast strain was grown on Sabouraud’s medium at 28±1 °C for 3 days, and the test cultures were grown on differential diagnostic media for each bacterial species (HiMedia Laboratories Pvt. Ltd, India) at 35±2 °C overnight. The analyses were done separately for each culture. A weak antimicrobial effect was observed with respect to L. casei and B. breve, the IGZ was 5.0 and 4.0 mm, respectively. The halos of inhibition against pathogenic bacteria were 7 mm for E. coli, 5 mm for S. typhimurium, 5 mm for P. vulgaris, 6 mm for S. fiexneri, 6 mm for S. aureus, and 5 mm for S. epidermidis. After incubation there was noted the absence of DZZR in the wild C. utilis CY 302 strain in relation to the C. albicans ATCC 10231 strain from the collection. As a result of the studies the antagonistic potential of the yeast isolate C. utilis CY 302 was revealed. The work was supported by the Ministry of Science and Higher Education of Russia.

Umbilical microflora and pediatric surgery.

In pediatric surgery, the umbilicus with humid environment prone to bacterial colonization has become the most common site of entrance into the peritoneum. However, the umbilical flora in children has never been reported. This study aimed to describe the characteristics of umbilical microflora in children before antiseptic skin preparation. We prospectively reviewed all children (age, ≤ 15years) undergoing surgical procedures using umbilical access between April 2020 and June 2021. Before antiseptic skin preparation, culture swabs were taken from the umbilicus. Data on age, sex, and surgical procedure as well as microflora results and clinical findings were analyzed. Overall, data on 123 children aged between 9days and 15years (median: 3years) were obtained. In the umbilicus, the most frequent colonizing bacteria were coagulase-negative Staphylococcus species and Corynebacterium. The isolation of intestinal bacteria from children aged ≤ 3years was significantly increased (P = 0.03). The results of the multidrug resistance test revealed that the bacteria from the umbilicus exhibited a high frequency of cefazolin (CEZ) resistance (46.1%). No postoperative surgical site infection was recorded in our study. This prospective study is the first report to investigate the umbilical microflora in over 100 children. In this study, a large spectrum of both resident and transient microflora was cultured from the umbilicus. This umbilical microflora was similar to previous reports of adult microflora except in children aged ≤ 3years. Our data suggest that in children ≤ 3years, preoperative and postoperative antibiotics should be chosen by considering CEZ resistance and intestinal bacteria. The result of umbilical microflora would be useful to select the antibiotics for treatment of surgical site infection (SSI), and the culture swabs from the umbilicus before the operation for the children at high risk for SSI are highly recommended.

Novel insight into physicochemical and flavor formation in koumiss based on microbial metabolic network

Koumiss, naturally fermented mare’s milk, has a unique flavor that is affected by its resident microflora. In this study, we analyzed time-dependent changes in the microbiota and volatile metabolite profiles of koumiss using high-throughput sequencing and liquid–liquid extraction–gas chromatography–mass spectrometry. The dominant microrganisms in mare milk were Enterobacter and Rhodotorula, which were gradually suppressed, then Lactobacillus and Dekkera became the core microorganisms of koumiss. 17 FAA and 52 volatile organic compounds (VOCs) were identified during koumiss fermentation, including 19 odor active compounds and 15 differential metabolites of VOCs, mainly acids and esters, and their anabolism showed significant positive correlations with Lactobacillus and Dekkera. The microbial metabolic network showed that Lactobacillus and Dekkera were identified as the most important flavor-producing microbiota due to their significantly positive correlation with the formation of 33 and 28 flavor metabolites, respectively. These microorganisms produce enzymes that catalyze a series of metabolic pathways that generate VOCs, including carbohydrate metabolism, amino acid metabolism, and fatty acid biosynthesis. Our findings provide new guidance for the screening and application of flavor-producing microorganisms inkoumiss.

Fecal Microbiota Functional Gene Effects Related to Single-Dose Antibiotic Treatment of Travelers' Diarrhea.

Travelers' diarrhea (TD) is common among military personnel deployed to tropical and subtropical regions. It remains unclear how TD and subsequent antibiotic treatment impact the resident microflora within the gut, especially given increased prevalence of antibiotic resistance among enteric pathogens and acquisition of multidrug-resistant organisms. We examined functional properties of the fecal microflora in response to TD, along with subsequent antibiotic treatment. Fecal samples from US and UK military service members deployed to Djibouti, Kenya, and Honduras who presented with acute watery diarrhea were collected. A sample was collected at acute presentation to the clinic (day 0, before antibiotics), as well as 7 and/or 21 days following a single dose of antibiotics (azithromycin [500 mg], levofloxacin [500 mg], or rifaximin [1650 mg], all with loperamide). Each stool sample underwent culture and TaqMan reverse transcription polymerase chain reaction analyses for pathogen and antibiotic resistance gene detection. Purified DNA from each sample was analyzed using the HumiChip3.1 functional gene array. In total, 108 day 1 samples, 50 day 7 samples, and 94 day 21 samples were available for analysis from 119 subjects. Geographic location and disease severity were associated with distinct functional compositions of fecal samples. There were no overt functional differences between pre- and postantibiotic treatment samples, nor was there increased acquisition of antibiotic resistance determinants for any of the antibiotic regimens. These results indicate that single-dose antibiotic regimens may not drastically alter the functional or antibiotic resistance composition of fecal microflora, which should inform clinical practice guidelines and antimicrobial stewardship. NCT01618591.

Effect of commonly used cosmetic preservatives on skin resident microflora dynamics

Human skin is populated by various microorganisms, the so-called microbiota, such as bacteria, viruses, yeasts, fungi, and archaea. The skin microbiota is in constant contact with the surrounding environment which can alter its eubiotic state. Recently it has been also observed that the application of cosmetic products can alter the balance of the skin microbiota. This effect may be attributed to many factors including the residual activity of the preservatives on the skin. In the present work, we studied the effect of eleven preservatives commonly found in cosmetic products on Propionibacterium acnes, Staphylococcus epidermidis, and Staphylococcus aureus in vitro using 3D skin models and culture-dependent methods. Also, the effect on Histone deacetylase 3 (HDAC3) has been investigated. Among tested combinations, three resulted as the best suitable for restoring a pre-existing dysbiosis since they act moderately inhibiting C. acnes and strongly S. aureus without simultaneously inhibiting the growth of S. epidermidis. The other four combinations resulted as the best suitable for use in topical products for skin and scalp in which it is necessary to preserve the eubiosis of the microbiota. Some of the tested were also able to increase HDAC3 expression. Taking together these data highlight the role of preservatives of skin resident microflora dynamics and could provide a reference for correctly choice preservatives and dosage in cosmetic formulations to preserve or restore homeostasis of skin microbiota.

MODERN MICROBIOLOGICAL TECHNOLOGIES IN DIAGNOSIS OF MYCOSES IN OTORHINOLARYNGOLOGICAL PRACTICE

The review article highlights the latest approaches to solve one of the important problems of modern otorhinolaryngology, the diagnosis of opportunistic mycoses of the upper respiratory tract and ear. Opportunistic mycoses of the ENT organs in recent decades have posed a significant challenge for modern clinical medicine not only in Ukraine but also throughout the world. According to the scientific literature, the share of fungal lesions of the ear and upper respiratory tract in the structure of chronic inflammation of these organs makes up 22.1%. The main causative agents of mycotic lesions of the ENT organs are opportunistic fungi of the genera Aspergillus, Рenicillium, Mucor and yeast-like fungi of the genus Сandida, which are characterized by a low level of pathogenicity and are a part of the resident microflora of the macroorganism. Classical methods of microbiological diagnosis have certain limitations in the identification of micromycetes. Therefore, the use of more reliable, fast and accurate methods in clinical practice will contribute to the timely and effective treatment of fungal diseases of the ENT organs. This review presents an analysis of modern microbiological diagnostic technologies, such as biochemical detection of microorganisms using identification plate test systems (ARI RapID, CrystalTM), semi-automatic and automatic microbiological identification using analyzers VITEK, VITEK 2, Walk Away. Identification of fungal pathogens can also be performed by performing direct protein profiling using mass spectrometry, polymerase chain reaction and sequencing. These methods with the highest level of reliability enable to identify the pathogen, as well as to assess its sensitivity to chemotherapeutic drugs. The combination of classical and modern microbiological technologies should become a standard for the diagnosis of fungal diseases, including the upper respiratory tract and ear.

1206. Association of Aging, Frailty and Place of Residence with Skin, Oral and Gut Microbiome Characteristics and Pathogenicity Reservoirs

BackgroundDespite their elevated risk for morbidity and mortality from infections, the microbiota of older adults remain understudied. While colonization resistance from resident microflora is a promising means to prevent infections, little is known about pathogenicity reservoirs and colonization resistance in this vulnerable population. Here we study the skin, oral, and gut microbiome dynamics of older adults in both community and Skilled Nursing Facility (SNF) settings, investigating relationships between age, frailty, environment, microbiota, and pathogenicity reservoirs.MethodsWe conducted a longitudinal metagenome survey of 47 adults age 65+ years of age; 22 residents of 3 different SNFs and 25 community dwelling individuals. We performed metagenomic whole genome shotgun sequencing on stool, oral, and skin samples from 8 sites, 1421 total. To correlate clinical and behavioral variables, we measured frailty, collected medical records, and interviewed participants on diet and lifestyle. We also draw comparisons with previous younger cohorts.Results• Compared to younger adults, the skin microbiota of older adults was characterized by ◦ High heterogeneity ◦ Decreased stability over time, suggesting increased susceptibility to colonization and pathogenicity ◦ Compositional differences including significantly lower levels of Cutibacterium acnes, with reciprocal increases in Staphylococci, Corynebacteria, and Malassezia• In older adults, Frailty (Rockwood) was found to have linear correlation with relative abundance of species relevant to infection risk including acnes, staphylococci, streptococci, E. coli, Akkermansia mucinophila, and Enterococcus faecalis.• The skin, oral, and gut microbiota of SNF residents had substantially elevated virulence factor and antibiotic resistance genes.ConclusionTo the best of our knowledge, this is largest report to date of the skin metagenome in older adults. We demonstrate distinct and significant differences between cohorts with clinically relevant implications. We believe these results may inform infection control and prevention by increasing our understanding of colonization resistance and pathogenicity reservoirs, as well as advance our knowledge of the relationship between aging, the microbiome, and infections.Disclosures All Authors: No reported disclosures

Sulfated vizantin causes detachment of biofilms composed mainly of the genus Streptococcus without affecting bacterial growth and viability

BackgroundSulfated vizantin, a recently developed immunostimulant, has also been found to exert antibiofilm properties. It acts not as a bactericide, but as a detachment-promoting agent by reducing the biofilm structural stability. This study aimed to investigate the mechanism underlying this activity and its species specificity using two distinct ex vivo oral biofilm models derived from human saliva.ResultsThe biofilm, composed mainly of the genus Streptococcus and containing 50 μM of sulfated vizantin, detached significantly from its basal surface with rotation at 500 rpm for only 15 s, even when 0.2% sucrose was supplied. Expression analyses for genes associated with biofilm formation and bacterial adhesion following identification of the Streptococcus species, revealed that a variety of Streptococcus species in a cariogenic biofilm showed downregulation of genes encoding glucosyltransferases involved in the biosynthesis of water-soluble glucan. The expression of some genes encoding surface proteins was also downregulated. Of the two quorum sensing systems involved in the genus Streptococcus, the expression of luxS in three species, Streptococcus oralis, Streptococcus gordonii, and Streptococcus mutans, was significantly downregulated in the presence of 50 μM sulfated vizantin. Biofilm detachment may be facilitated by the reduced structural stability due to these modulations. As a non-specific reaction, 50 μM sulfated vizantin decreased cell surface hydrophobicity by binding to the cell surface, resulting in reduced bacterial adherence.ConclusionSulfated vizantin may be a candidate for a new antibiofilm strategy targeting the biofilm matrix while preserving the resident microflora.

ОСОБЕННОСТИ КЛИНИКО-ЛАБОРАТОРНЫХ ПОКАЗАТЕЛЕЙ У ПАЦИЕНТОВ С РЕЦИДИВИРУЮЩИМИ АФТАМИ ПОЛОСТИ РТА НА ФОНЕ КИСЛОТОЗАВИСИМЫХ ЗАБОЛЕВАНИЙ ЖЕЛУДОЧНО-КИШЕЧНОГО ТРАКТА

Subject. Some features of the clinical picture, hygienic and microbiological state of the oral cavity, acidity and free-radical oxidation of the oral fluid in patients with recurrent aphthates on the background of acid-dependent diseases of the gastrointestinal tract are considered.
 The aim is to assess the relationship between clinical and laboratory parameters in patients with recurrent oral aphthae associated with acid-dependent diseases of the gastrointestinal tract.
 Methodology. A comprehensive clinical and laboratory examination of 58 young people with revealed recurrent aphthae of the oral cavity against the background of acid-dependent diseases of the gastrointestinal tract was carried out. The level of oral hygiene was assessed using the ONI-s index, the state of free-radical oxidation of the oral fluid - the "Chemilumimer-003" device, individual representatives of the normal flora were identified by inoculating the contents from the surface of aphthous elements on special differential diagnostic media of the HiCrome series. The acidity of the oral fluid was determined using a universal indicator litmus paper pH-1-14, short-term intragastric pHmetry was performed directly during fibrogastroduodenoscopy.
 Results. In 46.4 % of those examined, recurrent aphthae were found on the oral mucosa. In persons with chronic gastritis and duodenitis, the frequency of recurrences once a year was 37.9 %, 2-3 times a year ― 62.0, with the presence of chronic pancreatitis against the background of hyperacid gastric secretion ― 44.8 and 55.2, respectively. In patients with recurrent aphthae, a low level of oral hygiene, a violation of its microbiome, and a change in the acidity of the oral fluid were revealed.
 Conclusions. The presence of recurrent aphthae complicates individual oral hygiene, because accompanied by severe pain and paresthetic symptoms. This is manifested by an increase in the ONI-S index and a disturbance in the state of the normobiome in the form of an increase in the growth of resident microflora against the background of changes in the acidity of the oral fluid. Chemiluminescence indices correlate with the hygienic state of the oral cavity, qualitative changes in the normobiota and pH of the oral fluid in the studied clinical groups.

Metatranscriptome-based investigation of flavor-producing core microbiota in different fermentation stages of dajiang, a traditional fermented soybean paste of Northeast China

Dajiang, or naturally fermented soybean paste, has a unique flavor that is influenced by the resident microflora. However, the association between flavor and the core microbiota is unclear. Recent advances in RNA sequencing have identified genes that are actively expressed in complex microbial communities. To this end, we analyzed the time-dependent changes in the microbiota and the metabolite profiles of Dajiang using metatranscriptome sequencing, HS-SPME-GC-MS and amino acid analysis identified 10 volatile compounds that contribute to the development of soybean paste flavor. Further analysis of the correlation between the active microorganisms and the physicochemical characteristics and flavor substances in soybean paste indicated that Lactobacillus and Tetragenococcus were the core genera affecting chromaticity and flavor. These microorganisms produce enzymes that catalyze a series of metabolic pathways that generate flavor substances. Our findings provide new insights into the role of the microbiota in the development of flavor in fermented foods.

Comparative Transcriptomics of Shiga Toxin-Producing and Commensal Escherichia coli and Cytokine Responses in Colonic Epithelial Cell Culture Infections.

Ingestion of Shiga toxin-producing Escherichia coli (STEC) can result in a range of illness severity from asymptomatic to hemorrhagic colitis and death; thus risk assessment of STEC strains for human pathogenicity is important in the area of food safety. Illness severity depends in part on the combination of virulence genes carried in the genome, which can vary between strains even of identical serotype. To better understand how core genes are regulated differently among strains and to identify possible novel STEC virulence gene candidates that could be added to the risk assessment repertoire, we used comparative transcriptomics to investigate global gene expression differences between two STEC strains associated with severe illness and a commensal E. coli strain during in vitro intestinal epithelial cell (IEC) infections. Additionally, we compared a wide array of concomitant cytokine levels produced by the IECs. The cytokine expression levels were examined for a pattern representing STEC pathogenicity; however, while one STEC strain appeared to elicit a proinflammatory response, infection by the other strain produced a pattern comparable to the commensal E. coli. This result may be explained by the significant differences in gene content and expression observed between the STEC strains. RNA-Seq analysis revealed considerable disparity in expression of genes in the arginine and tryptophan biosynthesis/import pathways between the STEC strains and the commensal E. coli strain, highlighting the important role some amino acids play in STEC colonization and survival. Contrasting differential expression patterns were observed for genes involved in respiration among the three strains suggesting that metabolic diversity is a strategy utilized to compete with resident microflora for successful colonization. Similar temporal expression results for known and putative virulence genes were observed in the STEC strains, revealing strategies used for survival prior to and after initial adherence to IECs. Additionally, three genes encoding hypothetical proteins located in mobile genetic elements were, after interrogation of a large set of E. coli genomes, determined to likely represent novel STEC virulence factors.

Bidirectional alterations in antibiotics susceptibility in Staphylococcus aureus\u2014Pseudomonas aeruginosa dual-species biofilm

In mixed infections, the bacterial susceptibility differs significantly compared to monocultures of bacteria, and generally the concentrations of antibiotics required for the treatment increases drastically. For S. aureus and P. aeruginosa dual species biofilms, it has been numerously reported that P. aeruginosa decreases S. aureus susceptibility to a broad range of antibiotics, including beta-lactams, glycopeptides, aminoglycosides, macrolides, while sensitizes to quinolones via secretion of various metabolites. Here we show that S. aureus also modulates the susceptibility of P. aeruginosa to antibiotics in mixed cultures. Thus, S. aureus—P. aeruginosa consortium was characterized by tenfold increase in susceptibility to ciprofloxacin and aminoglycosides compared to monocultures. The same effect could be also achieved by the addition of cell-free culture of S. aureus to P. aeruginosa biofilm. Moreover, similar increase in antibiotics efficacy could be observed following addition of S. aureus suspension to the P. aeruginosa mature biofilm, compared to P. aeruginosa monoculture, and vice versa. These findings open promising perspectives to increase the antimicrobial treatment efficacy of the wounds infected with nosocomial pathogens by the transplantation of the skin residential microflora.

598 Identification of a human skin commensal bacterium that selectively kills cutibacterium acnes

Cutibacterium acnes (C. acnes) is one of the most common bacterial species on human skin and can promote acne vulgaris. Most current therapeutics for acne are antibiotics that fail to discriminate C. acnes from the other skin resident microflora, many of which have important roles in health. Such approaches have significant drawbacks due to poor antimicrobial activity on the skin surface and emergence of antibiotic resistance. The microbiome represents a vast resource for drug discovery as its members engage in constant conflict to outcompete one another by deploying diverse strategies for survival. Our aim was to identify and develop new antimicrobials from the skin microbiome, as a biotherapy for acne. To do this, we conducted a functional screen of coagulase-negative staphylococci collected from swabs of the face and arms of healthy individuals and recorded their antimicrobial activity against C. acnes. Amongst the antimicrobial hits, we found that Staphylococcus capitis (S. capitis E12) strain exhibited potent and selective activity against all acne- and health-associated C. acnes strains tested, but not against several other important commensal bacteria. HPLC and LC/MS analyses of S. capitis E12 supernatant identified the antimicrobial metabolites as four distinct amphipathic phenol soluble modulin β (PSMβ) peptides. These PSMβ peptides were found to act synergistically and were not toxic to human keratinocytes. Formulation and application of a purified S. capitis E12 extract onto ex vivo pig skin colonized with C. acnes, resulted in a 1-log decrease (p> 0.02) in CFU after 24 h. More impressively, the extract applied topically onto SKH1 mouse skin colonized with C. acnes, resulted in a 3-log decrease (p>0.01) in CFU after 48 h. Overall, these data show how some peptides from the skin microbiome can be highly selective against C. acnes and highlights their potential application as a biotherapy for acne vulgaris.

Shifts in Composition and Activity of Oral Biofilms After Fluoride Exposure.

Oral diseases are biofilm-mediated diseases caused by imbalances in the ecology of resident microflora. Among them, dental caries (tooth decay) is considered the most common disease worldwide, and toothbrushing, which physically eliminates the oral biofilm, is the most widespread prevention strategy. Although it is well established that fluoride increases enamel resistance to acidic pH and promotes tooth remineralization, its effect on the biofilm bacterial communities' composition and metabolism is not fully understood. We have grown in vitro oral biofilms and used 16S rRNA Illumina sequencing to study the effect of fluoride on DNA- and RNA-based bacterial populations. In addition, a metatranscriptomic approach has also been performed, in which total RNA has been sequenced to study gene expression profiles in the presence/absence of 500 ppm sodium fluoride. Our data show a lower pH drop and a clear shift in total and metabolically active bacterial composition after fluoride exposure. Streptococcus oralis was the species most affected, with a 10-fold reduction in both DNA and RNA samples, whereas Rothia mucilaginosa underwent an 8-fold increase in the DNA and S. salivarius a 4- and 5-fold increase in the RNA and DNA samples, respectively. The metatranscriptomes indicated that fluoride exposure induced a dramatic shutdown of sugar metabolism, including significant under-expression of different sugar transporters, fucosidases, and a pyruvate oxidase, among others. The reduction in saccharolytic organisms and the inhibition of sugar fermentation pathways by fluoride may therefore be considered instrumental for the beneficial effect of fluoride-containing oral hygiene products.

Helicobacter pylori Infection and Gastric Microbiota.

ABSTRACTOwing to its strong acid production, the stomach was known to be a bacteria-free organ for many years. On the other hand, the presence of Helicobacter pylori (H. pylori) and other acid-resistant microbiota that are to persist in the stomach challenged this. It is now recognized that the existence of H. pylori and non-H. pylori species have been linked to the improvement of gastric disease; despite this, there is little published data on the interaction of gastric bacterial flora and the resultant effect on gastric health. The stomach has a unique microbiota including five major phyla, such as Firmicutes, Proteobacteria, Actinobacteria, Fusobacteria and Bacteroidetes. These phyla are identified in both H. pylori-infected and uninfected persons. The resident gastric microflora may mediate the role of H. pylori in the gastric diseases. This article aims to review previous studies that examine the impact of H. pylori infection and the effect of resident gastric microbiota on gut health and disease conditions.How to cite this articleOzbey G, Sproston E, Hanafiah A. Helicobacter pylori Infection and Gastric Microbiota. Euroasian J Hepato-Gastroenterol 2020;10(1):36–41.

Effectiveness Of Gamma Radiation Technology For Abating Postharvest Losses Of Banana (Musa Sapientum) By Targeting The Residential Microflora

Banana (Musa spp.), member of the family Musaceae, is the perennial herb mostly grown in the tropical areas situated at latitude 30° above and below the equator covering the warmer countries of the world. This regional belt has wide seasonal variability with respect to temperature and rainfall. In the international trade, Banana holds 22% share in world fresh fruit production and is ranked as second amongst the most important fruits after citrus. Gamma irradiation of pre climacteric bananas at optimal doses, which vary with the variety of banana, delays the ripening process. For Pakistan, radiation processing technology supports remarkable potential not only for local food securities but also for greater exports earnings in agriculture sector. Farmers could be stimulated for greater production of fragile fruits like banana when putrefaction could be prevented by appropriate preservation practices resulting in increased and balanced consumption.

Bioinoculants play a significant role in shaping the rhizospheric microbial community: a field study with Cajanus cajan.

The present study is an attempt to understand the impact of bioinoculants, Azotobacter chroococcum(A), Bacillus megaterium(B), Pseudomonas fluorescens(P), on (a) soil and plant nutrient status, (b) total resident and active bacterial communities, and (c) genes and transcripts involved innitrogen cycle,during cultivation of Cajanus cajan. In terms of available macro- and micro-nutrients, triple inoculation of the bioinoculants (ABP) competed well with chemical fertilizer (CF). Their 'non-target' effects were assessed in terms of the abundance and activity of the resident bacterial community by employing denaturing gradient gel electrophoresis (DGGE). The resident bacterial community (16S rRNA gene) was stable, while the active fraction (16S rRNA transcripts) was influenced (in terms of abundance) by the treatments. Quantification of the genes and transcripts involved in N cycle by qPCR revealed an increase in the transcripts of nifH in the soil treated with ABP over CF, with an enhancement of 3.36- and 1.57- fold at flowering and maturity stages of plant growth, respectively. The bioinoculants shaped the resident microflora towards a more beneficial community, which helped in increasing soil N turnover and hence, soil fertility as a whole.

HEMATOLOGICAL AND BACTERIOLOGICAL PREDICTORS OF OCCUPATIONALLY AND INDUSTRIAL CONDITIONED DISEASES IN MEDICAL WORKERS

Introduction. Medical workers in the process of labor are exposed to a complex of adverse factors: neuro-emotional, physical stress of a chemical, physical and biological nature. A long stay in a hospital environment with medical workers can lead to the formation of various diseases of microbiological etiology or the carriage of pathogenic and conditionally pathogenic flora and the development of dysbiosis. Objective. Give hematological and bacteriological characteristics of the health status of medical workers. Material and methods. The results of laboratory studies of blood and mucous membranes of the upper respiratory tract of medical workers performed during the performing occupational medical examinations are analyzed. Results. Hematological indices were found to be characterized by an increased hemoglobin content in one-fourth of the examined workers, as well as lymphocytosis, eosinophilia, cholesterolemia. The microflora of the mucosa of the upper respiratory tract is mainly represented by Staphylococcus aureus, Candida albicans, Streptococcus pyogenes in 65.2% of the workers; the remaining 34.8% were revealed to have a resident microflora. With the experience of work, the frequency of insemination increases and doubles by 10 years of the work. The structure of microflora is also changing. In the nosological form of occupational morbidity, allergic diseases prevail and account for 60.8%, tuberculosis - 26.0%, and hepatitis - 8,6%. Conclusion. All these changes in the body are the basis of pathogenetic manifestations in the formation of allergic and pathogenic pathologies, which make up to 70.0% of all occupational morbidity.

P081 The structure of the proteomic profile and gut microbiota in patients with primary immunodeficiency, common variable immunodeficiency and inflammatory bowel disease

Abstract Background Infringement of intestinal microbiota in patients with primary immunodeficiency (PID), common variable immunodeficiency (CVID) and ulcerative colitis (UC) continues to be studied. The aim of the research is a comprehensive study of the structure of gut microbiota and mucosal protein profile (MPP) in patients with PID, CVID and UC in different phases of the disease course. Methods The research included 15 patients with PID, 22 CVID and 46 UC (28 with relapse and 18 with persistent remission of the disease), control group—20 healthy volunteers. Estimation of gut microbiota was performed by bacteriological seeding, hydrogen breath test (HBT) with lactulose. Content of short-chain fatty acids (SCFAs) and microbial lipid markers (MLM) in faeces and colon mucosa was determined by gas–liquid chromatographic and gas chromatography–mass spectrometry (GC–MS). MPP was based on isoelectric focusing techniques (SDS–PAGE, 2DGE). Mass spectrograms were obtained using MALDI-TOF-MS/MS (Bruker, USA). Results HBT showed a 4.5-, 8- and 11-fold increase in hydrogen production on a 150th-minute study. SCFA showed a 6-, 9- and 11-fold decrease in propionic and butyric acids, mainly in patients with PID and UC: 0.2 + 0.1, 0.14 + 0.03 and 0.04 + 0.02 mg/g, respectively. Microbiological analysis showed a decrease in titers of E. coli, bifido- and lactobacilli on average 4.6 + 0.8 Lg. The conditionally pathogenic microflora was represented by E. coli, lactose-negative strains (n = 51,105/g) and haemolytic strains (n = 34,104/g, Proteus spp. (n = 48,105/g), Staphylococcus spp. (n = 29,105–6/g), Candida (n = 51,104–6/g) and Clostridium spp. (n = 38,104/g) in stool culture. MLN GS-MS results showed a 5.5-, 3- and 6-fold increase in total bacterial load in patients with PID, CVID and UC, respectively, which was represented by resident anaerobic microflora: Streptococcus mutants, Clostridium difficile, Candida albicans. Results of MPP (detection rate of 75% or more) of the colon mucosa in patients with PID were detected: 1, 2, 4 okkludin, kininogen 1, interleukin-1B, interleukin 8, B2-glycoprotein, heat shock protein 27, in patients with CVID—translational elongation factor, apolipoprotein C-III. In patients with UC—NF-kB, alipoprotein C-III, TNF-α, interleukin-2 and 8 were presented. Conclusion In patients with PID, CVID and UC markers of colon excess growth and significant decrease in SCFA are recorded during the relapse period. MPP in patients with PID, CVID and UC was characterised by proteins characteristic of inflammation, apoptosis, proliferation and proteins reflecting the activity of the autoimmune process. Patients with UC in the remission have recovery trend of gut microbiota, production of SCFA, disappearance of specific components of altered MPP.