Genomic, metabolomic, and functional characterisation of beneficial properties of Pediococcus pentosaceus ST58, isolated from human oral cavity.

Bacteriocins produced by lactic acid bacteria are proteinaceous antibacterial metabolites that normally exhibit bactericidal or bacteriostatic activity against genetically closely related bacteria. In this work, the bacteriocinogenic potential of Pediococcus pentosaceus strain ST58, isolated from oral cavity of a healthy volunteer was evaluated. To better understand the biological role of this strain, its technological and safety traits were deeply investigated through a combined approach considering physiological, metabolomic and genomic properties. Three out of 14 colonies generating inhibition zones were confirmed to be bacteriocin producers and, according to repPCR and RAPD-PCR, differentiation assays, and 16S rRNA sequencing it was confirmed to be replicates of the same strain, identified as P. pentosaceus, named ST58. Based on multiple isolation of the same strain (P. pentosaceus ST58) over the 26 weeks in screening process for the potential bacteriocinogenic strains from the oral cavity of the same volunteer, strain ST58 can be considered a persistent component of oral cavity microbiota. Genomic analysis of P. pentosaceus ST58 revealed the presence of operons encoding for bacteriocins pediocin PA-1 and penocin A. The produced bacteriocin(s) inhibited the growth of Listeria monocytogenes, Enterococcus spp. and some Lactobacillus spp. used to determine the activity spectrum. The highest levels of production (6400 AU/ml) were recorded against L. monocytogenes strains after 24 h of incubation and the antimicrobial activity was inhibited after treatment of the cell-free supernatants with proteolytic enzymes. Noteworthy, P. pentosaceus ST58 also presented antifungal activity and key metabolites potentially involved in these properties were identified. Overall, this strain can be of great biotechnological interest towards the development of effective bio-preservation cultures as well as potential health promoting microbes.

[Forensic medical assessment of venous thromboembolic complications of mechanical injury of the lower limb after surgery]. / Sudebno-meditsinskaya otsenka venoznykh tromboembolicheskikh oslozhnenii mekhanicheskoi travmy nizhnei konechnosti posle operativnogo vmeshatel'stva.

In forensic medical practice, venous thromboembolic complications (VTEC) are relatively rare, due to hereditary and acquired factors. The issue of expert evaluation of the VTEC after the performed surgical intervention as an alleged defect in medical care causes discussion. The purpose of this publication is to demonstrate an expert case in the assessment of VTEC mechanical injury of the lower limb after surgery. The above case with the development of PATE after surgery clearly demonstrates the possibility of the appearance of a «medical case¼. The key to the correct expert assessment of the alleged defect of medical care during the forensic medical examination is not only a thorough and scrupulous study of medical documentation, but also a qualitatively performed forensic medical examination of the corpse.

Lactobacillus probiotics restore vaginal and gut microbiota of pregnant women with vaginal candidiasis.

The development of probiotics has now included the areas along the gut-vaginal axis. We thus aimed to investigate the effects of lactobacilli probiotic to modulate and restore vaginal and gut microbiota of pregnant women with vaginal candidiasis (VC). A randomised, double-blind and placebo-controlled study was performed in 78 pregnant women with VC. Patients were randomised to either the probiotic (SynForU-HerCare) or placebo which were administered at baseline and continued for 8-weeks (two capsules/day of 9.5 log cfu/capsule). Microbiota profiles were assessed at time points of weeks-0, 4 and 8 for high vaginal swab and faecal samples. Shannon diversity index showed that after 8-weeks amid VC, a shift in microbial community compositional changes occurred in the high vaginal region at both genus (P=0.025) and species (P=0.044) levels, where the administration of probiotic prevented such a shift. These changes were mainly attributed to a decreased in abundance of Lactobacillus (P=0.042) accompanied by increased abundance of Prevotella (P=0.002) and Atopobium (P=0.002) in the placebo group while the probiotic group remained unchanged over time. The administration of probiotics also prevented a reduced abundance of faecal phylum Firmicutes after 8-weeks as seen in the placebo group (P<0.0001), which also showed reduction at subsequent taxonomic levels of class, family, genera and species. VC has not only altered the microbiota of vagina regions but also gut microbiota profiles, causing lessening of gut microbiota that are crucial for gut nutrient availability, protection and immunity. The administration of lactobacilli probiotics has prevented such a shift, leading to better modulated gut and vaginal microenvironment amid VC. The study was registered at ClinicalTrials.gov: identifier number NCT03940612.

[Long-term inflammatory and neoplastic reaction of prostate tissues during its transurethral infection with uropathogens: evaluation of the results of animal model study].

INTRODUCTION: There is no convincing evidence of the persistence of acute or the development of chronic bacterial-induced prostatic inflammation in the long term when infected with various titers of the uropathogen. Along with this, controversial data are presented on the relationship between post-infectious chronic inflammation and neoplastic changes in prostate tissues. OBJECTIVE: To carry out, based on the experimental data: 1) assessment of the degree of bacterial contamination and the severity of histological changes in prostate tissues on the 60th follow-up day in case of transurethral infection with various uropathogens in titers of 102,3,5 CFU/ml; 2) fundamental comparative analysis between the indicators of the inoculated test-titer and microbial load with the severity of histological changes in prostate tissues; 3) verification of neoplastic transformations in the prostate tissues during a long-term persistent bacterial-induced inflammatory process. MATERIALS AND METHODS: Animal studies were conducted using FELASA protocols. Laboratory animals: 14 New Zealand rabbits. Tested uropathogens: aerobes - E. coli, S. haemolyticus, anaerobes - P. niger. Titers: 102,3,5 CFU/ml. Uropathogen inoculation technique: topical transurethral. RANDOMIZATION: all laboratory animals were divided into 5 groups according to the uropathogen (4 experimental, 1 control). Follow-up period: 60 days. Sacrification and autopsy of the animals were performed on day 60. Biopsies were taken from various parts of the prostate, as well as from the bladder neck and the edge of the membranous urethra. Cultural, histological and immunohistochemical (expression of p53 and Ki-67) studies of prostate tissues were conducted. Statistical data processing was performed using the GraphPad Prism 9.0 program (GraphPad Software Inc., Graphpad Holdings LLC, San Diego, CA, USA) applying descriptive and non-parametric statistics. RESULTS: Two individuals infected with S. haemolyticus + P. niger had a lethal outcome. The contamination of prostate tissue was determined in all cases of infection. In 88.9% of the cases, an increase in tissue microbial load was determined compared to the initial titer. Multivariate analysis of culture study values revealed the presence of intragroup differences in prostate contamination only between infection with E. coli 103 CFU/ml and E. coli 105 CFU/ml (p=0.006), as well as intergroup differences between infection with E. coli 105 CFU/ml and P. niger 105 CFU/ml (p=0.013). The histological study revealed moderate proliferative inflammation after inoculation with 102,3,5 CFU/ml in the E. coli and S. haemolyticus groups. In the case of S. haemolyticus, it was more pronounced due to the presence of persistent alterative lesion foci; in the P. niger group, mild proliferative transformations were observed in prostate tissues in all cases. The immunohistochemical study of changes determined p53 expression (10.0%) in some areas of the glandular epithelium of prostate glands (but without a positive internal control) only in case of infection with E. coli 105 CFU/ml. Areas of glandular epithelium with Ki-67 expression ( less or equal 25.0%) were visualized in all tested groups, mainly at titers of 103 and 105 CFU/ml, but the severity of proliferative activity was not high (1+). There were no foci of prostate tissue with simultaneous nuclear activity of p53 and Ki-67. CONCLUSION: Proliferative inflammation of different intensity in prostate tissues was observed after sixty days. Its severity was mainly determined by the type of infecting agent (S. haemolyticus > E. coli > P. niger) and was not dependent on the inoculated titer and the subsequent microbial load of prostate tissues. No areas of neoplastic transformation of prostate tissues were reliably identified in the case of a bacterial-induced inflammatory process in the estimated follow-up period.

Role of gastrointestinal microbial populations, a terra incognita of the human body in the management of intestinal bowel disease and metabolic disorders.

Intestinal bowel disease (IBD) is a chronic immune-mediated clinical condition that affects the gastrointestinal tract and is mediated by an inflammatory response. Although it has been extensively studied, the multifactorial aetiology of this disorder makes it difficult to fully understand all the involved mechanisms in its development and therefore its treatment. In recent years, the fundamental role played by the human microbiota in the pathogenesis of IBD has been emphasised. Microbial imbalances in the gut bacterial communities and a lower species diversity in patients suffering from inflammatory gastrointestinal disorders compared to healthy individuals have been reported as principal factors in the development of IBD. These served to support scientific arguments for the use of probiotic microorganisms in alternative approaches for the prevention and treatment of IBD. In a homeostatic environment, the presence of bacteria (including probiotics) on the intestinal epithelial surface activates a cascade of processes by which immune responses inhibited and thereby commensal organisms maintained. At the same time these processes may support activities against specific pathogenic bacteria. In dysbiosis, these underlying mechanisms will serve to provoke a proinflammatory response, that, in combination with the use of antibiotics and the genetic predisposition of the host, will culminate in the development of IBD. In this review, we summarised the main causes of IBD, the physiological mechanisms involved and the related bacterial groups most frequently associated with these processes. The intention was to enable a better understanding of the interaction between the intestinal microbiota and the host, and to suggest possibilities by which this knowledge can be useful for the development of new therapeutic treatments.

[Morphological and immunogistochemical characteristics of myocardial damage in miastenia].

The article presents a rare clinical case of a 45-year-old patient with generalized myasthenia with damages to the muscular apparatus of the extremities and the heart. A special feature of the case was myocardial damage evident as alterative-productive interstitial myocarditis with a peculiar immune phenotype of cell infiltrate, Cd3+, Cd4+, Cd8-, Cd68+. Furthermore, Cd68-expressing cells were presented by large macrophages with cytoplasmic granulation, which surrounded damaged cardiomyocytes. Around sites of cardiomyocyte alteration there were manifestations of neoangiogenesis with signs of Cd34 protein expression in thin-wall, capillary type blood vessels. These morphological and immunohistochemical changes in the myocardium supplement the concept of myasthenia morphogenesis.

Safety and beneficial properties of bacteriocinogenic Pediococcus acidilactici and Pediococcus pentosaceus isolated from silage.

The control of Listeria monocytogenes is a relevant goal for dairy products, a process that begins from the supply of feed and management of animals' health. In the present study, we evaluated the safety of two bacteriocinogenic Pediococcus strains and show that both can be considered as safe, based on their haemolytic activity, biogenic amine production and antibiotic resistance, all evaluated through phenotypical and biomolecular approaches. Both strains have shown potential as a producer of γ-aminobutiric acid (GABA) and carry an incomplete set of genes related to folate biosynthesis; both strains were able to adhere to Caco-2 cell lines with adhesion rates of 6·59% ± 3·73 and 0·84% ± 0·48. Laboratory prepared clover silage, inoculated with each bacteriocinogenic Pediococcus strain and contaminated with L. monocytogenes, proved the hypothesis for bioprotective effect of the tested strains, with the tested pathogen eliminated in the first 24 h of the experiment. These results indicate that evaluated strains can be potential beneficial candidates for application in silage production.

[Morphological and Molecular-Biological Changes in the Coronary Arteries after Stenting].

This review addresses morphological changes in coronary arteries following stenting, which result from damage to the vascular wall. These changes include 1) formation of a thrombus in the site of intimal injury; 2) inflammation; 3) proliferation and migration of smooth muscle cells; 4) formation of extracellular matrix. Each of these pathological processes has specific morpho-biological features. The review shows the role of von Willebrand factor in development of early thrombosis after intimal injury, which provokes activation of the inflammatory response followed by proliferation of smooth muscle cell that synthetize the extracellular matrix. These cellular and intercellular changes are based on overexpression of TGF-ß1 protein, which facilitates modulation of various types of smooth muscle cells, including contractile and secretory ones. Issues of fine regulation of cellular and intercellular interactions by apoptosis, activation of mTOR signaling molecules, and microRNA are still understudied. Dynamic changes in drug-coated stents during development of neoatherosclerosis and late thrombosis remain not elucidated. Current reports show that initial mechanisms triggering pathological regenerative and hyperplastic processes that result in coronary restenosis in the area of implanted stents may form early (first hours or days) after stenting. Most studies were performed on experimental rather than on autopsy material, which does not allow fully unbiased interpretation of obtained data. Studying dynamics of morphological and molecular changes in coronary arteries after stenting, including on autopsy material, will allow one to express an opinion on the risk of postoperative thrombosis and restenosis.

[Cystoscopic and morphological features of chronic recurrent papillomavirus cystitis].

INTRODUCTION: Chronic recurrent cystitis (CRC) is a common disease in the female population and a serious medical problem. There are not enough data about etiology of this desiase and effective treatment. OBJECTIVE: To increase the accuracy of the differential diagnosis of bacterial and papillomavirus chronic recurrent cystitis. MATERIALS AND METHODS: Analysis of endoscopic and morphological diagnostic methods of 118 patients with CRC, which, depending on the etiological factor, were divided into two groups. Group I (n=65) patients with CRC of HPV etiology and Group II (n=53) - CRC of bacterial etiology (E. coli). All patients were examined according to the EAU and RSU recommendations, and an endoscopic examination of the bladder (cystoscopy) was additionally included, followed by morphological examination of biopsy specimen of the urinary bladder. RESULTS: The endoscopic picture of bacterial CRC usually is caused by hyperemia of the mucous membrane of the bladder, hyperemia and injection of vessels with the participation of (small-puncture/diapedesic) hemorrhages without signs of proliferative changes. The endoscopic picture in papillomavirus CRC in all patients presented by focal or diffuse, slightly elevated, exophytic, often polypoid formations of a whitish or whitish-gray color, usually localized in the Leto triangle. Morphological analysis of biopsy tissue of the urinary bladder in patients with bacterial cystitis in the mucous and submucous layer showed edema, thickening of the urothelium, capillary abundance, inflammatory infiltration, represented by lymphocytes, plasma cells and leukocytes, among which neutrophilic macrophage leukocytes were predominant. Morphological analysis of the bladder mucosa in all examined patients with signs of HPV-related lesions showed a thickening of the urothelium, squamous metaplasia of the urothelium, chronic inflammation, microcirculatory vascular reaction and edema. An important distinctive pathomorphological sign of HPV within urinary bladder was the presence of koilocytic transformation of urothelium cells due to the cytopathic effect of the virus. CONCLUSIONS: Papillomavirus CR has characterized by an infectious-inflammatory process in the bladder paries, with lymphocytic-plasmatic infiltration and coylocytic transformation of the urothelium.

[Forensic expert evaluation problems of early and late complications of interventional methods of treating coronary heart disease (systematic literature review)].

This article focuses on current concepts of ischemic heart disease, its interventional treatment, pathomorphology of early and late postoperative complications, and forensic aspects in evaluation of restenosis of a stented blood vessel.

Bacteriocin production by Leuconostoc citreum ST110LD isolated from organic farm soil, a promising biopreservative.

AIMS: The objective of this study was to isolate a bacteriocin-producing strain and to characterize the expressed bacteriocin for the control of Listeria monocytogenes with aim of biopreservation application. METHODS AND RESULTS: Soil samples from a Korean organic farm were subjected to microbiological analysis for isolation of potential bacteriocinogenic LAB, based on a three-level approach, using L. monocytogenes ATCC 15313 as an indicator test micro-organism. From a total of 17 isolates with inhibitory potential, seven were confirmed to be bacteriocin producers. The selected isolates were differentiated based on their morphology, catalase reaction, sugar fermentation profile obtained by API50CHL and by RAPD-PCR generating two unique profiles. One of the isolates, ST110LD, a specific strong producer of anti-Listeria bacteriocins (12 800 AU ml-1 ) was identified as Leuconostoc citreum. The proteinaceous nature of the inhibitory compound produced by Leuc. citreum ST110LD was confirmed through treatment with pepsin and α-chymotrypsin. Bacteriocin activity was observed to be not affected by the presence of milk, NaCl, SDS, Tween 80 or glycerol. Bacteriocin ST110LD effectively inhibited the growth of exponentially growing L. monocytogenes ATCC 15313 during a 10-h incubation period in BHI at 37°C. In addition, this bacteriocin showed specific inhibition of only Listeria spp., but did not inhibit the growth of beneficial cultures included in the microbial test panel for assessment of the spectrum of activity. CONCLUSIONS: Leuconostoc citreum ST110LD was evaluated as safe bacterium strain, producing bacteriocin with high specificity against listerial and enterococcal species. Specificity of producer strain and expressed bacteriocin can be explored in biopreservation of different fermented food products or applied in biotherapy of antibiotic resistant listerial or enterococcal infections. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report of bacteriocin produced by Leuc. citreum strain with highly specific antimicrobial activity against Listeria sp. and Enterococcus sp.

[Are bacteriological and pathomorphological signs of bacterial prostatitis related between? (An experimental study)].

INTRODUCTION: In some cases, there is a dissociation between the severity of complaints, physical examination data, and levels of contamination of the biomaterial in the differential diagnosis of various categories of prostatitis (NIH-NIDDK, 1995). Patients note the presence of pronounced symptoms when verifying the threshold indicators of microbial load (104-105 CFU / ml) of prostate secretion or post-massage urine in a few observations. However, clinical manifestations and deviations in objective indicators are not so significant in some patients with higher titers of contamination. PURPOSE OF THE STUDY: To evaluate the relationship between the microbial load indices and the degree of pathomorphological changes in the prostate tissue during infection with the "reference" uropathogen E. coli in various titers under experimental conditions MATERIALS AND METHODS: Animal model was carried out using the FELASA protocols. Experimental individuals: 16 "New Zealand" rabbits, weight: 3580 [3480; 3695] (3300-4410), age: 25 [24; 26] (23-28) weeks. Uropathogen used: E. coli. Titers: 103 CFU / ml, 105 CFU / ml, 107 CFU / ml. Infection pathway: inoculation of the uropathogen was performed through urethra according to the modified technique of J.C. Nickel. Randomization: all lab animals were evenly divided into 4 groups of 4 animals, taking into account the initial titer of the uropathogen and the observation period - experimental groups 1 (103 CFU / ml), 2 (105 CFU / ml), 3 (107 CFU / ml); group 4 was control (Sol.NaCl 0.9%). Observation terms: 1, 3, 7 and 14 days, after which the animals were euthanized and dissected. 4 biopsies (1A-1D) for bacteriological (MacConkey agar - "HiMedia", India) and 2 biopsies (P1-P2) for morphological (Hematoxylin-eosin - "BlikMediklProduction", Russia; magn. 10x, 40x, 100x, 400x) studies were formed from various parts of the prostatic complex (prostate + proprostate + paraprostate). A three-point grading system was used for morphometric assessment of destructive changes in the prostate tissue in different groups. Analytical processing of the results was carried out using the software packages Microsoft 365 ("Microsoft", USA) and Statistica 10.2 ("StatSoft Inc.", USA) by methods of descriptive and nonparametric statistics. RESULTS: The maximum total absolute / median values of the seeding of biopsy specimens of the prostatic complex were revealed ( titer, 1A-1D) in group 1 and amounted to 76 lg CFU / ml / 5.00 [4.00; 5.25] lgCFU / ml, in group 2 defined as intermediate - 57 lgCFU / ml / 3.50 [3.00; 4.00] lgCFU / ml, and in group 3 were minimal - 48 lgCFU / ml / 3.00 [3.00; 3.25] lg CFU / ml according to the indicators of bacteriological research in the period from 1 to 14 days. Nevertheless, it was found that the cumulative maximum degree of histological changes in the prostate was recorded in group 2 ( p. - 84), in group 1 it was defined as intermediate ( p. - 68), and in group 3, it was the smallest ( p. - 64) according to the data of pathomorphological studies. When comparing daily changes in the seeding values and nominal indicators of inflammatory lesions in the prostate tissue, only in group 1, the trend graphs had a synchronous trend - an increase in microbial load led to the formation of more severe pathomorphological transformations on the day 7; in group 2, destructive changes were significant, and the rates of contamination, on the contrary, were minimal on the day 7; in group 3 a pronounced pathomorphological transformations in the tissues were noted on the day 3 and 7, but the microbial load of the tissues gradually decreased from the day 1. Also, differences in the increase in the "reactivity" of changes in the affected tissues were determined: in groups 1 and 3, a torpid reaction was revealed, accompanied by the development of moderate inflammatory changes on the day 1 and an "abrupt" increase in the intensity of destruction by the day 3; in group 2, in turn, a rapid reaction was determined, that characterized by the formation of a more pronounced destructive process already from the day 1. CONCLUSION: It was found that the parameters of contamination do not fully reflect the nature and severity of pathological changes in the prostate. Low titers of the uropathogen induce the development of an inflammatory process comparable in the severity of changes with that at higher concentrations. On the contrary, inoculation of obligate pathogenic titers does not always lead to a pronounced increase in microbial load but causes significant inflammatory changes in the prostate.

[Penile cavernous fibrosis: etiology, morphogenesis, erectile dysfunction].

Despite significant advances in andrology, the problem of penile cavernous fibrosis remains not fully understood. Many studies on the epidemiology of erectile dysfunction have been published, but consensus on the place and role of this pathology in the structure of sexual disorders has not yet been reached. The data obtained at different time intervals and in different geographical areas are strikingly different. Also, the role of organic disorders in the penis, including fibroplastic changes, in certain etiological factors has not been determined. In addition, the relationship between etiological factors and morphological changes in penile tissues is disputed due to the small amount of data obtained from the pathohistological study of human penis biopsies. This review is devoted to the systematization of epidemiological data and etiological factors of cavernous fibrosis, the definition of the relationship between them, the analysis of clinical and experimental studies, which study the relationship between the degree of severity of damaging agents and the formation of typical fibrogenic reactions.

[Forensic diagnosis of sudden death of young people based on morphological analysis of the cardiac conduction system]. / Sudebno-meditsinskaya diagnostika vnezapnoi smerti lits molodogo vozrasta na osnovanii morfologicheskogo analiza provodyashchei sistemy serdtsa.

The aim of study was to develop a comprehensive assessment of the central and peripheral parts of the nervous system with a determination of degree of severity of alterative changes in comparison with the condition of myocardial contractile apparatus. Own observations on modern methods of morphological study of the cardiac conduction system in the sudden death of young people are presented. Particular attention is paid to the technique of isolating and coloring the nervous structures of sympathetic and parasympathetic parts of the autonomic nervous system, including the medulla oblongata, sinus-atrial and atrial-ventricular nodes, atrioventricular bundle (the His bundle), Purkinje fibers. The features of morphological changes in the nerve structures of the heart, due to various etiological factors are highlighted.

[Renomedullary interstitial cell tumor: review of a rare clinical case].

Autopsy study of a 49-year-old patient demonstrates a rare kidney tumor that had a structure of a mesenchymal renomedullary interstitial tumor (RIO), which had specific characteristics. They include a predominance of the cellular component in the tumor, represented by cells with processes elements (fibroblasts, myofibroblasts), which were surrounded by collagen fibers of interstitium. Apparently, these cells can be involved in the regulation of renin and bicarbonates, the exchange of components of connective tissue (collagen fibers, proteoglycans) and hormones (estrogen, progesterone). Since these tumor cells regulate the synthesis and secretion of the substances, numerous clinical manifestations of RIO can be explained. Most likely, these include increased blood pressure, water-electrolyte disorders, hormonal disorders due to the presence of estrogen receptors, progesterone in the tumor tissue and metabolic disorders (diabetes mellitus). However, these assumptions require further clinical, morphological and immunohistochemical studies.

[Inguinal lymphadenitis due to invasion of Dirofilaria repens].

The paper presents the case of the right-sided inguinal inflammation of a lymph node as a result of invasion of Dirofilaria repens, the parasitic pathogen of subcutaneous dirofilariasis in animals of the canine family. The diagnosis was verified on the basis of the parallel application of morphological studies of cross sections of the nematode in histological samples and the molecular biological method polymerase chain reaction of scrapings of histological material. The localization of this helminth inside the cavities of the human body is extremely rare. Only isolated cases of atypical localization of D. repens are described: in the organs of the chest, cervical lymphatic node in the spermatic cord and epididymis, which led to pseudotumor formations that needed to be differentiated with neoplastic processes. This case is of great interest to experts of various fields (surgeons, oncologists, infectious disease specialists and pathologists), primarily in the differential diagnosis of malignant neoplasms of the lymphatic system.

Expression of coagulin A with low cytotoxic activity by Pediococcus pentosaceus ST65ACC isolated from raw milk cheese.

AIMS: We aimed to evaluate some specific conditions for growth of Pediococcus pentosaceus ST65ACC and its bacteriocin expression through ABC transporters; to purify the bacteriocin and determine its sequence; and to evaluate the cytotoxicity potential of the purified bacteriocin(s). METHODS AND RESULTS: The results presented for growth behaviour of P. pentosaceus ST65ACC showed that the bacterial growth was slightly influenced when cultured in MRS broth with different amounts of inoculum: 1, 2, 5 and 10%. The bacteriocin activity increased when 5 and 10% inocula were used. The carbon source (glucose) used in different amounts (1, 2, 3 or 4%) had no significant effect on growth and bacteriocin production. The studied strain P. pentosaceus ST65ACC was able to metabolize xylooligosaccharide (XOS) as the sole carbon source, resulting in the production of an antimicrobial peptide. The genes involved in the ABC transport system and sugar metabolism of P. pentosaceus ST65ACC were expressed at different levels. The bacteriocin produced by P. pentosaceus ST65ACC was partially purified by precipitation with ammonium sulphate (40% saturation), followed by reversed-phase liquid chromatography, resulting in the identification of an active bacteriocin. Tandem mass spectrometry was used to identify the partial sequence KYYGNGVTCGKHSCSVDWGK sharing high similarity to coagulin A. The semi-purified bacteriocin had low cytotoxicity based on estimated values for maximal nontoxic concentration (MNC) and cytotoxicity concentration (CC50 ). CONCLUSIONS: The bacteriocin produced by P. pentosaceus ST65ACC is similar to coagulin, with low cytotoxicity, strong antimicrobial activity and possible additional metabolite routes in the producer cell. In addition to MRS broth, bacteriocin was produced also in medium containing XOS (as the single carbon source). SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report of evaluation of the role of ABC transporters in the expression of bacteriocin by P. pentosaceus, cultured in MRS and XOS.

[Experimental evaluation of inflammatory process in the prostate caused by transurethral transmission of uropathogen in low counts].

BACKGROUND: According to the literature, bacterial count of uropathogens isolated from expressed prostate secretion and urine which is sufficient for a diagnosis of bacterial prostatitis I and II categories, remains contradictory. Undoubtedly, the identification of microorganisms from affected organ in high titers indicates the presence of a relevant infectious-inflammatory process. In turn, there is no consensus on the development of bacterial prostatitis at lower titers of uropathogens. Thus, the aim of our study was to identify and compare the potential features of the development and occurrence of an infectious inflammatory process in the prostate during the reproduction of bacterial prostatitis in an animal model using a low titer of causative uropathogens. MATERIALS AND METHODS: A total of 16 "New Zealand" mature male rabbits aged 24+/-2 weeks old with weight of 3.5+/-0.3 kg were examined. Inoculation was performed via transurethral route, according to the developed experimental technique. E. coli was used as bacterial agent with a count of 1 x 103 CFU/ml, 1 x 105 CFU/ml and 1 x 107 CFU/ml. All animals were randomized into 4 groups of 4 individuals depending on the titer of the inoculated microorganisms (groups 1-3, respectively), group 4 - control (with inoculation by Sol. NaCl 0.9%). Sacrification and vivisection were performed on days 1, 3, 7 and 14 of the control days. Biopsy specimens from the lower urinary tract and internal genital organs of laboratory animals (bladder, urethra, prostatic complex - 6 biopsies #1A-1D, 2A, 2B) were evaluated morphologically and bacteriologically. Analytical evaluation of the experimental data was presented using descriptive statistical methods. RESULTS: In experimental groups (Groups 1-3), bacteriological examination of prostatic complex biopsies showed growth of microflora in all samples in titers of 101-107 CFU / ml. In group 1, the maximum concentration of uropathogen was observed on day 7, compared to day 1 in both groups 2 and 3. In all observed cases, the highest degree of bacterial contamination was noted in the biopsy specimens from paraprostatic tissues and distal part of the prostate, which was 4.0+/-1.7 lg CFU/ml and 3.5+/-1.9 lg CFU/ml, respectively, and the smallest in proximal prostatic loci (1C) and bladder neck (2B) - 3.0+/-1.2 lg COE / ml and 3.0+/-1.7 lg COE / ml, respectively. According to the morphological study, a relevant progression of the suppurative and destructive inflammation (with foci of colliquation necrosis) was identified in group 1 in the biopsies from the prostate with a maximum degree of changes on day 7 with subsequent formation of loose connective tissue proliferation areas by 14 days. This indicates the conversion of the inflammatory process to the chronic stage. These changes corresponded with the results of histopathological studies in groups 2 and 3 where higher titers of bacterial agent were used. In group 4 (control) the commensal flora was bacteriologically determined in the biopsies, but there were no signs of inflammation, according to the results of the morphological study. CONCLUSION: In experimental model, we found that E. coli 103 CFU / ml induces the development of a phasic inflammatory process in the structures of the prostatic complex. These processes resulted in the formation of irreversible proliferative changes. As a consequence, it shold be recommended to consider these signs of contamination when evaluating the results of bacteriological examination of expressed prostate secretion/urine samples during planning treatment strategy.

Molecular screening of beneficial and safety determinants from bacteriocinogenic lactic acid bacteria isolated from Brazilian artisanal calabresa.

Despite of the beneficial relevance of several lactic acid bacteria (LAB) in the food industry, micro-organisms belonging to this group can determine spoilage in food products and carry a number of virulence and antibiotic resistance-related genes. This study aimed on the characterization of beneficial and safety aspects of five bacteriocinogenic LAB strains (Lactobacillus curvatus 12-named L. curvatus UFV-NPAC1), L. curvatus 36, Weissela viridescens 23, W. viridescens 31 and Lactococcus garvieae 36) isolated from an artisanal Brazilian calabresa, a traditional meat sausage. Regarding their beneficial aspects, all tested isolates were positive for mub, while EF226-cbp, EF1249-fbp and EF2380-maz were detected in at least one tested strain; none of the isolates presented map, EFTu or prgB. However, evaluated strains presented a variable pattern of virulence-related genes, but none of the strains presented gelE, cylA, efsA, cpd, int-Tn or sprE. Moreover, other virulence-related genes evaluated in this study were detected at different frequencies. L. curvatus 12 was generated positive results for ace, ccf, int, ermC, tetL, aac(6')-Ie-aph(2″)-Ia, aph(2″)-Ib, aph(2″)-Ic, bcrB, vanB and vanC2; L. curvatus 36: hyl, asa1, esp, int, ermC, tetK, aph(3')-IIIa, aph(2'')-Ic and vanC2; L. garvieae 32: asa1, ant(4')-Ia, aph(2'')-Ib, catA, vanA and vanC1; W. viridescens 23: esp, cob, ermB, aph(3')-IIIa, aph(2'')-Ic, vanA, vanB and vanC2; W. viridescens 31: hyl, esp, ermC, aph(3')-IIIa, aph(2'')-Ib, aph(2'')-Ic, catA, vanA and vanB. Despite presenting some beneficial aspects, the presence of virulence and antibiotic resistance genes jeopardize their utilization as starter or biopreservatives cultures in food products. Considering the inhibitory potential of these strains, an alternative would be the use of their bacteriocins as semi-purified or pure technological preparation. SIGNIFICANCE AND IMPACT OF THE STUDY: The food industry has a particular interest in using bacteriocinogenic lactic acid bacteria (LAB) as starter, probiotics and/or biopreservatives in different food products. Characterization of additional beneficial features is important to identify new, multifunctional potential probiotic strains. However, these strains can only be applied in food products only after being properly characterized according their potential negative aspects, such as virulence and antibiotic resistance genes. A wide characterization of beneficial and safety aspects of bacteriocinogenic LAB is determinant to guide the proper utilization of these strains, or their purified bacteriocins, by the food industry.