Regular Physical Activity Influences Gut Microbiota with Positive Health Effects

The gut microbiota is believed to have a major impact on human health and disease. It is involved in barrier functions and maintenance of homeostasis. It provides nutrients and metabolites, participates in a signaling network, regulates epithelial development, and influences the immune system as well as protects the intestinal mucosa from the aggression of pathogenic microorganisms. There is growing evidence that physical activity has an impact on the gut microbiota. Recent studies in animals and humans suggest that regular physical activity increases the presence of beneficial microbial species and improves host health. However, some specific differences should be noted: different forms of physical activity, frequency or intensity, aerobic or resistance training, and benefits and consequences for amateur or competitive athletes. Because the positive role of physical activity can have an impact on health and various types of diseases, the results of research studies in this area are increasingly becoming the focus of scientific interest. In addition, probiotic supplements modulate intestinal microbial flora, and the ability of probiotics to modulate perturbations in immune function after exercise highlights their potential for use in individuals exposed to high levels of physical activity

Sumnjivi i nepodudarni rezultati u dijagnostici bjesnoće testom imuno-fluorescencije.

A total of 7339 fox, dog, cat, cattle, sheep and other mammalian brains were tested to rabies virus antigen. Results of fluorescent antibody test (FAT) were evaluated by two microscopists. Doubtful and discordant results in FAT were analyzed again in virus isolation test (VIT) using mouse neuroblastoma (NA) cell line. Twenty-eight brain samples were determined as doubtful, while 9 brain samples were determined to be positive by one microscopist and negative by the other. Samples which were shown as doubtful and discordant in FAT were retested in VIT. Seventeen of these were positive in the VIT.Na prisutnost antigena virusa bjesnoće bilo je pretraženo ukupno 7339 uzoraka mozgova lisica, mačaka, goveda, ovaca i drugih vrsta životinja. Rezultate izravnog imunofluorescentnog testa vrednovala su dva stručnjaka. Uzorci koji su u testu imunofluorescencije davali sumnjive rezultate, ili se rezultati stručnjaka nisu slagali bili su pretraženi izdvajanjem virusa na kulturi stanica. Rabljena je stanična linija neuroblastoma miševa. Dvadeset osam uzoraka bilo je proglašeno sumnjivim na bjesnoću. Rezultat prvog mikroskopskog pregleda u devet uzoraka bio je pozitivan, a drugog negativan. Od ukupno 37 uzoraka, koji su bili ponovno pretraženi izdvajanjem virusa, 17 je bilo pozitivnih na bjesnoću

Molekularni pristup identifikaciji virusa influence ptica

The recently raised awareness of the threat of a new influenza pandemic has stimulated the interest in the detection of influenza A viruses in the secretions of a wide variety of birds and mammals. Influenza A viruses are subtyped conventionally according to the characteristics of the external glycoproteins, haemagglutinin (HA) and neuraminidase (NA). All 16 HA and 9 NA subtypes have been isolated from aquatic birds that are believed to be a natural host and reservoir for influenza viruses. Poultry, specifically chickens and turkeys, are not considered to be a normal host for the virus, although transmission from wild birds to poultry occurs routinely. Although most HA subtypes have been found in poultry, particular emphasis is placed on the H5 and H7 HA subtypes of avian influenza virus because only these subtypes are known to cause highly pathogenic avian influenza (HPAI) in poultry. Recently developed molecular methods enable fast, accurate and reliable approach to identify influenza virus and its subtypes.Svijest o prijetnji nove pandemije influence potaknula je zanimanje za dokaz virusa influence A u sekretima različitih vrsta ptica i sisavaca. Virusi influence A uobičajeno se tipiziraju na osnovi osobina njihovih površinskih glikoproteina, hemaglutinina (HA) i neuraminidaze (NA). Svih 16 HA i devet NA podtipova izdvojeno je iz vodenih ptica koje su prirodni domaćin i rezervoar virusa influence. Perad, posebice kokoši i pure, ne smatra se uobičajenim domaćinom virusa iako se virus redovito s divljih ptica prenosi na perad. Premda su mnogi HA podtipovi izdvojeni iz peradi, posebna pozornost pridaje se podtipovima H5 i H7 virusa influence ptica, jer samo oni pripadaju visoko patogenim sojevima za perad. Nedavno razvijene molekularne metode omogućuju brzu, točnu i pouzdanu identifikaciju virusa influence i njegovih podtipova

Sumnjivi i nepodudarni rezultati u dijagnostici bjesnoće testom imuno-fluorescencije.

A total of 7339 fox, dog, cat, cattle, sheep and other mammalian brains were tested to rabies virus antigen. Results of fluorescent antibody test (FAT) were evaluated by two microscopists. Doubtful and discordant results in FAT were analyzed again in virus isolation test (VIT) using mouse neuroblastoma (NA) cell line. Twenty-eight brain samples were determined as doubtful, while 9 brain samples were determined to be positive by one microscopist and negative by the other. Samples which were shown as doubtful and discordant in FAT were retested in VIT. Seventeen of these were positive in the VIT.Na prisutnost antigena virusa bjesnoće bilo je pretraženo ukupno 7339 uzoraka mozgova lisica, mačaka, goveda, ovaca i drugih vrsta životinja. Rezultate izravnog imunofluorescentnog testa vrednovala su dva stručnjaka. Uzorci koji su u testu imunofluorescencije davali sumnjive rezultate, ili se rezultati stručnjaka nisu slagali bili su pretraženi izdvajanjem virusa na kulturi stanica. Rabljena je stanična linija neuroblastoma miševa. Dvadeset osam uzoraka bilo je proglašeno sumnjivim na bjesnoću. Rezultat prvog mikroskopskog pregleda u devet uzoraka bio je pozitivan, a drugog negativan. Od ukupno 37 uzoraka, koji su bili ponovno pretraženi izdvajanjem virusa, 17 je bilo pozitivnih na bjesnoću

Real-time PCR applications in clinical research and diagnostics

Polymerase chain reaction (PCR) has enabled enormous progress in the field of molecular biology in the last twenty-five years. It became popular due to its high sensitivity and specificity and was introduced to numerous scientific fields. Today many different variants of PCR exist, with real-time PCR being the most common. This method is being successfully used at our institution for a variety of different applications, among which gene expression analyzes and development of protocols for clinical diagnostics currently dominate. But the extreme flexibility and customizability of real-time PCR, coupled with our team’s expertise, ensures efficient application of this technique to various new research projects

Nadzor influence u dvorišne peradi i ptica selica u Sloveniji

Blood samples and clocal swabs were taken from free-living migratory birds and backyard flock birds. Enzyme linked immunosorbent assays (ELISA) and haemagglutination inhibition test (HI) was used to screen free-living migratory birds and backyard flocks for antibodies against avian influenza viruses (AIV). Pools of cloacal swabs were used for the attempt of isolation of AIV. Samples were inoculated of 9 to 11 day-old chicken embryos. Amnioallantoic fluid were collected and analysed for haemagglutination activity. Pools of cloacal swabs were used for the detection of AIV by RT-PCR. Sera samples of four out of twenty three geese were positive on presence of specific antibodies against AIV. All attempts of virus isolation were negative. No AIV nucleic acid was detected by RT-PCR.Uzeti su uzorci krvi i obrisci nečisnica slobodnoživućih ptica selica i dvorišne peradi. Imunoenzimni test i inhibicija hemaglutinacije korišteni su kako bi se u slobodnoživućih ptica selica i dvorišne peradi utvrdila nazočnost protutijela za viruse influence ptica. Skupni uzorci briseva nečisnica korišteni su za izdvajanje virusa influence ptica. Uzorci su inokulirani u 9 do 11 dana stare kokošje embrije. Sakupljena je amnioalantoisna tekućina i pretražena na sposobnost hem-aglutinacije. Skupni uzorci brisova nečisnica korišteni su za dokaz virusa influence ptica pomoću RT-PCR. Četiri od 23 seruma gusaka bila su pozitivna na nazočnost specifičnih protutijela za virus influence ptica. Svi pokušaji izdvajanja virusa dali su negativan rezultat. Postupkom RT-PCR nije dokazana nazočnost nukleinske kiseline virusa influence ptica

Molecular characterization of a new porcine rotavirus P genotype found in an asymptomatic pig in Slovenia

AbstractRotaviral RNA was detected in the stool sample of an asymptomatic fattening pig at a Slovenian pig farm. To characterize the rotavirus, RT-PCR was used, employing primers specific for the VP7, VP4 and NSP4 genes. Specific products were purified and the sequencing reaction was performed for the molecular analysis of amplified genes. Nucleotide and amino acid sequences of the VP7 gene were found highly identical (85.3–88.1% and 90.7–91.6%) to G1 genotype strains. Phylogenetic and molecular analyses of the VP7 antigen regions revealed the sample to be from a new lineage of G1 genotype. In the molecular analysis of the VP4 gene, only 70.9% nucleotide (76.2% amino acid) identity was found with the most related rotavirus VP4 gene from GenBank. Following this, the NSP4 gene was also analyzed. After the phylogenetic analysis, it clustered with the NSP4 B genotype, but also seemed to represent a new lineage of this genotype. This new rotavirus strain, named P21-5, differed greatly from all rotaviruses characterized so far in all three genes analyzed. The virulence of this strain is not clear yet and has to be investigated

Effect of Composting Under Semipermeable Film on the Sewage Sludge Virome

The addition of compost from sewage sludge to soils represents a sustainable option from an environmental and economic point of view, which involves the valorisation of these wastes. However, before their use as a soil amendment, compost has to reach the quality levels according to the normative, including microbial parameters. Viruses are not included in this regulation and they can produce agricultural problems and human diseases if the compost is not well sanitised. In this study, we carried out the analysis of the viral populations during a composting process with sewage sludge at an industrial scale, using semipermeable cover technology. Viral community was characterised by the presence of plant viruses and bacteriophages of enteric bacteria. The phytopathogen viruses were the group with the highest relative abundance in the sewage sludge sample and at 70 days of the composting process. The diversity of bacterial viruses and their specificity, with respect to the more abundant bacterial taxa throughout the process, highlights the importance of the interrelations between viral and bacterial communities in the control of pathogenic communities. These results suggest the possibility of using them as a tool to predict the effectiveness of the processThis research was conductedwith funding from Junta de Andalucía [Research project RNM-7370]. E. A. would like to thank the Ministry of Economy and Competitiveness (MINECO) and European Regional Development Fund (ERDF) funds [RYC-2013-12481]. We acknowledge financial support from the Slovenian Research Agency (research core funding no. P4-0092)

Antioxidant and Antimicrobial Properties of Helichrysum italicum (Roth) G. Don Hydrosol

(1) Background: According to the emergence and spread of antibiotic resistance, there is an urge for new promising substances. The purpose of the study was to test the antioxidant, cytotoxic and antimicrobial properties of the Helichrysum italicum (Roth) G. Don essential oil (EO) and hydrosol. (2) Methods: The antioxidant potential was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The cytotoxicity for human skin and intestinal cells was tested using primary and immortalized cell line models. The minimum inhibitory concentration (MIC) of hydrosol was then determined for six bacterial strains covering four commonly reported food pathogens. Further on, the hydrosol at a concentration of 1/8 MIC was used to test the antiadhesive effect by the crystal violet (CV) staining method. (3) Results: the EO showed a 100-times higher antioxidant and 180- to 25.000-times higher cytotoxic activity, when compared to hydrosol. Nevertheless, all bacterial strains, with the exception of Staphylococcus aureus, were sensitive to hydrosol in the range of 12.5% (V/V) for Campylobacter jejuni, to MIC values of 100% (V/V) for Escherichia coli and Pseudomonas aeruginosa. The antiadhesive potential of hydrosol was also shown. (4) Conclusions: Even though hydrosols are a by-product of the EO distillation process, they possess valuable biological activities