RESUMEN
Bacteriocin-producing bacteria with probiotic character are known as nutritional supplements mainly for livestock. Among those beneficial bacteria we also found enterococci. Because the species strains Enterococcus mundtii also can produce bacteriocins, this study was focused on fecal strains E. mundtii from horses and their bioactivity with a view to their possible future use in breeding. Rectal removal from 47 horses (40 mares and 7 stallions), the Norik breed from Murán were sampled in eastern Slovakia during November 2019 year. Horses age ranged from five months up to 23 years. Using MALDI-TOF mass spectrometry and 16S rRNA sequences analysis, 14 strains were allotted to the species E. mundtii. Bacteriocin substances produced by the strains EMKD 38/1, EMKD 40/2, EMKD 34/2 and EMKD 41/3 showed inhibitory activity against the most susceptible (principal) indicator strain Enterococcus avium EA5 and against listeriae as well (inhibitory activity from 100 up to 1 600 AU/mL). Only strain EMKD 41/3 possess Ent P and Mundticin KS genes and showed the broadest inhibitory activity. Ent B gene possessing strain EMKD 24/1 inhibited a growth of only indicator strain EA5. Identified E. mundtii tolerate low pH 3 and oxgall/bile. They were hemolysis, gelatinase and DNase negative and mostly susceptible to clinical antibiotics which are properties requested for application potential of strain. Substance from the strain with the broadest antimicrobial spectrum showed its practical/application potential, e.g. for optimizing the host microbiota which is important regarding the maintenance of animal`s health status.
Asunto(s)
Bacteriocinas , Animales , Antibacterianos/farmacología , Bacterias/genética , Bacteriocinas/genética , Enterococcus , Heces , Femenino , Caballos , Masculino , ARN Ribosómico 16S/genéticaRESUMEN
1. The effects of supplementation of broiler chicken diets with pea meal, carbohydrase enzymes and a probiotic were investigated for potential performance improvement. 2. Raw or extruded pea meal (cv Model, grown in Poland) was included in a wheat-soybean meal-based diet at 250 g/kg. The diets were unsupplemented (control) or supplemented with either carbohydrase enzymes (200 U/kg xylanase and 10 U/kg ß-glucanase in feed) or a probiotic (Bacillus subtilis), or both. The diets were fed to Ross 308 broilers aged 9-28 days. 3. After two additional days, chick gastrointestinal tracts were excised and analysed for the presence of Bacillus subtilis biofilm; and the ileal and caecal digesta were analysed for bacterial enzyme activities and to determine the concentration of short-chain fatty acids (SCFAs). 4. Feeding the pea-based diet supplemented with the probiotic compromised feed utilisation, due to higher feed intake. The addition of enzymes to the raw, but not the extruded, pea containing diet partially ameliorated this effect (pea form × additives; P < 0.002). 5. In the ileal digesta, interactions between the dietary treatments were observed for the activities of all bacterial glycolytic enzymes and for SCFA concentrations. ß-glucosidase, α-galactosidase and ß-glucuronidase were highest in birds fed the diet containing extruded pea supplemented with the probiotic and enzymes (pea form x additives; P = 0.018 to P < 0.006). In the caecal digesta, interactions were observed for bacterial enzyme activities, but not for total SCFA concentration. Biofilm formation in the caecum indicated that the probiotic strain was metabolically active in the broiler gut. 6. In conclusion, supplementation of diets containing raw or extruded pea meal with enzymes and a Bacillus subtilis spore-based probiotic modulated microbiota activity but had no clear effects on broiler performance. Probiotic administration did not cause excessive fermentation in the ileum and caecum but enhanced Bacillus subtilis spp. biofilm formation in the caecum, which may be indicative of a beneficial effect on gut health.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Pollos/microbiología , Pollos/fisiología , Glicósido Hidrolasas/administración & dosificación , Pisum sativum , Probióticos/administración & dosificación , Alimentación Animal , Animales , Bacillus subtilis/fisiología , Dieta/veterinaria , Femenino , Tracto Gastrointestinal/microbiología , Microbiota/fisiología , Pisum sativum/química , Semillas/química , Glycine max , TriticumRESUMEN
For an overview on the occurrence of Giardia assemblages in children in Eastern Slovakia, we examined 259 faecal samples of children from the segregated settlement in Medzev, 30 samples of children from the orphanage in Medzev and 40 samples of children with autism from the Special Elementary School in Kosice. Thirty-eight samples (14.67 %) from the segregated settlement, 19 samples (63.33 %) from the orphanage and two samples (5.0 %) from the Special Elementary School were positive for Giardia by flotation. The initial microscopic diagnostics were completed by the genotyping of the triosephosphate isomerase-gene loci (tpi genes) which revealed the existence of two Giardia assemblages in Slovak population, namely Giardia duodenalis (assemblage A) and Giardia enterica (assemblage B). These results represent the first evidence of A and B assemblages in children in Slovakia. Epidemiological significance and the impact on the public health of Giardia infection are highlighted.
Asunto(s)
Giardia/clasificación , Giardiasis/epidemiología , Giardiasis/parasitología , Animales , Niño , Heces/parasitología , Genotipo , Giardia/genética , Humanos , Eslovaquia/epidemiologíaRESUMEN
The study of biofilm function in vivo in various niches of the gastrointestinal tract (GIT) is rather limited. It is more frequently used in in vitro approaches, as an alternative to the studies focused on formation mechanisms and function of biofilms, which do not represent the actual in vivo complexity of microbial structures. Additionally, in vitro tests can sometimes lead to unreliable results. The goal of this study was to develop a simple approach to detect bacterial populations, particularly Lactobacillus and Bifidobacterium in biofilms, in vivo by the fluorescent in situ hybridisation (FISH) method. We standardised a new Histo-FISH method based on specific fluorochrome labelling probes which are able to detect Lactobacillus spp. and Bifidobacterium spp. within biofilms on the mucosal surface of the GIT embedded in paraffin in histological slices. This method is also suitable for visualisation of bacterial populations in the GIT internal content. Depending on the labelling probes, the Histo-FISH method has the potential to detect other probiotic strains or pathogenic bacteria. This original approach permits us to analyse bacterial colonisation processes as well as biofilm formation in stomach and caecum of BALB/c and germ-free mice.
Asunto(s)
Bifidobacterium/fisiología , Biopelículas/crecimiento & desarrollo , Histocitoquímica/métodos , Hibridación Fluorescente in Situ/métodos , Mucosa Intestinal/microbiología , Lactobacillus/fisiología , Animales , Bifidobacterium/genética , Bifidobacterium/crecimiento & desarrollo , Vida Libre de Gérmenes , Lactobacillus/genética , Lactobacillus/crecimiento & desarrollo , Ratones Endogámicos BALB CRESUMEN
Antirabies virus neutralization antibodies in sera and/or transudates modified RFFIT method by Smith et al. (1973). Sera were titrated on Lab-Tek 8 chamber TC slides. Sera and/or transudates (content of pleural cavity) as well as the challenge virus strain (vaccination strains of the rabies virus Vnukovo-32/107th passage and/or CVS 11/Paris) were incubated at 37 degrees C during 90 minutes subsequently BHK-21/C13 cell culture was added. The cultures were fixed after 24 to 48 hours and stained with antirabic fluorescent conjugate (Bioveta a.s., Ivanovice n. H., Czech Republic). The highest dilution of the virus was used as the challenge dose where 50 percent of the cells in the examined range of view were infected (fluorescent inclusions can be observed). The antirabic reference serum was used as a control in RFFIT in each examined serum. To ensure a good control, the serum was diluted to contain 0.5 IU/ml of antirabic virus neutralization antibodies. Sera and/or transudates which were sent to our Laboratory were examined in this way. We examined 40 sera or pleural transudates of orally vaccinated foxes by those methods. These sera were sent to National reference laboratories for rabies (NRPB) in Kosice. Samples were examined for the monitoring of efficiency of oral antirabic vaccination. The parallel quantification of antirabic antibodies by virus neutralization test (VNT) in vivo was applied to mice and indirect haemagglutination test (NHT). The results of these three tests are comparable or in correlation. RFFIT has many advantages. When using highly attenuated strain Vnukovo-32/107th passage as the challenge virus in RFFIT method the potential risk of laboratory exposition is absent.