RESUMO
Wild ruminants are an interesting topic for research because only limited information exists regarding their microbiota. They could also be an environmental reservoir of undesirable bacteria for other animals or humans. In this study faeces of the 21 free-living animals was sampled (9 Cervus elaphus-red deer, adult females, 12 Capreolus capreolus-roe deer, young females). They were culled by selective-reductive shooting during the winter season of 2014/2015 in the Strzalowo Forest District-Piska Primeval Forest (53° 36 min 43.56 sec N, 21° 30 min 58.68 sec E) in Poland. Buttiauxella sp. is a psychrotolerant, facultatively anaerobic, Gram-negative rod anaerobic bacte- rial species belonging to the Phylum Proteobacteria, Class Gammaproteobacteria, Order Entero- bacteriales, Family Enterobacteriacae and to Genus Buttiauxella. Buttiauxella sp. has never previ- ously been reported in wild ruminants. In this study, identification, antimicrobial profile and sensitivity to enterocins of Buttiauxella strains were studied as a contribution to the microbiota of wild animals, but also to extend knowledge regarding the antimicrobial spectrum of enterocins. Five strains were identified using the MALDI-TOF identification system (evaluation score value was up to 2.224) and allotted to the genus Buttiauxella including the species Buttiauxella gaviniae, B. ferragutiae, B. agrestis. Strains were DNase negative, and they hydrolysed esculin; fermentation of L-arabinose, D-mannitol and D-mannose was positive. Dulcitol, inositol reaction, urea and indol were negative. Buttiauxella strains did not form biofilm. They were resistant to at least one of the 13 antibiotics tested. B. agrestis 2/109/1 was resistant to amdinocillin, clindamycin and pen- icillin. However, Buttiauxella strains were sensitive to the enterocins used (inhibition activity ranged from 100 to 25 600 AU/ml).
Assuntos
Antibacterianos/farmacologia , Cervos/microbiologia , Farmacorresistência Bacteriana , Enterobacteriaceae/efeitos dos fármacos , Fezes/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , AnimaisRESUMO
With an increasing number of pheasants as gamebirds being reared each year, these species are becoming a more prominent part of the workload of many veterinary practices. Only limited information can be found concerning the microflora of common pheasants. A significant part of the obligate microflora consists of lactic acid bacteria, including enterococci. In this study, faeces were sampled from 60 pheasants aged 16-17 weeks. Enterococcal counts reached 5.48±1.9 (log10) CFU/g. Strains (17) were taxonomically classified to the genus Enterococcus using the Maldi-Tof identification system; they were allotted to the species E. hirae (58.8%), E. faecium (23.5%) and E. faecalis (17.7%) by highly probable species identification or by secure genus identification/probable species identication. Species allocation was also confirmed using conventional biochemical tests. Most strains formed ß-hemolysis. Gelatinase active phenotype was found in three E. faecalis strains. Enterococci were ß-glucuronidase negative, mostly trypsin negative with slight or moderate production of α-chymotrypsin. EH52b and EF42 strains possessed the highest potential for pathogenicity. Average value of lactic acid was 1.78±0.33 mmo/L. Most strains were tetracycline resistant (82.4%). Polyresistant E. faecalis strains with positive gelatinase phenotype and possessing virulence factor genes confirmed using PCR (gelE, efaAfs, ccf cob, cpd) were sensitive to enterocins (activity 1600-25,600 AU/mL).
Assuntos
Enterococcus/efeitos dos fármacos , Galliformes , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Doenças das Aves Domésticas/microbiologia , Fatores de Virulência/metabolismo , Animais , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Fezes/microbiologia , Fatores de Virulência/genéticaRESUMO
1. This study evaluated the duodenal wall integrity, antioxidant status as well as some immunological parameters of broiler chickens supplemented with 0.5 g Thymus vulgaris essential oil (EO)/kg diet and 0.4 mg Se/kg DM (dry matter) derived from sodium selenite. 2. A total of 192 one-d-old randomly divided chickens of both sexes (Ross 308 hybrid broilers) were divided into 4 treatment groups of 48 birds each. 3. The first group was fed on a nutritionally balanced basal diet (BD). The other three groups received BD supplemented with 0.5 g/kg thyme oil, or 0.4 mg Se/kg DM, or both feed additives together. 4. The results for the evaluated feed additives were (1) thyme oil - decreased malondialdehyde (MDA) concentration in duodenal mucosa and kidney, increased immunoglobulin A (IgA) concentration in duodenal mucosa, stimulated phagocytic activity in blood, improved intestinal barrier integrity (2) selenium - increased glutathione peroxidase (GPx) activity in blood and liver as well as thioredoxin reductase (TrxR) activity in duodenal mucosa, liver and in the kidney, (3) EO with selenium - increased thioredoxin reductase (TrxR) activity in duodenal mucosa. 5. These results demonstrated that thyme oil alone showed more effective potential to improve intestinal barrier integrity and antioxidant status as well as evoking an immune response in chickens, than if diets were supplemented with both thyme oil and selenium.
Assuntos
Galinhas/microbiologia , Galinhas/fisiologia , Intestinos/microbiologia , Óleos Voláteis/farmacologia , Selênio/farmacologia , Thymus (Planta)/química , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Contagem de Colônia Microbiana/veterinária , Dieta/veterinária , Suplementos Nutricionais , Impedância Elétrica , Feminino , Imunidade Inata/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Masculino , Microbiota/efeitos dos fármacos , Óleos Voláteis/administração & dosagem , Distribuição Aleatória , Selênio/administração & dosagem , Selenito de Sódio/administração & dosagem , Selenito de Sódio/farmacologiaRESUMO
The effect of Enterococcus faecium CCM7420 (EF) - enterocin-producing and probiotic strain of rabbit origin, Eleutherococcus senticosus extract (ES) and their combination (ES+EF) was determined on selected bacteria in faeces and caecum content, leukocytes phagocytosis, blood biochemistry and growth performance. Ninety-six weaned rabbits were divided into 3 experimental (ES, EF, ES+EF) and control group (CG). The rabbits in the groups ES and EF+ES were fed commercial diet enriched with E. senticosus extract (30 g/100 kg feed), rabbits in groups EF and CG were fed untreated diet. The rabbits in the EF and ES+EF groups were administered with an overnight culture of E. faecium CCM7420 strain (500 microl/animal/day into water, 109 CFU/ml). The treatment period lasted 21 days. The microbiological examinations in faecal samples confirmed the presence of E. faecium CCM7420 strain. In groups EF and ES+EF, the reduction of faecal coliforms, Pseudomonas-like sp., Clostridium-like sp. and S. aureus was recorded. Leucocyte phagocytosis significantly increased in all experimental groups (P < 0.0001) compared to CG. The lowest GPx values were measured in the ES+EF group. Higher total protein, triglycerides and calcium concentrations were detected in experimental groups compared to CG. The cholesterol concentration decreased in the ES group. The highest average daily gain was recorded in EF group; in ES+EF the better feed conversion ratio and no mortality was recorded. These results indicated that the dietary supplementation with the E. faecium CCM7420 and E. senticosus extract stimulate the leukocytes phagocytosis and reduces the potential pathogens in rabbits digestive tract without oxidative stress and improve the growth performance.
Assuntos
Bacteriocinas/metabolismo , Eleutherococcus/fisiologia , Enterococcus faecium/classificação , Enterococcus faecium/metabolismo , Probióticos/farmacologia , Animais , Feminino , Masculino , CoelhosRESUMO
Enterocin (Ent) 4231, produced by non-rabbit origin strain Enterococcus faecium CCM 4231 was used in combination with sage plant extract in rabbits with the aim to check their antimicrobial activity against microbiota, their effect on immunological, biochemical blood parameters, values of volatile fatty acids in caecum, Eimeria sp. oocysts occurrence and selected parameters of rabbits meat. The animals were divided into three experimental groups (EG1-Ent 4231; EG2- sage; EG3- Ent 4231 with sage) and control group (CG); 24 rabbits in each. Natural substances (NS) were administered for 21 days. The experiment lasted for 42 days. The reduction of microbiota in faeces was observed in EG3 at day 21 by a decrease in the numer of coagulase-positive staphylococci (P<0.01) in comparison with that determined in CG. The bacterial counts in the caecum were lower than those found in faeces. A decrease in the numer of Pseudomonas-like sp. in caeca of the experimental groups was observed at days 21 and 42 (difference in range 0.40-1.87 log cycles) comparing with that determined in CG. At day 21, a significant increase in phagocytic activity (PA, P<0.001) was found in blood of rabbits from EG2 comparing with that observed in CG. At day 42, a significant increase in PA (P<0.001) was determined in all experimental groups in comparison with CG. At day 21, in caecal content of EG3 significantly higher values of lactic acid were observed (P<0.05) in comparison with those found in CG. The reduction of Eimeria sp. oocysts was demonstrated after application of each of NS. Addition of NS did not influence biochemical parameters, meat quality of the animals and does not influence negatively the health status of rabbits.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Enterococcus faecium/classificação , Coelhos , Salvia officinalis , Ração Animal/análise , Animais , Bacteriocinas/química , Dieta/veterinária , Enterococcus faecium/química , Fezes/microbiologia , Carne/normasRESUMO
The present study investigates the effects of enterocin Ent M and durancin Ent ED26E/7 applied separately and in combination on the intestinal microbiota, caecal enzymatic activity, and fermentaion of rabbits. Eighty rabbits (M91 meatline, aged 5 weeks, both sexes) were divided into groups E (Ent M; 50 µL/animal/day), D (Ent ED26E/7; 50 µL/animal/day), E + D (Ent M + Ent ED26E/7), and control (C). The additives were administered in drinking water for 21 days. Antimicrobial activity of Ent M and Ent ED26E/7 on coliforms (E, E + D: P < 0.001) and pseudomonads (D: P < 0.05) in feces was noted, compared to C. Ent M and Ent ED26E/7 application stimulated caecal enzymatic activity in rabbits. Pectinolytic (E vs. D, E + D: P < 0.01), inulolytic (E vs. E + D: P < 0.01; E vs. C: P < 0.05), and amylolytic (E vs. D, E + D. P < 0.001; E vs. C: P < 0.01) activities were influenced by Ent M, while cellulolytic (D vs. E + D: P < 0.01) and inulolytic (D vs. E + D, C: P < 0.01) activities by Ent ED26E/7 treatment. The cellulolytic and pectinolytic acitivities changed with time. Treatment × time interaction was detected for cellulose and xylan degradation. During Ent M and Ent ED26E/7 treatment, increased ammonia, lactic, butyric and iso-valeric acid, and lower acetic, propionic, iso-butyric, valeric, and caproic acid concentrations were noted. It can be concluded that Ent M and Ent ED26E/7 application can improve rabbit health due to reduced spoilage microbiota and enhanced caecal enzymatic activity.
Assuntos
Ceco , Dieta/veterinária , Microbioma Gastrointestinal , Coelhos , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Hidrocarbonetos Aromáticos com Pontes/administração & dosagem , Ceco/enzimologia , FermentaçãoRESUMO
Although the physiological and nutritional properties of many probiotic strains are well understood, the study of their antiprotozoal potential is only at the beginning. This study describes the in vitro effect of four probiotic/bacteriocin-producing strains towards poultry Eimeria sp. oocysts. The oocyst reduction (P < 0.05) was detected from 4 h and/or 6 h of the cultivation in broth till the end of the experiment (32 h) without species or origin dependence. No differences in the reductive effect of bacteriocin-producing and non-producing strains were found. The character of substances with anticoccidial effect remains to be determined.
Assuntos
Bacteriocinas/metabolismo , Eimeria/efeitos dos fármacos , Eimeria/fisiologia , Lactobacillus/fisiologia , Probióticos/farmacologia , Bacteriocinas/farmacologia , Técnicas de CoculturaRESUMO
Our current knowledge of microbiota in wild ruminants is limited. The goal of this study was to evaluate staphylococcal species in red and roe deer for various attributes (haemolysis, DNase, and urease activities; lactic acid and biofilm production; and antibiotic profile) and their susceptibility to gallidermin. Sixteen staphylococcal strains were identified from faeces of 21 free-living animals (9 adult female Cervus elaphus-red deer and 12 young female Capreolus capreolus-roe deer) sampled by the Polish colleagues in the Strzalowo Forest District, Piska Primaeval Forest. The variability in the species of staphylococci was determined. Seven species (Staphylococcus capitis, S. epidermidis, S. haemolyticus, S. hominis, S. pseudintermedius, S. vitulinus and S. warneri) and five clusters/groups of coagulase-negative staphylococci (CoNS) were identified. The strains were generally not haemolytic and Dnase negative; did not form biofilms or only produced low-grade biofilms; exhibited high levels of lactic acid; were urease positive; and were generally susceptible to antibiotics (only two strains were resistant to multiple antibiotics). However, all of the strains were susceptible to the lantibiotic bacteriocin gallidermin, with a minimal inhibitory concentration of 0.0156 µg (up to 6400 AU/ml in arbitrary units). This is the first study to perform a detailed study of the properties of CoNS from roe and red deer.
Assuntos
Bacteriocinas/farmacologia , Cervos/microbiologia , Fezes/microbiologia , Staphylococcus , Animais , Antibacterianos/farmacologia , Feminino , Testes de Sensibilidade Microbiana , Polônia , Staphylococcus/efeitos dos fármacos , Staphylococcus/isolamento & purificação , Staphylococcus/metabolismoRESUMO
The effects of enterocin (Ent) M and sage extract applied separately and in combination were investigated. EntM (E 50 µL/animal/day in water) and sage extract (S 10 µL/animal/day in water) were applied individually and in combination (E+S) to rabbits during 21 days of treatment. The rabbits' growth was not significantly influenced by the additives. Lower feed conversion (FC) was noted in the experimental groups compared with controls, with the lowest data detected in E. The antimicrobial activity of EntM was noted (in E+S: lactic acid bacteria-P < 0.01; in E, E+S: enterococci, enterobacteria-P > 0.05; in E: clostridia-P > 0.05). The most significant changes in fermentation between weaned and older rabbits were noted in amylolytic activity at day 21 (E P < 0.05; E + S P < 0.05); prolonged reduction effect of sage extract on amylolytic activity was observed. The activity of cellulase, pectinase and xylanase was higher in older than in younger animals. Decrease in lactic acid and volatile fatty acids was noted during EntM administration, with significant effect on propionic acid concentration (E P < 0.05; E+S P < 0.001). The sage extract reduced propionic acid (S P < 0.001) and butyric acid levels (S P < 0.05) and increased the concentrations of butyric, iso-valeric, valeric, caproic acids and lactic acid (P < 0.001). It seems to be that EntM and sage supplementation may improve the economy of rabbit farms (increased FC) and the health status of rabbits (reduction of spoilage microbiota, enhanced enzymatic activities in caecum).
Assuntos
Ceco , Microbioma Gastrointestinal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Salvia officinalis/química , Animais , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Ceco/efeitos dos fármacos , Ceco/metabolismo , Suplementos Nutricionais , Enzimas/metabolismo , Feminino , Masculino , Coelhos , DesmameRESUMO
The intestinal microbiota has enormous impact on the health and performance of horses. Staphylococci belong in the phylum Firmicutes, and their occurrence, especially of methicillin-resistant strains and species, has been reported in horses previously. Moreover, biofilm formation is one of the virulence factors; it has been not completely studied in fecal coagulase-negative staphylococci (CoNS) from horses. Therefore, this study was focused on biofilm formation by various species of fecal CoNS from horses because it has been never reported before. In addition, their antibiotic profile was tested. Horses (42) of various breeds from Slovakia/Poland were sampled. Variability in the species of CoNS was detected in feces of horses. Thirty-two strains were identified by using the MALDI-TOF system and classified into nine species and three subspecies of CoNS: Staphylococcus capitis, S. cohnii subsp. cohnii, S. cohnii subsp. urealyticus, S. cohnii subsp. casei, S. epidermidis, S. haemolyticus, S. pasteuri, S. sciuri, S. vitulinus, S. warneri, and S. xylosus. The most frequent species was S. vitulinus. Twenty-two strains showed high biofilm production; 10 strains showed low-grade biofilm production. The highest biofilm formation was measured in the species S. xylosus. Eleven strains (of 32) were methicillin-resistant; the others were susceptible to methicillin.
Assuntos
Coagulase/deficiência , Fezes/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Microbioma Gastrointestinal , Cavalos , Ácido Láctico/metabolismo , Resistência a Meticilina , Testes de Sensibilidade Microbiana , Polônia/epidemiologia , Eslováquia/epidemiologia , Especificidade da Espécie , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologiaRESUMO
Enterococci isolated from 28 different commercially available feeds (10-1000 CFU/mL) were identified and their probiotic potential was determined. Species identification of 22 selected strains was performed by intergenic length-polymorphism analysis (tRNA-PCR); PCR products were analyzed using capillary electrophoresis. Six strains were allotted to the species Enterococcus faecium, four to E. faecalis, one to E. hirae; the remaining strains were not classed. The strains were sensitive to vancomycin, ampicillin, tetracycline and rifampicin. They were able to adhere to human as well as canine intestinal mucus. They produced lactic acid (0.99-1.04 mmol/L) and most of them were urease-positive with sufficient survival in 5 % Oxgall-bile. They did not show any inhibitory activity due to antimicrobial substances. Plasmid DNA was detected in 8 strains, the bands responding to small molecular size (10 kbp). Considering all probiotically important properties, E. faecium strain EE3 was suggested as potential feed additive.
Assuntos
Ração Animal/microbiologia , Enterococcus/isolamento & purificação , Probióticos/isolamento & purificação , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Cães , Enterococcus/química , Enterococcus/efeitos dos fármacos , Enterococcus/metabolismo , Humanos , Intestinos/microbiologia , Ácido Láctico/metabolismo , Probióticos/química , Urease/metabolismoRESUMO
Enterococci from different free-living rodents were isolated and selected. The strains were genotyped and their antibiotic sensitivity and /or resistance, production of lactic acid, urease activity, the presence of enterocin structural genes, plasmid detection, and their binding ability to proteins were determined. Among 24 enterococcal strains, 17 strains were allotted to the species Enterococcus faecalis by tDNA-PCR, 7 strains being not yet identified. Only Enterococcus sp. ES66 possessed the structural genes for the production of enterocins (Ent) A and P. E. faecalis EE61 had the gene for EntL50B. The other isolates were Ent-gene-free. Enterococci were mostly sensitive to antibiotic treatment . The plasmid DNA was detected only in the strain EE97. The average value of lactic acid production reached 896 +/- 90 micromol/L. Most of the strains possessed low ureolytic activity. Enterococci bound very well sub-epithelial proteins, reaching the value of 3 by the particle agglutination assay, especially concerning the heparin, bovine lactoferrin and porcine fibronectin.
Assuntos
Animais Selvagens/microbiologia , Enterococcus faecalis/isolamento & purificação , Roedores/microbiologia , Animais , Antibacterianos/farmacologia , Bovinos , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/metabolismo , Fibronectinas/metabolismo , Heparina/metabolismo , Lactoferrina/metabolismo , Testes de Sensibilidade Microbiana , Polônia , Ligação Proteica , SuínosRESUMO
Antibiotic susceptibility or resistance, urease activity, detection of the structural genes for bacteriocin production, bacteriocin activity as well as sensitivity of the isolates to enterocins (Ent) A and M were determined in 23 isolates of new species Enterococcus haemoperoxidus and E. moraviensis. The majority of the strains were antibiotic sensitive and exhibited low urease activity (< 10 nkat/mL). The most frequently detected genes for Ent were entA and entP. However, only the strain 466 of E. haemoperoxidus produced an antibacterial substance with inhibitory activity against 21 G+ indicators. It was partially purified reaching an activity of up to 12 800 AU/mL. This bacteriocin active strain also possessed the genes for EntA and EntP. The other strains did not inhibit the indicator strains. The substance produced by the 466 strain was active even after a 5-months storage at +4 and -20 degrees C. This substance has proteolytic and hydrophilic character, pH optimum of bacteriocin production by this strain being between 4 and 7. While E. moraviensis strains showed sensitivity to EntA (produced by E. faecium EK13) and to EntM (produced by E. faecium AL41), E. haemoperoxidus strains were sensitive to EntA (except strain 382) but less sensitive to the treatment by EntM.
Assuntos
Bacteriocinas/metabolismo , Farmacorresistência Bacteriana , Enterococcus/genética , Enterococcus/metabolismo , Bacteriocinas/genética , Enterococcus/efeitos dos fármacos , Água Doce/microbiologia , Testes de Sensibilidade Microbiana , Urease/metabolismo , Microbiologia da ÁguaRESUMO
The ability of canine strain Enterococcus faecium EE3 to survive in healthy dogs and its effect on microbiological and biochemical parameters was determined. The strain was individually applied to 11 dogs per os at a dose of 10(9) CFU/mL (differed from 2 to 3 mL) for 1 week and persisted in feces for 3 months after cessation of its administration (reaching average concentration of 6.83 +/- 0.95 log CFU/g). Seven d after administration, a decrease in staphylococci and a significant decrease in Pseudomonas-like bacteria was observed. On the other hand, concentration of lactic acid bacteria increased but the growth of E. coli was not influenced. In the blood samples of dogs after 0-1 d (before application) and the blood samples 1 week after application, total lipids decreased in 8 dogs; the total protein also decreased. The levels of cholesterol were brought to the physiological level, i.e. in blood samples with low cholesterol values it increased to the physiological level and in those with high levels it decreased; cholesterol was not influenced in 3 dogs.
Assuntos
Doenças do Cão/prevenção & controle , Enterococcus faecium/crescimento & desenvolvimento , Intestinos/microbiologia , Probióticos/administração & dosagem , Animais , Contagem de Colônia Microbiana , Doenças do Cão/microbiologia , Cães , Feminino , Lipídeos/sangue , Masculino , Obesidade/prevenção & controle , Obesidade/veterináriaRESUMO
The AD 2 strain isolated from feces of a healthy dog in Slovakia was characterized phenotypically by the conventional tests and commercial identification kits API Staph and ID32 Staph. Results of biochemical tests identified the strain as S. piscifermentans, fully corresponding with the species description. Further characterization by whole-cell protein profile analysis (SDS-PAGE) confirmed the identification based on biochemical tests and showed that the AD 2 strain is S. piscifermentans; lactic acid production, urease activity, bacteriocin production and the antibiotic susceptibility of it were also determined. S. piscifermentans AD 2 isolated first from an animal source was deposited in the Czech Collection of Microorganisms as Staphylococcus piscifermentans CCM 7165.
Assuntos
Cães/microbiologia , Fezes/microbiologia , Staphylococcus/classificação , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/análise , Técnicas de Tipagem Bacteriana , Fermentação , Peixes/microbiologia , Microbiologia de Alimentos , Testes de Sensibilidade Microbiana , Kit de Reagentes para Diagnóstico , Staphylococcus/química , Staphylococcus/efeitos dos fármacos , Staphylococcus/isolamento & purificaçãoRESUMO
Forty strains of enterococci and forty strains of lactobacilli isolated from feces of 10 healthy dogs were tested for the antimicrobial activity, tolerance to bile and adhesion activity. The total count of fecal enterococci reached 5.5 log CFU/g and of lactobacilli 7.6 log CFU/g. Screening for production of bacteriocin-like substances showed an to partly inhibit the growth of Enterobacter sp. (hazy zones of inhibition). Ten strains of Enterococcus sp. and nine strains of Lactobacillus sp. were found without any inhibitory activity against all indicators used. Seven enterococcal strains and six lactobacilli with the broadest antimicrobial spectrum were selected for further probiotic assays. In the presence of 1% bile, the survival rate of selected enterococci (71.7-97.5%) was higher than that of lactobacilli (66.7-75.4%). The adhesion of strains to human intestinal mucus (5.1-8.2% by enterococci, 2.7-8.3% by lactobacilli) was found to be similar as adhesion to canine intestinal mucus (3.7-10.6% by enterococci, 2.1-6.0% by lactobacilli). Strain AD1, one lactobacillus isolate, reduced the higher level of serum cholesterol and alanine aminotransferase after oral administration to dogs suffering from diseases of the gastrointestinal tract.
Assuntos
Cães/microbiologia , Enterococcus/isolamento & purificação , Lactobacillus/isolamento & purificação , Probióticos , Alanina Transaminase/sangue , Animais , Antibacterianos/biossíntese , Antibacterianos/farmacologia , Antibiose , Aderência Bacteriana , Bacteriocinas/biossíntese , Bacteriocinas/farmacologia , Ácidos e Sais Biliares/farmacologia , Colesterol/sangue , Contagem de Colônia Microbiana , Enterobacter/efeitos dos fármacos , Enterococcus/efeitos dos fármacos , Enterococcus/crescimento & desenvolvimento , Enterococcus/metabolismo , Fezes/microbiologia , Gastroenteropatias/terapia , Inibidores do Crescimento/farmacologia , Lactobacillus/efeitos dos fármacos , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/metabolismo , Muco/metabolismoRESUMO
The adhesive capacity of selected enterococci to human, canine and porcine intestinal mucus was investigated in order to select for potential probiotic strains with good adhesive properties for human or animal use. The adhesion to the human intestinal mucus of the tested strains was found to range from log10 3.8 to log10 8.6 cfu per microtitre plate well. The highest adhesion to the human intestinal mucus was found among strains from horse faeces, dog faeces and dog feed. The adhesion to canine mucus was observed to range from log10 3.8 to log10 8.3 cfu/well, with the highest adhesive capacity among strains from dog faeces, horse faeces and dog's feed; on average log10 7.9, 7.3 and 7.0 cfu/well, respectively. Isolates from dogs did not bind at higher levels to canine mucus than to human mucus. A strong correlation was observed for the adhesion to human and canine intestinal mucus (p < 0.0001) and also between porcine and canine or human mucus (p < 0.05 for both). When comparing the adhesion of Enterococcus faecium and E. faecalis, no statistical significant differences were observed for any of the tested mucus types. The tested Enterococcus strains were found to exhibit a strain dependent on in vitro adhesion to human, canine and porcine intestinal mucus and did not exhibit host specificity in their adhesion, though some sources appeared to contain more adhesive strains than others. To our knowledge, this is the first report on the in vivo adhesion to intestinal mucus of a large number of enterococci from different sources.
Assuntos
Aderência Bacteriana/fisiologia , Enterococcus/fisiologia , Mucosa Intestinal/microbiologia , Probióticos , Ração Animal/microbiologia , Animais , Células Cultivadas , Contagem de Colônia Microbiana , Cães , Enterococcus faecalis/fisiologia , Enterococcus faecium/fisiologia , Fezes/microbiologia , Cavalos , Humanos , Enteropatias/prevenção & controle , Enteropatias/veterinária , Especificidade da Espécie , SuínosRESUMO
Sixty-two animal enterococci were examined for their binding of bovine fibrinogen, porcine fibronectin, bovine lactoferrin, bovine apotransferrin and human holotransferrin in the particle agglutination assay (PAA). Individual strains expressed binding of selected glycoproteins to various degrees (0, 1, 2, 3), whereas bovine fibrinogen binding of enterococci from goats, rabbits and rodents was the strongest (3) in general. Porcine fibronectin was bound weakly (1 or 2) by enterococci from horses, dogs, poultry, rabbits and rodents, while most of the goat isolates and half of the dog feed isolates did not bind fibronectin (0). Bovine lactoferrin was bound especially by the isolates from rodents and rabbits. Bovine apotransferrin was bound very weakly (1) by only a few isolates. Human holotransferrin was bound to a greater extent than apotransferrin by some isolates from rabbits and rodents. Since multiresistant strains are preferred in our binding studies, enterococci were also examined for their antibiotic resistance pattern. Almost all investigated isolates were resistant at least to one antibiotic. However, some strains displayed resistance to five or six antibiotics of 10 antibiotics tested. In a study of the inhibitory effect of heparin, porcine mucin and hyaluronic acid, the greatest effect was observed after heparin treatment of bacterial cells. These observations, as well as the expression of heparin binding by most strains, may suggest that at least one mode of enterococcal attachment utilizes glycosaminoglycan chains present on the surface of adherent cells.
Assuntos
Enterococcus/fisiologia , Matriz Extracelular/microbiologia , Fibrinogênio/metabolismo , Fibronectinas/metabolismo , Lactoferrina/metabolismo , Transferrina/metabolismo , Animais , Antibacterianos/farmacologia , Aderência Bacteriana/fisiologia , Farmacorresistência Bacteriana , Enterococcus/isolamento & purificação , Matriz Extracelular/metabolismo , Humanos , Testes de Fixação do Látex/veterinária , Testes de Sensibilidade Microbiana/veterinária , MicroesferasRESUMO
The impact of probiotic supplementation of canine-derived strain Lactobacillus fermentum AD1-CCM7421 in freeze-dried form on quantitative composition of microbiota and short-chain fatty acid profile in feces of dogs was demonstrated by two independent studies (straightforward repeated-measures model; study I: a dose of 2 g per dog for 2 weeks, 10(8) CFU/g, n = 12; study II: 1 g per dog for 1 week, 10(7) CFU/g, n = 11. The results revealed a significant increase of lactic acid bacteria population persisting also after the cessation of probiotic application in both studies. A reduction of clostridia (study I, p (sum) < 0.01) and tested Gram-negative bacterial genera (coliforms, Aeromonas sp., Pseudomonas sp., study II, p < 0.05) was also detected. The strain AD1-CCM7421 colonized the canine digestive tract in sufficient numbers (10(5)-10(6) CFU/g) and it persisted in the majority of dogs after cessation of probiotic application. An increase of short-chain fatty acid concentrations (study I: butyric, succinic, valeric, formic acid) especially in the early post-treatment phase (p < 0.05) most likely led to a decrease of fecal pH value (p < 0.05) without negative influence on fecal consistency throughout the studies.
Assuntos
Doenças do Cão/prevenção & controle , Limosilactobacillus fermentum/fisiologia , Probióticos/química , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Suplementos Nutricionais/análise , Doenças do Cão/metabolismo , Doenças do Cão/microbiologia , Cães , Ácidos Graxos/metabolismo , Fezes/química , Fezes/microbiologia , Liofilização , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Limosilactobacillus fermentum/química , MetagenomaRESUMO
Fifty-five strains of enterococci isolated from the piglet intestine were characterized in vitro for probiotic activity. Identification of the isolates revealed Enterococcus faecium as the predominant species (84 %). Forty strains (73 %) were found to produce bacteriocin-like substances (only into solid media) with activity almost only toward Gram-positive genera. Thirty-eight % of strains were resistant to tetracycline, 27 % to chloramphenicol, 18 % to erythromycin and 16 % to vancomycin. In addition to control of strain safety, 6 % of isolates were beta-hemolytic and 16 % produced gelatinase. Seven strains selected for further probiotic assays exhibited sufficient survival rate at pH 3.0 after 3 h, in the presence of 1 % ox-bile and lysozyme after 1 d (over 107 CFU/mL in all tests). The adhesion of tested strains to porcine and human intestinal mucus was found in a similar range (1.4-14.0 % and 1.4-17.6 %, respectively). In accordance with current research effort to use and/or to combine various health promoting substances, the sensitivity of all isolates toward plant extracts and toward bacteriocins produced by animal and environmental strains was determined. All enterococci were sensitive toward oregano and sage extracts and toward one (E. faecium EF55--chicken isolate, activity of 25 600 AU/mL) of ten bacteriocin substances. It means that a similar anti-enterococcal potential of some bacteriocin substances may be observed as for certain plant extracts.