Microbial status and quality of rabbit meat after rabbits feed supplementation with phyto-additives

Nowdays, there is an increasing  interest in public health issue due to consumption of healthy and nutritive food, e.i. rich in proteins, low in cholesterol and lipid contents. The aim of this study was to examine if oregano, sage and Eleutherococcus senticosus extracts, commercial Xtract as well can influence microbial status of rabbit meat after their supplementation of rabbit feed. Reduction of enterococci, coagulase-negative staphylococci and Staphylococcus aureus after feed supplementation by the sage and oregano was more detectable at the end of the experiment (at day 42) than after their application (at day 21). In animals with Xtract addition, antibacterial effect has been noted already at day 21. Our in vivo results confirmed in vitro antibacterial effect of the tested extracts and showed that maintaining of rabbit meat quality by plant extracts is very promising

Enterococci from pannon white rabbits: detection, identification, biofilm and screening for virulence factors

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.The results achieved were financially supported by the bilateral Slovak-Hungarian project APVV:SK-HU-0006-08 and partially by the projects Probiotech ITMS 26220220204 and Vega 2/0006/2017.Lauková, A.; Strompfová, V.; Szabóová, R.; Bónai, A.; Matics, Z.; Kovács, M.; Pogány Simonová, M. (2019). Enterococci from pannon white rabbits: detection, identification, biofilm and screening for virulence factors. World Rabbit Science. 27(1):31-39. https://doi.org/10.4995/wrs.2019.10875SWORD3139271Bagóne Vántus V., Dalle Zotte A., Cullere M., Bónai A., Dal Bosco A., Szendro Zs., Tornyos G., Pósa R., Bóta B., Kovács M., Zsolnai A. 2018. Quantitative PCR with 16S rRNA-genetargeted specific primers for analysis of caecal microbial community in growing rabbits after dietary supplementation of thyme (Thymus vulgaris) and spirulina (Arthrospira platensis). Ital. J. Anim. Sci., 17: 657-665.Bino E., Lauková A., Kandričáková A., Nemcová R. 2018. Assessment of biofilm formation by faecal strains of Enterococcus hirae from different species of animals. Polish J. Vet. Sci., 21: 747-754. https://doi.org/10.24425/124314Bruker Daltonics Biotyper 2.0.2011. Software for microorganismsidentification and classification user manual.Clinical and Laboratory Standards Institute (CLSI). 2012. In Performance Standards for Antimicrobial Disk Susceptibility T; Approved Standards-Eleventh Edition CLSI=NCCLS M02-A11. Clinical and Laboratory Standards Institute, Wayne, Pensylvania 19087, USA.Dalle Zotte A. 2002. Perception of rabbit meat quality and major factors influencing the rabbit carcass and meat quality. Liv. Prod. 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Med., 15: 308-320. https://doi.org/10.1177/154411130401500506Kubašová I., Strompfová V., Lauková A. 2017. Safety assessment of commensal enterococci from dogs. Folia Microbiol., 62: 491-498. https://doi.org/10.1007/s12223-017-0521-zLatasa C., Solano C., Penades J.R., Lasa I. 2006. Biofilmassociated proteins. C. R. Biol., 329: 849-857. https://doi.org/10.1016/j.crvi.2006.07.008Lauková A. 1995. Characteristics of streptococci and enterococci isolated from rumen of mouflons and European bisons. Asian Austral. J. Anim., 8: 37-41. https://doi.org/10.5713/ajas.1995.37Lauková A. 1999. Vancomycin-resistant enterococci isolates from rumen content of deer. Microbios, 97: 95-101.Lauková A., Bomba A., Kmeť V. 1990. Enterococcus occurrence and urease activity in rumen content of claves after dieteticomicrobial stimulation. (in Slovak) Anim. Prod. (Czech), 11: 971-975.Lauková A., Simonová M., Strompfová V., Štyriak I., Ouwehand A.O., Várady M. 2008. Potential of enterococci isolated from horses. Anaerobe, 14: 234-236. https://doi.org/10.1016/j.anaerobe.2008.04.002Lauková A., Chrastinová Ľ., Pogány Simonová M., Strompfová V., Plachá I., Čobanová K., Formelová Z., Chrenková M., Ondruška Ľ. 2012. Enterococcus faecium AL41: Its Enterocin M and their beneficial use in rabbits husbandry. Prob. Antimicro. Prot., 4: 243-249. https://doi.org/10.1007/s12602-012-9118-7Lauková A., Strompfová V., Kandričáková A., Ščerbová J., Semedo-Lemsaddek T., Miltko R., Belzecki G. 2015. Virulence factors genes in enterococci isolated from beavers (Castor fiber). Folia Microbiol., 60: 151-154. https://doi.org/10.1007/s12223-014-0357-8Lauková A., Kandričáková A., Ščerbová J., Strompfová V. 2016. Enterococci isolated from farm ostriches and their relation to enterocins. Folia Microbiol., 61: 275-281.https://doi.org/10.1007/s12223-015-0435-6Lauková A., Pogány Simonová M., Chrastinová Ľ., Gancarčíková S., Kandričáková A., Plachá I., Chrenková M., Formelová Z., Ondruška Ľ., Ščerbová J., Strompfová V. 2018. Assessment of lantibiotic type bacteriocin gallidermin application in model experiment with broiler rabbits. Int. J. Anim. Sci., 2: 1028.Linaje J., Coloma M.D., Pérez-Martínez G., Zuniga M. 2004. Characterization of faecal enterococci from rabbits for selection of probiotic strains. J. Appl. Microbiol., 96: 761-771. https://doi.org/10.1111/j.1365-2672.2004.02191.xMarounek M., Březina P., Baran M. 2000. Fermentation of carbohydrates and yield of microbial protein in mixed cultures of rabbit caecal microorganisms. Arch. Anim. Nutr., 53: 241-252. https://doi.org/10.1080/17450390009381950Pogány Simonová M., Lauková A., Plachá I., Čobanová K., Strompfová V., Szabóová R., Chrastinová Ľ. 2013. Can enterocins affect phagocytosis and gluthationperoxidase in rabbits? Cent. Eur. J. Biol., 8: 730-734.Pogány Simonová M., Lauková A. 2017. Virulence factor genes possessing Enterococcus faecalis from rabbits and their sensitivity to enterocins. World Rabbit Sci., 25: 63-71. https://doi.org/10.4995/wrs.2017.5694Ribeiro T., Oliveira M., Fraqueza M.J., Lauková A., Elias M., Tenreiro R., Barreto A.S., Semedo-Lemsaddek T. 2011. Antibiotic resistance and virulence factors among Enterococci isolated from chourico, a traditional Portuguesee dry fermented sausage. J. Food Prot., 74: 465-469.https://doi.org/10.4315/0362-028X.JFP-10-309Rice L.B., Eliopoulos G.M., Wennerstein C.H., Goldman D, Jacoby G.A., Moellering R.C. jr. 1991. Chromosomally mediated beta-lactamase production and gentamicin resistance in Enterococcus faecalis. Antimicrob. Agents Chemother., 2: 272-276. https://doi.org/10.1128/AAC.35.2.272Semedo T., Santos M.A., Lopes M.F., Figueirdo Marques J.J., Barreto Crespo M.T., Tenreiro R. 2003. Virulence factors in food, clinical and reference Enterococci: a common trait in the genus? Syst. Appl. Microbiol., 26: 13-22. https://doi.org/10.1078/072320203322337263Simonová M., Lauková A. 2004. 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Beneficial effects of Enterococcus faecium EF9a administration in rabbit diet

[EN] Forty-eight rabbits aged five weeks (Hycole breed, both sexes) were divided into experimental (EG) and control (CG) groups, 24 animals in each, and fed a commercial diet with access to water ad libitum. Rabbits in EG had Enterococcus faecium EF9a probiotic strain added to their drinking water (1.0×109 colony forming units/mL 500 μL/d/animal) for 28 d (between 35 and 63 d). The experiment lasted for 42 d. The animals remained in good health condition throughout the experiment, and no morbidity and mortality was noted. There was a higher live weight at 63 d of age (+34 g; P<0.0001), final live weight at 77 d of age (+158 g; P=0.0483), and average daily weight gain between 63 and 77 d of age in the EG group rabbits than in CG group rabbits (+8 g/d; P<0.0001). No significant changes in caecal lactic acid and total volatile fatty acid concentrations, jejunal morphological parameters and phagocytic activity were noted during the treatment. The tested serum parameters were within the range of the reference values. EF9a strain sufficiently established itself in the rabbit’s gastrointestinal tract. At 63 d of age, a significant decrease in coliforms (P<0.05), coagulase-positive staphylococci (P<0.01), pseudomonads (P<0.01) and coagulasenegative staphylococci (CoNS, P<0.001) was noted in the faeces of the EG group rabbits compared to the CG rabbits. Antimicrobial effects of EF9a strain in the caecum against coliforms (P<0.001), CoNS (P=0.0002) and pseudomonads (P=0.0603) and in the appendix (coliforms, P<0.05) were detected.Slovak – Hungarian project APVV:SK-HU-0006-08 and the national VEGA project 2/0006/17 This work was financially supported by the bilateral Slovak – Hungarian project APVV:SK-HU-0006-08 and the national VEGA project 2/0006/17. Part of the preliminary results was presented in the Proceedings from the Conference in Kaposvár, Hungary, 30.05.2012, pp. 89-92. We are grateful to Mrs. M. Bodnárová and Mr. P. Jerga for their skilful technical assistance. We are also grateful to Dr. V. Párkányi and Dr. R. Jurčík, from the National Agricultural and Food Centre in Nitra for blood sampling and Mr. J. Pecho for slaughtering. All care and experimental procedures involving animals followed the guidelines stated in the Guide for the Care and Use of Laboratory Animals and the trials were accepted by the Ethic Commission of the Institute of Animal Physiology in Košice and by the Slovak Veterinary and Food Administration. We would like to thank to Mr. A. Billingham for English language correction.Pogány Simonová, M.; Lauková, A.; Chrastinová, Ľ.; Plachá, I.; Szabóová, R.; Kandričáková, A.; Žitňan, R.... (2020). Beneficial effects of Enterococcus faecium EF9a administration in rabbit diet. World Rabbit Science. 28(4):169-179. https://doi.org/10.4995/wrs.2020.11189OJS16917928

Detection of structural genes for enterocins production among enterococci isolated from french traditional fermented sausages

National audienceIn European countries, the demand for traditional food products has increased. Dry fermented sausages, mainly manufactured with pork and/or lean beef and pork fat, account for a significant part of traditional meat products. Many studies have dealt with the microbiology of traditional fermented sausages but there are few detail studies of properties of individual genera. The genus Enterococcus involved about 55 validated species. Its representants are mostly lactic acid producing; they can possess probiotic properties or produce bacteriocins (antimicrobial substances); on the other side, some strains can posess virulence factors genes. Our study has been focused on positive property-on detection of structural genes for enterocins production among enterococci isolated from French traditional fermented sausages. Enterocins are proteinaceous antimicrobial substances produced mostly by enterococci with inhibition spectrum against Gram-positive but also against Gram-negative bacteria. This study was performed in the framework of the EU project TRADISAUSAGE-QLK1-CT2002-02240; that is why also French traditional fermented meat products were analysed. Samples of products from 10 low-capacity processing units (placed in Massif-central-France) were collected. Sampling was performed from the initial meat (Z), during processing (M) and from the final product (F). Sampling and analytical methods for the microbiota analysis were based on the International Organisation for Standardisation (ISO). Enterococci were counted on M-Enterococcus agar (Merck, Germany). Fifty–three pure colonies were identified by PCR (primers) confirmed by Maldi-Tof identification system. Enterocins genes were tested by primers and PCR. Most of strains belonged to the species Enterococcus faecium, followed by E. faecalis and other not specified species. Strains survived in Oxgall bile evironment in high %. The most detected was Ent P gene (34%) followed by Ent A gene (27%) and Ent L50B (20%). The least was detected Ent B gene (19%)

Effect of Enterococcus faecium EF 55 on morphometry and proliferative activity of intestinal mucosa in broilers infected with Salmonella Enteritidis

Introduction: The present study aimed to investigate the effect of Enterococcus faecium EF55 on chickens, as well as its influence on proliferative activity of epithelial intestinal cells after infection with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4). Moreover, the length and area of duodenal and jejunal villi of the birds were examined

Effects of Dietary Zinc and/or an Herbal Mixture on Intestinal Microbiota and Barrier Integrity in Lambs

The purpose of this experiment was to determine the impact of feed supplementation with organic zinc and/or a medicinal plants mixture on the composition and enzymatic activity of intestinal microflora as well as on the duodenal and jejunal barrier integrity in lambs. A total of 28 lambs were randomly allocated into 4 dietary treatments (n = 7) and were fed an unsupplemented basal diet (BD), or the BD enriched with organic Zn (Zn, 70 mg/kg diet), an herbal mixture (Herbmix, 100 g/day) or a combination of both additives (Zn+Herbmix). The Herbmix contained 33% each of Fumaria officinalis, Malva sylvestris, Matricaria chamomilla and 1% Artemisia absinthium. No significant effect on the fecal microbiota composition was observed due to the 35-day or 70-day dietary treatment. The intake of Zn alone resulted in decreased bacterial enzyme activities, such as β-glucuronidase, N-acetyl-glucosaminidase, β-galactosidase and β-glucosidase. The transepithelial electrical resistance of the small intestinal mucosa was not influenced by the dietary treatment, whereas simultaneous feeding of Zn and Herbmix exhibited higher claudin-1 and occludin levels in the jejunal mucosa. These results indicate that dietary intake of organic zinc and/or medicinal plants in the mentioned dosage did not alter the diversity of intestinal bacteria in growing lambs but did significantly influence bacterial enzyme activity. Supplementing the zinc and herbs combination showed the potential to regulate intestinal permeability by increasing the level of tight junction proteins in the jejunal mucosa

Antimicrobial activity of Enterococcus faecium EF 55 against Salmonella Enteritidis in chicks

The protective effect of Enterococcus faecium EF 55 against Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was studied in 1-day-old chicks. The EF 55 strain (isolated and characterised by the authors earlier) was applied daily (1.109 CFU/0.2 ml PBS) for 7 days. Oral inoculation of the SE PT4 strain was performed on day 8 in a single dose of 5.108 CFU/0.2 ml PBS. The experiment lasted for 21 days. Samples were collected on day 1 of the experiment to verify the absence of Salmonella, on day 8 to check colonisation of EF 55 and immunological status in experimental birds, and on days 2, 4, 6, 8 and 14 after SE PT4 infection of chicks. Strain EF 55 sufficiently colonised the digestive tract of chicks after 7 days of application. The highest numbers of EF 55 in the faeces of chicks were observed before SE infection and persisted to day 6 post infection (p.i.) in both the EF and EF+SE groups. PCR confirmed the identity of the EF 55 strain. The counts of SE PT4 strain in faeces of the EF+SE group were significantly reduced in comparison to those in the SE group on days 2 and 14 p.i. (P < 0.01). The significant reduction of salmonellae in the caecum was recorded at the end of the experiment (day 14 p.i.) in the EF+SE group in comparison to the SE group (P < 0.01). At day 4 p.i., colonies of S. Enteritidis PT4 were found in the liver of chicks of the SE group in a higher concentration than in chicks of the EF+SE group (P < 0.001). Salmonellae were isolated from the liver until days 8 and 6 p.i. in the SE and EF+SE groups, respectively. The mean values of actual lymphocyte subpopulations in the blood and the relative percentage of caecal intraepithelial lymphocyte subpopulations (CD4, CD8, CD44, TCR, MHC II and IgM) were not influenced at a statistically significant level by the application of the EF 55 and/or the SE PT4 strain. The results demonstrate the antimicrobial effect of E. faecium EF 55 against S . Enteritidis PT4