Lactic acid bacteria isolated from young calves – Characterization and potential as probiotics
Introduction
The gastrointestinal microbiota of farm animals is of fundamental importance and can affect the host in the digestion of fibre (in ruminants) and act as a barrier against pathogens and toxic substances, mainly in young or neonates and those experiencing stress and unfavourable environmental conditions. The indigenous microbiota established after birth interacts with the digestive and immune systems of the host. The colonization of the different compartments of the intestinal tract by specific commensal bacteria is facilitated initially by their association with the mucus layer or epithelial cells of the mucosal surfaces to produce a first barrier against invading microorganisms (Nousiainnen et al., 2004). Lactic acid bacteria (LAB) and related microorganisms are microbial groups that initially colonise the intestinal tract of newborns, coming from the vaginal tract during parturition, and later from the mammary gland and the environment (Long and Swenson, 1977).
In healthy animals, each segment of the intestine is colonized by a typical microbiota, which are adapted to grow in symbiosis with the host. The indigenous microbiota is stable for some months of life, and can be modified by exogenous and endogenous factors, exert many different effects in the host, and some scientists suggest that it acts as a “different organ” interacting permanently with the host. Also, microbial imbalances during growth might permit the colonization by pathogens and the appearance of many different syndromes with deleterious effects on the host (Bauer et al., 2006, O’Hara and Shanahan, 2006, Guarner and Malagelada, 2003). Diarrhoea is one and causes the calf death and financial loss in dairy farms (Torsein et al., 2011, Gulliksen et al., 2009, Morrell et al., 2008, Virtala et al., 1996, Azzam et al., 1993, Bellinzoni et al., 1990, Bellows et al., 1987). Probiotic microorganisms have been proposed as adjuvant to promote health status in newborn calves (Ewaschuk et al., 2004, Magalhăes et al., 2008, Timmerman et al., 2005). Probiotics are defined as “live microorganisms administered to the host to produce a beneficial physiological effect” (Reid et al., 2003) and are recognized as GRAS (generally recognized as safe) microorganisms. Many probiotic products are available in the market for different purposes, but the bacteria they contain are sometimes uncharacterized (Saxelin, 2008, Brink et al., 2005).
One of the objectives of our research group was to identify the lactic acid bacteria to administer to newborn calves, isolated from this group of animals. We initially isolated bacteria classified as GRAS from faeces and oral cavity of newborn calves, supported by the host specificity phenomenon described and exerted by members of the indigenous microbiota (Kotarski and Savage, 1979, Zoetendal et al., 2006). Screening of properties relating to beneficial characteristics (potential to adhere to the intestinal tract or production of inhibitory substances) was performed based in previous experience (Ocaña et al., 1999, Ocaña and Nader Macías, 2002, Otero et al., 2006). The resistance to the intestinal tract conditions was also evaluated. Selected bacteria were identified by molecular techniques, included in an international Culture Collection and assessed for suitability as probiotics for newborn calves.
Section snippets
Animals, samples collection and enumeration of aerobic population
Fifty-one samples were obtained from the oral cavities and rectums of 42 healthy young calves (less than four months of age) from dairy and beef farms in Tucumán and Jujuy provinces (Argentina). Animals sampled were clinically healthy (based on body weight gain, daily intake, natural behavior, lack of diarrhoea symptoms) and had not received antibiotics for the last month prior to sample collection.
The samples were collected with sterile swabs in MRS 1% agar pH 4.5 (De Man et al., 1969) (Merck,
Isolation of microorganisms
The mesophilic bacteria recovered from faecal samples of calves of different age was from 107 to 109 CFU/g, without any significant differences between them (p ≤ 0.05, Kruskal Wallis test) as shown in Fig. 1. There were differences statistically significant between the numbers of LAB and enterobacteria isolated from newborn calves (from 0 to 1 month age) and those from older animals (2–4 months age) as indicated in the same Fig. 1.
Ninety-six isolates were identified as LAB: 81% of the bacilli and
Discussion
The increased use of uncharacterised probiotics in animals to modify weight gain and resistance to infections led us to study the characteristics of LAB strains that might be included in the design of a probiotic for calves. This product could increase the health status of this group of animals, and also might be used for the prevention of diarrhoea responsible for high mortality and morbidity in neonates (Morrell et al., 2008, Azzam et al., 1993, Bellinzoni et al., 1990, Bellows et al., 1987).
Acknowledgements
This work was supported by PICT 0543 from ANPCYT (Agencia Nacional de Promoción Científica y Tecnológica Agencia, Argentina) and PIP 0632 from CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina)
We thank Lic. Elena Bru for the statistical evaluation of the data.
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