RESUMEN
Post-weaning diarrhoea in piglets remains an important cause of economic losses for swine producers. Feed supplementation with probiotics is one of the alternatives to antibiotics used to reduce the impact of such gastrointestinal disease. The aim of the present study was to evaluate the effect of Ligilactobacillus salivarius PS21603 supplementation on the intestinal structure and the gut microbiota composition of weaned piglets. Safety and tolerance of L. salivarius PS21603 were previously evaluated in a 28-days study using 384 weaned piglets (28 ± 2 days old and 7.5 ± 1.5 kg) divided in three treatment groups: T1: Basal diet + L. salivarius PS21603 109 cfu/day, T2: Basal diet + L. salivarius PS21603 107 cfu/day, and T3: Basal diet (control group). For the present study, 16 piglets per treatment group were randomly selected and faecal samples were collected on day 0 (weaning) and 28 of study. At the end of study, three males and three females per treatment were euthanised. Intestinal morphometric values were measured after necropsy. Faecal counts of Escherichia coli were evaluated by culture techniques, and faecal microbiota composition was assessed by high-throughput sequencing. All data were analysed and compared between treatment groups. Supplementation with L. salivarius PS21603 caused an increase in the intestine length of piglets from T1 and in the villous height:crypt ratio of piglets from T2 (P < 0.05) compared to T3 on day 28. According to the Shannon Diversity Index, microbiota diversity increased on day 28 compared to day 0, with no significant differences observed between treatments. The main changes in the relative abundance of bacteria at the phylum, family, and genus levels were observed between different sampling time points. However, piglets from T1 and T2 had lower faecal E. coli counts than T3 on day 28 (P < 0.05). Moreover, supplementation with L. salivarius PS21603 modulated gut microbiota through a more optimal composition, reducing Escherichia and increasing Bifidobacterium relative abundance in piglets from T1 (P < 0.05) from the beginning to the end of the study. Therefore, the strain L. salivarius PS21603 has shown probiotic properties to be used as feed additive in the pig industry, along with good hygiene and farm management practices, for the prevention and/or treatment of post-weaning diarrhoea in piglets.
Asunto(s)
Alimentación Animal , Suplementos Dietéticos , Heces , Microbioma Gastrointestinal , Ligilactobacillus salivarius , Probióticos , Destete , Animales , Microbioma Gastrointestinal/efectos de los fármacos , Porcinos/microbiología , Probióticos/administración & dosificación , Probióticos/farmacología , Masculino , Femenino , Ligilactobacillus salivarius/fisiología , Alimentación Animal/análisis , Heces/microbiología , Intestinos/microbiología , Diarrea/veterinaria , Diarrea/microbiología , Diarrea/prevención & control , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/prevención & control , Escherichia coliRESUMEN
The present study aimed to characterise in vitro properties of the strain Ligilactobacillus salivarius PS21603 and evaluate in vivo piglets' tolerance for its use as feed additive in swine. The ability of L. salivarius PS21603 of inhibiting enteropathogens' growth in vitro was evaluated using a co-culture assay. Low pH tolerance, bile tolerance, and resistance to osmotic changes were evaluated. The antibiotic susceptibility profile of L. salivarius PS21603 was assessed through broth microdilution method. Whole genome sequencing (WGS) was performed to exclude the presence of antibiotic resistance genes. L. salivarius PS21603 showed a high antimicrobial activity in vitro, reducing in a mean of 6.16 log cfu/ml eight different enterotoxigenic Escherichia coli strains. Moreover, L. salivarius PS21603 showed resistance to osmotic changes and was able to survive to a pH above 3.5 during 24 h and up to pH 2 at least during 2 h. In addition, WGS revealed that L. salivarius PS21603 did not harbour any resistance genes and thus there was no risk of transmissibility. Finally, an in vivo 28-days safety and tolerance study was performed. For that, 384 healthy piglets (28±2 days old and 7.5±1.5 kg, at weaning) were divided into three treatment groups receiving a different dose of L. salivarius PS21603: T1, 109 cfu/day; T2, 107 cfu/day; T3, control. Piglet's health status was daily controlled. Individual bodyweight and feed intake per pen were weekly recorded to determine performance parameters. Blood samples were collected in 16 piglets from each treatment group on days 0 and 28 for determination of cytokine profiles. L. salivarius PS21603 was safe and well tolerated by piglets, there were no differences in performance nor cytokine profile between treatment groups. In conclusion, L. salivarius PS21603 is a potential candidate for a probiotic prevention strategy against pig diarrhoea.