This study examines the effects of the dietary supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) (LA) on productive performances, incidence of foot pad dermatitis and caecum microbioma in broiler chickens. A total of 1,100 one-day old male Ross 308 chicks were divided into 2 groups of 16 replicates with 25 birds each and reared from 1–41 d. One group was fed a basal diet (CON) and the other group the same diet supplemented with LA. Caecum contents were collected from 4 selected birds at day one and 5 selected birds at the end of the rearing period. Then, they were submitted to DNA extraction and whole DNA shotgun metagenomic sequencing. Overall, the LA supplementation produced a significant beneficial effect on body weight gain between 15–28 d and improved feed conversion rate in the overall period. On the contrary, litter moisture, pH and incidence of the foot pad lesions were not affected by LA. Birds treated with LA showed a lower occurrence of pasty vent at both 14 and 28 d. At the end of the rearing period, Lachanospiraceae were significantly higher in LA birds in comparison to CON (17.07 vs 14.39%; P = 0.036). Moreover, Ruminococcus obeum, Clostridium clostridioforme, Roseburia intestinalis, Lachnospiraceae bacterium 14-2T and Coprococcus eutactus were significantly higher in LA birds in comparison to CON. The relative abundance of Lactobacillus acidophilus was comparable between LA and CON groups. However, a positive effect was observed in relation to the metabolic functions in the treated group, with particular reference to the higher abundance of β-glucosidase. In conclusion, the LA supplementation improved broiler productive performances and metabolic functions promoting animal health.

In order to evaluate the effects of dietary addition of probiotic strain Lactobacillus acidophilus D2/CSL on hen performance and egg quality, 160 commercial Hy-Line Brown pullets, 17 weeks old, were divided in control group (C) (N=80) and treated group (T) (N=80), with 4 alternate replicates of 20 animals each per group. C was fed with a "standard diet", whereas the T group received the same diet with an inclusion of 1x10 9 CFU kg -1 of Lactobacillus acidophilus D2/CSL (freeze dried cells). The experimental trial lasted 39 weeks after one week of acclimatization. Hen performance and egg quality (egg production, FCR, egg specific gravity, shell thickness, Haugh Units) were recorded. The results show a higher overall egg production (P<0.01) and better FCR (Kg feed intake/Kg saleable eggs) (P<0.05) in the T birds, but no statistically significant differences were observed in egg weight. The eggs from the T birds were characterized by a higher specific gravity (ESG) (P<0.01) and albumen viscosity (Haugh Units) (P<0.05). No significant differences in egg shell thickness were recorded. In conclusion, Lactobacillus acidophilus D2/CSL improved some important parameters in laying hen performance and egg quality. RIASSUNTO EFFETTO DEL LACTOBACILLUS ACIDOPHILUS D2/CSL SULLE PERFORMANCE DELLA GALLINA OVAIOLAAl fine di verificare gli effetti del ceppo probiotico denominato Lactobacillus acidophilus D2/CSL, 160 pollastre commerciali Hy-Line Brown di 17 settimane di vita sono state divise in due gruppi omogenei di 80 animali ciascuno, controllo (C) e trattamento (T). Ogni gruppo era composto da 4 ripetizioni, con 20 animali/ripetizione. Il gruppo C era alimentato con la "dieta standard", mentre il gruppo T riceveva la stessa dieta additivata con 1x10 9 UFC kg -1 di Lactobacillus acidophilus D2/CSL (cellule liofilizzate). La raccolta dei dati sperimentali è iniziata dopo una settimana di acclimatamento degli animali ed è proseguita per 39 settimane.

In this study we gained insights into the effects of the supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) in the chicken drinking water on crop and caeca microbiomes. The probiotic was supplemented at the concentrations of 0.2 g Lactobacillus acidophilus/day/bird and 0.02 g Lactobacillus acidophilus/day/bird and its effect on the crop and caeca microbiomes was assessed at 14 and 35 days of rearing. The results showed that mean relative abundance of Lactobacillus acidophilus in the caeca did not show significative differences in the treated and control birds, although Lactobacillus acidophilus as well as Faecalibacterium prausnitzii, Lactobacillus crispatus and Lactobacillus reuteri significantly increased over time. Moreover, the treatment with the high dose of probiotic significantly increased the abundance of Clostridium asparagiforme, Clostridium hathewayi and Clostridium saccharolyticum producing butyrate and other organic acids supporting the chicken health. Finally, at 35 days, the Cell division protein FtsH (EC 3.4.24.-) and the Site-specific recombinase genes were significantly increased in the caeca of birds treated with the high dose of probiotic in comparison to the control group. The results of this study showed that Lactobacillus acidophilus D2/CSL (CECT 4529) supplementation in the drinking water at the concentrations of 0.2 and 0.02 g Lactobacillus acidophilus/day/bird improved beneficial microbes and functional genes in broiler crops and caeca. Nevertheless, the main site of action of the probiotic is the crop, at least in the early stage of the chicken life. Indeed, at 14 days Lactobacillus acidophilus was significantly higher in the crops of chickens treated with the high dose of LA in comparison to the control (14.094 vs 1.741%, p = 0.036).

Twenty-nine strains of mastitis pathogens were used to study the antibacterial activity of the cell-free supernatants (CFS) of 25 strains of Lactococcus lactis ssp. lactis. Out of the tested strains, only the CFS of L. lactis LL11 and SL153 were active, inhibiting and killing most of the pathogens. By means of ultra-performance liquid chromatography/high resolution mass spectrometry, they were shown to produce nisin A, a class I bacteriocin. A variable sensitivity to nisin A-containing CFS was observed among Streptococcus uberis and Enterococcus faecalis strains. Nonetheless, Streptococcus agalactiae, Strep. uberis, and E. faecalis displayed high minimum inhibitory concentration values, reaching 384 arbitrary units/mL. Interestingly, the minimum inhibitory values and the bactericidal concentrations were almost identical among them for each of the 2 stains, LL11 and SL153. Staphylococci were, on average, less sensitive than streptococci, but the 2 CFS inhibited and killed, at different dilutions, strains of methicillin-resistant Staphylococcus aureus. The immune response to nisin A-containing CFS was tested using the bovine mammary epithelial cell line BME-UV1. Application of CFS did not damage epithelial integrity, as demonstrated by the higher activity of N-acetyl-β-d-glucosaminidase (NAGase) and lysozyme inside the cells, in both treated and control samples. On the other hand, the increase of released NAGase after 15 to 24h of treatment with LL11 or SL153 live cultures demonstrated an inflammatory response of epithelial cells. Similarly, a significantly higher lysozyme activity was detected in the cells treated with LL11 live culture confirming the stimulation of lysosomal activity. The treatment of epithelial cells with SL153 live culture induced a significant tumor necrosis factor-α downregulation in the cells, but did not influence IL-8 expression. The control of tumor necrosis factor-α release could be an interesting approach to reduce the symptoms linked to clinical intramammary infections. Due to their antibacterial activity and to the stimulation of lysosomal activity of mammary epithelial cells, the L. lactis strains SL153 and LL11 could be of interest for the development of alternative intramammary treatments to control cow mastitis.

BackgroundThe present study aimed to evaluate the effects of the probiotic strain Lactobacillus acidophilus D2/CSL (CECT 4529) on nutritional condition and faecal quality in cats.MethodsTen healthy adult cats from the same cattery were included (aged >9 months; male:female sex ratio=3:7). The animals were randomly assigned to a control group (CTR; n=5; male:female=1:4; room 1: 16 m2) and to a treated group (LACTO; n=5; male:female=2:3; room 2: 16 m2) receiving the same commercial dry diet. The LACTO group diet was supplemented with the probiotic (5 x 109 cfu/kg feed at least). A five-week experimental period was applied, and nutritional status was monitored by bodyweight (BW) and body condition score (BCS). Faecal quality was evaluated using faecal score (FS) and faecal moisture (FM) parameters. Plate counts of some faecal bacteria species were carried out. The data obtained were analysed using MIXED, GLM and NPAR1WAY procedures (SAS V.9.4; P≤0.05).ResultsThe two groups did not show differences in BW and BCS data. A clear effect of the probiotic supplementation on FM was recorded (LACTO 44 per cent v CTR group 46 per cent; P=0.04). FS in the LACTO group (3.35) was close to ideal values (2–3) in comparison with the CTR group (3.75). Positive effects of L acidophilus D2/CSL have been recorded in terms of increase in faecal lactobacilli counts and reduction in faecal coli counts.ConclusionsThis study’s preliminary results describe how inclusion of L acidophilus D2/CSL (CECT 4529) probiotic strain in cats’ diets could effectively improve faecal quality parameters and consequently gut health in adult healthy cats.

The present research tested the effects of dietary nisin-producing Lactococcus lactis on growth performance, feed utilization, intestinal morphology, transcriptional response, and microbiota in gilthead sea bream (Sparus aurata). A feeding trial was conducted with fish weighting 70–90 g. Fish were tagged with passive, integrated transponders and distributed in nine 500 L tanks with 40 fish each. Fish were fed for 12 weeks with either a control (diet A) or experimental diets (diets B and C) in triplicate (3 tanks/diet). Extruded pellets of diets B and C were supplemented with a low (2 × 109 CFU/kg) and a high (5 × 109 CFU/kg) dose of probiotic, respectively. No significant differences were found between groups for the feed conversion ratio or specific growth rates. However, the final body weight of fish fed diet C was significantly higher than the control group with intermediate values for fish fed diet B. Histological analysis conducted using a semi-quantitative scoring system showed that probiotic did not alter the morphology of the intestine and did not trigger inflammation. With regard to the transcriptomic response, a customized PCR array layout was designed to simultaneously profile a panel of 44 selected genes. Significant differences in the expression of key genes involved in innate and acquired immunity were detected between fish fed probiotic and control diets. To analyze the microbiota associated to the feeds and the gut autochthonous microbial communities, we used the Illumina MiSeq platform for sequencing the 16S rRNA gene and a metagenomics pipeline based on VSEARCH and RDP databases. The analysis of gut microbiota revealed a lack of colonization of the probiotic in the host’s intestinal mucosa. However, probiotic did modulate the fish gut microbiota, confirming that colonization is not always necessary to induce host modification. In fact, diets B and C were enriched with Actinomycetales, as compared to diet A, which instead showed a higher percentage of Pseudomonas, Sphyngomonas, and Lactobacillus genera. These results were confirmed by the clear separation of gut bacterial community of fish fed with the probiotic from the bacterial community of control fish group in the beta-diversity and PLS-DA (supervised partial least-squares discriminant analysis) analyses.

Background The aim of the present study was to investigate the effects of Lactobacillus acidophilus D2/CSL (CECT 4529) probiotic strain on nutritional status and faecal and microbiological parameters in a group of purebred boxers. Methods Forty healthy adult boxer dogs were randomly assigned to a treated (LACTO) group receiving a commercial diet supplemented with L acidophilus D2/CSL (CECT 4529) to a final concentration of 5.0 x 10 9 colony-forming unit/kg of food, and a control (CTR) group receiving the same diet but without the probiotic (placebo). Nutritional status (body weight, skinfold thickness, body condition score) and faecal quality parameters were analysed. Results No differences in body weight and skin thickness were found during the whole experimental period. Dogs in the LACTO group showed a significantly higher body condition score than those in the CTR group (4.86±0.55 v 4.65±0.65), and no significant differences were recorded in body weight and skinfold thickness. The LACTO group showed a significantly lower faecal moisture (in per cent) compared with the CTR group (0.67±0.007 v 0.69±0.007). Faecal hardness (in kg) was higher in the LACTO group than in the CTR group (0.86±0.047 v 0.70±0.051), and faecal score also improved in the LACTO group (3.78±0.95 v 4.25±0.91). A significant difference in total Escherichia coli counts as well as in lactobacilli counts between the CTR and LACTO groups was only detected at 28 days. Conclusion Supplementation of L acidophilus D2/CSL (CECT 4529) significantly improved the nutritional status and faecal parameters of dogs.

The aim of our case-control study is to evaluate the effects of a diet integrated with the probiotic Lactobacillus acidophilus D2/CSL (CECT 4529) on the nutritional status and faecal consistency of healthy dogs belonging to the English Cocker Spaniel (ECS) and Labrador Retriever (LR) breeds. A total of 30 dogs were enrolled in this study, and they were randomly assigned to a Control (CTR, n ¼ 14) and a Treated group (LACTO, n ¼ 16). The trial consisted in a 7-days adaptation period where all the animals received the same commercial food, followed by a 35-days of data collection period where the LACTO group received the food supplemented with Lactobacillus acidophilus D2/CSL. We evaluated Body weight (BW), Body Condition Score (BCS) and Skinfold thickness, Faecal Score (FS) and Faecal Moisture (FM). All dogs in the LACTO group maintained an ideal BCS score during the whole experimental period compared to the CTR group. A significant decrease in skin thickness was found throughout the trial in the LACTO group. A significant improvement of the FM was recorded in the LACTO compared to the CTR group in the overall period for both dog breeds, and the FS significantly decreased in the LACTO group. Our results showed good maintenance of the nutritional conditions in dogs that are prone to overweight and a significant improvement of faecal parameters, meaning that even in healthy dogs with no gastrointestinal disorder the addition of this supplement to the diet helps to maintain the optimal balance of their intestinal microbiota. HIGHLIGHTS Probiotics help to maintain and promote the optimal gastro-intestinal health and well-being. Lactobacillus acidophilus D2/CSL (CECT 4529) improve faecal parameters and nutritional status of dogs.