Seed-specific expression of porcine verotoxigenic Escherichia coli antigens in tobacco plants as a potential model of edible vaccines.

Vaccines can reduce the use of antibiotics by preventing specific infective diseases in pigs. Plant-based edible vaccines are particularly attractive because, upon oral ingestion via feed, they can elicit the local immune system against a foreign disease-causing organism. The aim of this study was to engineer two different independent lines of tobacco plants for the seed-specific expression of immunogenic proteins of VTEC as a model of an edible vaccine. For each antigen, fifty Nicotiana tabacum L. cv Xanthi leaf disks were transformed by agroinfection for the seed-specific expression of the structural parts of the fimbrial subunit FedF of F18 and the B-subunit of Vt2e genes. The synthetic genes, optimized by the codon adaptation index for their expression in tobacco, were inserted into expression cassettes under the control of ß-conglycinin promoter. Regenerated tobacco plants (T0) were characterized by molecular and immunoenzymatic techniques. Our results showed that both FedF and Vt2eB genes were integrated into tobacco genome efficiently (> 80%) and they are also maintained in the second generation (T1). Western blotting analyses carried out on the positive producing lines, showed the tissue-specific expression in seeds and the temporal protein accumulation in the mid-late maturation phases. The enzyme-linked immunosorbent assay showed seed expression levels of 0.09 to 0.29% (from 138 to 444 µg/g of seeds) and 0.21 to 0.43% (from 321 to 658 µg/g of seeds) of total soluble protein for the FedF and Vt2eB antigens, respectively. This study confirmed the seed-specific expression of the selected antigens in plant seeds. The expression level is suitable for seed-based edible vaccination systems, which could represent a cost-effective way to prevent VTEC infection. Our findings encourage further in vivo studies focused on the activation of the local immune response.

Effect of Dietary Supplementation of Chestnut and Quebracho Tannin Supplementation on Neonatal Diarrhoea in Preweaning Calves.

Neonatal calf diarrhoea (NCD) poses a significant health challenge in cattle herds, resulting in considerable economic losses and antimicrobial use. In response to the escalating threat of antimicrobial resistance, viable alternatives are imperative, aligning with European policies. This study evaluated the in-milk supplementation of the chestnut and quebracho tannin extract in preweaning calves on performance, diarrhoea occurrence, Cryptosporidium spp. shedding, protein digestibility, and intestinal health. Twenty newborn calves were divided, after colostrum administration, into two experimental groups for 30 days as follows: the control (CTRL) was fed with whole milk and solid feed, and tannins (TAN) were fed whole milk supplemented with 6/g day of tannin extract and solid feed. Faecal samples were collected on days 0, 3, 7, 14, and 30 for the evaluation of Cryptosporidium oocyst shedding and protein digestibility. Faecal consistency was evaluated during the sampling using the faecal score scale (0-3 scale, considering diarrhoea > 1). The results showed a significant reduction in diarrhoea frequency in the TAN compared to the CTRL group (p < 0.05) over 30 days of the trial. The prevalence of Cryptosporidium spp. was generally low (12%), considering all analysed samples. Protein digestibility revealed comparable values for the TAN and CTRL groups, suggesting that tannins did not negatively affect milk protein availability. In conclusion, the in-milk supplementation of 6/g day of the chestnut and quebracho tannin extract could be considered a valuable functional feed additive to decrease NCD occurrence, thus supporting animal health and decreasing antibiotic use in livestock.

A Commercial Clay-Based Material as a Carrier for Targeted Lysozyme Delivery in Animal Feed.

The controlled supply of bioactive molecules is a subject of debate in animal nutrition. The release of bioactive molecules in the target organ, in this case the intestine, results in improved feed, as well as having a lower environmental impact. However, the degradation of bioactive molecules' in transit in the gastrointestinal passage is still an unresolved issue. This paper discusses the feasibility of a simple and cost-effective procedure to bypass the degradation problem. A solid/liquid adsorption procedure was applied, and the operating parameters (pH, reaction time, and LY initial concentration) were studied. Lysozyme is used in this work as a representative bioactive molecule, while Adsorbo®, a commercial mixture of clay minerals and zeolites which meets current feed regulations, is used as the carrier. A maximum LY loading of 32 mgLY/gAD (LY(32)-AD) was obtained, with fixing pH in the range 7.5-8, initial LY content at 37.5 mgLY/gAD, and reaction time at 30 min. A full characterisation of the hybrid organoclay highlighted that LY molecules were homogeneously spread on the carrier's surface, where the LY-carrier interaction was mainly due to charge interaction. Preliminary release tests performed on the LY(32)-AD synthesised sample showed a higher releasing capacity, raising the pH from 3 to 7. In addition, a preliminary Trolox equivalent antioxidant capacity (TEAC) assay showed an antioxidant capacity for the LY of 1.47 ± 0.18 µmol TroloxEq/g with an inhibition percentage of 33.20 ± 3.94%.

The Effects of Milk Replacer Supplemented with Ascophyllum nodosum as a Novel Ingredient to Prevent Neonatal Diarrhea in Dairy Calves and Improve Their Health Status.

Nutrition and health during pre-weaning affect the calves' future fertility, calving age, production, and carrier length. Calves are highly susceptible to neonatal calf diarrhea (NCD), which can be fatal. NCD is due to hypovolemia and acidosis, which may involve anorexia and ataxia. The One Health principle calls for a drastic reduction in antimicrobial use. One approach is to improve animal health and reduce the use of antibiotics and functional ingredients that have beneficial effects due to bioactive compounds. Several functional ingredients and additives can be considered, and, in particular for this study, Ascophyllum nodosum was considered. The present study aimed to evaluate the role of A. nodosum as a functional ingredient implemented into the milk replacer in neonatal calves. Twelve pre-weaned Holstein Frisian calves, housed in twelve individual pens in the same environmental conditions, were divided into two groups of six animals: a control group (CTRL, n = 6) fed with a milk replacer, and a treatment group receiving milk enriched with 10 g of A. nodosum in their diet (TRT, n = 6) for 42 days. The fecal score was evaluated daily (3-0 scale) to monitor the incidence of diarrhea in the two groups. The body weight was evaluated weekly, and every two weeks feces were collected for microbiological evaluation using a selective medium for plate counting of total, lactic acid, and coliform bacteria. To verify the presence of Lactobacillus, Bifidobacterium, and Escherichia coli, real-time qPCR was used. At the beginning and at the end of the trial, blood samples were obtained for serum metabolite analysis. The growth performance did not differ in either of the two groups, but significant differences were observed in the incidence of moderate diarrhea (p-value < 0.0113), where the TRT group showed a lower incidence of cases during the 42-day period. Serum analysis highlighted higher contents of albumin, calcium, phosphorus, and total cholesterol in the TRT group compared to CTRL (p-value < 0.05). In conclusion, implementation of A. nodosum in the diet of calves can lead to better animal welfare and may reduce the use of antibiotics.

Evaluation of Beeswax Supplementation on Productive Performance of Growing Assaf Lambs.

The aim of this work was to assess the effects of beeswax supplementation on growth rate, feed intake, nutrient digestion, rumen fermentation, blood parameters, and economic sustainability in Assaf lambs. Eighteen growing Assaf (5 months old) lambs were separated into three experimental groups (n = 6 lambs/group). The lambs were fed a basal diet without supplementation (G1) or supplemented with 2 and 4 g beeswax/head/day in G2 and G3 groups, respectively. Zootechnical performance was evaluated over a 90 day period. Feed digestibility was assessed in faeces through the acid insoluble-ash method, and rumen liquor was collected to measure ammonia (NH3-N) and total volatile fatty acid (TVFA) levels. Blood samples were obtained for the titration serum metabolites by colorimetric tests. The findings showed that G3 had an improved performance compared to the other groups (p < 0.01). The lambs in G3 revealed the highest nutrient digestibility and feed use, followed by G2, and G1. G3 recorded the highest economic efficiency followed by G2 and G1 (p < 0.01). The TVFA, acetate, and propionate concentrations were higher and the pH values, NH3-N, and butyrate concentrations were lower in G3 compared to G2 and particularly to G1 (p < 0.01). The concentrations of total protein, globulin, and glucose were significantly higher with 4 g beeswax (p < 0.05). However, albumin, cholesterol, total lipids, urea, creatinine, glutamic oxaloacetic transaminase (GOT), and glutamate pyruvate transaminase (GPT) concentrations as well as the albumin to globulin ratio decreased significantly with both levels of beeswax (p < 0.05). The addition of beeswax at the level of 4 g/head/day for growing Assaf lambs significantly improved the growth performance, digestibility, rumen fermentation, and blood serum parameters in addition to the economic efficiency.

Evaluation of tributyrin supplementation in milk replacer on diarrhoea occurrence in preweaning Holstein calves.

Neonatal calf diarrhoea is one of the most important health challenges in cattle herds causing substantial economic losses and antimicrobial use. Due to the raising problem of antimicrobial resistance, effective alternatives are urgently required, in line with European policies. The aim of this study was to evaluate the effect of tributyrin supplementation in milk replacer on diarrhoea, performance and metabolic status in preweaning Holstein calves. Twelve newborn calves, after colostrum administration, were randomly allotted in two experimental groups for 42 days: control (CTRL) fed milk replacer, tributyrin (TRIB) fed milk replacer supplemented with 0.3% of liquid tributyrin on milk powder weight. Calves BW was recorded on a weekly basis from day 7 to day 42, and feed intake was recorded daily to calculate zootechnical performance. Faecal consistency was assessed daily through the faecal score (0-3 scale; considering diarrhoea moderate = 2 and severe = 3). Faecal samples were collected weekly from rectal ampulla for microbiological analysis by plate counting method evaluating the number of total bacteria, lactic acid bacteria and coliform bacteria. On day 0 and day 42, individual blood samples were collected from jugular vein for metabolic profile analysis. Serum samples of day 42 were also evaluated for the antioxidant barrier using a colorimetric test, while glucagon-like peptide 2 and diamine oxidase concentrations were measured through immunoenzymatic assays. Tributyrin supplementation did not influence the zootechnical performance of calves over 42 days of trial. Diarrhoea frequency was significantly lower in TRIB compared to CTRL group (27.91 and 38.37%; P < 0.01) considering the whole experimental period. In particular, the major effect was observed for moderate diarrhoea in TRIB group that showed a significantly reduced frequency compared to CTRL (P < 0.01) thus suggesting a preventive effect of tributyrin. Faecal total bacterial, lactic acid and coliform bacteria counts did not show differences between groups. Urea serum concentrations tended to be lower in TRIB compared to CTRL, indicating an efficient utilisation of dietary protein. Antioxidant barrier and glucagon-like peptide 2 were comparable between CTRL and TRIB on day 42. Diamine oxidase concentrations were significantly decreased in TRIB compared to CTRL group after 42 days of trial (P < 0.01), suggesting a higher gut epithelial integrity probably due to lower diarrhoea frequency and the nourish effect of tributyrin on enterocytes. In conclusion, tributyrin could be considered as a valuable bioactive feed additive to decrease the neonatal diarrhoea occurrence and support intestinal integrity in preweaning calves.

Tobacco Seed-Based Oral Vaccination against Verocytotoxic O138 Escherichia coli as Alternative Approach to Antibiotics in Weaned Piglets.

Post-weaning diarrhoea and enterotoxaemia caused by Escherichia coli are serious threats in the pig (Sus scrofa domesticus) livestock industry and are responsible for economic losses related to mortality, morbidity and stunted growth. The aim of this study was to evaluate the effect of an engineered tobacco seeds-based edible vaccine in O138 Escherichia coli-challenged piglets throughout a multidisciplinary approach. Thirty-six weaned piglets were enrolled and randomly divided into two experimental groups, a control (C; n = 18) group and a tobacco edible vaccination group (T, n = 18), for 29 days of trial. At days 0, 1, 2, 5 and 14, piglets of the T group were fed with 10 g of the engineered tobacco seeds line expressing F18 and VT2eB antigens, while the C group received wild-type tobacco seeds. After 20 days, 6 piglets/group were orally challenged with the Escherichia coli O138 strain (creating four subgroups: UC = unchallenged control, CC = challenged control, UT = unchallenged tobacco, CT = challenged tobacco) and fed with a high protein diet for 3 consecutive days. Zootechnical, clinical, microbiological, histological and immunological parameters were assayed and registered during the 9 days of post-challenge follow up. At 29 days post-challenge, the CT group displayed a lower average of the sum of clinical scores compared to the CC group (p < 0.05), while the CC group showed a higher average sum of the faecal score (diarrhoea) (p < 0.05) than the CT group. A decreased number of days of shedding of the pathogenic strain was observed in the CT compared to the CC group (p < 0.05). Specific anti-F18 IgA molecules were significantly higher in the CT group compared to the CC group's faecal samples during the post-challenge period (p < 0.01). In conclusion, edible vaccination with engineered tobacco seeds showed a protective effect on clinical symptoms and diarrhoea incidence during the post-challenge period, characterized by a limited time of pathogenic strain shedding in faeces.

Swine intestinal segment perfusion model for the evaluation of nutrients bioaccessibility.

Nutrition science requires more science-based evidences for the development of effective functional diets. To reduce animals for experimental purposes innovative reliable and informative models, simulating the complex intestinal physiology, are needed. The aim of this study was to develop a swine duodenum segment perfusion model for the evaluation of nutrient bioaccessibility and functionality across time. At the slaughterhouse, one sow intestine was harvested following Maastricht criteria for organ donation after circulatory death (DCD) for transplantation purposes. Duodenum tract was isolated and perfused in sub-normothermic conditions with heterologous blood after cold ischemia induction. Duodenum segment perfusion model was maintained under controlled pressure conditions through extracorporeal circulation for 3 hours. Blood samples from extracorporeal circulation and luminal content samples were collected at regular intervals for the evaluation of glucose concentration by glucometer, minerals (Na+, Ca2+, Mg2+, K+) by ICP-OES, lactate-dehydrogenase and nitrite oxide by spectrophotometric methods. Dacroscopic observation showed peristaltic activity caused by intrinsic nerves. Glycemia decreased over time (from 44.00±1.20 mg/dL to 27.50±0.41; p < 0.01), suggesting glucose utilization by the tissue confirming the organ viability in line with histological examinations. At the end of the experimental period, intestinal mineral concentrations were lower than their level in blood plasma suggesting their bioaccessibility (p < 0.001). A progressive increase of LDH concentration over time was observed in the luminal content probably related to a loss of viability (from 0.32±0.02 to 1.36±0.02 OD; p < 0.05) confirmed by histological findings that revealed a de-epithelization of the distal portion of duodenum. Isolated swine duodenum perfusion model satisfied the criteria for studying bioaccessibility of nutrients, offering a variety of experimental possibilities in line with 3Rs principle.

Protective effect of phytogenic plus short and medium-chain fatty acids-based additives in enterotoxigenic Escherichia coli challenged piglets.

Post Weaning Diarrhea (PWD) is the most important multifactorial gastroenteric disease of the weaning in pig livestock. Phytogenic (PHY) natural extracts are largely studied as alternatives to antibiotic treatments in combating the global concern of the antimicrobial resistance. The aim of this study was to evaluate the protective effect of innovative phytogenic premix with or without short and medium chain fatty acids (SCFA and MCFA) in O138 Escherichia coli challenged piglets. Twenty-seven weaned piglets were allotted into four groups fed different diets according to the following dietary treatments: CTRL (n = 13) group fed basal diet, PHY1 (n = 7) fed the basal diet supplemented with 0.2% of phytogenic premix, PHY2 (n = 7) fed the basal diet supplemented with 0.2% of phytogenic premix added with 2000 ppm of SCFA and MCFA. After 6 days of experimental diet feeding, animals were challenged (day 0) with 2 × 109 CFU of E. coli and CTRL group was divided at day 0 into positive (challenged CTRL + ; n = 6) and negative control group (unchallenged CTRL-; n = 7). Body weights were recorded at -14, -6, 0, 4 and 7 days and the feed intake was recorded daily. E. coli shedding was monitored for 4 days post-challenge by plate counting. Fecal consistency was registered daily by a four-point scale (0-3; diarrhea > 1) during the post-challenge period. Tissue samples were obtained for gene expression and histological evaluations at day 7 from four animals per group. Lower average feed intake was observed in CTRL + compared to PHY2 and CTRL during the post-challenge period. Infected groups showed higher E. coli shedding compared to CTRL- during the 4 days post-challenge (p < 0.01). PHY2 showed lower frequency of diarrhea compared to PHY1 and CTRL + from 5 to 7 days post-challenge. No significant alterations among groups were observed in histopathological evaluation. Duodenum expression of occludin tended to be lower in challenged groups compared to CTRL- at 7 days post-challenge (p = 0.066). In conclusion, dietary supplementation of PHY plus SCFA and MCFA revealed encouraging results for diarrhea prevention and growth performance in weaned piglets.

Antioxidant and Antimicrobial Activity of Algal and Cyanobacterial Extracts: An In Vitro Study.

Algae and cyanobacteria, other than their nutritional value, possess different beneficial properties, including antioxidant and antimicrobial ones. Therefore, they can be considered functional ingredients in animal feed and natural substitutes for antibiotics. The aim of this study was to evaluate the antioxidant and antimicrobial capacity against porcine O138 E. coli of Ascophyllum nodosum, Chlorella vulgaris, Lithotamnium calcareum, Schizochytrium spp. as algal species and Arthrospira platensis as cyanobacteria. The antioxidant capacity was determined by ABTS Radical Cation Decolorization Assay testing at three different concentrations (100%; 75%; 50%). The growth inhibition effect of the extracts at concentrations of 25%, 12.5%, 6%, 3% and 1.5% against porcine O138 E. coli was genetically characterized by PCR to detect the presence of major virulence factors; this was evaluated by following the microdilution bacterial growth method. The ABTS assay disclosed that Ascophyllum nodosum was the compound with the major antioxidant properties (57.75 ± 1.44 percentage of inhibition; p < 0.0001). All the extracts tested showed growth inhibition activity at a concentration of 25%. Among all extracts, A. nodosum was the most effective, showing a significant growth inhibition of E. coli; in particular, the log10 cells/mL of E. coli used as a control resulted in a significantly higher concentration of 25% and 12.5% after 4 h (8.45 ± 0.036 and 7.22 ± 0.025 log10 cells/mL, respectively; p < 0.005). This also suggests a dose-dependent relationship between the inhibitory activity and the concentration. Also, a synergistic effect was observed on antioxidant activity for the combination of Ascophyllum nodosum and Lithotamnium calcareum (p < 0.0001). Moreover, to determine if this combination could affect the viability of the IPEC-J2 cells under the normal or stress condition, the viability and membrane integrity were tested, disclosing that the combination mitigated the oxidative stress experimentally induced by increasing the cell viability. In conclusion, the results obtained highlight that the bioactive compounds of algal species are able to exert antioxidant capacity and modulate O138 E. coli growth. Also, the combination of Ascophyllum nodosum and Lithotamnium calcareum species can enhance their bioactivity, making them a promising functional feed additive and a suitable alternative to antibiotics.

Effect of a Blend of Essential Oils, Bioflavonoids and Tannins on In Vitro Methane Production and In Vivo Production Efficiency in Dairy Cows.

Two trials were performed to evaluate the efficacy of a blend of essential oils, bioflavonoids and tannins on methane (CH4) emissions (in vitro) and on the production efficiency of dairy cows (in vivo). The in vitro trial tested the production of total gas and CH4 at 16, 20 and 24 h of incubation, and volatile fatty acids (VFA) at 16 and 24 h, through biochemical methane potential (BMP) assays. In the in vivo trial, milk yield, dry matter intake (DMI), feed conversion rate (FCR), milk quality and apparent total tract digestibility (aTTD) were evaluated in 140 lactating Holstein Friesian cows. Animals were allocated into two groups: (i) Control, standard diet; (ii) Treatment, standard diet plus 10 g/head/d of a powder with a 10% concentration of a blend of essential oils, bioflavonoids and tannins. Statistical analysis was performed using the mixed procedure of SAS either for single or repeated measures. For all the parameters a p-value ≤ 0.05 was considered statistically significant. The blend significantly reduced the in vitro total gas and CH4 emissions at 16, 20 and 24 h of incubation (p < 0.001). In addition, acetic acid was reduced (p < 0.001), while propionic acid concentration was increased (p < 0.001) at 16 h and 24 h. In the in vivo trial, the Treatment group showed significantly raised milk yield, DMI, FCR (p < 0.001), and of the aTTD of cellulose and starch (p ≤ 0.002), while the milk quality traits were not affected. Overall, the results from the study indicated that the blend of essential oils, bioflavonoids, and tannins significantly reduced in vitro total gas and CH4 production and improved the production efficiency of lactating dairy cows in vivo.

Trends and potential human health risk of trace elements accumulated in transplanted blue mussels during restoration activities of Flekkefjord fjord (Southern Norway).

The monitoring of contaminants represents a priority to preserve the integrity of marine ecosystems, as well as to plan and to manage restoration activities in order to protect environmental and human health. In the present study, a 6-months active biomonitoring was performed to explore the levels of eighteen trace and toxic elements, including heavy metals (TEs; i.e. Al, As, Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Sr, Ti, and Zn), accumulated in soft tissues of blue mussel (Mytilus edulis Linnaeus, 1758) individuals transplanted at different depths (5- and 15-m depth) in five locations within the Flekkefjord fjord (Southern Norway). As this area suffered a long-lasting contamination due to both organic and inorganic contaminants, a series of restoration activities were activated to tackle and to prevent potential risks for ecosystem and local population. Our results demonstrated that the levels of TEs accumulated in edible tissues of transplanted mussels in the Flekkefjord fjord were generally low before the beginning of the restoration activities. However, location- and time-specific differences in the accumulation of TEs were noted after the implementation of such activities. Interestingly, the levels of Fe and Mn significantly increased after the beginning of the restoration activities, likely because the release of these TEs from the slag used in such operations and/or resuspension of contaminated sediments. However, assuming that native mussels can accumulate the same TEs at levels measured in transplanted individuals, our results suggest a substantial safety for human consumption of native mussels from the Flekkefjord fjord, regardless of restoration activities.

Nutritional and Functional Properties of Colostrum in Puppies and Kittens.

The present review aims toward a better understanding of the nutrition of newborn puppies and kittens. The post-natal period is very sensitive in dogs and cats, as in other animal species. During the first two weeks of life, puppies and kittens are at high risk of dehydration, hypothermia, and hypoglycemia, as well as infectious diseases as they start to acquire the physiological functions of the adult. Neonatal hepatic glycogen storage is low, and newborns depend on colostrum intake to survive. Colostrum provides immunoglobulins and other important substances such as lipids and carbohydrates. Immunoglobulins are central to the immunological link that occurs when the mother transfers passive immunity. The mechanism of transfer varies among mammalian species, but in this review, we focused our attention on dogs and cats. Furthermore, there are components of colostrum which, although their presence is not absolutely necessary, play an important role in nutrition. These components have received considerable interest because of their presumed safety and potential nutritional and therapeutic effects both in humans and animals; however, unfortunately, there are few recent studies in companion animals. Here, we have gathered the published articles that describe studies involving different species of animals, emphasizing companion animals. In particular, the purpose of this narrative of the nutritional and functional proprieties of queens' and bitches' colostrum.

Supplementation of Live Yeast, Mannan Oligosaccharide, and Organic Selenium during the Adaptation Phase of Newly Arrived Beef Cattle: Effects on Health Status, Immune Functionality, and Growth Performance.

The effect of a nutraceutical mixture, based on live yeast (LY), mannan oligosaccharides (MOS) and organic selenium (Se) on health status, as well as immune functionality and growth performance in the fattening of newly received beef cattle, was evaluated. A total of 1036 Charolaise heifers were allocated into two experimental groups: (i) control group, without any nutraceutical support (n = 487; initial weight = 325 ± 21 kg); and (ii) treatment group, supplementation during the first 30 days, with LY (5 g/head/day), organic Se (3 mg/head/day), and MOS (10 g/head/day) (n = 549; initial weight = 323 ± 23 kg). The incidence of bovine respiratory disease (BRD) and other health issues was monitored, as well as the mortality rate. Blood samples were taken at d0 and d30 to evaluate the immune functionality and the inflammatory status. Growth performances, feces chemical composition, and carcass characteristics were recorded. The BRD occurrence tended to be reduced (p = 0.06) in the Treatment group. The BHV-1 antibody production after vaccination was significantly improved (p = 0.031), as well as the bactericidal activity (p = 0.0012) in the Treatment group. No differences were found in the inflammatory status parameters. The final weight (p = 0.006) and the average daily gain at d30 (p < 0.0001) were significantly improved by the treatment. No differences were found in terms of carcass characteristics, while the fecal content of NDF (p < 0.0001), ADF (p = 0.0003), and starch (p < 0.0001) were significantly reduced by the treatment. The result of the present study suggests that the nutraceutical mixture used can support the animal's immune systems, improving its ability to react against pathogens, as well as feed efficiency and growth performances during the whole fattening period.

Effect of Slow-Release Urea Administration on Production Performance, Health Status, Diet Digestibility, and Environmental Sustainability in Lactating Dairy Cows.

The effects of partially replacing soybean meal (SBM) with a slow-release urea source (SRU) on production performance, feed efficiency, digestibility, and environmental sustainability of dairy cows were evaluated. A total of 140 lactating Holstein Frisian cows were allocated into two study groups: (i) control (diet entirely based on SBM), and (ii) treatment (diet of 0.22% on dry matter basis (d.m.)) of SRU. Milk yield, dry matter intake (DMI), feed conversion rate (FCR), body condition score (BCS), reproductive parameters, and milk quality were evaluated. The chemical composition of the feeds and feces were analyzed to calculate the in vivo digestibility of the two diets. The carbon footprint (CFP) and predicted methane (CH4) emissions were evaluated. The inclusion of SRU significantly increases milk yield, DMI, and FCR (p < 0.0001), whereas milk quality, BCS, and reproductive indicators were not affected (p > 0.05). In the treatment group, the digestibility of crude protein (CP) (p = 0.012), NDF (p = 0.039), and cellulose (p = 0.033) was significantly higher, while the other nutritional parameters weren't affected. All the environmental parameters were significantly improved in the treatment group (p < 0.0001). Replacing SBM with SRU can be a strategy to enhance dairy cows' sustainability due to improved production efficiency, reduced feed CFP, and predicted CH4 production.

Translational Approach to Induce and Evaluate Verocytotoxic E. coli O138 Based Disease in Piglets.

Pig livestock was influenced by several global concerns that imposed a re-thinking of the farming system, which included the reduction in chemical dependency and the development of antimicrobial alternatives. Post-weaning diarrhea and enterotoxaemia caused by Escherichia coli, are serious threats that are responsible for the economic losses related to mortality, morbidity and stunted growth in weaning piglets. The aim of the study was to set up experimental conditions to simulate the simultaneous outbreak of post-weaning diarrhea and enterotoxaemia in weaned piglets, through verocytotoxic O138 Escherichia coli challenge, with a multidisciplinary approach. Eighteen piglets susceptible to F18 VTEC infection were selected by polymerase chain reaction for polymorphism on the fucosyltransferase 1 gene and randomly divided in two experimental groups, non-infected controls (C; n = 6) and infected ones (I; n = 12) and housed into individual pens at the same environmental conditions for 29 days. At day 20, I pigs were orally inoculated with Escherichia coli O138 and fed a high protein ration for 3 days. Zootechnical, clinical, microbiological, histological and immunological parameters were evaluated along the follow up (3 and 9 days). Experimental infection, confirmed by bacteria faecal shedding of the I group, significantly affected the clinical status. The I group showed significantly higher total scores, corresponding to medians of the sum of daily scores from days 1 to 3 (Σ3) and 1 to 9 (Σ9) post infection, epiphora, vitality, hair irregularity, oedema and depression. Histological examination showed evident inflammatory infiltrate of lymphocytes, and follicular hyperplasia in I pigs; in the same group, the immunohistochemical and immunological assays revealed an increase in IgG in the intestinal crypts and CD3-positive T cells in intestinal epithelium. The experimental Escherichia coli infection in controlled conditions is crucial for both the evaluation of innovative compounds and the elucidation of the mechanisms associated with the persistence of antibacterial resistant strains. In conclusion, the adopted infection model, carried out on receptor-mediated susceptible piglets, allowed us to identify a discriminative panel of clinical symptoms related to Escherichia coli O138 infection, and could be used to assess the protective effect of antibiotic alternatives.

Effects of Heated Drinking Water on the Growth Performance and Rumen Functionality of Fattening Charolaise Beef Cattle in Winter.

The effects of heated drinking water on growth performance and rumen functionality in fattening beef cattle during winter were evaluated. Newly received Charolaise bulls (n = 224) were allocated to two experimental groups: (i) water at room temperature (RTW) (weight 408 ± 34 kg); (ii) constantly heated water (25 °C) (HW) (weight 405 ± 38 kg). Growth performances, feed intake, feed conversion rate, water intake and carcass characteristics were evaluated. Internal reticuloruminal wireless boluses were used to collect rumen pH and temperature values every 10 min. Bodyweight was not affected by the water temperature, but the overall average daily gain (ADG) was significantly higher in the HW group (1.486 vs. 1.438 kg/head/day in the RTW; p = 0.047). Dry matter intake was significantly higher in the HW group (p = 0.001), even though the final feed conversion rate (FCR) was not influenced. There was also a tendency for better cold carcass weight (CCW) and carcass yield (CY) in the HW group. Drinking heated water reduced the time (min/day) during which the ruminal pH was below pH 5.8 or 5.5, and the time during which the temperature was lower than 37 or 39 °C (p < 0.001). The use of heated drinking water is a plausible a strategy for enhancing ruminal stability and the overall production efficiency in fattening beef cattle, which will lead to both better growth performance and higher ruminal stability.

Evaluation of Adhesive Characteristics of L. plantarum and L. reuteri Isolated from Weaned Piglets.

Limosilactobacillus reuteri and Lactiplantibacillus plantarum strains, previously isolated from weaned piglets, were considered for the evaluation of their adhesive characteristics. Lactobacilli were treated with LiCl in order to remove the surface protein layer, and probiotic activity was compared with those of untreated strains. The autoaggregation, co-aggregation to E. coli F18+, and adhesive abilities of LiCl-treated Limosilactobacillus reuteri and Lactiplantibacillus plantarum were significantly inhibited (p < 0.05) compared with the respective untreated strain. The hydrophobic and basic phenotypes were observed due to the strong affinity to chloroform and low adherence to ethyl acetate. In particular, L. plantarum showed higher hydrophobicity compared to L. reuteri, which may reflect their different colonizing ability. After treatment with LiCl to remove surface proteins, the adherence capabilities of L. reuteri and L. casei on IPEC-J2 cells decreased significantly (p < 0.001) and L. reuteri adhered more frequently. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that both L. reuteri and L. plantarum had several bands ranging from 20 to 100 kDa. Two-dimensional gel electrophoresis showed an acidic profile of the surface-layer polypeptides for both bacterial strains, and more studies are needed to characterize their profile and functions. The results confirm the pivotal role of surface proteins in the probiotic potential of L. reuteri and L. plantarum.

Lactobacillus plantarum and Lactobacillus reuteri as Functional Feed Additives to Prevent Diarrhoea in Weaned Piglets.

The effects of Lactobacillus plantarum and Lactobacillus reuteri and their combination were assessed in weaned piglets. Three hundred and fifty weaned piglets (Landrace × Large White), balanced in terms of weight and sex, were randomly allotted to four experimental groups (25 pens, 14 piglets/pen). Piglets were fed a basal control diet (CTRL, six pens) and a treatment diet supplemented with 2 × 108 CFU/g of L. plantarum (PLA, 6 pens), 2 × 108 CFU/g L. reuteri (REU, six pens) and the combination of both bacterial strains (1 × 108 CFU/g of L. plantarum combined with 1 × 108 CFU/g of L. reuteri, P+R, 7 pens) for 28 days. Body weight and feed intake were recorded weekly. Diarrhoea occurrence was assessed weekly by the faecal score (0-3; considering diarrhoea ≥ 2). At 0 and 28 days, faecal samples were obtained from four piglets per pen for microbiological analyses and serum samples were collected from two piglets per pen for serum metabolic profiling. Treatments significantly reduced diarrhoea occurrence and decreased the average faecal score (0.94 ± 0.08 CTRL, 0.31 ± 0.08 PLA, 0.45 ± 0.08 REU, 0.27 ± 0.08 P+R; p < 0.05). The PLA group registered the lowest number of diarrhoea cases compared to other groups (20 cases CTRL, 5 cases PLA, 8 cases REU, 10 cases P+R; p < 0.01). After 28 days, the globulin serum level increased in PLA compared to the other groups (24.91 ± 1.09 g/L CTRL, 28.89 ± 1.03 g/L PLA, 25.91 ± 1.03 g/L REU, 25.31 ± 1.03 g/L P+R; p < 0.05). L. plantarum and L. reuteri could thus be considered as interesting functional additives to prevent diarrhoea occurrence in weaned piglets.

Evaluation of Tannin Extracts, Leonardite and Tributyrin Supplementation on Diarrhoea Incidence and Gut Microbiota of Weaned Piglets.

The effects of the dietary administration of a combination of Quebracho and Chestnut tannins, leonardite and tributyrin were evaluated in weaned piglets. A total of 168 weaned piglets (Landrace × Large White) were randomly allotted to two experimental groups (6 pens/group, 14 piglets/pen). Animals were fed a basal control diet (CTRL) and a treatment diet (MIX) supplemented with 0.75% tannin extracts, 0.25% leonardite and 0.20% tributyrin for 28 days. Individual body weight and feed intake were recorded weekly. Diarrhoea incidence was recorded by a faecal scoring scale (0-3; considering diarrhoea ≥ 2). At 0 and 28 days, faecal samples were obtained from four piglets/pen for microbiological and chemical analyses of faecal microbiota, which were then assessed by V3-V4 region amplification sequencing. At 28 days, blood from two piglets/pen was sampled to evaluate the serum metabolic profile. After 28 days, a reduction in diarrhoea incidence was observed in the MIX compared to CTRL group (p < 0.05). In addition, compared to CTRL, MIX showed a higher lactobacilli:coliform ratio and increased Prevotella and Fibrobacter genera presence (p < 0.01). The serum metabolic profile showed a decreased level of low-density lipoproteins in the treated group (p < 0.05). In conclusion, a combination of tannin extract, leonardite and tributyrin could decrease diarrhoea incidence and modulate the gut microbiota.