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1. The aim of this study was to evaluate the production of equol (4ʹ,7-isoflavandiol; a bacterial polyphenol metabolite which is an isoflavandiol oestrogen metabolised from daidzein from plants) enriched eggs from free-range hens fed different pasture species. Four species were tested: red clover, white clover, ryegrass and chicory. 2. The study was conducted from June to September 2017 on eight free range, outdoor areas, each containing fifteen laying hens and sown with a single pasture species 3. Precursors of equol (daidzein, formononetin) were analysed every fortnight from the fresh pasture cover in each area, as well as equol and daidzein levels in eggs. 4. Daidzein and formononetin concentrations in the fresh pasture samples differed significantly according to species (P

Tannins are secondary plant compounds which have been extensively studied in order to improve the nitrogen use efficiency (NUE) of ruminants. A meta-analysis was performed of 58 in vivo experiments comparing milk yield, composition and nitrogen metabolism of lactating dairy cows fed diets with or without tannins. The meta-analysis shows that tannins have no impact on corrected milk yield, fat and protein content or NUE (p > .05). However, tannins reduce ruminal ammoniacal nitrogen (N) production by 16% (from 10.95 to 8.47 mg/dl on average), milk urea by 9% (from 15.82 to 14.03 mg/dl) and urinary N excretion (-11%; p < .05). This is compensated for by a lower apparent N digestibility (61.51% with dietary tannins compared to 66.17% without). The effect of tannin on N metabolism parameters increases with tannin dose (p < .05). The shift from urinary to faecal N may be beneficial for environment preservation, as urinary N induces more harmful emissions than faecal N. From a farmer's perspective, tannins seem unable to increase fat- and protein-corrected milk yield or reduce feed protein requirements and thus have no direct economic benefit. Potentially less costly than tannin extracts, forage or by-products naturally rich in tannins could still be useful to reduce the environmental impact of ruminant protein feeding.

The objective of this study was to investigate the effects of xylo-oligosaccharides (XOSs) supplementation on growth performance, serum parameters, small intestinal morphology, intestinal mucosal integrity, and immune function in weaned piglets. A total of 240 weaned piglets with an average body weight (BW) of 8.82 ± 0.05 kg (28 d of age) were assigned randomly to four dietary treatments in a 28-d trial, including a control (CON) diet and three diets with XOS supplementation at the concentration of 100 (XOS100), 500 (XOS500), and 1,000 (XOS1000) mg/kg. There were four replicates per treatment with 15 pigs per pen. From day 1 to 14, there were no differences (P > 0.05) in average daily gain (ADG), average daily feed intake, and gain to feed ratio (G:F) during the different treatments. The different doses of XOSs showed a quadratic effect on BW on day 28, ADG, and G:F on day 1 to 28 of piglets (P < 0.05). From day 15 to 28, ADG of pigs fed the XOS500 diet was higher (P < 0.05) than pigs fed the CON diet. During the overall period (day 1 to 28), pigs fed the XOS500 diet had a higher BW, ADG, and G:F than pigs fed the CON diet (P < 0.05). In addition, compared with the CON group, the XOS500 group had significantly higher serum total antioxidant capacity, total superoxide dismutase and catalase levels, and lower malondialdehyde levels on days 14 and 28 (P < 0.05). The serum immunoglobulin G (IgG) concentration in the XOS500 group was also significantly higher compared with the CON group on days 14 and 28 (P < 0.05). However, serum immunoglobulin A and immunoglobulin M were not affected by the dietary treatments. Supplementation of XOS500 to the feed significantly increased the villus height (VH) and VH to crypt depth ratio in the jejunum and ileum in comparison with the CON and XOS1000 groups. Moreover, the XOS500 group significantly elevated the expression levels of occludin and zonula occludens protein-1 in the ileum compared with the CON group. The ileal interleukin (IL)-1β, IL-8, and interferon (IFN)-γ mRNA expression levels in the XOS100 and XOS500 groups were markedly lower than in the CON group. In contrast, the ileal IL-10 mRNA expression levels were remarkably higher in the XOS500 than in the CON group. In conclusion, XOSs have a beneficial effect on growth performance by improving serum antioxidant defense system, serum IgG, small intestinal structure, and intestinal barrier function in weaned piglets.

Climate change (CC) is expected to increase temperatures and the frequency of extreme weather events, which renewed interest in heat stress (HS) effects on dairy cattle farms. Resilience is a key concept that should be considered to better understand the dairy farms exposure to HS and to combat CC-related risks. Thus, this study aimed to investigate the aspects of HS vulnerability for Tunisian dairy cattle farming systems. Historical milk test-day records from official milk recording were merged with temperature and humidity data provided by public weather stations. Firstly, different models relying in two heat load indices were applied for HS exposure assessment. Secondly, broken line models were used to estimate HS thresholds, milk losses, and rates of decline of milk production associated with temperature-humidity index (THI) across parities. Thirdly, individual cow responses to HS estimated using random regression model were considered as key measures of dairy farming system sensitivity assessment to HS. Dairy farms are annually exposed for 5 months to high THI values above 72 in Tunisia. The tipping points, at which milk yield started to decline over parities with 3-day average THI, ranged between 65 and 67. The largest milk decline per unit of THI above threshold values was 0.135 ± 0.01 kg for multiparous cows. The milk losses estimated due to HS in the Holstein breed during the summer period (June to August) ranged between 110 and 142 kg/cow in north and south, respectively. A high HS sensitivity was proved especially in dairy farms characterized by large herd size and high milk production level. Hence, providing knowledge of dairy farms vulnerability to HS may provide the basis for developing strategies to reduce HS effects and plan for CC adaptation.

Dairy farming systems are evolving. This study presents dairy producers' perceptions of their ideal future farm (IFF) to ensure revenue, and attempts to determine the reasons for this choice, the environmental aspects related to this choice, the proximity between the current farm and the IFF and the requirements for reaching this IFF. Just before the end of the European milk quota, a total of 245 Walloon dairy producers answered a survey about the characteristics of their IFF and other socio-environmental-economic information. A multiple correspondence analysis (MCA) was carried out using seven characteristics of the IFF (intensive vs. extensive, specialised vs. diversified, strongly vs. weakly based on new technologies, managed by a group of managers vs. an independent farmer, employed vs. familial workforce, local vs. global market, standard vs. quality-differentiated production) to observe the relationships between them. Based on the main contributors to the second dimension of the MCA, this axis was defined as an IFF gradient between the local-based extensive (LBE) producers (26%) and the global-based intensive (GBI) producers (46%). The differences of IFF gradient between modalities of categorical variables were estimated using generalised linear models. Pearson correlations were calculated between the scores on the IFF gradient and quantitative variables. Finally, frequencies of IFF characteristics and the corresponding characteristic for the current situation were calculated to determine the percentages of "unhappy" producers. Some reasons for the choice of IFF by the producers have been highlighted in this study. Environmental initiatives were more valued by LBE than GBI producers. Low similarity was observed between the current farm situation of the respondents and their IFF choice. LBE and GBI producers differed significantly regarding domains of formation (technical and bureaucratic vs. transformation and diversification respectively) and paths of formation (non-market vs. market respectively). Two kinds of farming systems were considered by dairy producers and some socioeconomic and environmental components differed between them.

1. The grain drying process may affect the feeding value of maize but until now, no general consensus has been reached. This knowledge is essential to manage maize nutritional value in feed and ensure optimal growth performance of broiler chickens. 2. A total of 72 male Ross 308 were used in a complete randomised block design to assess the effect of initial moisture content (MC) at harvest (high or low MC after the appearance of the black layer) and drying temperature (54°C, 90°C or 125°C) on the apparent faecal digestibility and the AMEn value of two maize grain types (flint and flint-dent varieties). Moreover, the relationship between

1. This study assessed the impact of drying temperature (54, 90, and 130°C) and maize grain moisture content at harvest (36% and 29%) on in vitro digestibility, the growth performance and ileal digestibility of broiler chickens. 2. In contrast to the results from the in vitro digestibility, apparent ileal digestibility of starch and energy decreased when the drying temperature was raised from 54 to 130°C, and this effect was more pronounced in maize grain harvested at high initial moisture content (36%). Ileal protein digestibility of maize grain decreased significantly when dried at the intermediate temperature (90°C) and with a high harvest moisture content (36%). Drying temperature and initial moisture content did not significantly affect AMEn. 3. When maize was dried at 130°C, the particle sizes of flour recovered after standard milling procedures decreased significantly, which would influence animal growth performance and in vivo digestibility through animal feed selection.

The calving interval (CI) can potentially impact the economic results of dairy farms. This study highlighted the most profitable CI and innovated by describing this optimum as a function of the feeding system of the farm. On-farm data were used to represent real farm conditions. A total of 1832 accounts of farms recorded from 2007 to 2014 provided economic, technical and feeding information per herd and per year. A multiple correspondence analysis created four feeding groups: extensive, low intensive, intensive and very intensive herds. The gross margin and some of its components were corrected to account for the effect of factors external to the farm, such as the market, biological status, etc. Then the corrected gross margin (cGMc) and its components were modelled by CI parameters in each feeding system by use of GLM. The relationship between cGMc and the proportion of cows with CI

An experiment was conducted to uncover the effects of increasing dietary grain levels on expression of thiamine transporters in ruminal epithelium, and to assess the protective effects of thiamine against high-grain-induced inflammation in dairy cows. Six rumen-fistulated, lactating Holstein dairy cows (627 ± 16.9 kg of body weight, 180 ± 6 d in milk; mean ± standard deviation) were randomly assigned to a replicated 3 × 3 Latin square design trial. Three treatments were control (20% dietary starch, dry matter basis), high-grain diet (HG, 33.2% dietary starch, DM basis), and HG diet supplemented with 180 mg of thiamine/kg of dry matter intake. On d 19 and 20 of each period, milk performance was measured. On d 21, ruminal pH, endotoxic lipopolysaccharide (LPS), and thiamine contents in rumen and blood, and plasma inflammatory cytokines were detected; a rumen papillae biopsy was taken on d 21 to determine the gene and protein expression of toll-like receptor 4 (TLR4) signaling pathways. The HG diet decreased ruminal pH (5.93 vs. 6.49), increased milk yield from 17.9 to 20.2 kg/d, and lowered milk fat and protein from 4.28 to 3.83%, and from 3.38 to 3.11%, respectively. The HG feeding reduced thiamine content in rumen (2.89 vs. 8.97 μg/L) and blood (11.66 vs. 17.63 μg/L), and the relative expression value of thiamine transporter-2 (0.37-fold) and mitochondrial thiamine pyrophosphate transporter (0.33-fold) was downregulated by HG feeding. The HG-fed cows exhibited higher endotoxin LPS in rumen fluid (134,380 vs. 11,815 endotoxin units/mL), and higher plasma concentrations of lipopolysaccharide binding protein and pro-inflammatory cytokines when compared with the control group. The gene and protein expression of tumor necrosis factor α (TNFα), IL1B, and IL6 in rumen epithelium increased when cows were fed the HG diet, indicating that local inflammation occurred. The depressions in ruminal pH, milk fat, and protein of HG-fed cows were reversed by thiamine supplementation. Thiamine supplementation increased thiamine contents in rumen and blood, and also upregulated the relative expression of thiamine transporters compared with the HG group. Thiamine supplementation decreased ruminal LPS (49,361 vs. 134,380 endotoxin units/mL) and attenuated the HG-induced inflammation response as indicated by a reduction in plasma IL6, and decreasing gene and protein expression of pro-inflammatory cytokines in rumen epithelium. Western bottling analysis showed that thiamine suppressed the protein expression of TLR4 and the phosphorylation of nuclear factor kappa B (NFκB) unit p65. In conclusion, HG feeding inhibits thiamine transporter expression in ruminal epithelium. Thiamine could attenuate the epithelial inflammation during high-grain feeding, and the protective effects may be due to its ability to suppress TLR4-mediated NFκB signaling pathways.

International audience; The objective of this study was to evaluate the effects of oak tannin extract (OTE) added in forage before ensiling on dairy cows fed at 92% of their digestible protein requirements. Six multiparous lactating Holstein cows were used in a crossover design (two treatments × two periods). The control treatment (CON) was based on a diet including 50% of grass silage, whereas the experimental treatment (TAN) included grass silage sprayed with OTE (26 g/kg DM) just before baling. Milk yield (on average 24 kg fat protein corrected milk per day) was not affected, but both milk and rumen fatty acids profiles were impacted by OTE. Nitrogen intake (415 g N per cow per day) and nitrogen use efficiency (NUE; 0.25 on average) were not affected, but a shift from urine (-8% of N intake relatively to control, P = 0.06) to faecal N (+5%; P = 0.004) was observed with the TAN diet (P ≤ 0.05). Nitrogen apparent digestibility was thus reduced for TAN (-3%; P ≤ 0.05). The effect of OTE on ruminal and milk FA profiles suggests an impact on rumen microbiota. Nitrogen isotopic discrimination between animal proteins and diet (Δ15N) was evaluated as a proxy for NUE. While no differences in NUE were observed across diets, a lower Δ15N of plasma proteins was found when comparing TAN v. CON diets. This finding supports the concept that Δ15N would mainly sign the N partitioning at the metabolic level rather than the overall NUE, with the latter also being impacted by digestive processes. Our results agree with a N shift from urine to faeces, and this strategy can thus be adopted to decrease the environmental impact of ruminant protein feeding

Two experiments were conducted to reveal the effects of grain-induced subacute rumen acidosis (SARA) on thiamine status in blood and rumen fluid in dairy cows. In both experiments, 6 multiparous, rumen-fistulated Holstein dairy cows were used in a 2-treatment, 2-period crossover design. Each experimental period consisted of 21d (total of 42d). Experiment 1 was to investigate the effects of SARA on thiamine status in blood and rumen fluid. Treatments were either control (20% starch, dry matter basis) or SARA-inducing diet (SAID, 33.2% starch, dry matter basis). In experiment 2, the effects of dietary thiamine supplementation on attenuating SARA and ruminal fermentation characteristics in dairy cows were studied. All cows received the same SAID diet during the whole experimental period; treatments were with or without thiamine (180mg of thiamine/kg of dry matter intake). In both experiments, rumen fluid samples were collected at 0, 3, 6, 9, and 12h after morning feeding on d 21 and 42 of the experiments for measurement of pH, thiamine, volatile fatty acid, and lactate contents. Peripheral blood was also collected at 3h after morning feeding on d 21 and 42 to measure thiamine, carbohydrate metabolites, and enzyme activities. In experiment 1, cows fed the SAID diet had lower ruminal and plasma thiamine concentrations and higher lactate than cows fed the control diet. The ruminal thiamine contents were positively related to pH and the concentrations of acetate in the rumen, and negatively correlated with the lactate contents. Experiment 2 demonstrated that ruminal pH and the concentrations of thiamine, acetate, and total volatile fatty acids in the rumen were increased, whereas ruminal lactate contents were reduced by thiamine supplementation. The concentrations of lactate and the activity of lactate dehydrogenase in blood were reduced in the thiamine supplemented group, and the opposite was true for the nonesterified fatty acids and α-ketoneglutarate dehydrogenase contents. In conclusion, the thiamine status was affected by SARA in dairy cows and ruminal infusion of thiamine could help attenuate SARA by improving theproportions of ruminal volatile fatty acids and reducing lactate contents in rumen fluid and blood.

Heat stress (HS) adversely influences dairy cattle welfare and productivity. This study aimed to investigate the effects of HS on production and physiological parameters of Holstein cows. Two experiments each lasted 6 weeks were conducted in four Tunisian farms, firstly during summer under HS (n = 80, THI = 77) and later during autumn under thermo-neutral (n = 80, THI = 54) conditions. Respiration rate (RR), skin temperature (ST), rectal temperature (RT) and milk yield were measured, and milk samples were collected on 2 days every week during each experimental period. Temperature and relative humidity were measured inside the barn to calculate the temperature-humidity index (THI). Mixed models were used to evaluate the effects of period and the relationships between THI and physiological and production traits. Reaction norm models were applied to quantify the individual responses of cows across the trajectory of THI during the HS period. A clustering methodology was developed to identify tolerant and sensitive cows to HS based on their slope for response of physiological and production traits during HS period. In summer, RR (61 breaths/min) and ST (37.7 °C) were 2.3- and 1.3-fold higher, whereas milk yield per milking was 24% lower compared with thermo-neutral conditions. Linear relationship between THI and RR, ST and RT was observed and showed increases by 2 breaths/min, 0.5 °C and 0.04 °C per increase in one THI unit, respectively. Inversely, milk, fat and protein yields showed a drop of 0.13 kg, 0.4 g and 0.3 g per milking per increase in one THI unit, respectively. Cows qualified to be heat tolerant by our work tended to have higher RR, ST, and RT and lower to almost no decay in milk yield compared to cows qualified to be heat sensitive. Specifically, RR could be used as a reliable indicator for thermotolerance. The results of this study deepen our understanding of different aspects of HS resilience.

Inulin and wheat bran were added to broiler diets during the starter period or during the entire rearing period to investigate whether the effects of using these ingredients remained until slaughter age. Diets containing no inulin and no wheat bran (CON), 2% inulin (IN), 10% wheat bran (WB), or 2% inulin + 10% wheat bran (IN+WB) were provided until day 11. Thereafter, each dietary treatment was further divided into a continued diet with supplementation or a control diet, resulting in 7 groups (CON, IN/IN, IN/CON, WB/WB, WB/CON, IN+WB/IN+WB, or IN+WB/CON). On day 40, 12 chickens per group were euthanized. The IN/IN group increased the cecal molar ratio of butyrate but had a lower relative abundance of Lactobacillus (P0.05). Additionally, the cecal molar ratio of propionate was higher in the IN/CON group compared to the IN/IN group (P = 0.034). The WB/CON group had the best results on BW and feed conversion ratio (FCR) (P0.05). Only the cecal molar ratio of iso-butyrate was higher in the WB/WB group (P = 0.013). Moreover, compared to the CON group, both WB/WB and WB/CON groups reduced the relative abundances of Bifidobacterium and Escherichia coli, and only the WB/WB group reduced the relative abundance of Enterobacteriaceae (P0.05). Both IN+WB/IN+WB and IN+WB/CON groups increased BW until day 21 and lowered the relative abundance of Bifidobacterium (P0.05). The IN+WB/IN+WB group increased the cecal molar ratio of butyrate but reduced the molar ratio of propionate with a higher relative abundance of Enterobacteriaceae (P0.05). In conclusion, the lack of positive effects induced by inulin might be explained by the dose being too high. The beneficial effects on BW, FCR, and microbiota induced by wheat bran during the starter period were lasting when supplementation was stopped, suggesting that wheat bran could be a favorable ingredient during the starter period.

The chemical characteristics of co-products recovered during a laboratory-scale wet milling procedure as well as that of whole corn flour were characterised and their digestibility and fermentability value determined using a 2 steps in vitro digestibility and fermentation model of the pig digestive tract. Five co-products differing in their chemical composition were collected and analysed. These co-products differed in their in vitro dry matter Digestibility and in their kinetic of fermentation. High coefficients of digestibility were observed for starchy samples, while low coefficients of digestibility were observed for samples rich in lignocellulosic components. Fermentation patterns of samples analysed were different as well as the profile of volatile fatty acids produced during the fermentation. The production of straight-chain fatty acids produced was significantly correlated with the proportion of starch in the sample, while branched-chain fatty acids were correlated to proteins concentration of samples.

Inulin and wheat bran were added to the starter diets of broiler chickens to investigate the potential of these ingredients to improve the host's health and growth performance, as well as the underlying mechanisms of their effects. A total of 960 1-day-old chicks were assigned to 4 treatments: control (CON), 2% inulin (IN), 10% wheat bran (WB), and 10% wheat bran +2% inulin (WB+IN). On day 11, 6 chicks per treatment were euthanized. A general linear model procedure with Tukey's multiple range test was performed to compare a series of parameters between treatments. The WB-containing treatments improved BW on day 7, day 11, day 35, and BW gain until day 11 (P0.05), but only the WB+IN treatment showed a lower feed conversion ratio than the CON treatment (P = 0.011). Furthermore, the WB+IN treatment showed the highest villus height in the jejunum and ileum (P0.05), and the highest jejunal ratio villus height/crypt depth (P = 0.035). The concentration of acetate in the ceca was higher in the CON treatment compared to the IN treatment (P = 0.040). The IN treatment increased the concentration (P = 0.003) and ratio (P = 0.004) of iso-butyrate compared to the WB+IN and the CON treatments (P0.05). A clustering result exhibited similar intestinal microbiota profiles in the chicks receiving the IN and the WB+IN diets (P0.05), but these profiles were different from those found in chicks receiving the WB and the CON diets (P0.05). In conclusion, wheat bran and the combination of wheat bran and inulin ameliorated the growth performance and gut morphology of the starter chicks, which resulted in a higher BW until day 35. Inulin, on the other hand, had a greater ability to influence the microbiota profile. The beneficial results found in relation to BW and gut morphology during the starter period suggested a synergistic effect of inulin and wheat bran.

Culicoides (Diptera: Ceratopogonidae) are biological vectors of bluetongue virus (BTV). Bluetongue is a viral disease that affects domestic and wild ruminants. Since its recent emergence in northern Europe, this disease has caused considerable economic losses to the sheep and cattle industry. The biotopes, and more particularly the chemical characteristics which are suitable for larval development of the main vector species, are still relatively unknown. This study shows that the larvae of biting midges belonging to the species Culicoides obsoletus and Culicoides scoticus are able to breed in different types of silage residue (maize, grass, sugar beet pulp and their combinations). The chemical composition of substrates strongly influences the presence of the immature stages of these biting midges. Higher lignin and insoluble fibre contents seem to favour their presence and could play the role of a physical support for semi-aquatic larvae. In contrast, higher concentrations of magnesium and calcium are negatively correlated with the presence of these two species. These data will help to locate and monitor the breeding sites of these species and could contribute to the control of these insects on farms.

This study aimed to evaluate the fermentation in the large intestine of indigestible dietary protein sources from animal, insect, and plant origin using an in vitro model of the pig’s gastrointestinal tract. Protein sources were used raw and after a cooking treatment. Results showed that the category of the ingredient (meats, insects, or grain legumes) exerts a stronger impact on enzymatic digestibility, fermentation patterns, and bacterial metabolites such as short-chain fatty acids (SCFA) and hydrogen sulfide (H2S) than the cooking treatment. The digestibility and the fermentation characteristics of insects were more affected by the cooking procedure than the other categories. Per gram of consumed food, ingredients from animal origin, namely, meats and insects, were associated with fewer fermentation end-products (gas, H2S, SCFA) than ingredients from plant origin, which is related to their higher small intestinal digestibility.

Salmonella is a human pathogen that frequently infects poultry flocks. Consumption of raw or undercooked contaminated poultry products can induce acute gastroenteritis in humans. Faced with the public health concerns associated with salmonellosis, the European Union has established a European regulation forcing member states to implement control programs aimed at reducing Salmonella prevalence in poultry production, especially at the primary production level. The purpose of the present review article is to summarize the current research and to suggest future developments in the area of Salmonella control in poultry, which may be of value to the industry in the coming years. The review will focus especially on preventive strategies that have been developed and that aim at reducing the incidence of Salmonella colonization in broiler chickens at the farm level. In addition to the usual preventive hygienic measures, other strategies have been investigated, such as feed and drinking water acidification with organic acids and immune strategies based on passive and active immunity. Modification of the diet by changing ingredients and nutrient composition with the intent of reducing a bird's susceptibility to Salmonella infection also has been examined. Because in ovo feeding accelerates small intestine development and enhances epithelial cell function, this approach could be an efficient tool for controlling enteric pathogens. Feed additives such as antibiotics, prebiotics, probiotics, and synbiotics that modify the intestinal microflora are part of another field of investigation, and their success depends on the additive used. Other control methods such as the use of chlorate products and bacteriophages also are under study.

Anti-Salmonella spp. egg yolk antibodies (IgY) simultaneously directed against Salmonella Enteritidis and Salmonella Typhimurium were tested to determine if their inclusion in feed decreased Salmonella spp. cecal colonization in experimentally infected broiler chickens. Egg yolk powder (EYP) was obtained by freeze-drying egg yolks containing anti-Salmonella spp. Immunoglobin Y was included in feed at 5 levels of concentration (0 to 5%). Feeds were formulated to similar nutrient levels and provided for ad libitum intake from d 1 to 28. Three days after initiation of feed treatments (d 4), chickens were co-challenged with equal numbers of Salmonella Enteritidis and Salmonella Typhimurium (2x10(6) cfu/bird). Cecal samples were recovered weekly over the experimental period (d 7 to 28) to enumerate Salmonella spp. The effect of anti-Salmonella spp. IgY feed supplementation on growth performance of infected chickens was also evaluated during the same period. In comparison with the positive control treatment (PC), treatments involving EYP (T1, T2, T3, T4, and T5), whether containing anti-Salmonella spp. IgY or not, significantly improved (P

The antibody production in the egg yolks of immunized laying hens is seen as a way of improving animal welfare compared with conventional production by mammals. Immunoglobulin Y (IgY) technology, however, has still to address welfare issues linked to the widespread use of an adjuvant in vaccines. Currently, Freund's adjuvants, complete (FCA) or incomplete (FIA), remain the standard. This study sought to evaluate various approaches used to enhance egg yolk antibody production in terms of both productivity and avian welfare. The outer membrane protein (OMP) of Salmonella Typhimurium was used as the prototype antigen. At 20 weeks of age, 56 ISA Brown hens, with specific-Salmonella-free status, were divided into seven groups (n=8) and received an initial intramuscular immunization. Hens in the two negative control groups received phosphate buffered saline (PBS) or FIA alone. Hens in the other groups received 80μg of Salmonella OMP emulsified with one of the following adjuvants: 200μl of FIA alone (T1); 200μl of FIA supplemented with 8μg of C-phosphate-guanosine oligodeoxynucleotides (CpG-ODN) (T2); and 280μl of Montanide ISA 70 VG (T4). Birds in the T3 group received the antigen in emulsion with FIA and were given the tested immunostimulatory component (l-carnitine) via their feed (100mg/kg). A positive control group (PC) received FCA for the first and final immunizations and FIA for the other boosters. Immunization was repeated after 20, 46, 82 and 221 days. Eggs were collected regularly until 242 days after the first immunization and the anti-Salmonella Typhimurium activities in the yolk were determined by ELISA. After 242 days, the birds were euthanized and the injection sites were evaluated for gross and microscopic lesions. Among the tested immunostimulatory approaches, supplementation of FIA with CpG-ODN led to a significant and long-lasting enhancement of the specific antibody response. This treatment was even higher than the positive benchmark using FCA in the first immunization. The study results showed that a clinical examination of injection sites is insufficient for drawing conclusions about the local tolerance of vaccines. Tissue damage was noticeable in all treatment groups. The birds receiving the Montanide adjuvant, however, had fewer and less severe lesions. Given these limited side-effects, Montanide ISA 70 VG could provide the depot effect needed to ensure the immunomodulatory efficiency of CpG-ODN. The association of these two adjuvants could prove a promising alternative to Freund's adjuvants (FA).

In this study, Salmonella enterica serovar Typhimurium challenge models were tested to identify the best conditions under which to perform the experimental infection of 3-wk-old broilers. Such a model would be useful to study the efficiency of therapeutic treatments that could take place at the end of the grow-out period. Salmonella-free chicks were obtained from a breeder flock vaccinated with Salmonella. Intestinal maternal immunity was monitored by ELISA analyses at 2, 9, and 16 d of age. Data indicated that protection of maternal origin was not maintained over time and was drastically reduced at 9 d of age (P < 0.01). At 21 d of age, chickens were orally inoculated with Salmonella Typhimurium. The effects of the oral challenge dose (0, 3 × 10(3), 3 × 10(6), and 3 × 10(9) cfu/bird) and vancomycin pretreatment (no administration or 25 mg/bird) on intestinal immune responses, growth performance, and Salmonella colonization of chickens were investigated. After infection, the mucosal immune response was rapid, with increased (P < 0.01) anti-Salmonella Typhimurium IgA titers measured at 8 d postinfection in intestinal contents. A linear relationship (P < 0.05) existed between specific IgA levels in intestinal and cecal contents and the challenge dose inoculated. None of the challenge protocols caused mortality or clinical symptoms after infection. Nevertheless, the experimental infection induced a significant deterioration of growth performance. The pretreatment with 25 mg of vancomycin at 3 h before Salmonella inoculation was able to establish stable infection rates among the population of 3-wk-old infected chickens. Nevertheless, Salmonella shedding was not stable over the rearing period, and the bacteria seemed to be naturally eliminated from most birds at 22 d postinfection. This natural clearance of the gut, which was related, at least in part, to the intestinal immune response, should limit the usability of the created mature challenge model within 1 to 2 wk after inoculation.

The protective effects of powder preparation of egg yolk immunoglobulin Y (IgY), specific to Salmonella Enteritidis and Salmonella Typhimurium outer membrane proteins (OMP), against these two Salmonella sp. serovars were investigated in vitro in two different assays: adhesion-prevention and growth-inhibition. The adhesion-prevention assay was conducted using polarized monolayers of the human intestinal epithelial Caco-2 cell line. First, the conditions of Salmonella adherence to Caco-2 cells were optimized, and interferences of bacteria with the transepithelial electrical resistance (TER) of fully differentiated Caco-2 cell monolayers and the lactate dehydrogenase release upon exposure of the cells to Salmonella were evaluated. Both Salmonella sp. serovars were able to adhere to Caco-2 cells and decreased TER. Results from the adhesion-prevention assay demonstrated that specific IgY reduced the decrease in TER of the infected Caco-2 cell monolayers and blocked the Salmonella sp. adhesion in a concentration-dependent manner (p < 0.05). Nonspecific IgY also exhibited an inhibitory effect on these two parameters, but to a lesser extent than that of the specific IgY (p < 0.05). The protective effect of nonspecific IgY could be attributed to the low-density lipoprotein component of the water-soluble fraction of egg yolks that may not have been eliminated during ultrafiltration. The growth-inhibition assay revealed that specific IgY had an inhibitory effect on the bacterial growth, markedly during the late exponential phase, whereas nonspecific IgY failed to do so. Taken together, these results suggest that the in vitro growth inhibitory effect of specific IgY on Salmonella spp. resulted from the specific binding activity of these IgY to Salmonella sp. OMP. Passive immunization with Salmonella sp. OMP-specific IgY could thus be useful to prevent Salmonella colonization in broiler chickens and the subsequent carcass contamination during processing.

The present study was an attempt to raise hen egg yolk Ig (IgY) simultaneously directed against Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in the same egg yolk. The immunopotentiating effect of 2 different adjuvants -- Freund's adjuvants (FA) and immunostimulating complexes matrix (IM) -- on antibody response was also evaluated. Bacterial outer membrane proteins (OMP) were selected as target antigens. The ISA Brown hens, specific-Salmonella spp.-free status, divided into 6 groups were intramuscularly injected with a mono-compound antigen preparation: SE-OMP (treatment SE-FA or SE-IM) or ST-OMP (treatment ST-FA or ST-IM), or a combined antigen preparation: (1/2) SE-OMP and (1/2) STOMP (treatment SEST-FA or SEST-IM). Titers of antibodies in yolk were evaluated biweekly with ELISA. There was no antigen x adjuvant interaction on antibody titers. Anti-SE IgY titers in hens that received treatment SEST-FA or SEST-IM were statistically similar (P > 0.05) as compared with those obtained from hens immunized with treatment SE-FA or SE-IM. Anti-ST IgY titers in hens immunized with treatment SEST-FA or SEST-IM were slightly lower than those of hens that received treatment ST-FA or ST-IM. The cross-reactivity of anti-SE IgY, induced by treatment SE-FA or SE-IM, with ST-OMP antigen and that of anti-ST IgY, induced by ST-FA or ST-IM, with SE-OMP antigen were arbitrarily assessed on d 43 and 155 by ELISA. The average cross-reactivity of anti-SE IgY with ST-OMP antigen was 71.7%. The average cross-reactivity of anti-ST IgY with SE-OMP antigen was 78.8%. In FA groups, antibody titers were found higher (P < 0.05) than those in IM groups. Furthermore, no extensive lesions or clinical abnormalities were detected in hens injected with FA. These findings showed the opportunity to raise IgY antibody against 2 Salmonella serovars in the same yolk and that FA was more efficient than IM in mediating antibody response.

The three-step technique was used to determine the requirements of total amino acids (TAA) and the first-limiting amino acid (AA) in growing double-muscled Belgian Blue bulls (BBb). In Exp. 1, three double-muscled BBb weighing initially 306 +/- 28 kg received a basal diet consisting of 30% meadow hay and 70% concentrate that was poor in digestible protein but had adequate NE because of continuous infusion of dextrose into the duodenum. The intestinal apparent digestibility of essential AA (EAA) was defined according to their duodenal and ileal flows. It averaged 72% but varied between 60% for Met and 79% for Arg. In Exp. 2, five double-muscled BBb (334 +/- 22 kg) received the same diet supplemented with duodenal infusions of dextrose and four doses of Na-caseinate (28, 56, 84, and 112% of intestinal digestible dietary AA) in a 4 x 4 Latin square design with one additional animal. Nitrogen retention for the basal diet alone and the four increasing supplements of Na-caseinate reached 49, 61, 70, 80, and 86 g/d, respectively. Nitrogen utilization improved from 34.3% without Na-caseinate supplementation to a maximum of 40.6%, with the third dose supplying 788 g/d of apparently digestible AA. Based on patterns of plasma concentrations, Met, Phe, and Arg were probably the limiting AA when animals optimized N utilization. In Exp. 3, six double-muscled BBb (315 +/- 25 kg) fed the basal diet received duodenal infusions of dextrose and AA, equivalent to the third dose in Exp. 2, except for digestible Met (9.3, 14.4, 18.4, 22.4, 26.4, and 30.4 g/d) in a 6 x 6 Latin square design. The Met requirement was close to 26.4 g/d on the basis of N retention.

Stable 15 N was used to evaluate the influence of bacterial contamination on in situ DM and N degradabilities (Dg) of meat and bone meal (MBM), soybean meal (SBM), and wheat bran (WB) in two steers. Bacterial DM and N contamination ranged from 2.4 to 28.6% and 2.1 to 56.8% of residual DM and N, respectively. Effective N degradability increased when bacterial contamination was taken into account (P .05) and different for WB (P< .05). In a second experiment, five treatments were applied to incubated feeds to remove bacteria fixed to the residues and consequently to determine directly the Dg of DM and N corrected for the bacterial contamination without the need for a marker. These treatments involved chilling for 6 h at 4 o C in saline solution alone (T1) or with a commercial detergent (T2), or with sodium dodecyl sulfate (T3) or with methylcellulose (T4), followed by pummeling in a stomacher for 5 min. The last treatment was only machine washing twice (T5). The Dg of DM can be directly determined following the first four treatments, nevertheless their application to MBM and SBM led to higher Dg of N than that corrected for the bacterial contamination determined in the first experiment (P

In many dietary conditions, methionine (Met) and lysine (Lys) are the most limiting amino acids (AA) for ruminants. The AA protected from ruminal fermentation are not commercially avail- able, with the exception of Met which is not always economical, especially for meat production. This study measured ruminal escape of free Met and Lys supplemented intraruminally to fast grow- ing bulls. Six double-muscled Belgian Blue bulls, fed a high concentrate diet and fitted with a rumen cannula, received free Met (40 g .d -1 ) and free Lys (60 g.d -1 ), individually or simultaneously, in a dupli- cated Latin square design. The mean ruminal escape of Met and Lys reached 37 and 45% respectively, and did not differ if administered separately or together. Plasma Lys and Met concentrations were increased by 504 and 126%, respectively. Substantial proportions of free AA escaped ruminal fer- mentation and were available for absorption from the small intestine when they were administered at physiologically high levels. ruminant / methionine / lysine / rumen escape / plasma