Methane (CH4) production is a ubiquitous, apparently unavoidable side effect of fermentative fibre digestion by symbiotic microbiota in mammalian herbivores. Here, a data compilation is presented of in vivo CH4 measurements in individuals of 37 mammalian herbivore species fed forage-only diets, from the literature and from hitherto unpublished measurements. In contrast to previous claims, absolute CH4 emissions scaled linearly to DM intake, and CH4 yields (per DM or gross energy intake) did not vary significantly with body mass. CH4 physiology hence cannot be construed to represent an intrinsic ruminant or herbivore body size limitation. The dataset does not support traditional dichotomies of CH4 emission intensity between ruminants and nonruminants, or between foregut and hindgut fermenters. Several rodent hindgut fermenters and nonruminant foregut fermenters emit CH4 of a magnitude as high as ruminants of similar size, intake level, digesta retention or gut capacity. By contrast, equids, macropods (kangaroos) and rabbits produce few CH4 and have low CH4 : CO2 ratios for their size, intake level, digesta retention or gut capacity, ruling out these factors as explanation for interspecific variation. These findings lead to the conclusion that still unidentified host-specific factors other than digesta retention characteristics, or the presence of rumination or a foregut, influence CH4 production. Measurements of CH4 yield per digested fibre indicate that the amount of CH4 produced during fibre digestion varies not only across but also within species, possibly pointing towards variation in microbiota functionality. Recent findings on the genetic control of microbiome composition, including methanogens, raise the question about the benefits methanogens provide for many (but apparently not to the same extent for all) species, which possibly prevented the evolution of the hosting of low-methanogenic microbiota across mammals.

To further understand the contribution of feedstuff ingredients to gut health in swine, gut histology and intestinal bacterial profiles associated with the use of two high-quality protein sources, microbially enhanced soybean meal (MSBM) and Menhaden fishmeal (FM) were assessed. Weaned pigs were fed one of three experimental diets: (1) basic diet containing corn and soybean meal (Negative Control (NEG)), (2) basic diet + fishmeal (FM; Positive Control (POS)) and (3) basic diet + MSBM (MSBM). Phase I POS and MSBM diets (d 0 to d 7 post-wean) included FM or MSBM at 7.5%, while Phase II POS and MSBM diets (d 8 to d 21) included FM or MSBM at 5.0%. Gastrointestinal tissue and ileal digesta were collected from euthanised pigs at d 21 (eight pigs/diet) to assess gut histology and intestinal bacterial profiles, respectively. Data were analysed using Proc Mixed in SAS, with pig as the experimental unit and pig (treatment) as the random effect. Histological and immunohistochemical analyses of stomach and small intestinal tissue using haematoxylin–eosin, Periodic Acid Schiff/Alcian blue and inflammatory cell staining did not reveal detectable differences in host response to dietary treatment. Ileal bacterial composition profiles were obtained from next-generation sequencing of PCR generated amplicons targeting the V1 to V3 regions of the 16S rRNA gene. Lactobacillus-affiliated sequences were found to be the most highly represented across treatments, with an average relative abundance of 64.0%, 59.9% and 41.80% in samples from pigs fed the NEG, POS and MSBM diets, respectively. Accordingly, the three most abundant Operational Taxonomic Units (OTUs) were affiliated to Lactobacillus, showing a distinct abundance pattern relative to dietary treatment. One OTU (SD_Ssd_00001), most closely related to Lactobacillus amylovorus, was found to be more abundant in NEG and POS samples compared to MSBM (23.5% and 35.0% v. 9.2%). Another OTU (SD_Ssd_00002), closely related to Lactobacillus johnsonii, was more highly represented in POS and MSBM samples compared to NEG (14.0% and 15.8% v. 0.1%). Finally, OTU Sd_Ssd-00011, highest sequence identity to Lactobacillus delbrueckii, was found in highest abundance in ileal samples from MSBM-fed pigs (1.9% and 3.3% v. 11.3, in POS, NEG and MSBM, respectively). There was no effect of protein source on bacterial taxa to the genus level or diversity based on principal component analysis. Dietary protein source may provide opportunity to enhance presence of specific members of Lactobacillus genus that are associated with immune-modulating properties without altering overall intestinal bacterial diversity.

Ruminants living in seasonal environments face a two-fold challenge during winter. The energetic cost of maintaining a high body temperature is higher at lower ambient temperatures, and this is compounded by poor availability and quality of feed. Wild ruminants acclimatize to this energetic challenge by hypothermia, that is, reduced endogenous heat production and abandoning the maintenance of a high body temperature, particularly in peripheral body parts. Further but lesser contributions to lower energy expenditure during winter are reduced foraging activity; lower heat increment of feeding; and reduced maintenance cost of size-reduced organs. Altogether, metabolic rate, estimated by the continuous measurement of heart rate, during winter is downregulated to more than half of the summer level, as is voluntary food intake, even in animals fed ad libitum. The transformation from the summer into the thrifty winter phenotype is also evident in the physiology of digestion. Microbial protein synthesis is less facilitated by diminished phosphorus secretion into the shrunk rumen during winter. In line with this result, the concentration of ammonia, the end-product of protein digestion in the rumen, peaks in rumen liquid in spring, whereas the molar proportion of acetate, an indicator of fermentation of a diet rich in fiber, peaks in winter. In contrast to reduced stimulation of growth of ruminal microbes during winter, active transport of nutrients across the intestinal epithelium is increased, resulting in more efficient exploitation of the lower amount and quality of ingested winter feed. Nevertheless, the energy balance remains negative during winter. This is compensated by using fat reserves accumulated during summer, which become a major metabolic fuel during winter.

Large ham weight losses (WL) in dry-curing are undesired as they lead to a loss of marketable product and penalise the quality of the dry-cured ham. The availability of early predictions of WL may ease the adaptation of the dry-curing process to the characteristics of the thighs and increase the effectiveness of selective breeding in enhancing WL. Aims of this study were (i) to develop Bayesian and Random Forests (RFs) regression models for the prediction of ham WL during dry-curing using on-site infrared spectra of raw ham subcutaneous fat, carcass and raw ham traits as predictors and (ii) to estimate genetic parameters for WL and their predictions (P-WL). Visible-near infrared spectra were collected on the transversal section of the subcutaneous fat of raw hams. Carcass traits were carcass weight, carcass backfat depth, lean meat content and weight of raw hams. Raw ham traits included measures of ham subcutaneous fat depth and linear scores for round shape, subcutaneous fat thickness and marbling of the visible muscles of the thigh. Measures of WL were available for 1672 hams. The best prediction accuracies were those of a Bayesian regression model including the average spectrum, carcass and raw ham traits, with R2 values in validation of 0.46, 0.55 and 0.62, for WL at end of salting (23 days), resting (90 days) and curing (12 months), respectively. When WL at salting was used as an additional predictor of total WL, the R2 in validation was 0.67. Bayesian regressions were more accurate than RFs models in predicting all the investigated traits. Restricted maximum likelihood (REML) estimates of genetic parameters for WL and P-WL at the end of curing were estimated through a bivariate animal model including 1672 measures of WL and 8819 P-WL records. Results evidenced that the traits are heritable (h2 ± SE was 0.27 ± 0.04 for WL and 0.39 ± 0.04 for P-WL), and the additive genetic correlation is positive and high (ra = 0.88 ± 0.03). Prediction accuracy of ham WL is high enough to envisage a future use of prediction models in identifying batches of hams requiring an adaptation of the processing conditions to optimise results of the manufacturing process. The positive and high genetic correlation detected between WL and P-WL at the end of dry-curing, as well as the estimated heritability for P-WL, suggests that P-WL can be successfully used as an indicator trait of the measured WL in pig breeding programs.

Adipogenesis, myogenesis and fibrogenesis are related processes that can contribute to meat quality. Therefore, extending the knowledge of these processes would facilitate the identification of molecular markers that predict intramuscular fat accretion. The main purpose of this work, based on previous results, was to further study the expression of key genes related to adipogenic, myogenic, fibrogenic processes and some cytokines in Longissimus thoracis (LT) and Masseter (MS) muscles of Pirenaica and Holstein young bulls. Longissimus thoracis and MS muscles from Pirenaica (n = 4) and Spanish Holstein (n = 4) were sampled for proximate analysis, determination of adipocyte size distribution and expression of key candidate genes. Fat percentage was lower in LT than in MS muscle in Pirenaica young bulls (P = 0.023) and was higher in LT muscle in Holstein than in Pirenaica young bulls (P = 0.007). Gene expression analysis revealed that the mRNA level of myogenic differentiation 1 (MYOD) was higher in LT than in MS muscles in both groups of animals (P < 0.001) and that myostatin (MSTN) expression was also higher in LT than in MS muscle in Holstein bulls (P = 0.001). On the other hand, MSTN and PPARG showed higher expression in LT and MS in Pirenaica young bulls (P = 0.026), while the expression of fatty acid-binding protein 4 (FABP4) was higher in Holstein young bulls, also in both muscles (P < 0.001). The results suggested that the development of intramuscular adipose depot was directly related to the expression of adipogenic genes, such as FABP4, but inversely related to the expression of the cytokine MSTN and the myogenic gene MYOD, genes which showed a muscle-specific expression.

Piglet birth weight and within-litter birth weight variation are important for piglet survival and growth. Pre-mating diets may improve IGF-1 and follicle development during the weaning-to-oestrus interval (WEI) and subsequent piglet birth weight. The objective of this study was to modulate IGF-1 concentration during late lactation and the WEI of young sows by using specific pre-mating diets supplemented with microfibrillated cellulose (MF), l-carnitine (LC) or l-arginine (AR). A further objective was to investigate the relationship between IGF-1 and subsequent follicle development and oestrus and ovulation characteristics. In total, 56 first-parity and 20 second-parity sows in three consecutive batches were used for this experiment. Sows received daily either wheat (CON) or wheat plus MF, LC or AR at one of two supplementation levels (low and high) during last week of lactation and WEI. From weaning onwards, follicle and corpus luteum (CL) diameters were repeatedly measured with ultrasound. Blood samples were collected during the WEI for IGF-1 and on day 21 of pregnancy for progesterone analyses, respectively. Insulin-like growth factor-1 concentration, follicle diameter, oestrus and ovulation characteristics and CL diameter were not affected by pre-mating diets. Low IGF-1 class (≤156 ng/ml, N = 22) sows had smaller follicles at weaning (3.5 v. 3.8 mm, P < 0.05) and a longer weaning-to-ovulation interval (147.2 v. 129.8 h, P < 0.05) than high IGF-1 class sows. In first-parity sows, high loin muscle depth (LM) loss sows (≥8%, N = 28) had lower IGF-1 concentrations at weaning (167 v. 214 ng/ml, P < 0.05) compared to low LM loss sows (<8%, N = 28). However, after weaning, IGF-1 concentrations increased and did not differ between high LM loss and low LM loss sows. In conclusion, the different supplemented compounds in pre-mating diets did not improve IGF-1 concentrations around weaning in young sows. Furthermore, high body condition loss caused lower IGF-1 concentrations at weaning, but these levels rapidly recovered after weaning and were related to follicle development and the interval from weaning to ovulation.

For their glucose supply, ruminants are highly dependent on the endogenous synthesis in the liver, but despite the numerous studies that evaluated hepatic glucose production, very few simultaneously measured hepatic glucose production and uptake of all precursors. As a result, the variability of precursor conversion into glucose in the liver is not known. The present study aimed at investigating by meta-analysis the relationships between hepatic glucose net release and uptake of precursors. We used the FLuxes of nutrients across Organs and tissues in Ruminant Animals database, which gathers international results on net nutrient fluxes at splanchnic level measured in catheterized animals. Response equations were developed for intakes up to 41 g DM intake/kg BW per day of diets varying from 0 to 100 g of concentrate/100 g DM in the absence of additives. The net hepatic uptake of propionate, α-amino-N and l-lactate was linearly and better related to their net portal appearance (NPA) than to their afferent hepatic flux. Blood flow data were corrected for lack of deacetylation of the para-aminohippuric acid, and this correction was shown to impact the response equations. To develop response equations between the availability of precursors (portal appearance and hepatic uptake) and net glucose hepatic release, missing data on precursor fluxes were predicted from dietary characteristics using previously developed response equations. Net hepatic release of glucose was curvilinearly related to hepatic supply and uptake of the sum of precursors, suggesting a lower conversion rate of precursors at high precursor supply. Factors of variation were explored for the linear portion of this relationship, which applied to NPA of precursors ranging from 0.99 to 9.60 mmol C/kg BW per h. Hepatic release of glucose was shown to be reduced by the portal absorption of glucose from diets containing bypass starch and to be increased by an increased uptake of β-hydroxybutyrate indicative of higher body tissue mobilization. These relationships were affected by the physiological status of the animals. In conclusion, we established equations that quantify the net release of glucose by the liver from the net availability of precursors. They provide a quantitative overview of factors regulating hepatic glucose synthesis in ruminants. These equations can be linked with the predictions of portal absorption of nutrients from intake and dietary characteristics, and provide indications of glucose synthesis from dietary characteristics.

In teleosts, vitellogenin (Vtg) is a phospholipoglycoprotein synthesized by the liver, released into the blood circulation and incorporated into the oocytes via endocytosis mediated by the Vtg receptor (VTGR) to form the yolk granules. The VTGR is crucial for oocyte growth in egg-laying animals but is also present in non-oviparous vertebrates, such as human. The VTGR belongs to the low-density lipoprotein receptor superfamily (LDLR) and is also named very-low-density lipoprotein receptor (VLDLR). In this study, we identified and phylogenetically positioned the VTGR of a basal teleost, the European eel, Anguilla anguilla. We developed quantitative real-time PCR (qRT-PCR) and investigated the tissue distribution of vtgr transcripts. We compared by qRT-PCR the ovarian expression levels of vtgr in juvenile yellow eels and pre-pubertal silver eels. We also analyzed the regulation of ovarian vtgr expression throughout vitellogenesis in experimentally matured eels. The Vtg plasma level was measured by homologous ELISA experimental maturation. Our in silico search and phylogenetical analysis revealed a single vtgr in the European eel, orthologous to other vertebrate vtgr. The qRT-PCR studies revealed that vtgr is mainly expressed in the ovary and also detected in various other tissues such as brain, pituitary, gill, fat, heart, and testis, suggesting some extra-ovarian functions of VTGR. We showed that vtgr is expressed in ovaries of juvenile yellow eels with no higher expression in pre-pubertal silver eels nor in experimentally matured eels. This suggests that vtgr transcription already occurs during early pre-vitellogenesis of immature eels and is not further activated in vitellogenic oocytes. European eel Vtg plasma level increased throughout experimental maturation in agreement with previous studies. Taken together, these results suggest that vtgr transcript levels may not be a limiting step for the uptake of Vtg by the oocyte in the European eel.

The objectives of this study were to analyse the differences in the genetic determination of functional longevity in five Spanish lines of rabbits and to check how different systematic factors might affect this genetic determination. Four of the lines were maternal (lines A, V, H and LP), these lines were established selecting base generation animals according to different criteria, but in the subsequent generations all of them were selected for litter size at weaning. The other is the paternal line R, this line was constituted by selecting animals with an outstanding daily growth rate. The trait analysed, length of productive life, was the time in days between the date of the first positive pregnancy test and the date of culling or death of a doe. Four models extended from the Cox proportional hazard model were used to analyse data of each line separately and jointly. The complete model (Model 1) included the fixed effect of year-season (YS) combination, positive palpation order (OPP), that is, reproductive cycle, physiological status of the doe (PS) at service and number of kits born alive (NBA) in each kindling as time-dependent factors. The inbreeding coefficient was fitted as a continuous covariate and the animal’s additive genetic effect was also fitted to the model (Model 1). The other models were identical to Model 1 but excluding OPP (Model 2) or PS (Model 3) or NBA (Model 4), which were explored to assess the consequence on additive variance estimates of not correcting for these animal-dependent factors. Estimated effective heritabilities of longevity were 0.07 ± 0.03, 0.03 ± 0.02, 0.14 ± 0.09, 0.05 ± 0.04, 0.02 ± 0.01 and 0.04 ± 0.01 for lines A, V, H, LP, R and for the merged data set, respectively. Removing the PS from the model led to an increase in the estimated additive genetic variance in all lines (0.17 ± 0.05, 0.05 ± 0.03, 0.29 ± 0.19, 0.29 ± 0.20, 0.07 ± 0.04 and 0.05 ± 0.02 for lines A, V, H, LP, R and the merged data set, respectively). The highest hazard of death and/or culling was observed during the first two parities and decreased as the order of parity progressed. Does non-pregnant-non-lactating had the highest risk of death or culling. The does that had zero kits born alive incurred the highest risk, and this risk decreased as the NBA increased. In conclusion, the consideration of longevity as selection criterion for the studied rabbit lines is not recommended.

Feed form is well recognized to improve broiler performance, specially by increasing feed intake (FI). However, when different diet energy levels are used, the results differ in the literature. Therefore, this experiment was conducted to evaluate the influence of feed form and dietary metabolizable energy (ME) levels on broiler performance, carcass yield and on the digestibility of DM, CP, starch and gross energy. In total, 1152 male Cobb 500 broilers were evaluated between 35 and 47 days. The birds were distributed according to a completely randomized design in a 2 × 4 factorial arrangement, consisting of two feed forms (mash or pellet) and four ME levels (12.73, 13.06, 13.40 or 13.73 MJ/kg), totaling eight treatments with eight replicates of 18 birds. Broilers fed the lowest ME level presented the lowest weight gain (WG) and worst feed per unit gain (P < 0.01). Metabolizable energy intake increased (P < 0.01) with progressive increments of ME, which, however, did not affect caloric conversion (CC, P > 0.05). Pelleted diets promoted higher FI, WG, ME intake (P < 0.01) and better feed per unit gain and CC (P < 0.05) compared with mash. In mash diets, increasing dietary ME levels promoted a linear increase in WG (P < 0.01) and reduced feed per unit gain (P ≤ 0.05), but did not affect FI (P > 0.05). In pelleted diets, on the other hand, increasing ME levels linearly reduced FI (P < 0.05) and feed per unit gain (P < 0.01). Broilers fed pelleted diets presented higher abdominal fat deposition than those fed mash (P < 0.05). Increasing ME levels reduced the coefficients of ileal apparent digestibility of DM (P < 0.01) and total starch (P < 0.05) but did not affect the digestibility of other evaluated nutrients. The digestibility of all nutrients was lower when pelleted diets were fed compared with mash. Increasing inert material inclusion in the diets at the expense of soybean oil to reduce dietary ME levels promoted higher pellet durability index values (P < 0.05) and the percentage of fines (P < 0.01). Overall, the results suggest that pelleted diets promote better broiler performance because they increase FI, since the digestibility of dietary fractions is reduced. Chickens consuming low-energy pelleted diets may increase FI to compensate for energy deficit. In contrast, broilers fed mash diets may have reached their maximum intake capacity and did not regulate FI by changing feed energy density. When feeding pelleted diets, dietary energy reduction should be considered to reduce feed costs and to improve the carcass quality of broilers.

The growing interest in rotational crossbreeding in Western countries is due to its potential to improve reproductive and health performances of cows. Although a large amount of research focuses on assessing crossbred cows’ performances, how to manage the transition from purebred to rotational crossbred herds is under-explored. Based on a retrospective analysis of French dairy herd case studies, we aimed to identify and characterise technical pathways to make such a transition. In 2018, we performed semi-directive interviews on 26 commercial dairy farms. Data were collected to describe changes in breeding, replacement and culling management practices from the first crossbred mating with purebred cows to the management of a mainly crossbred herd in 2018. Based on a multivariate analysis, we identified two main guidelines structuring technical pathways to move towards rotational crossbred herds: (i) the depth and scale of change (i.e. farm v. herd) associated with the introduction of rotational crossbreeding in the whole-farm dynamics and (ii) the changes in herd replacement and breeding practices to adapt to the evolution of herd demographics induced by the evolution of the dairy crossbred mating rate over time (high from the beginning v. distributed over time). Hierarchical clustering discriminated three groups of farmers differing in their technical pathway to move towards a rotational crossbred herd. In pathway 1, farmers customised one or several rotational crossbreeding schemes to support whole-farm transition towards an organic or grass-based system. Once the scheme stabilised, they quickly implemented it and had to readjust replacement and culling practices to regulate imbalance in herd demographics induced by the improvement in cow fertility. In pathway 2, farmers also customised one or several rotational crossbreeding schemes to support whole-farm redesign but they implemented it more gradually in the herd, which induced no major imbalance in herd demographics. In pathway 3, farmers predefined a relatively well-known rotational crossbreeding scheme to correct fertility issues of purebred cows without any changes at the farm level. They implemented it quickly from the beginning and had to adapt herd replacement and culling to regulate imbalance in herd demographics induced by the improvement in cow fertility. These first empirical evidences on how dairy farmers manage the transition from a purebred to rotational crossbred herd provide original scientific and operational contributions.

Overfeeding in ducks is questioned because forced introduction of food into the animal and metabolic overload may induce damage to health and discomfort. In this context, the objective of our experiment was to measure the impact of dietary strategy on the progression of animal status evaluated through 28 health and behavioural indicators in ducks reared for fatty liver production. To do this, 320 ducks were forced-fed twice a day from 70 to 90 days of age including 10 days of overfeeding (87 to 96 days). They were divided into two groups differing in the feeding strategy during overfeeding period: a moderate feed intake (MI, an average of 376 g of maize flour per meal, n = 160 ducks) or at high feed intake (HI, 414 g/meal, n = 160 ducks). We evaluated 28 indicators related to Good feeding (n = 3), Good housing (n = 4), Good health (n = 10) and Appropriate behaviour (n = 11) principles, taken from the European Welfare Quality Consortium® at four stages: before overfeeding (BEF; 80 days), at the beginning (88 days), the middle (MID; 92 days) or the end of overfeeding (END; 96 days). Animals were slaughtered at 93 and 97 days to measure fatty liver weight at MID and END stages (n = 80 per group). The results showed that dietary strategy influenced the fatty liver weight at MID (+23% in HI v. MI group; P < 0;05) and END stage (+23%; P < 0.05). Assessment stage influenced 13 of the 28 indicators measured. Among these 13 indicators, (i) BEF differed from END stage for 7 indicators and (ii) the dietary strategy degraded all the indicators chosen to evaluate the Good feeding (2/2) principle, but had no effect on the indicators related to the Good health (0/4) principle while (iii) most of the indicators that evaluated Good housing (2/3) and Appropriate behaviour (2/4) principles were affected by an interaction between both factors. Our results suggest that (i) duck fattening status, including the fatty liver weight, and several welfare indicators progressed during the fatty liver production process; and (ii) feeding strategy influenced or even interacted with this progression.

Intramuscular connective tissue (IMCT) is mainly composed of several fibrils (known as total collagen (TCol)) linked between each other by different chemical cross-links (CLs), the whole being embedded in a matrix of proteoglycans (PGs). In the field of beef quality, there is limited information on the role of CLs and PGs. Accordingly, several authors suggest that, to investigate the role of IMCT, it is important to investigate them just like TCol and insoluble collagen (ICol). In muscle, there are two other components, the muscle fibres and intramuscular fat (IMF) content. There are limited data on the relationships between these three components of muscle and then on possibility to independently manipulate these characteristics in order to control the final quality of meat. The present study aimed to investigate whether consistent relationships exist between these different components of muscle. Therefore, the present study compared four muscles of two cattle types (dairy and beef) to determine associations between TCol, ICol, CLs and PGs. Data were analysed across and within muscle (M) and animal type (AT) based on residuals. There was a strong M and AT effect for all muscle characteristics and an interaction M × AT for type I muscle fibres and IMF. Correlations between TCol, ICol and their CLs were M- and AT-independent. Total proteoglycans were positively correlated with TCol and ICol in a muscle-dependent manner irrespective of AT, but no correlation was found with CLs. On the contrary, CLs were negatively correlated with the ratio TPGs : TCol in an M-dependent manner, irrespective of AT. TCol, ICol and CLs were positively and negatively correlated with type IIA and IIB+X muscle fibres only in longissimus thoracis (LT) muscle, regardless the AT. Insoluble collagen was the only parameter of IMCT to be correlated with type I muscle fibres but only in LT muscle, irrespective of AT. There was no correlation between PGs and muscle fibre types, but PGs were the only IMCT component to be related with IMF in an M-dependent manner, irrespective of AT. Finally, there was no correlation between muscle fibre types and IMF content within M and AT. This study revealed that there is a strong relationship between IMCT components irrespective of M, an M-dependent relationship between the IMCT components and muscle fibre types and few (only with PGs) or no relationship between IMF and IMCT and muscle fibres.

Wheat bran (WB) is an important side product of the milling industry and can serve as dietary fiber compound for monogastric animals. The aim of this study was to evaluate the influence of native or fermented WB on the gut physiology and microbiology of laying hens. To accomplish this, 24 laying hens were fed the following diets: conventional diet without WB; 15% native WB in the diet; 15% WB fermented with Pleurotus eryngii; and 15% WB fermented with P. eryngii and a lactic acid bacterial culture. Immediately after slaughtering, digesta samples were taken from the jejunum, ileum and cecum, respectively. Total DNA was extracted and subsequently investigated with 16S DNA amplicon sequencing. Neither native nor fermented WB supplementations negatively affected the feed conversion ratio, laying performance or the relative abundances and alpha-diversity of microbiota in the intestine. Effects of WB-based diets on gut morphology were only recognized in the jejunum (reduced villum height and mucosa thickness). Likewise, WB supplementation decreased the digestibility of DM and starch. Based on these findings, it was demonstrated that different WB variants are applicable without exerting practically negative consequences on performance or on gut microbiota. Fermentation improved the digestibility/retention of dietary fat and phosphorus. However, no further beneficial effects were observed. This study also allowed a more in-depth view on the laying hens’ gut microbiome and its variation within the gut segments.

Sample sizes of welfare assessment protocols must warrant to reflect prevalences on-farm properly – regardless of farm size. Still, solely a fixed sample size was specified for the Welfare Quality® protocol for sows and piglets. The present study investigated whether animals may be assessed from only one body side as applied in the protocol and whether the pre-set sample size of 30 animals mirrors the prevalences of the animal-based indicators on-farm in the gestation unit considering different farm sizes. All indicators were assessed for both sides of an animal’s body by one observer on 13 farms in Germany, which were visited five times within 10 months. The farm visits were treated as independent since different animals were housed in the gestation units. The number of sows in the gestation units varied between 18 and 549 animals. The comparison of sides was carried out calculating exact agreement between animals’ sides and a Wilcoxon signed-rank test (W). The results signified that it is sufficient to assess the animal from one side (exact agreement: 88.3% to 99.5%, except for bursitis (70.0%); W: P-values 0.14 to 0.92). However, if side preferences existed in the indicator bursitis a potential bias must be considered. In the following, the sample size was evaluated by comparing samples’ prevalences against true prevalence, that is, the prevalence of all observed animals in the gestation unit in each farm visit. Therefore, subsets of data were generated by applying simple random sampling without replacement. The samples randomly included the animals’ right or left sides. Linear regression was rated as appropriate provided: coefficient of determination R2 ≥ 0.90, slope = 1 and intercept = 0 signifying exact agreement. The results revealed that the sample size required by the protocol and the application of calculation formulas are solely appropriate to mirror the prevalences of frequent indicators in the gestation unit, for example, bursitis (mean prevalence 34.4%). Using a proportion of animals, for example, a sample of 30% of all observed animals in a farm visit, pointed out that proportions must increase with indicators’ underlying prevalence narrowing 0.00%. Local infections (mean prevalence 13.3%) needed samples including 60% of all observed animals in each farm visit, whereas vulva lesions (mean prevalence 7.28%) only reached accuracy with the inclusion of 70% of the animals. Indicators with a mean prevalence of <1% were not analysed but can most likely only be ascertained by the assessment of all animals.

Brominated flame retardants (BFR) are primarily used as flame retardant additives in insulating materials. These lipophilic compounds can bioaccumulate in animal tissues, leading to human exposure via food ingestion. Although their concentration in food is not yet regulated, several of these products are recognised as persistent organic pollutants; they are thought to act as endocrine disruptors. The present study aimed to characterise the occurrence of two families of BFRs (hexabromocyclododecane (HBCDD) and polybrominated diphenyl ethers (PBDE)) in hen eggs and broiler or pig meat in relation to their rearing environments. Epidemiological studies were carried out on 60 hen egg farms (34 without an open-air range, 26 free-range), 57 broiler farms (27 without an open-air range, 30 free-range) and 42 pig farms without an open-air range in France from 2013 to 2015. For each farm, composite samples from either 12 eggs, five broiler pectoral muscles or three pig tenderloins were obtained. Eight PBDE congeners and three HBCDD stereoisomers were quantified in product fat using gas chromatography–high-resolution mass spectrometry, or high-performance liquid chromatography–tandem mass spectrometry, respectively. The frequencies of PBDE detection were 28% for eggs (median concentration 0.278 ng/g fat), 72% for broiler muscle (0.392 ng/g fat) and 49% for pig muscle (0.403 ng/g fat). At least one HBCDD stereoisomer was detected in 17% of eggs (0.526 ng/g fat), 46% of broiler muscle (0.799 ng/g fat) and 36% of pig muscle (0.616 ng/g fat). Results were similar in concentration to those obtained in French surveillance surveys from 2012 to 2016. Nevertheless, the contamination of free-range eggs and broilers was found to be more frequent than that of conventional ones, suggesting that access to an open-air range could be an additional source of exposure to BFRs for animals. However, the concentration of BFRs in all products remained generally very low. No direct relationship could be established between the occurrence of BFRs in eggs and meat and the characteristics of farm buildings (age, building materials). The potential presence of BFRs in insulating materials is not likely to constitute a significant source of animal exposure as long as the animals do not have direct access to these materials.