Higher accumulation of skatole in the fat of male pigs compared with female pigs might be due to gender-related differences in the rate of skatole degradation. In the present study, skatole metabolites and cytochrome P450 (CYP450) isoforms involved in skatole metabolism were for the first time investigated in hepatic S9 fractions from six male and four female pigs (crossbred Landrace×Yorkshire dams and Duroc boar). Surprisingly, the rates of production of major skatole metabolites were similar in male and female pigs. The most abundant metabolite of skatole was 3-hydroxy-3-methyloxindole (HMOI) followed by 3-methyloxindole and indole-3-carbinol in both male and female S9 fractions. Concentrations of formed HMOI and 3-methyloxindole did not differ between the genders (P=0.124 for HMOI, and P=0.575 for 3-methyloxindole). Indole-3-carbinol formation was higher in S9 fractions from the females compared with male pigs (P=0.0001). Enzyme kinetic parameters were similar for both genders (P>0.05). In both male and female pigs, ellipticine, diallyl sulphide (DAS) and quercetin inhibited HMOI formation, confirming the involvement of CYP1A1 and CYP2E1. The formation of 3-methyloxindole was reduced in the presence of the CYP2E1 inhibitor DAS, and formation of indole-3-carbinol was reduced in the presence of CYP1A1 and CYP2A19 inhibitors. We found only minor differences in skatole metabolism between male and female pigs, particularly the involvement of CYP2C and CYP3A in indole-3-carbinol formation in female but not in male pigs. This is a very essential finding, suggesting the involvement of larger number of CYP450 isoforms in female pigs. On the other hand, indole-3-carbinol is a minor skatole metabolite, and the physiological significance of CYP2C and CYP3A involvement in its formation in female pigs, but not in male pigs, needs to be elucidated. Our results, however, should be interpreted with caution because of the low number of animals and possibility of breed and age effects on skatole metabolism.

The effects of early life events on average daily weight gain from birth to day 21 (ADG) of suckling pigs kept at different room temperatures (15°C, 20°C and 25°C) from birth to weaning were investigated. Data were collected from litters born by 61 sows in a loose housing system. The ADG for piglets with low birth weight (estimated for birth weights below the 10% percentile) was estimated to be 20 to 30 g higher per day at room temperature 20°C to 25°C compared with 15°C. In contrast, the ADG during the lactation period decreased for larger piglets (estimated for birth weights above the 10% percentile) by 28 g/day at room temperature 25°C compared with 15°C. Thus, high ambient temperatures (20°C to 25°C) are favourable for the growth in smaller piglets during lactation. Neither latency to first suckle nor birth-induced hypoxia, measured as concentration of umbilical cord lactate, affected the growth rate of the piglets. Lowest rectal temperature during the first 24 h after birth had a long-term negative effect on ADG (P<0.05), so that piglets with a lowest rectal temperature of 32.8°C (10% percentile) had an ADG which was on average 19 g lower per day than piglets with a rectal temperature of 37.3°C (90% percentile). Our results showed that hypothermia at birth, low birth weight and high number of suckling piglets lead to reduced ADG during the suckling period. The results suggest that keeping the room temperature at 20°C during lactation to some extent could compensate for the otherwise negative effects of low birth weight on ADG in piglets without decreasing the ADG of high birth weight piglets. However, to avoid hypothermia in the smallest piglets it may be beneficial to increase the room temperature above 20°C during the farrowing period of loose housed sows.

To evaluate effects of different concentrations of nanosilver colloid on the cell culture of Sertoli cells, the proportion of lipid peroxidation, antioxidant capacity, nitric oxide (NO) production and genes expression of superoxide dismutases (SOD1 and SOD2) and nitric oxide synthases (eNOS and iNOS) were measured. Sertoli cells were incubated at concentrations of 25, 75 and 125 ppm nanosilver for 48 h. There was progressive lipid peroxidation in treatments according to increasing of nanosilver. Lipid peroxidation, as indicated by malondialdehyde levels, was significantly elevated by the highest concentration of silver colloid (125 ppm), although antioxidant capacity, as measured by ferric ion reduction, was unaffected. Nitrite, as an index of NO production was reduced only in 125 ppm of nanosilver. Expression of SOD1 gene was reduced in nanosilver-treated cells at all concentrations, whereas expression of SOD2 gene was reduced only in cells treated with 125 ppm nanosilver. Expression of iNOS gene was progressively increased with higher concentrations of nanosilver. Expression of eNOS gene was also increased in 125 ppm of nanosilver. In conclusion, toxic effects of nanosilver could be due to high lipid peroxidation and suppression of antioxidant mechanisms via reduced expression of SOD genes and increased expression of NOS genes.

Myogenesis is facilitated by four myogenic regulatory factors and is significantly inhibited by myostatin. The objective of the current study was to examine embryonic gene regulation of myostatin/myogenic regulatory factors, and subsequent manipulations of protein synthesis, in broiler embryos under induced hyperammonemia. Broiler eggs were injected with ammonium acetate solution four times over 48 h beginning on either embryonic day (ED) 15 or 17. Serum ammonia concentration was significantly higher (P<0.05) in ammonium acetate injected embryos for both ED17 and ED19 collected samples when compared with sham-injected controls. Expression of mRNA, extracted from pectoralis major of experimental and control embryos, was measured using real-time quantitative PCR for myostatin, myogenic regulatory factors myogenic factor 5, myogenic determination factor 1, myogenin, myogenic regulatory factor 4 and paired box 7. A significantly lower (P<0.01) myostatin expression was accompanied by a higher serum ammonia concentration in both ED17 and ED19 collected samples. Myogenic factor 5 expression was higher (P<0.05) in ED17 collected samples administered ammonium acetate. In both ED17 and ED19 collected samples, myogenic regulatory factor 4 was lower (P⩽0.05) in ammonium acetate injected embryos. No significant difference was seen in myogenic determination factor 1, myogenin or paired box 7 expression between treatment groups for either age of sample collection. In addition, there was no significant difference in BrdU staining of histological samples taken from treated and control embryos. Myostatin protein levels were evaluated by Western blot analysis, and also showed lower myostatin expression (P<0.05). Overall, it appears possible to inhibit myostatin expression through hyperammonemia, which is expected to have a positive effect on embryonic myogenesis and postnatal muscle growth.

Based on the potential benefits for long-term human health, there is interest in developing sustainable nutritional strategies for lowering medium-chain saturated fatty acids (FA) and increasing specific unsaturated FA in ruminant milk. Dietary supplements of extruded linseeds (EL), fish oil (FO) or a mixture of EL and FO increase cis-9,trans-11 CLA and long-chain n-3 polyunsaturated FA in bovine milk. Supplements of FO cause milk fat depression in lactating cows, but information for dairy goats is limited. A total of 14 Alpine goats were used in a replicated 3×3 Latin square with 28-days experimental periods to examine the effects of EL alone or in combination with FO on animal performance, milk fat synthesis and milk FA composition. Treatments comprised diets based on natural grassland hay supplemented with no additional oil (control), 530 of EL or 340 g/day of EL and 39 g/day of FO (ELFO). Compared with the control, ELFO tended (P=0.08) to lower milk fat yield, whereas EL increased (P<0.01) milk fat content and yield (15% and 10%, respectively). Relative to EL, ELFO decreased (P<0.01) milk fat content and yield (19% and 17%, respectively). Relative to the control and ELFO, EL decreased (P<0.05) milk 10:0 to 16:0 and odd- and branched-chain FA content and increased 18:0, cis-18:1, trans-13 18:1 (and their corresponding ∆-9 (desaturase products), trans-12,cis-14 CLA, cis-13,trans-15 CLA, cis-12,trans-14 CLA and trans-11,cis-13 CLA and 18:3n-3 concentrations. ELFO was more effective for enriching (P<0.05) milk cis-9, trans-11 CLA and trans-11 18:1 concentrations (up to 5.4- and 7.1-fold compared with the control) than EL (up to 1.7- and 2.5-fold increases). Furthermore, ELFO resulted in a substantial increase in milk trans-10 18:1 concentration (5.4% total FA), with considerable variation between individual animals. Relative to the control and EL, milk fat responses to ELFO were characterized by increases (P<0.05) in milk trans-16:1 (Δ9 to 11), trans-18:1 (Δ6 to 11), trans-18:2, CLA (cis-9,trans-11, trans-9,cis-11, trans-8,trans-10 and trans-7,trans-9) and 20- and 22-carbon FA concentrations. Overall, EL resulted in a relatively high cis-9 18:1 concentration and an increase in the 18:3n-3/18:2n-6 ratio, whereas combining EL and FO resulted in substantial increases in trans-FA, marginal enrichment in 20:5n-3 and 22:6n-3 and lower 16:0 concentration changes associated with a decrease in milk fat content. In conclusion, data provide further evidence of differential mammary lipogenic responses to diet in the goat compared with the cow and sheep.

Orexin A and B are hypothalamic peptides derived from the prepro-orexin (PPO) precursor. Orexins stimulate food intake and arousal. Those peptides bind and activate two G protein-coupled receptors: orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R). Numerous authors have suggested that orexins play an important role in the regulation of the reproductive functions. The objective of the present study was to analyse the presence of and changes in the gene and protein expression pattern of the orexin system in the porcine uterus, conceptus and trophoblast (chorioallantois) during early pregnancy. In the endometrium, the highest PPO and OX1R gene expression was detected on days 15 to 16 of gestation. The OX2R mRNA content in the endometrium was higher on days 10 to 11 and 15 to 16 than on days 12 to 13 and 27 to 28. In the trophoblasts, PPO gene expression was higher on days 30 to 32 than on days 27 to 28. The highest PPO protein content in the endometrium was noted on days 12 to 13. The highest OX1R protein content in the endometrium was detected on days 10 to 11, whereas OX2R protein on days 15 to 16. In the trophoblasts, PPO and OX1R protein levels were more pronounced on days 27 to 28 than on days 30 to 32, but OX2R expression was higher on days 30 to 32. The expression of PPO, OX1R and OX2R was different in the conceptuses and trophoblasts during early pregnancy. Local orexin production and the presence of the specific orexin receptors suggest that the orexin system may participate in the control of porcine reproductive functions by exerting endocrine and auto/paracrine effects on the uterus, conceptuses and trophoblasts during early pregnancy. This study provides the first evidence for the presence of orexins and their receptors in the uteri, conceptuses and trophoblasts in pigs during early pregnancy. The local orexin system is dependent on the stage of pregnancy.

Increasing the concentration of dietary lipid is a promising strategy for reducing methane (CH4) emissions from ruminants. This study investigated the effect of replacing grass silage with brewers’ grains on CH4 emissions of pregnant, non-lactating beef cows of two breeds. The experiment was a two×two factorial design comprising two breeds (LIMx, crossbred Limousin; and LUI, purebred Luing) and two diets consisting of (g/kg diet dry matter (DM)) barley straw (687) and grass silage (301, GS), or barley straw (763) and brewers’ grains (226, BG), which were offered ad libitum. Replacing GS with BG increased the acid-hydrolysed ether extract concentration from 21 to 37 g/kg diet DM. Cows (n=48) were group-housed in equal numbers of each breed across two pens and each diet was allocated to one pen. Before measurements of CH4, individual dry matter intake (DMI), weekly BW and weekly body condition score were measured for a minimum of 3 weeks, following a 4-week period to acclimatise to the diets. CH4 emissions were subsequently measured on one occasion from each cow using individual respiration chambers. Due to occasional equipment failures, CH4 measurements were run over 9 weeks giving 10 observations for each breed×treatment combination (total n=40). There were no differences between diets for daily DMI measured in the chambers (9.92 v. 9.86 kg/day for BG and GS, respectively; P>0.05). Cows offered the BG diet produced less daily CH4 than GS-fed cows (131 v. 156 g/day: P<0.01). When expressed either as g/kg DMI or kJ/MJ gross energy intake (GEI), BG-fed cows produced less CH4 than GS-fed cows (13.5 v. 16.4 g/kg DMI, P<0.05; 39.2 v. 48.6 kJ/MJ GEI, P<0.01). Breed did not affect daily DMI or CH4 expressed as g/day, g/kg DMI or kJ/MJ GEI (P>0.05). However, when expressed as a proportion of metabolic BW (BW0.75), LUI cows had greater DMI than LIMx cows (84.5 v. 75.7 g DMI/kg BW0.75, P<0.05) and produced more CH4 per kg BW0.75 than LIMx cows (1.30 v. 1.05 g CH4/kg BW0.75; P<0.01). Molar proportions of acetate were higher (P<0.001) and propionate and butyrate lower (P<0.01) in rumen fluid samples from BG-fed compared with GS-fed cows. This study demonstrated that replacing GS with BG in barley straw-based diets can effectively reduce CH4 emissions from beef cows, with no suppression of DMI.

The objective of this study was to determine whether rabbits fed in a restricted regimen (75%) showed increased competition for feeding, drinking and use of specific areas of the cages as compared with those provided feed ad libitum. This evaluation was carried out by measuring their space utilisation in the cage, the incidence of agonistic behaviour and rates of mortality. In total, 504 rabbits between 31 and 66 days of age were used in this study. A total of 200 heavy-weight rabbits and 56 light-weight rabbits were randomly housed in 32 cages, each cage containing eight rabbits: 25 cages housing heavy rabbits and seven cages housing the light-weight ones. They were all fed ad libitum (AD). In addition, a total of 208 heavy-weight rabbits and 40 light-weight rabbits were randomly housed in 31 cages, each of them containing eight rabbits: 26 cages housing heavy weight rabbits and five cages housing light-weight ones. They were all fed a restricted diet (R) regimen. The restriction was calculated to be 75% of the feed consumed by the AD group. The total space available in the cage was 3252 cm2, with a stocking density of 24.6 animals/m2. Animals between 32 and 60 days of age from 20 different cages were observed nine times per week (morning or afternoon) by means of scan and focal sampling by one observer. During each period, cages were assessed for 5 min, registering every minute the position of all the animals in relation to Area A (feeder), Area B (central part) or Area C (back and drinker area). The incidence of agonistic behaviour such as displacement, biting and jumping on each other was also assessed. Performance variables such as daily gain and feed conversion ratio, in addition to general health status and mortality rates, were recorded for all rabbits. When the rabbits were under restricted feeding, the competition for feed and drink increased with clear signs of agonistic behaviour such as biting, displacement and animals jumping on top of each other. Although this competition was maintained during the entire growing period, the BW homogeneity between animals in the same cage was similar in both cases, suggesting that all animals could consume similar quantities of feed. The possible advantages of a restricted diet, such as better feed conversion ratio, were observed in this study only in the last few weeks of the growing period.

In fish, in vitro fertilization is an important reproductive tool used as first step for application of others biotechniques as chromosome and embryo manipulation. In this study, we aimed to optimize gamete quality and their short-term storage from the yellowtail tetra Astyanax altiparanae, for future application in laboratory studies. Working with sperm, we evaluated the effects of spawning inducers (carp pituitary gland and Ovopel® [(D-Ala6, Pro9-NEt) – mGnRH+metoclopramide]) and the presence of female on sperm motility. Additionally, we developed new procedures for short-term storage of sperm and oocytes. Briefly, sperm motility was higher when male fish were treated with carp pituitary gland (73.1±4.0%) or Ovopel® (79.5±5.5%) when compared with the control group treated with 0.9% NaCl (55.6±27.2%; P=0.1598). Maintenance of male fish with an ovulating female fish also improved sperm motility (74.4±7.4%) when compared with untreated male fish (42.1±26.1%; P=0.0018). Storage of sperm was optimized in modified Ringer solution, in which the sperm was kept motile for 18 days at 2.5°C. The addition of antibiotics or oxygen decreased sperm motility, but partial change of supernatant and the combination of those conditions improve storage ability of sperm. Fertilization ability of oocytes decreased significantly after storage for 30, 60 90 and 120 min at 5, 10, 15 and 20°C when compared with fresh oocytes (P=0.0471), but considering only the stored samples, the optimum temperature was 15°C. Those data describe new approaches to improve semen quality and gametes short-term storage in yellowtail tetra A. altiparanae and open new possibilities in vitro fertilization.

The aim of this research was to determine the influence of dietary replacement of n-6 with n-3 polyunsaturated fatty acids on cellular immunity and oxidative stress in the transition period dairy cows. The experiment was conducted on 20 dairy Holstein cows from 3±1 weeks before parturition until the 6th week of lactation. Both groups were fed an iso-energetic and iso-nitrogenous diet. Soybean meal from control (C) group was replaced with linseed in the experimental (LS) group. Cellular immunity and oxidative stress were measured on days −10, 1, 21 and 42 relative to parturition. During the entire experimental period, the proportion of CD45+ cells was lower (P<0.05) in LS group compared with the C group. The phagocytosis ability and phagocytosis index of cows fed with n-3 fatty acids were significantly reduced (P<0.05) compared with the group of cows fed with n-6 fatty acids. The most severe decrease in phagocytosis ability was on day −10 and 1 relative to parturition. The activity of superoxide dismutase (P<0.05) and plasma glutathione peroxidase (P<0.05) increased around calving, although activities were not influenced by dietary treatment. Increased malondialdehyde concentration (P<0.05) was influenced by dietary n-3 fatty acids and the time relative to parturition. The immune suppression was most pronounced during periparturient period. In that matter we can conclude that not only dietary n-3 fatty acids but also oxidative stress, which reached peak at time of parturition, contributed to the reduced cellular immunity during the periparturient period.

Factors affecting calving interval (CI) in double-muscled Belgian Blue (DMBB) beef cows were investigated with regard to the BW yield (BWY) of the cow–calf pair, using 834 CI records from 386 females with parities 1 to 6. The effect of parity and CI on BWY was also studied. Cow–calf pair BWY was defined as calf birth weight plus dam BWY per CI. CI (mean±s.e.: 404±1.9 days) was affected by parity, calving season, suckling and calf birth weight/dam weight. Primiparous cows had a shorter CI than cows with three or more calvings (P<0.05), with an intermediate CI for second-calf cows. Spring calvings resulted in a shorter CI than summer and autumn calvings, with intermediate values for winter calvings. Suckling dams had longer CIs than non-suckling dams. There were interactions (P<0.05) between calving season and suckling, and between calving season and mating system. Shortest CIs were observed for spring calvings in case of non-suckling and for summer calvings in case of suckling. Longest CIs were observed for autumn calvings in case of natural service (NS) and for winter calvings in case of artificial insemination (AI). Calf birth weight/dam weight of 6% to 10% resulted in shorter CI than a ratio of <6% (P<0.05). Body condition and mating system (NS v. AI) did not affect CI. Daily cow–calf pair BWY was affected by parity (P<0.001) and CI (P=0.013), with a tendency for an interaction (P=0.094). Daily cow–calf pair BWY did not differ for CIs of <12 to 16 months in primiparous cows and was lowest for a CI of 13 to 15 months in second-calf cows, whereas the effect of CI was more variable in older cows. Dam contribution to cow–calf pair BWY was larger than calf birth weight in first- and second-calf cows, and increased with increasing CI. Dam contribution to cow–calf pair BWY was smaller than calf birth weight in older cows, varying from 0.2 to 1.0 depending on CI. A short CI is advised for DMBB cows because of a larger BWY and more efficient nutrient utilisation.

In organic pig production one of the major challenges is to be able to fulfil amino acid requirements based on organic and locally grown protein feed crops. The pig is an opportunistic omnivore with a unique capacity for foraging above and below the soil surface. It is hypothesized that direct foraging in the range area can pose an important contribution in terms of fulfilling nutrient requirements of growing pigs. Foraging activity, lucerne nutrient intake and pig performance were investigated in 36 growing pigs, foraging on lucerne or grass and fed either a standard organic pelleted feed mixture (HP: high protein) or a grain mixture containing 48% less CP (LP: low protein) compared with the high protein feed mixture, from an average live weight of 58 kg to 90 kg in a complete block design in three replicates. The pigs were fed 80% of energy recommendations and had access to 4 m2 of pasture/pig per day during the 40 days experimental period from September to October 2013. Behavioural observations were carried out 12 times over the entire experimental period. For both crops, LP pigs rooted significantly more compared with HP pigs but the effect of CP level was more pronounced in grass (44% v. 19% of all observations) compared with lucerne (28% v. 16% of all observations). Feed protein level turned out not to have any significant effect on grazing behaviour but pigs foraging on lucerne grazed significantly more than pigs foraging on grass (10% v. 4% of all observations). Daily weight gain and feed conversion ratio were significantly affected by feed protein and forage crop interactions. Compared to HP pigs, LP treated pigs had 33% lower daily weight gain (589 v. 878 g) and 31% poorer feed conversion ratio (3.75 v. 2.59 kg feed/kg weight gain) in grass paddocks, whereas in lucerne paddocks LP pigs only had 18% lower daily weight gain (741 v. 900 g) and a 14% poorer feed conversion ratio (2.95 v. 2.54 kg feed/kg weight gain) compared with HP pigs. LP pigs foraging on lucerne used 169 g less concentrate CP/kg weight gain, compared with HP pigs, indicating the nitrogen efficiency of the system. The results indicate that direct foraging of lucerne may be a valuable strategy in terms of accommodating CP and lysine requirements of organic growing pigs.

In cattle, the detection of very early endometrial responses is considered to be hampered by the presence of only a single embryo. Therefore, we have previously developed a model of multiple embryo transfer to circumvent this hindrance. In this work, we analysed embryo–maternal interactions in the bovine uterus on day 8 of development while comparing the presence of multiple v. single embryos using embryo transfer and artificial insemination, respectively. Concentration of proteins (β-actin, NFkB, clusterin and immunoproteosome 20S β5i subunit–i20S), by western blot, and hexoses (glucose and fructose) were measured in paired samples of uterine fluid (UF) from the same animal with and without embryos in the uterus and were compared with UF obtained after artificial insemination. Prostaglandin (PG) F2α and PGE2 concentrations were also analysed in blood plasma. The four proteins analysed and hexoses were unaffected by the presence of one or more embryos in the uterus. However, blood PGF2α showed similar, significant increases with one or more embryos over cyclic animals; such changes were not observed in blood PGE2. Although multiple embryo transfer may appear to be non-physiological, we showed that the uterus, at the very early embryonic stages, does exhibit physiological reactions. Multiple embryo transfer can, therefore, be used for studies of very early embryo–maternal interactions in vivo in monotocous species.

The factors influencing the priority of access to food and the effects of the priority of access to food on their carcass traits were analyzed for Japanese Black (Wagyu) cattle in a semi-intensive fattening production system. The records of 96 clinically healthy steers and heifers were analyzed. The calves at ∼3 to 4 months of age were allocated to pens with four animals per pen; all four animals in the same pen were of the same sex and of similar body size. The ranking of the animals’ priority of access to food (1st, 2nd, 3rd and 4th), which was determined by the farm manager, was used as an indicator of social dominance in the present study. Four models including sire line, maternal grandsire line and the difference in the animals’ birth dates as fixed effects were used to analyze factors influencing the priority of access to food. Ranking was represented by ordinal scores (highest=4, lowest=1) in Model 1, and the binary scores were assigned in Model 2 (highest=1; 2nd, 3rd and 4th=0), Model 3 (1st and 2nd=1; 3rd and 4th=0) and Model 4 (1st, 2nd and 3rd=1; lowest=0). The results showed that the difference in the animals’ birth dates had a significant effect on the establishment of the priority of access to food in Model 3 (P<0.05), suggesting that animals born earlier may become more dominant in the pen. The maternal grandsire line tended to affect the social rank score in Models 2 and 3 (P<0.10). Our results indicated that the maternal grandsire line may affect the temperament of calves through their mothers’ genetic performance and thereby more aggressive calves may be more dominant and have higher priority of access to food. On the other hand, there was a significant effect of the priority of access to food on beef marbling score (BMS; P<0.05), and the priority of access to food also tended to influence the carcass weight (P=0.09). The highest BMS was observed for animals with the first rank of the priority of access to food (P<0.05), and the higher-ranking animals had the tendency to be heavier carcass than the lower-ranking animals. Our findings emphasized the importance of information about the priority of access to food determined by farmers’ own observation on implementing best management practices in small-scaled semi-intensive beef cattle production systems.

Androstenone is a steroid pheromone occurring in the pubertal Leydig cells. Breeding against androstenone can decrease pheromone odour in swine meat but appears to cause unwanted side effects such as delayed onset of puberty. To study causality, global gene expression in developing boar testes at 12, 16, 20 and 27 weeks was investigated using a porcine cDNA microarray. The morphological status and androgenic levels of the same individuals have been described in a previous publication. In the present paper, expression of genes and pathways has been analysed with reference to these findings. Nine clusters of genes with significant differential expression over time and 49 functional charts were found in the analysed testis samples. Prominent pathways in the prepubertal testis were associated with tissue renewal, cell respiration and increased endocytocis. E-cadherines may be associated with the onset of pubertal development. With elevated steroidogenesis (weeks 16 to 27), there was an increase in the expression of genes in the MAPK pathway, STAR and its analogue STARD6. A pubertal shift in genes coding for cellular cholesterol transport was observed. Increased expression of meiotic pathways coincided with the morphological onset of puberty. Puberty-related change in Ca(2+) pathway transcripts, neurosteroids, neuronal changes and signalling in redox pathways suggested a developmental-specific period of neuromorphogenesis. Several growth factors were found to increase differentially over time as the testis matured. There may be interactions between MAPK, STAR and growth factors during specific periods. In conclusion, pathways for neurogenesis, morphological pathways and several transcripts for growth factors, which have known modulating effects on steroidogenesis and gonadotropins in humans and rodents, act at specific ages and developmental stages in the boar testis. The age dependency and complexity shown for development-specific testis transcripts must be considered when selecting phenotypic parameters for genetic selection for low androstenone. The results of selection based on measurement of phenotypic maturation and androstenone (or other steroid) levels at one specific age may differ depending on the age used. More research is necessary to find the optimal phenotype to use in order to reduce the unwanted side effects.

Exercise has been shown to increase mRNA expression of a growing number of genes. The aim of this study was to assess if mRNA expression of the metabolism- and oxidative stress-related genes GLUT4 (glucose transporter 4), COX2 (cyclooxygenase 2), SOD1 (superoxide dismutase 1) and HSP70 (heat shock protein 70) in saliva changes following acute exercise stress in dogs. For this purpose, 12 avalanche dogs of the Italian Military Force Guardia di Finanza were monitored during simulation of a search for a buried person in an artificial avalanche area. Rectal temperature (RT) and saliva samples were collected the day before the trial (T0), immediately after the descent from a helicopter at the onset of a simulated avalanche search and rescue operation (T1), after the discovery of the buried person (T2) and 2 h later (T3). Expressions of GLUT4, SOD1, COX2 and HSP70 were measured by real-time PCR. The simulated avalanche search and rescue operation was shown to exert a significant effect on RT, as well as on the expression of all metabolism- and oxidative stress-related genes investigated, which peaked at T2. The observed expression patterns indicate an acute exercise stress-induced upregulation, as confirmed by the reductions in expression at T3. Moreover, our findings indicate that saliva is useful for assessing metabolism- and oxidative stress-related genes without the need for restraint, which could affect working dog performance.

Using behavioural indicators of thermal discomfort, that is, shade seeking, panting scores (PS) and respiration rate (RR), we evaluated the effect of hot summer conditions and shade, for a herd of adult Holstein dairy cows and a herd of Belgian Blue beef cows kept on pasture in a temperate area (Belgium). During the summer of 2012, both herds were kept on pasture without access to shade (NS). During the summers of 2011 and 2013 each herd was divided into one group with (S) and one without (NS) access to shade. Shade was provided by young trees with shade cloth (80% reduction in solar radiation) hung between them. For S cows, we investigated how shade use was related to hot conditions as quantified by six climatic indices. The heat load index (HLI), which incorporates air temperature and humidity, solar radiation and wind speed, was the best predictor of the six indices tested. In 2011, there was a relatively high threshold for use of shade. When HLI=90, shade use probability reached 17% for dairy cows and 27% for beef cows. In 2013, however, at HLI=90, shade use probability reached 48% for dairy cows and 41% for beef cows. For animals from the NS treatment we determined the effect of hot summer conditions on RR and PS (with 0=no panting and 4.5=extreme panting). In both types of cattle, an increase in black globe temperature was the best predictor for increasing RR and PS. Furthermore, we determined how the effect of hot summer conditions on RR and PS was affected by the use of shade. Under hot conditions (black globe temperature ⩾30°C), >50% of the animals under shade retained normal PS and RR (PS<1 and RR<90 breaths per minute), whereas normal RR and PS were significantly less prevalent for animals outside shade. Our findings suggest that, even in temperate summers, heat can induce thermal discomfort in cattle, as evidenced by increases in shade use, RR and PS, and that shade increases thermal comfort.

Insulin-like growth factor binding protein-5 (IGFBP-5) is a key molecule in mammary gland development, which facilitates the removal of mammary epithelial cells (MECs) by apoptosis that takes place during remodeling of the mammary gland during involution. IGFBP-5 binds with IGFs for their bioavailability. IGFBP-5 has been reported to perform pleiotropic roles such as cellular apoptosis, proliferation and differentiation. To understand the role of IGFBP-5 during lactation and clinical mastitis, expression profiling of IGFBP-5 at the protein level was performed in both indigenous cows (Bos indicus) and buffaloes (Bubalus bubalis) belonging to two different breeds – Sahiwal cows and Murrah buffaloes. Reverse-transcriptase PCR (RT-PCR) of IGFBP-5 mRNA confirmed its expression in milk somatic cells and MECs of Sahiwal cows. ELISA was performed for quantitative measurement of IGFBP-5 concentrations in milk during different days (0, 50, 100, 150, 200, 250 and 300) of lactation, during the involution period and in animals exhibiting short lactation and clinical mastitis. The highest concentration of IGFBP-5 in milk was observed during the involution period followed by colostrum, late and early lactation, respectively, in both cattle and buffaloes. No significant difference in the concentration of IGFBP-5 was observed during the first 150 days of lactation between cows and buffaloes. However, higher concentration of IGFBP-5 was observed in cows during late lactation (200 to 300 days) in comparison with buffaloes. To validate the ELISA data, quantitative real-time PCR was performed in MECs of Sahiwal cows. The relative mRNA abundance of IGFBP-5 was found to be significantly (P<0.05) higher on day 15 than between 50 and 150 days of lactation in case of Sahiwal cows. Highest mRNA expression of IGFBP-5 was observed around 300 days of lactation followed by 200 and 250 days (P<0.05), respectively. Murrah buffaloes showed low levels of IGFBP-5 protein in milk as compared with Sahiwal cows during lactation in ELISA. Animals having history of short lactation length (short lactating animals) showed higher levels of IGFBP-5 expression (at protein level) in comparison with normal lactating animals. We propose that higher level IGFBP-5 expression may have functional significance in lactation persistency. As a pro-apoptotic molecule, higher expression of IGFBP-5 was observed to be inversely related to lactation length and milk production.

The aim of this work was to establish the response of growing sheep, goats and cattle to different nutritional environments. Data from 590 publications representing 2225 treatments were analysed. The results showed that each 10% increase in NDF was accompanied by 0.11 g/kg live weight (LW) and 0.32 g/kg metabolic live weight (LW0.75) decreases in DMI. Otherwise, the response of DMI to CP (CP%DM) content was curvilinear (P<0.01), without any significant difference in the slope between species. The percentage of concentrate (% CC) affected DMI curvilinearly, without any significant difference between species. This meta-analysis demonstrated the negative linear effect of NDF and the quadratic effect of CP concentration on organic matter digestibility (OMd). For growth performance, the three species responded curvilinearly to variations in metabolisable energy intake (MEI MJ/kg LW0.75) and digestible CP (DCPI g/kg LW0.75) intake (P<0.01). At the same level of MEI, average daily gain (ADG) varied with CP contents of the diet, and only the intercept differences were significant between the three levels (P=0.07). At the same level of DCPI, ADG varied with energy level (below maintenance (LE−−), 1 to 1.2×maintenance (LE−), 1.2 to 1.4× maintenance (ME+−), and >1.4, corresponding to maximum growth (HE+)). No significant difference was observed between LE−− and LE−, and no significant difference was observed between ME+− and HE+. For nitrogen balance, no difference was observed between species for a given level of nitrogen intake.

Newly hatched chicks may be held longer than 48 h and experience long periods of fasting in commercial hatcheries. Limited information is known about the physiological status of chicks in such situations, due to the difficulty of precisely recording time of hatch. This study investigated the effect of the time from hatch to pulling (holding period) on physiological measures/parameters in 109 broiler chicks. Fertile Ross 308 eggs were incubated in a custom built small-scale incubator. The individual hatching time of each focal chick was determined using eggshell temperature monitoring. At ‘pulling’ (512 h of incubation time), the quality of focal chicks was assessed using the chick scoring method and physiological parameters were measured including BW, organ (heart, liver and stomach) weights, blood values and plasma corticosterone level. The time from hatch to pulling varied from 7.58 to 44.97 h. Egg weight at setting was significantly correlated with chick BW and weight of organs at pulling, but had no effect on chick quality, blood values and plasma corticosterone. Relative BW at pulling was negatively associated with the duration of holding period (P=0.002). However, there was a positive correlation between relative stomach weight and the duration of the holding period (P<0.001). As the holding period duration increased, there was a trend that blood partial pressure of oxygen, haematocrit and haemoglobin also increased, and blood partial pressure of carbon dioxide, total carbon dioxide and bicarbonate decreased (P<0.05). A wide range of plasma corticosterone was observed from chicks that had experienced different durations of holding period. We conclude that shortening the hatch window and minimising the number of chicks that experience a long holding period before pulling may improve chick quality and physiological status, which may be due to unfavourable environmental conditions that include feed and water deprivation.