Effect of animal mixing as a stressor on biomarkers of autophagy and oxidative stress during pig muscle maturation

The objective of this work was to study the postmortem evolution of potential biomarkers of autophagy (Beclin 1, LC3-II/LC3-I ratio) and oxidative stress (total antioxidant activity, TAA; superoxide dismutase activity, SOD and catalase activity, CAT) in the Longissimus dorsi muscle of entire male ((Large White×Landrace)×Duroc) pigs subjected to different management treatments that may promote stress, such as mixing unfamiliar animals at the farm and/or during transport and lairage before slaughter. During the rearing period at the farm, five animals were never mixed after the initial formation of the experimental groups (unmixed group at the farm, UF), whereas 10 animals were subjected to a common routine of being mixed with unfamiliar animals (mixed group at the farm, MF). Furthermore, two different treatments were used during the transport and lairage before slaughter: 10 pigs were not mixed (unmixed group during transport and lairage, UTL), whereas five pigs were mixed with unfamiliar animals on the lorry and during lairage (mixed group during transport and lairage, MTL). These mixing treatments were then combined into three pre-slaughter treatments – namely, UF-UTL, MF-UTL and MF-MTL. The results show that MF-UTL and MF-MTL increased significantly the muscle antioxidant defense (TAA, SOD and CAT) at short postmortem times (4 and 8 h; P<0.001), followed by an earlier depletion of the antioxidant activity at 24 h postmortem (P<0.05). We also found that mixing unfamiliar animals, both at the farm and during transport and lairage, triggers postmortem muscle autophagy, which showed an earlier activation (higher expression of Beclin 1 and LC3-II/LC3-I ratio at 4 h postmortem followed by a decreasing pattern of this ratio along first 24 h postmortem) in the muscle tissues of animals from the MF-UTL and MF-MTL groups, as an adaptive strategy of the muscle cells for counteracting induced stress. From these results, we propose that monitoring the evolution of the main biomarkers of autophagy (Beclin 1, LC3-II/LC3-I ratio) and muscle antioxidant defense (TAA, SOD, CAT) in the muscle tissue within the first 24 h postmortem may help the detection of animal stress and its potential effect on the postmortem muscle metabolism.     

Activation of caspase-9 and its influencing factors in beef during conditioning

To study the activation of caspase-9 and its potential influence in conditioning, longissimus thoracis (LT), semitendinosus (STN) and psoas minor (PMi) muscles were used to analyze the ratio of pro-apoptotic bax to anti-apoptotic bcl-2 in fresh tissues and observe the changes in ATP, cytosolic cytochrome c and caspase-9 activity levels during storage at 4°C. Caspase-9 activity at 5 h is higher than the activity at 0 and 24 h in the muscles (P<0.001). The ATP content decreased between 0 and 3 h, between 8 and 14 h in the PMi and LT muscles (P<0.0001), whereas between 0 and 5 h, between 8 and 14 h in the STN muscle (P<0.0001). There is 60.2%, 55.3% and 43.1% available ATP in the STN, LT and PMi muscles at 5 h, respectively. The cytosolic cytochrome c level increased during 5 and 24 h storage in the LT and PMi muscles (P<0.0001), during 5 and 96 h in the STN muscle (P<0.0001). The cytosolic cytochrome c at 24 h (P<0.001) and ratio of bax to bcl-2 (P<0.05) was higher in the PMi than in other muscles. We concluded that the increase in cytosolic cytochrome c and available intracellular ATP should be responsible for the increase in caspase-9 activity; the activation of caspase-9 could be limited by the subsequent depletion of ATP; the postmortem release level of cytochrome c could be determined by the ratio of bax to bcl-2 in fresh tissues.     

Influence of production system in local and conventional pig breeds on stress indicators at slaughter,muscle and meat traits and pork eating quality

Sensory quality of pork is a complex phenotype determined by interactions between genetic and environmental factors. This study aimed at describing the respective influences of breed and production system on the development of pork quality. Plasma stress indicators and Longissimus muscle (LM) composition, physicochemical and sensory quality traits were determined in two contrasted breeds – the conventional Large White (LW, n=40) and the French local Basque (B, n=60). Pigs were reared in either a conventional (C; n=20 per breed), alternative (A; sawdust bedding and outdoor area, n=20 per breed) or extensive system (E; free-range, n=20 B). All the pigs from A and C systems were slaughtered at the same slaughterhouse, whereas B pigs from the E system were slaughtered at a local commercial abattoir. Major breed differences were found for almost all traits under study. LM from B pigs exhibited higher lipid, lower water and collagen concentrations, as well as lower collagen thermal solubility (P<0.001). Although plasma stress indicators at slaughter did not differ between breeds, except lower (P<0.05) lactate levels in B pigs, they exhibited higher LM pH1 and pHu values, and lower meat lightness, hue angle, water (drip, thawing and cooking) losses, glycolytic potential and shear force. Sensory analyses highlighted higher redness, marbling, tenderness, juiciness and flavour scores (P<0.01) of meat from B compared with LW pigs. Within both LW and B breeds, compared with C, the A system did not (P>0.05) influence plasma stress indicators, LM chemical composition and physicochemical or sensory traits of pork. In contrast, within the B pigs, the E system affected the meat quality more. Lower plasma cortisol levels (P<0.05), but higher plasma lactate, creatine kinase and lactate dehydrogenase activities, and more skin lesions (P<0.05), indicating higher muscular activity during pre-slaughter handling, were found in pigs produced in the E compared with the C system. E pigs exhibited higher meat pH1 and pHu values and shear force (P<0.01) and exhibited lower lightness, hue angle and drip and thawing losses (P<0.01) compared with the C pigs, whereas LM lipid, protein or collagen concentrations were not affected. Regarding sensory traits, the E system produced redder meat, but did not impact the eating quality of pork. Altogether, this study demonstrates that differences in meat quality between B and LW breeds can be modulated by extensive pig production system.     

Pig cognitive bias affects the conversion of muscle into meat by antioxidant and autophagy mechanisms

Slaughter is a crucial step in the meat production chain that could induce psychological stress on each animal, resulting in a physiological response that can differ among individuals. The aim of this study was to investigate the relationship between an animal’s emotional state, the subsequent psychological stress at slaughter and the cellular damage as an effect. In all, 36 entire male pigs were reared at an experimental farm and a cognitive bias test was used to classify them into positive bias (PB) or negative bias (NB) groups depending on their decision-making capabilities. Half of the animals, slaughtered in the same batch, were used for a complete study of biomarkers of stress, including brain neurotransmitters and some muscle biomarkers of oxidative stress. After slaughter, specific brain areas were excised and the levels of catecholamines (noradrenaline (NA) and dopamine (DA)) and indoleamines (5-hydroxyindoleacetic acid and serotonin (5HT)) were analyzed. In addition, muscle proteasome activity (20S), antioxidant defence (total antioxidant activity (TAA)), oxidative damage (lipid peroxidation (LPO)) and autophagy biomarkers (Beclin-1, microtubule-associated protein I light chain 3 (LC3-I) and LC3-II) were monitored during early postmortem maturation (0 to 24 h). Compared with PB animals, NB pigs were more susceptible to stress, showing higher 5HT levels (P<0.01) in the hippocampus and lower DA (P<0.001) in the pre-frontal cortex. Furthermore, NB pigs had more intense proteolytic processes and triggered primary muscle cell survival mechanisms immediately after slaughter (0 h postmortem), thus showing higher TAA (P<0.001) and earlier proteasome activity (P<0.001) and autophagy (Beclin-1, P<0.05; LC3-II/LC3-I, P<0.001) than PB pigs, in order to counteract the induced increase in oxidative stress, that was significantly higher in the muscle of NB pigs at 0 h postmortem (LPO, P<0.001). Our study is the first to demonstrate that pig’s cognitive bias influences the animal’s susceptibility to stress and has important effects on the postmortem muscle metabolism, particularly on the cell antioxidant defences and the autophagy onset. These results expand the current knowledge regarding biomarkers of animal welfare and highlight the potential use of biomarkers of the proteasome, the autophagy (Beclin-1, LC3-II/LC3-I ratio) and the muscle antioxidant defence (TAA, LPO) for detection of peri-slaughter stress.     

Intrauterine crowding impairs formation and growth of secondary myofibers in pigs

There are indications that intrauterine crowding may cause intrauterine growth retardation with the possibility of an impaired myofiber hyperplasia. The aim of the study was to confirm this by generating large differences in uterine space using sows that were unilaterally hysterectomized-ovariectomized (HO; crowded) or unilaterally oviduct ligated (OL; non-crowded). In the study, seven HO and seven OL Swiss Large White third parity sows were used. At farrowing, litter size and litter birth weight were determined. Subsequently, within each litter two male and two female progenies each with the respectively lowest (L) and highest (H) birth weight were sacrificed. Internal organs and brain were weighed, and longissimus (LM) and semitendinosus muscle (SM) samples were collected. Histological analyses were performed in both muscles using mATPase staining after preincubation at pH 4.3 and 10.2. Myosin heavy chain (MyHC) polymorphism was determined in the LM by means of SDS-PAGE. The number of piglets born alive was similar in both sow groups, but litter size expressed per uterine horn was lower (P < 0.05) in OL than HO sows. Consequently, OL progeny were markedly heavier (P < 0.01). Regardless of gender, the organs, the brain and the SM were heavier (P < 0.001) in OL and H compared with HO and L offspring, respectively. Compared with HO pigs, the SM of OL offspring tended (P < 0.1) to have more myofibers, which were of larger (P < 0.05) size. However, myofiber density appeared to be lower (P < 0.1) in the SM of OL than HO pigs. The impact of birth weight on myofiber characteristics was limited to the lower (P < 0.05) myofiber density in the SM and the larger (P < 0.01) myofiber size in the light portion of the SM of H than L offspring, whereas myofiber hyperplasia did not differ between birth weight categories. The SM, but not the LM, of male offspring had a greater (P < 0.05) myofiber density. This did not affect total SM myofiber number. The relative abundance of fetal and type I MyHC in the LM was lower (P < 0.05) and that of type II MyHC was greater (P < 0.001) in OL than HO pigs. The current data suggest that regardless of birth weight and gender, in the LM and SM of individuals born from a crowded environment, not only hyperplasia but also hypertrophy of myofibers is impaired and their maturity seems delayed.     

Growth,slaughter and intra-muscular collagen characteristics in Garganica kids

Growth performance, ossification of metacarpal growth plate, and intra-muscular collagen characteristics in three muscles of 10 intact male Garganica kids were studied. Each week, starting from the 47th day of age to slaughter (75th day of age), kids were weighed and blood samples taken for plasma non-protein hydroxyproline analysis. At slaughter, metacarpal, metatarsal bones and metacarpal growth plate thickness were measured. The semitendinosus (ST), semimembranosus (SM), and rectus medialis (RM) muscle samples were analysed for collagen concentration and hydroxylysyl pyridinoline (HP) cross-links. ST muscle had a collagen amount higher than SM and RM (P<0.01), and RM muscle had the highest (P<0.01) HP concentration. Tenderness index was the highest (P<0.01) in RM muscle. Animal growth rates slowed, starting from 61st day of age (P<0.01); this trend coincided with a marked reduction in concentration of plasma non-protein hydroxyproline (P<0.01). In addition, plasma non-protein hydroxyproline concentration was positively correlated (r=0.61, P=0.001) with growth rate of animals, suggesting that collagen turnover decreases during a slow growth phase. Based on the metacarpal growth plate width, the rate of bone ossification appears to be slower in goats than sheep.     

Prevalence and risk factors for gastric ulceration in pigs slaughtered at 170 kg

Oesophago-gastric ulcers (OGU) are a production and welfare problem in pigs. Stomach condition was scored for 22 551 pigs in 228 batches over a 7-month period at an abattoir in Italy processing heavy pigs for ham production. Mild or severe ulceration was observed in 20.7% of pigs, of which 13% had scar tissue. Variation between batches was high (0% to 36% prevalence of severe ulcers) and showed a significant effect of farm of origin (P<0.001). Overnight lairage increased the prevalence of mild ulcers (P<0.001), but not severe or scarred ulcers. Scarred ulcers increased in the hottest summer months. Prevalence of ulcers showed only few and weak correlations at batch level with pathologies of the pleura, lungs and liver, but a strong correlation with on-farm mortality of the batch. Analysis of farm risk factors for OGU was assessed by questionnaire with a response rate of 17% of farms. Risk factors retained in a multivariable model included a protective effect of anthelmintic treatment (risk ratio (RR)=5.1, P=0.03), increased risk in farms using Mycoplasma vaccination (RR=5.6, P=0.04) and a tendency for association with type of flooring (P=0.06). Univariable analyses also highlighted possible influences of other stress-inducing factors including lack of enrichment objects and mixing of pigs during fattening, suggesting that the role of on-farm stressors merits further investigation. It is concluded that abattoir screening of OGU in future programmes for the assessment of well-being on farm should encompass only severe lesions and scarring, and results be returned to farmers to facilitate improvement of production and welfare.     

Meat and fat quality of Krškopolje pigs reared in conventional and organic production systems

Data on production traits of the only Slovenian autochthonous pig breed, the Krškopolje pig, is very scarce. Krškopolje pigs are reared in conventional and organic production systems, which were compared in the present study. After weaning, 24 barrows were assigned within litter to either conventional (CON) or organic (ECO) rearing system. Group CON (n=12) was housed indoors in two pens (7.5 m²) with partly slatted floor. Group ECO (n=12) was held in a sty with sheltered area (concrete floor, bedded with straw, 16 m²) and outdoor paddock area (100 m²). The trial started when pigs had 68±8 kg BW and 157±6 days of age. Two diets were formulated with equivalent ingredients and composition. For ECO diet the ingredients used were ecological. Group ECO received a diet with 12.4 MJ metabolisable energy (ME) per kilogram and 12.9% CP and group CON a diet with 12.7 MJ ME/kg and 13.6% CP. Feed distribution was limited to 3.5 kg per pig daily. In line with the rules for organic production, ECO pigs were additionally given alfalfa hay ad libitum. After 73 days on trial, the pigs were slaughtered and carcass, meat and fat quality was evaluated. Meat quality traits (pH, colour, water holding capacity), fatty acid composition, lipid and protein oxidation, collagen content and solubility were analysed in longissimus lumborum (LL) muscle. Fatty acid composition, lipid oxidation, and vitamins A and E concentrations were determined in backfat. There were no significant differences in growth rate and carcass traits between ECO and CON pigs, however, ECO pigs tended (P<0.10) to have higher daily gain and lower dressing percentage, higher (P<0.001) pH 45 min and lower (P<0.01) pH 24 h postmortem, affecting (P<0.10) also water holding capacity and objective colour parameters (P<0.05) of LL muscle. There were no differences in intramuscular fat (IMF) content of LL muscle, however, IMF of ECO pigs had lower (P<0.05) proportion of saturated and higher (P<0.01) proportion of monounsaturated fatty acids accompanied by higher (P<0.001) values of thiobarbituric reactive substances (TBARS). In backfat, ECO pigs showed lower (P<0.05) vitamin E content, higher (P<0.001) TBARS, higher (P<0.01) degree of unsaturation (percentage of polyunsaturated fatty acids), and also higher (P<0.05) vitamin A concentration than CON pigs, which can be related to alfalfa hay supplementation of ECO pigs. In brief, organic rearing of Krškopolje pigs did not affect performances but had an effect on meat and fat quality.     

Tear staining in pigs: a potential tool for welfare assessment on commercial farms

Tear staining or chromodacryorrhea refers to a dark stain below the inner corner of the eye, caused by porphyrin-pigmented secretion from the Harderian gland. It has been shown to be a consistent indicator of stress in rats and to correlate with social stress and a barren environment in pigs. The current study was, to our knowledge, the first to test it on commercial pig farms as a potential welfare indicator. The study was carried out on three commercial farms in Finland, in connection to a larger study on the effects of different types of manipulable objects on tail and ear biting and other behavioural parameters. Farm A was a fattening farm, on which 768 growing-finishing pigs were studied in 73 pens. Farm B had a fattening unit, in which 656 growing-finishing pigs were studied in 44 pens, and a farrowing unit, in which 29 sows and their litters totalling 303 piglets were studied in 29 pens. Farm C was a piglet-producing farm, on which 167 breeder gilts were studied in 24 pens. Data collection included individual-level scoring of tear staining; scoring of tail and ear damage in the growing-finishing pigs and breeder gilts; a novel object test for the piglets; and a novel person test for the growing-finishing pigs on Farm B and the breeder gilts on Farm C. On Farm A, tear staining was found to correlate with tail damage scores (n=768, r_s=0.14, P<0.001) and ear damage scores (n=768, r_s=0.16, P<0.001). In the growing-finishing pigs on Farm B, tear staining of the left eye correlated with tail damage (n=656, r_s=0.12, P<0.01) and that of the right eye correlated with ear damage (n=656, r_s=0.10, P<0.01). On Farm A, tear-staining sores were lower in the treatment with three different types of manipulable objects as compared with controls (mean scores 3.3 and 3.9, respectively, n=31, Fs₂₉=4.2, P<0.05). In the suckling piglets on Farm B, tear staining correlated with the latency to approach a novel object (n=29, r_p=0.41, P<0.05). Although correlations with tail and ear damage were low, it was concluded that tear staining has promising potential as a new, additional welfare indicator for commercial pig farming. Further research is needed on the mechanisms of tear staining.     

Effects of resistant starch on behaviour,satiety-related hormones and metabolites in growing pigs

Resistant starch (RS) has been suggested to prolong satiety in adult pigs. The present study investigated RS-induced changes in behaviour, satiety-related hormones and metabolites in catheterized growing pigs to explore possible underlying mechanisms for RS-induced satiety. In a cross-over design with two 14-day periods, 10 pigs (initial BW: 58 kg) were assigned to two treatments comprising diets containing either 35% pregelatinized starch (PS) or 34% retrograded starch (RS). Diets were isoenergetic on gross energy. Pigs were fed at 2.8× maintenance. Postprandial plasma response of satiety-related hormones and metabolites was measured at the end of each period using frequent blood sampling. Faecal and urinary energy losses were measured at the end of each period. Behaviour was scored 24 h from video recordings using scan sampling. Energy digestibility and metabolizability were ~6% lower in RS compared with PS diet (P<0.001), and metabolizable energy (ME) intake was ~3% lower in RS-fed than in PS-fed pigs (P<0.001). RS-fed pigs showed less feeder-directed (P=0.001) and drinking (P=0.10) behaviours than PS-fed pigs throughout the day. Postprandial peripheral short-chain fatty acid (SCFA) levels were higher in RS-fed than in PS-fed pigs (P<0.001). Postprandial glucose and insulin responses were lower in RS-fed than in PS-fed pigs (P<0.001). Triglyceride levels were higher in RS-fed than in PS-fed pigs (P<0.01), and non-esterified fatty acid levels did not differ between diets (P=0.90). Glucagon-like peptide-1 (GLP-1) levels were lower in RS-fed than in PS-fed pigs (P<0.001), and peptide tyrosine tyrosine (PYY) levels did not differ between diets (P=0.90). Blood serotonin levels were lower (P<0.001), whereas monoamine oxidase activity (P<0.05) and tryptophan (P<0.01) levels were higher in RS-fed than in PS-fed pigs. Despite a lower ME intake, RS seemed to prolong satiety, based on behavioural observations. Possible underlying mechanisms for RS-induced satiety include increased 24 h plasma SCFA levels, and decreased postprandial glucose and insulin responses. GLP-1 and PYY seemed not to play a role in RS-induced satiety. Low blood serotonin levels in RS-fed pigs suggested a difference in intestinal serotonin release between treatments. Increased postprandial plasma triglyceride levels corresponded with increased SCFA levels, but it is unclear whether triglycerides may have signalled satiety in RS-fed pigs.     

The rearing system modulates biochemical and histological differences in loin and ham muscles between Basque and Large White pigs

Conventional pork production, based on highly selected breeds for growth efficiency and carcass leanness, is generally considered to decrease pork quality. In contrast, non-selected breeds produced in extensive systems are associated with high pork quality, which is generally attributed to higher intramuscular fat (IMF) content and less glycolytic muscle metabolism. The present study aimed to determine biochemical, histological and quality traits of loin and ham muscles of pigs from selected Large White (LW) and local French, non-selected Basque (B) breeds. Pigs were reared in a conventional indoor (C, slatted floor), alternative (A, indoor bedding and outdoor area) or extensive system (E, free range, B pigs only). A total of 100 castrated males were produced in 2 replicates, each containing 5 groups of 10 pigs based on breed and system: LWC, LWA, BC, BA and BE. The glycolytic longissimus muscle (LM) and semimembranosus muscle (SM), and the deep red (RSTM) and superficial white (WSTM) portions of semitendinosus muscle (STM) were studied at 145 kg BW. Overall, breed induced stronger effects on muscle traits than the rearing system, among which the E system induced greater changes. The lower muscle growth of B pigs was associated with fewer muscle fibers and a smaller cross-sectional area (CSA) of glycolytic fibers (P < 0.01). The SM was less glycolytic and more oxidative in B than in LW pigs (P < 0.001). The WSTM followed a similar trend, with a larger relative area of type I fibers in B pigs. In contrast, the LM and RSTM were more oxidative in LW pigs. B pigs had higher IMF content and ultimate pH in all muscles, along with lower glycolytic potential, less light and redder meat in the LM and SM (P < 0.001). Compared to the C system, the A system induced only a shift towards a more oxidative metabolism in the LM and a smaller fiber CSA in the RSTM of LW pigs (P < 0.05), without influencing pork quality traits. Compared to BC pigs, BE pigs had a more oxidative and less glycolytic muscle metabolism, along with higher ultimate pH, lower lightness and redder meat (P < 0.01), but similar IMF content. Overall, results indicate that influences of breed and rearing system on muscle properties depend on muscle type, and that IMF content and fiber-type composition are unrelated traits that can be modified independently by genetic or rearing factors.     

Hot-water spraying is a sensitive test for signs of life before dressing and scalding in pig abattoirs with carbon dioxide (CO2) stunning

This study investigated the benefits of hot-water spraying (HWS) as a diagnostic test to verify the absence of signs of life (SOL) before scalding in pigs slaughtered with carbon dioxide (CO₂) stunning. A total of 37 108 finishing pigs from five German abattoirs (A to E) operating at 55 to 571 pigs per hour were assessed. Suspended pigs were sprayed onto the muzzle, head and front legs (143 to 258 s post sticking for 4 to 10 s, 57°C to 72°C). Any active movements during HWS were rated as positive test outcomes. In comparison, SOL were considered to be absent if a subsequent manual examination was negative and no active movements were observed following HWS. The incidence of pigs with activity during hot-water spraying (PWA) was restricted to two abattoirs (B: 0.25%; D: 0.02%; A, C, E: 0.00%). PWA showed movements of facial muscles (88%), mouth opening (78%), righting reflex (63%), isolated leg movements (35%) and vocalization (4%). The manual examination was positive in 71% of PWA (corneal/dazzle reflex: 67%/53%, nasal septum pinch: 33%), whereas all inactive pigs tested negative (P<0.001). The sensitivity for HWS as a test for SOL was calculated as 100%, dropping to 75% when only obvious and strong movements were taken into account. The specificity was >99.9% in either case. Any positive manual findings as well as any respiratory activity were instantly terminated using a penetrating captive bolt. Active movements triggered by the shot were shown to be an indicator for SOL (P<0.001). Video analyses revealed that spontaneous movements (SM) following sticking were present in 100% of PWA as opposed to 3.1% in pigs without such activity (controls). Results for different categories of SM in PWA v. controls were as follows: 100% v. 2.6% for mouth opening, 16.0% v. 0.1% for righting reflex and 22.0% v. 0.9% for isolated leg movements (all P<0.001). First mouth opening after sticking was observed later in PWA (28±24 v. 10±7 s), but mouth openings were observed for a longer period of time (141±44 v. 27±25 s) (both P<0.001). PWA with shorter mouth-opening intervals showed higher movement intensities during HWS and more positive manual findings (P<0.05). We conclude that HWS is a promising test for SOL. SM and sustained mouth opening in particular are indicators for compromised animal welfare and affected pigs should be shot by captive bolt.     

Management factors affecting mortality,feed intake and feed conversion ratio of grow-finishing pigs

The aim of this study was to determine the effect of animal management and farm facilities on total feed intake (TFI), feed conversion ratio (FCR) and mortality rate (MORT) of grower-finishing pigs. In total, 310 batches from 244 grower-finishing farms, consisting of 454 855 Pietrain sired pigs in six Spanish pig companies were used. Data collection consisted of a survey on management practices (season of placement, split-sex by pens, number of pig origins, water source in the farm, initial or final BW) and facilities (floor, feeder, ventilation or number of animals placed) during 2008 and 2009. Results indicated that batches of pigs placed between January and March had higher TFI (P=0.006), FCR (P=0.005) and MORT (P=0.03) than those placed between July and September. Moreover, batches of pigs placed between April and June had lower MORT (P=0.003) than those placed between January and March. Batches which had split-sex pens had lower TFI (P=0.001) and better FCR (P<0.001) than those with mixed-sex in pens; pigs fed with a single-space feeder with incorporated drinker also had the lowest TFI (P<0.001) and best FCR (P<0.001) in comparison to single and multi-space feeders without a drinker. Pigs placed in pens with <50% slatted floors presented an improvement in FCR (P<0.05) than pens with 50% or more slatted floors. Batches filled with pigs from multiple origins had higher MORT (P<0.001) than those from a single origin. Pigs housed in barns that performed manual ventilation control presented higher MORT (P<0.001) in comparison to automatic ventilation. The regression analysis also indicated that pigs which entered to grower-finisher facilities with higher initial BW had lower MORT (P<0.05) and finally pigs which were sent to slaughterhouse with a higher final BW presented higher TFI (P<0.001). The variables selected for each dependent variable explained 61.9%, 24.8% and 20.4% of the total variability for TFI, FCR and MORT, respectively. This study indicates that farms can increase growth performance and reduce mortality by improving farm facilities and/or modifying management practices.     

Resistance to gastrointestinal parasite infection in Djallonké sheep

Gastrointestinal parasitism places serious constraints on small ruminant production. The situation has been exacerbated by development of drug resistance in many parasite populations, leading to interest in identification of animals with genetically mediated resistance or tolerance to nematode infections. This study assessed the response to natural infection with gastrointestinal nematodes (GIN) in Djallonké sheep during the rainy season in the Sudan-Guinea Savannah region of Burkina Faso. Haemonchus contortus is the most prevalent GIN at this site and time. Djallonké lambs (n=434) were sampled from 40 households and evaluated at a common location in southern Burkina Faso. Lambs were dewormed with levamisole at 2 to 6 months of age and returned to infected pastures. Fecal egg counts (FEC), packed cell volumes (PCV), and FAffa Malan CHArt (FAMACHA©) scores were determined 28 and 35 days after deworming. Lamb mortality was monitored throughout the experiment. Least-squares means for BW increased from 13.8±0.2 kg at 28 days to 14.0±0.2 kg at 35 days (P<0.01). Simple means and medians for FEC were 615 and 100, respectively, at 28 days and 850 and 175, respectively, at 35 days. The FEC exhibited strong right skewness. Following logarithmic transformation and back-transformation of resulting least-squares means to the original scale, FEC were higher (P<0.01) for males (208±27) than females (122±10). Least-squares means for PCV decreased (P<0.001) from 28 (36.3±0.5%) to 35 days (33.7±0.5%), and were higher (P<0.01) for females (36.0±0.4%) than males (33.9±0.7%). Correlations (r) between repeated measurements of BW, FEC, PCV and FAMACHA scores at 28 and 35 days were all positive (P<0.001). The correlation between FAMACHA scores and PCV was negative at 28 (r=−0.14) and 35 days (r=−0.18) (P<0.001). This study revealed that BW was an easily measured predictor of the ability of the lamb to resist infection with GIN and maintain PCV, and confirmed that FAMACHA scores are useful indicators of differences in FEC. Approximately 40% of female and 30% of male lambs did not show detectable levels of infection (i.e. FEC=0) under field conditions. The great variability that was observed in FEC and PCV suggests potential to use Djallonké sheep in breeding programs to enhance resistance to GIN.     

A novel feeding behavior index integrating several components of the feeding behavior of finishing pigs

Describing the feeding behavior of pigs is difficult given the large day-to-day variations observed for a given animal. The objectives of this study were to create an index that integrates the information from several components of feeding behavior in order to account for intra-animal variation within a day and between days, and to evaluate the capability of this index to study the impact of relevant nutritional factors affecting feeding behavior. Feed intake information from 160 pigs during the last 28 d of the growing phase from three studies was used. For each pig, the sum of the absolute values of the deviation areas between the regression line of the relative cumulative feed intake and the observed cumulative feed intake was used to calculate the weekly index measuring the irregularity of feed intake (IIFI). Spearman’s correlations of IIFI with the number of daily meals (r = −0.42; P < 0.001), meal duration (r = 0.38; P < 0.01), and feed intake per meal (r = 0.41; P < 0.01) indicate that pigs with high IIFI have fewer meals of longer duration and higher feed intake compared to pigs with low IIFI. This shows that IIFI captures information from several components of feeding behavior. Analysis of variance showed no effect of treatment on feeding behavior for datasets 1 and 2. However, the correlation between IIFI and dietary levels of CP (r = 0.34) indicates that diets high in CP were associated with pigs having more irregular meals. In dataset 3, pig feeding behavior was more regular in control diets than in pigs fed fibrous diets (IIFI; 164 vs. 197, respectively; P < 0.05). Additionally, IIFI was smaller in pigs fed canola by-product diets than in pigs fed wheat by-product diets, indicating that the source of fiber may also influence the feeding behavior of pigs. In most cases, IIFI was more effective at identifying differences in feeding behavior between dietary treatments than conventional feeding behavior variables. These results show the ability of IIFI to integrate information from several conventional components of the feeding behavior of pigs and its potential to successfully evaluate the effect of nutritional factors on feeding behavior.     

The influence of azaperone treatment at weaning on reproductive function in sows: ovarian activity and endocrine profiles during the weaning-to-ovulation interval

Azaperone can reduce stress caused by weaning and relocation of breeding sows, but its effects on reproductive processes are still poorly understood. The primary aim of this study was to describe and compare the endocrine and ovarian activity in ultrasonographically monitored second parity sows, with or without azaperone treatment at weaning (2 mg/kg BW i.m.). The intervals from weaning to the onset of estrus and ovulation were both greater (P<0.05) in azaperone-treated (n=12) than in control sows (n=12) by ~12 h. Mean daily growth rates of identified antral follicles were less (P<0.05) in azaperone-treated than in control sows (1.08±0.17 v.1.23±0.18 mm/day; mean±SD) and treated animals exceeded (P<0.05) controls in the mean ovulation rate (13.7±1.3 v. 12.6±1.2). A transient suppression of cortisol release was observed in the treatment group (at 10 and 30 min after azaperone injections) but circulating cortisol concentrations were variable in both groups of sows for the remainder of the study. The preovulatory rise in LH and estradiol secretion was delayed (P<0.05), and the duration of the LH surge was greater (P<0.001) in azaperone-treated sows compared with their control counterparts. The amplitude of episodic fluctuations in serum cortisol concentrations was correlated with the number of stillborn piglets in control sows (r=0.63, P=0.04). The amplitude and concentration of the preovulatory rise in estradiol secretion were negatively correlated with ovulatory response and litter size (r=−0.63 to −0.82, P<0.05), whereas the time at which the LH surge ended was directly related to the number of live-born piglets (r=0.82, P=0.002) in azaperone-treated animals. The present results indicate that administration of azaperone at weaning had a profound effect on preovulatory LH secretion as well as growth kinetics and estrogenicity of ovarian antral follicles. However, the causative associations among various characteristics of the preovulatory LH discharge, ovarian and adrenal steroid secretion post-weaning, and reproductive variables in sows remain equivocal.     

The effect of immunocastration and a diet based on granulated barley on growth performance and carcass,meat and fat quality in heavy gilts

A total of 48 Duroc×(Landrace×Large White) gilts of 33.2 kg BW were used to investigate the influence of immunocastration and diet on growth performance and carcass, meat and fat quality. Four treatments were arranged factorially (2×2) with two sexes (immunocastrated gilts: IG v. entire gilts: EG) and two dietary treatments (a commercial feedstuff as control v. granulated barley as a single major ingredient) provided during the finishing period (from 103 to 126 kg BW). There were four replicates of three pigs per treatment. At the end of the trial, the IGs grew faster (P<0.05) and ate more feed (P<0.05) than the EGs. Carcasses from the IGs had thicker backfat depth than those from the EGs (P<0.01) and carried out a lower percentage of rejected carcasses (P<0.05) at slaughterhouse owing to lack of fat. Meat from the IGs tended to have higher intramuscular fat (IMF) content in the Longissimus thoracis (LT) muscle than that from the EGs (P=0.09). In addition, immunocastration increased the total saturated fatty-acid proportion in subcutaneous fat and IMF (P<0.001) and decreased the total monounsaturated fatty acid (MUFA) and total polyunsaturated fatty acid (PUFA) percentages in subcutaneous backfat (P<0.05 and P<0.001, respectively) and in IMF (P<0.01 and P=0.06, respectively). The use of a diet based on granulated barley during the finishing period had no effect on growth performance but tended to increase IMF content in the LT muscle (P<0.06), and increased MUFA (P<0.05) and decreased PUFA (P<0.01) proportions in omental fat. It is concluded that immunocastration of gilts intended for dry-cured ham industry improved some aspects of growth performance and carcass and meat quality, whereas granulated barley had scarce effect on productive traits and fatty-acid profile but tended to improve IMF content.     

Is high prevalence of vitamin D deficiency a correlate for attention deficit hyperactivity disorder?

The aim of the study was to determine the association between vitamin D and attention deficit hyperactivity disorder (ADHD), and difference in the level of vitamin D in ADHD children and control. This a case–control study carried out in school health and primary health care clinics. A total of 1,331 children and adolescents who were diagnosed with ADHD based on clinical criteria and standardized questionnaires were enrolled in this study and were matched with 1,331 controls, aged 5–18 years old. Data on body mass index (BMI), clinical biochemistry variables including serum 25-hydroxyvitamin D were collected. The study found significant association between ADHD and vitamin D deficiency after adjusting for BMI and sex (adj. OR 1.54; 95 % CI 1.32–1.81; P < 0.001). Majority of the ADHD children were in the age group 5–10 years (40.7 %), followed by 11–13 years (38.4 %). The proportion of BMI <85th percentile was significantly over represented in ADHD group as compared to healthy control (87.8 vs. 83 %; P < 0.001, respectively), while on the other hand, BMI >95th percentile was over represented in the control than ADHD group (7.6 vs. 4.6 %; P < 0.001, respectively). Mean values of vitamin D (ng/mL) were significantly lower in ADHD children (16.6 ± 7.8) than in healthy children (23.5 ± 9.0) (P < 0.001). There was significant correlation between vitamin D deficiency and age (r = ?0.191, P = 0.001); calcium (r = 0.272, P = 0.001); phosphorous (r = 0.284, P = 0.001); magnesium (r = 0.292, P = 0.001); and BMI (r = 0.498, P = 0.001) in ADHD children. The vitamin D deficiency was higher in ADHD children compared to healthy children.     

Contrasting cellularity on fat deposition in the subcutaneous adipose tissue and longissimus lumborum muscle from lean and fat pigs under dietary protein reduction

The production of pork with high amounts of intramuscular fat (IMF) without an increase in subcutaneous fat is highly desirable for the pig industry and consumers. Herein, we question the impact of dietary protein reduction (18% v. 13%) on fat deposition in the subcutaneous adipose tissue (SAT) and longissimus lumborum (LL) muscle using genetically diverse pigs for body fatness (lean v. fat). A clear effect of genotype was observed on plasma insulin (P=0.004) and leptin (P<0.001), as well as on backfat thickness (P<0.001), with the fat pigs having higher values. Accordingly, IMF was higher in the fat pigs, when compared with their lean counterparts (P=0.003), which was supported by enlarged adipocytes (P<0.001). The area of lipid droplets within the LL fibres (P=0.039) and extramyocellular lipids number (P=0.017) were increased in pigs fed reduced protein diets, regardless of genotype, which is consistent with higher levels of plasma triacylglycerols (P=0.002). The gene-expression pattern of lipogenic factors in the SAT was distinct from the LL muscle. In the SAT, PPARG expression was similar among genotypes (P>0.05), whereas in the LL muscle it was higher in the lean pigs (P=0.023), especially when fed on low protein diet (P=0.057). The CEBPA and FABP4 mRNA levels were increased in the SAT of fat pigs (P<0.001), without changes in the LL muscle (P>0.05). The influence of diet on FABP4 expression in the SAT was dependent on pig’s genetic background (P=0.005). In conclusion, fat deposition was clearly influenced by genotype and, to a lesser extent, by dietary protein level, the SAT being more sensitive than the LL muscle. One can speculate that the pathways involved in lipid metabolism are downregulated in intramuscular adipocytes when compared with SAT fat cells. This result might be a direct consequence of the relatively low proportion of adipocytes found in the LL muscle.     

Circulating microparticles in patients with coronary heart disease and its correlation with interleukin-6 and C-reactive protein

Microparticles (MPs) are vesicles released from activated or apoptotic cells. MP derive from various cells, most notably platelets, but also leucocytes, lymphocytes, erythrocytes, and endothelial cells. The aim of this study was to investigate endothelial MP (EMP), platelet MP (PMP), lymphocyte MP and monocyte MP and TF-positive MPs (TF⁺ MPs) in patients with coronary heart disease (CHD), and to evaluate the correlation of these MPs with Interleukin-6 (IL-6) and C-reactive protein (CRP). Different cell-derived MPs and TF⁺ MPs were analyzed by flow cytometry in 40 patients with myocardial infarction (MI), 30 unstable angina (UA), 20 stable angina (SA) and 20 healthy individuals, and IL-6 and CRP were determined by ELISA and special protein analyzer, respectively. Compared with SA and control, EMP and PMP was significantly elevated in MI and UA (P < 0.001), and TF⁺ MPs was significantly elevated in MI and UA (P < 0.001). EMP and PMP correlated with IL-6 (r = 0.822, P < 0.001 and r = 0.567, P < 0.001; respectively) or CRP level (r = 0.597, P < 0.001 and r = 0.66, P < 0.001; respectively). Different cell-derived MPs in CHD may indicate the different pathophysiological changes in vessels, and MPs may both participate in the development of thrombosis and enhance the vascular inflammation.