Sevoflurane inhibits equine myeloperoxidase release and activity in vitro

Objective To investigate the effects of the volatile anaesthetic sevoflurane on the release of total and active myeloperoxidase (MPO) by non‐stimulated and stimulated polymorphonuclear neutrophils (PMNs) in whole blood from healthy horses. Study design In vitro experimental study. Animals Adult healthy horses. Methods Samples of whole venous blood were collected and incubated in air or in air plus 2.3% or 4.6% sevoflurane for 1 hour. PMNs were stimulated with N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP), with a combination of cytochalasin B (CB) and fMLP or with phorbol myristate acetate (PMA). Total and active MPO contents released by PMNs in blood were measured by enzyme‐linked immunosorbent assay (ELISA) and specific immunological extraction followed by enzymatic detection (SIEFED) respectively. Additional experiments were performed to assess the effect of sevoflurane on the peroxidase and chlorination cycles of purified equine MPO using Amplex Red and 3’‐(p‐aminophenyl) fluorescein as fluorogenic substrates respectively. Results As compared with air alone, 1 hour exposure of whole blood to 4.6% sevoflurane in air significantly inhibited the release of total and active MPO by unstimulated and both fMLP‐ and CB + fMLP‐stimulated PMNs but not by PMA‐stimulated PMNs. Although 2.3% sevoflurane had no effect on total MPO release by unstimulated and stimulated PMNs, it significantly reduced the release of active MPO by unstimulated and fMLP‐stimulated PMNs. Additionally, sevoflurane reversibly inhibited the activity of MPO, especially the peroxidase cycle of the enzyme. Conclusions and clinical relevance Although our experimental study was not designed to assess the effects of sevoflurane in vivo, this inhibition of MPO release and activity may have relevance for anaesthetized horses and deserves further studies to examine the clinical importance of these findings.     

Modulating effects of acepromazine on the reactive oxygen species production by stimulated equine neutrophils

ObjectiveTo investigate the effect of acepromazine (ACP) on reactive oxygen species (ROS) production by stimulated equine neutrophils.Study designEx vivo biochemical experiments.AnimalsIsolated neutrophils from healthy untreated horses.MethodsNeutrophils were incubated with ACP at concentrations of 10^?4, 10^?5 or 10^?6 m and then stimulated with phorbol-myristate-acetate (PMA) before measurement of lucigenin-enhanced chemiluminescence (CL). In a second experiment neutrophils were incubated in the presence of α-keto-γ methylthiobutyric acid (KMB) and treated with ACP at concentrations of 10^?4, 10^?5 or 10^?6 m. Subsequent PMA stimulation lead to neutrophilic ROS production and decomposition of KMB to ethylene, which is measured by gas chromatography. Electron paramagnetic resonance-spin trapping (EPR) analysis was performed with PMA-stimulated neutrophils in the presence of ACP (10^?4, 10^?5 or 10^?6 m) directly added to the cell suspension. In the second experiment, the same concentrations of ACP were pre-incubated with neutrophils, then centrifuged to eliminate the excess of ACP and re-suspended in phosphate buffer before stimulation with PMA. In all experiments, the results of ACP-treated and ACP-untreated stimulated neutrophils were compared.ResultsOverall, results obtained with lucigenin-enhanced CL and KMB oxidation were in agreement with those seen in electron paramagnetic resonance spectroscopy. Acepromazine induced a dose-dependent inhibitory effect on neutrophilic ROS production. Electron paramagnetic resonance also showed, at high ACP concentration, the appearance of a cation radical derived from ACP. In contrast, electron paramagnetic resonance study performed with pre-incubated neutrophils showed an important dose-dependent inhibitory effect of ACP.ConclusionThe results indicate that ACP can neutralize O^˙?₂ or its by-products during the stimulation of neutrophils.Clinical relevanceThese findings may have a therapeutic relevance when phenothiazines are used in horses suffering from inflammatory diseases in which neutrophil activation and ROS production are implicated.     

Polyphenols from Silybum marianum inhibit in vitro the oxidant response of equine neutrophils and myeloperoxidase activity

A recent study showed that silymarin, a standardized extract of S. marianum might be used in the prevention of equine laminitis. We investigated the effects of quercetin and some compounds found in silymarin (silybin, taxifolin and dehydrosilybin) on reactive oxygen species (ROS) production and myeloperoxidase (MPO) release by stimulated equine neutrophils (PMNs) and on MPO activity. All compounds (tested between 100 nm and 100 μm ) inhibited superoxide anion production by stimulated PMNs in a dose‐dependent manner. Dehydrosilybin and quercetin inhibited superoxide production and MPO release from 10 μm . Classical MPO assay showed quercetin as the most potent inhibitor, followed by taxifolin, dehydrosilybin and silybin. SIEFED MPO assay highlighting the binding of tested compounds to MPO showed that only quercetin and taxifolin maintained an efficient inhibition above 90% at 10 μm . Altogether, our results showed a strong inhibition of PMN activation by planar compounds such as quercetin and dehydrosilybin and a strong inhibition of MPO activity by the smallest molecules, quercetin and taxifolin. In conclusion, the compounds from silymarin may be useful for modulating the oxidative response of PMNs, involved in the pathogenesis of laminitis, but further in vivo studies are needed.     

The use of multivariate analysis to characterize carcass and meat quality of goat kids protected by the PGI “Cabrito de Barroso”

Fifty five suckling kids from three genotypes and two sexes protected by the PGI “Cabrito de Barroso” European quality label were used in this experiment. Carcass quality was assessed using indices from carcass measurements, dressing percentages, refrigerated losses, higher priced joints proportion and tissue composition of the carcass. Meat pH, colour, total pigment, fat, dry matter, collagen determinations (total and soluble), cooking losses and shear force estimated in longissimus thoracis et lumborum (LTL) and gluteobiceps (GB) muscles were used to characterize meat quality. Principal component (PC) analysis was performed in order to examine carcass quality traits (n = 16) and meat quality (n = 16) traits. The five first principal components (PCs) explained about 86% of the total variability for carcass quality and 75% of the total variability for meat quality. Compactness indices of carcass and leg, carcass weight and subcutaneous fat were the most effective variables for the PC1, whereas the higher priced joints proportion, muscle proportions of the higher priced joints and of the carcass and the muscle and bone ratio were useful to define the PC2. The first PC of the meat quality parameters was characterized by colour traits (L, b, a, C, H and total pigment) whereas collagen determinations (total collagen and collagen solubility) defined the second PC. When the carcass quality data were projected on the plane defined by the first two PCs, two separate groups of points appeared, corresponding to the animals with slaughter live weight higher or lower than 10 kg. The distribution of the meat quality data on the plane defined by the first two PCs allowed the identification of two separate groups, corresponding to the muscles GB and LTL. The differences between genotypes tend to be small and related to slaughter live weight, which implies certain constancy in carcass and meat quality of the PGI “Cabrito de Barroso”.     

Investigations on the water intake of growing bulls

The voluntary water intake was evaluated in 62 growing Holstein bulls (mean body weight range = 193 to 550 kg) during a whole fattening period. The diet was composed of corn silage and concentrates. Water was offered for ad libitum consumption. A total of 17,772 measurements of water intake were recorded over 282 days. The average daily water consumption was 18 kg/animal (S.D. = 6.7 kg). Applying a multiple regression analysis to the data set yielded the following equation: voluntary water intake (kg/day) = − 3.85 + 0.507  average ambient temperature (°C) + 1.494  dry matter intake (kg/day) − 0.141  roughage part of the diet (%) + 0.248  dry matter content of roughage (%) + 0.014  body weight (kg). The incorporation of the variables body weight gain, relative humidity, maximum ambient temperature, Na intake, and K intake into the equation did not increase the accuracy of the prediction.     

Antioxidant activity of hyaluronic acid investigated by means of chemiluminescence of equine neutrophil bursts and electron paramagnetic resonance spectroscopy

Activated neutrophils (PMNs), the ROS/RNS released by PMNs and the derived inflammatory processes are involved in the pathogenesis and progression of human inflammatory airway diseases. Similar diseases are also present in horses which suffer from recurrent airway obstruction (RAO), exercise‐induced pulmonary haemorrhage (EIPH) and inflammatory airway diseases (IAD). Hyaluronic acid (HA) plays numerous roles in modulating inflammatory processes. The aim of this study was to examine whether a preparation of HA (MW 900 000 Da) interferes with ROS/RNS during the course of equine PMN respiratory bursts, and to establish the lowest concentration at which it still has antioxidant activity by means of luminol‐amplified chemiluminescence (LACL). Electron paramagnetic resonance (EPR) spectroscopy was also used to investigate the direct antiradical activity of HA. The hydroxyl radical was significantly scavenged in a concentration‐dependent manner at HA concentrations ranging from 2.5 to 0.16 mg/mL. Superoxide anion, Tempol radical and the ABTS^{? +} were significantly inhibited at concentrations ranging from 2.5 to 0.62 mg/mL. The LACL of stimulated equine neutrophils showed that HA induced a statistically significant concentration–effect reduction from 5 mg/mL to 1.25 mg/mL. These findings were confirmed also when l ‐Arg was added to investigate the inhibition of the resulting peroxynitrite anion. Our findings indicate that, in addition to the human use, HA can also be used to antagonize the oxidative stress generated by free radicals in horses peripheral blood mononuclear cells (PBMCs). In order to achieve therapeutic concentrations, a direct aerosol administration to horses with horse respiratory diseases can be considered, as this route of application is also recommended in human medicine.     

Calcium homeostasis and its relationship to superoxide production in blood and milk neutrophils of lactating goats

Polymorphonuclear neutrophils (PMN), which comprise over 70% of the somatic cells in goat milk, are a major cellular component of innate immunity in the goat mammary gland. However, the function of milk PMNs is modified after diapedesis compared to PMNs in blood. As many aspects of PMN activity depend directly on intracellular Ca²⁺ concentration ((Ca²⁺)_i), the present study aimed to determine the changes in Ca²⁺ homeostasis of milk PMNs from lactating goats compared to autologous blood PMNs, and to examine the significance of these variations to the immuno-competency of milk PMNs. The intracellular Ca²⁺ store of freshly prepared milk cells was estimated from the elevation of (Ca²⁺)_i after ionomycin treatment, which was found to be significantly less than blood PMNs. Replenishment of the intracellular Ca²⁺ store in milk cells after intracellular Ca²⁺ depletion by Bapta-AM followed by spiking with 2.5 mM Ca²⁺ for 20 min was also compared to that of blood PMNs, showing that after depletion/spiking the intracellular Ca²⁺ store in milk cells was much less than blood PMNs. The production of superoxide anion (O₂^?) in vitro in response to (Ca²⁺)_i-dependent or (Ca²⁺)_i-independent modulators was used to evaluate the relevance of altered Ca²⁺ homeostasis on the immuno-competency of milk cells compared to blood PMNs. The results indicated that milk cells produced similarly low levels of O₂^? as blood PMNs when treated with ionomycin. However, the amount of O₂^? produced by milk cells in response to phorbol 12-myristate 13-acetate (PMA) stimulation, although greater than ionomycin treatment, was significantly less than that of blood PMNs. The capacity for O₂^? production by both cell types in response to PMA reverted to the resting state with use of the protein kinase C (PKC) inhibitor, staurosporine. In conclusion, the current study demonstrated an irreversible shortage of intracellular Ca²⁺ in the milk PMNs of lactating goats compared to blood PMNs. It also showed that preliminary O₂^–production, primed by ionomycin treatment, remained unchanged in milk PMNs, despite the shortage in intracellular Ca²⁺, but decreased O₂^? production capacity, mediated via the PKC pathway, in milk PMN. It is suggested that the defects in Ca²⁺ homeostasis in milk PMNs of lactating goats is partially attributable for the post-diapedesis functionality modifications.     

Optimization of conditions for in vitro production of radical oxygen species and expression of tissue factor by canine mononuclear cells and granulocytes for use in high-throughput assays

The purpose of this study was to optimize conditions for high throughput measurement of radical oxygen species (ROS) production and expression of tissue factor, also termed procoagulant activity, by canine leukocytes. Granulocytes and mononuclear cells were separated by density gradient centrifugation from peripheral blood collected on several occasions from three healthy large breed dogs. To determine optimal conditions for ROS production, granulocytes were incubated for 1 or 3h in PBG (PBS containing 0.5% BSA and 5mM glucose) or RPMI containing 10% fetal bovine serum (FBS); lipopolysaccharide (LPS), zymosan, peptidoglycan (PGN) and phorbol myristate acetate (PMA) were used as stimuli. ROS was assessed by conversion of the nonfluorescent dye dihydrorhodamine 123 to fluorescent rhodamine 123 by radical species released into the media. To identify optimal conditions for expression of tissue factor, mononuclear cells were incubated for 5h in RPMI containing different concentrations of heat-inactivated FBS (HI-FBS), and LPS, zymosan, PGN or PMA as stimuli. Expression of tissue factor was determined using a one-stage recalcification assay performed in an automated nephelometric coagulation analyzer. Neither LPS nor zymosan increased ROS production by granulocytes incubated in PBG media. In contrast, granulocytes incubated in RPMI had dose-dependent increases in ROS production in response to zymosan and PGN. ROS production was significantly increased by incubation with concentrations of LPS of 0.01microg/ml or greater, and by zymosan concentrations of 0.1microg/ml or greater. ROS production in response to incubation with PMA was significantly increased starting at 10(-7)M, and was significantly greater for cells incubated in RPMI than cells incubated in PBG. LPS-, zymosan- and PGN-stimulated procoagulant activity increased in a dose-dependent manner, whereas PMA-stimulated procoagulant activity peaked at 10(-7)M. Increasing concentrations of HI-FBS significantly increased LPS-, zymosan- and PGN-induced procoagulant activity of mononuclear cells. Results obtained in this study indicate production of ROS by canine granulocytes is optimal when these cells are incubated for 3h in RPMI with LPS (0.1microg/ml), zymosan (10 microg/ml), PGN (10 microg/ml), and PMA (10(-7)M). Furthermore, canine mononuclear cells express procoagulant activity in response to LPS, zymosan, PGN, and PMA, and responses to LPS, zymosan and PGN are enhanced by the addition of HI-FBS. These findings suggest that HI-FBS retains important serum proteins that facilitate interactions between each of these bacterial or yeast derived products and the mononuclear cells. Consequently, future studies regarding the regulation of procoagulant activity by canine mononuclear cells should be performed in the presence of HI-FBS. Both assays utilized in this study allow high throughput of samples, and therefore are appropriate choices for rapid screening of conditions and/or therapeutic interventions affecting the canine inflammatory system.     

A comparative study on certain enzymes of the granulocyte from different ruminant species

In the present study the level of enzyme hydrolases (alkaline phosphatase, myeloperoxidase, elastase, arginase, lysozyme and β-galactosidase) of polymorphonuclear cell (PMN) granules in different ruminant species and their release in response to activation was studied. Buffalo PMN alkaline phosphatase activity was higher (P < 0.01) than in PMNs of cattle and goats. Interestingly, myeloperoxidase was higher in cattle PMNs and least in goat PMNs (P < 0.01), a similar pattern was observed in the distribution of enzyme arginase. As far as lysozyme is concerned, its activity was significantly higher (P < 0.01) in PMNs of buffaloes than in the case of cattle and goat PMNs. On activation, these cells released MPO and elastase, in all the species studied, while lysozyme was secreted only in buffalo PMN cells. Activity of certain enzymes related to oxidant defence systems such as glutathione peroxidase and glutathione reductase were higher in cattle and goats compared to that in buffaloes. These observations are likely to have bearing on immunodefense roles played by PMNs and reflected differences among the ruminant species studied.     

Differential effects of alpha1-acid glycoprotein on bovine neutrophil respiratory burst activity and IL-8 production

During bacterial-mediated diseases, neutrophils (PMNs) play a critical role in defending the host against invading pathogens. PMN production of reactive oxygen species (ROS) contributes to the bactericidal capabilities of these cells. ROS are produced intracellularly and can be released extracellularly. The aberrant extracellular release of ROS, however, has been reported to induce injury to host tissues during mastitis and other inflammatory-mediated diseases of cattle. The acute phase response, which occurs shortly after infection or tissue injury, is characterized by the induction of a large number of plasma proteins referred to as acute phase proteins (APP). alpha1-Acid glycoprotein (AGP) is an APP that increases in response to infection or injury in cattle and humans. The precise function of AGP is unknown, but it has been reported to possess anti-inflammatory properties. The objective of this study was to evaluate the effects of bovine AGP on PMN pro-inflammatory responses, including respiratory burst activity and cytokine production. Bovine AGP dose-dependently inhibited zymosan-induced PMN extracellular release of superoxide anion and hydrogen peroxide without affecting the capacity of PMN to engulf and kill Staphylococcus aureus. Moreover, AGP exerted its effect on ROS production regardless of whether PMNs were exposed to AGP prior to or after activation. In contrast to respiratory burst activity, AGP enhanced PMN production of IL-8. The precise mechanism by which AGP regulates PMN functions remains unknown, but data presented in this study suggest that AGP may have a complex role by differentially regulating PMN pro-inflammatory activities.     

Crossbreeding parameters of general immune response traits in White Leghorn chickens

Crossbreeding parameters of immune response traits were estimated from a set of well characterized crossbred populations derived from three chicken lines selected over 12 generations for three different general immune response traits and their F1, F2 and backcrosses. The three traits investigated were the selection criteria from each of the lines, i.e. antibody response to the Newcastle disease virus vaccine 3 weeks after vaccination (ND3), cell-mediated immune response (response to phytohemagglutinin, PHA) and phagocytic activity measured as carbon clearance (CC). Crossbreeding parameters included direct and maternal additive line effects, direct and maternal heterosis as well as direct epistatic recombination loss. They were estimated as linear combinations of genetic group effects estimated using animal model methodology. Significant line differences were obtained for ND3 and, to a lesser extent, CC. They were mainly due to direct effects, maternal effects being significant for none of the 3 traits. Significantly negative direct heterosis effects were also observed for ND3 and CC, but not for PHA. Maternal heterosis effects were not estimated for CC. They were non significant for PHA, and negative and significant (− 0.78 ± 0.24) for ND3. The significant favourable recombination gain estimated for ND3 (3.21 ± 0.88) indicates that epistatic interactions could be important for this trait.The present work shows that it was worthwhile to complete second generation crosses to be able to assess to what extent immunity gained by selection is maintained in advanced crossbred generations, and to compare the transmission of immune traits implicated in different aspects of immunity.     

In vitro modulatory effect of ω-3 polyunsaturated fatty acid (EPA and DHA) on phagocytosis and ROS production of goat neutrophils

An in vitro study was carried out to examine the influence of two fish-oil-derived long chain ω-3 polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on goat polymorphonuclear leukocytes (PMN). Twelve Saanen healthy goats were used as blood donors. Neutrophils were isolated from blood and incubated with increasing concentration of EPA and DHA (25, 50, 100, 200 μM). Control samples were incubated in the absence of ω-3 PUFAs. Phagocytosis was evaluated by fluorescein-labeled Escherichia coli incorporation, while extracellular Reactive Oxygen Species (ROS) production was determined by cytochrome c reduction assay, which was selected among the others due to its specificity for extracellular superoxide anion release. Phagocytic activity was significantly increased by EPA (P < 0.05) and DHA (P < 0.01). Treating PMN with EPA does not affect extracellular ROS production which is, on the contrary, down-regulated by DHA. This effect was increased in experimental conditions which mimic pro-inflammatory challenges (stimulation with PMA).This study demonstrates that EPA and DHA may have beneficial effect on neutrophil function by increasing their phagocytosis activity and, in the meanwhile, decreasing the tissue damages due to extracellular release of ROS.     

The Effect of Activation of Granulocytes on Enzyme Release and Hydrogen Peroxide and Superoxide Production in Buffaloes

Singh, V.K., More, T. and Singh, S., 1997. The effect of activation of granulocytes on enzyme release and hydrogen peroxide and superoxide production in buffaloes. Veterinary Research Communications, 21 (4), 241-247Polymorphonuclear cells kill microorganisms by the stock of antibiotic proteins and peptides stored in their lysosomal granules and have the ability to produce reactive oxygen intermediates (ROI) such as H₂O₂, O ₂ ^– , and HOCl. Since the components involved in the microbicidal functions of buffalo (Bos bubalis) polymorphonuclear cells (PMN) have not been characterized, an assessment was made of the levels of various enzymes, the extent of extracellular release of these enzymes, and also their ability to produce H₂O₂/O ₂ ^– upon activation with opsonized zymosan (OZ) or lipopolysaccharide (LPS). Using GPC-HPLC, OZ was shown to be a more potent secretagogue than LPS, causing a significantly greater release of low-molecular-weight components. Varying levels of the enzymes (myeloperoxidase, lactate dehydrogenase, acid and alkaline phosphatases, -galactosidase, -D-glucuronidase, elastase and lysozyme) were recorded in the buffalo PMN and both the activators (OZ and LPS) caused significant release of all the enzymes except alkaline phosphatase. Both the activators also caused a significant increase in H₂O₂/O ₂ ^– production by the PMN. However, OZ caused a more pronounced activation than LPS. The studies revealed the presence of oxygen-dependent and oxygen-independent microbicidal systems with buffalo PMN, which responded more effectively to zymosan activation.     

Simulation of foot-and-mouth disease spread within an integrated livestock system in Texas, USA

We used a simulation study to assess the impact of an incursion of foot-and-mouth disease (FMD) virus on the livestock industries in an 8-county area of the Panhandle region of Texas, USA. The study was conducted in a high-density livestock area, with an estimated number of cattle on-feed of approximately 1.8 million. We modified an existing stochastic, spatial simulation model to simulate 64 scenarios for planning and decision-making. Our scenarios simulated four different herd types for the index herd (company feedlot, backgrounder feedlot, large beef, backyard) and variations in three mitigation strategies (time-of-detection, vaccine availability, and surveillance during disease control). Under our assumptions about availability of resources to manage an outbreak, median epidemic lengths in the scenarios with commercial feedlot, backgrounder feedlot, large beef and backyard index herd types ranged from 28 to 52, 19 to 39, 18 to 32, and 18 to 36 days, respectively, and the average number of herds depopulated ranged from 4 to 101, 2 to 29, 1 to 15 and 1 to 18, respectively. Early detection of FMD in the index herd had the largest impact on reducing (13–21 days) the length of epidemics and the number of herds (5–34) depopulated. Although most predicted epidemics lasted only 1–2 months, and <100 herds needed to be depopulated, large outbreaks lasting 8–9 months with up to 230 herds depopulated might occur.     

Characterization of certain inflammatory variables in the peripheral blood of clinically healthy dogs

Many laboratory techniques have been developed to study and quantify the inflammatory response, including the release of acid hydrolase enzymes, leukotriene B(4) (LTB(4)) production, reactive oxygen species (ROS) production and complement conversion studies. Although extensively studied in human health and disease, the relevance of such tests in the dog is largely unknown. After isolation of the peripheral blood mononuclear cell (PBMC) and polymorphonuclear cell (PMN) fractions from the peripheral blood of 38 clinically healthy dogs, values for ROS production were similar for both cell fractions when measured by luminol-enhanced chemiluminescence (17,853+/-9,695 U/10(6) cells versus 19,138+/-14,569 U/10(6) cells for the PBMC (n=38) and PMN (n=18) fractions, respectively). However, the mean time taken to reach maximum chemiluminescence was noticeably shorter in the PBMC fraction (5.1+/-3.3 versus 10.7+/-2.5 min for PBMCs (n=36) and PMNs (n=18), respectively). Intracellular concentrations of beta-glucuronidase, beta-galactosidase and N-acetyl-beta-glucosaminidase were assayed by spectrofluorometry. Mean values for all three enzymes were higher in PBMCs (n=31-35) than in PMNs (n=10-14). Both cell fractions released 20% of the intracellular enzyme concentration when stimulated with opsonized zymosan. Following incubation with A23187 (1 microM), mean LTB(4) production was higher in PBMCs (4.45+/-2.92 ng/10(6) cells; n=27) than in PMNs (0.96+/-2.22 ng/10(6) cells; n=13) using a validated high performance liquid chromatography (HPLC) assay. Immunoprecipitation studies revealed that the mean percentage conversion of C3 to C3b following stimulation with opsonized zymosan was 57.3+/-13.4% (n=36). The results provide normal values for clinically healthy dogs that may subsequently be used in future studies investigating dogs with various inflammatory disorders.     

Interactions between the naked neck gene, sex, and fluctuating ambient temperature on heat tolerance, growth, body composition, meat quality, and sensory analysis of slow growing meat-type broilers

Influence of the Naked Neck gene, NA, on heat tolerance was evaluated in slow growing meat-type chickens in interaction with sex. Standard male Ross broilers were used as rapid growth controls. Fluctuating temperature was used to simulate day–night variations, i.e. 17 °C to 23 °C in normal and 27 °C to 33 °C in hot conditions. Male and female chickens were weighed twice and once a week, respectively, and Gompertz function was fitted to our data to calculate theoretical age at 1 kg (A1K) and 2 kg (A2K). Carcass, abdominal fat, breast and leg yields were measured (CY, AFY, BRY, and LY). Meat quality was evaluated 24 h post-mortem with pH and colour (L, a, b) of the breast, and 72 h post-mortem with breast meat drip loss (DL). Rectal temperature and its variation were measured at the maximum and minimum ambient temperature at 1 and 2 kg (RT2_min, RT2_max, and ΔBT_2 kg). Hematocrit (HCT, ΔHCT) were measured at the same stages. Organoleptic characteristics of breast and leg muscles were studied for females from both ambient temperatures.Significant genotype × sex × temperature interaction was observed for A2K, AFY, RT2_max, ΔBT_2 kg, and ΔHTC_2 kg. Hot condition did not affect ΔBT_2 kg and A2K in homozygous NA birds; ΔBT_2 kg was markedly increased in all other genotypes for males but not for females. Significant genotype × environment interactions were found only for A2K and CY. Sex × temperature interactions were found for all traits except for A1K, b, pH, and ΔHCT_2 kg. In both conditions, males reached 2 kg at the same age (69 d) while females reached this weight 11 d latter in hot than in normal condition. Heat decreased CY in males (− 1.0%) and increased it in females (+ 1.4%). Meat was paler in males and darker in females in the hot condition leading to a difference in meat brightness between males and females only in the hot condition. Concerning sensory analysis, genotype × temperature interaction was significant for meat consistence, both in leg and breast muscles.Effects of the NA gene on susceptibility to heat stress were smaller in slow growing animals than in broilers. However, heat tolerance was still improved in homozygous NA slow growing birds, as shown by the limited change in diurnal variation of body temperature. Furthermore, the NA gene improved breast meat percentage.In contrast to broilers, where females should be recommended for production in hot climates, the present study would rather suggest that naked neck males from slow-growing meat-type ‘label’ chicken lines should be preferred.     

Effects of transportation during the hot season and low voltage electrical stimulation on histochemical and meat quality characteristics of sheep longissimus muscle

The effect of transportation during the hot season (42 °C) and low voltage electrical stimulation on physiological, histochemical and meat quality characteristics of Omani sheep was studied. Forty intact male sheep were divided into two equal groups: 3 h transported or non-transported. The non-transported group remained in holding pens for 48 h prior to slaughter, while the transported group was transported 300 km (approximately 3 h) in an open truck under solar radiation on the day of slaughter. Blood samples were collected from the animals before loading and prior to slaughter. Fifty percent of the carcasses from each group were randomly assigned to low voltage (90 V) at 20 min postmortem. Temperature and pH decline of the left longissimus thoracis muscle were monitored. Ultimate pH, WB-shear force, sarcomere length, myofibrillar fragmentation index (MFI), expressed juice, cooking loss and color L, a, b were measured on samples from both sides muscles collected at 24 h postmortem at 3–4 °C. The transported sheep had significantly higher plasma cortisol (P < 0.01), adrenaline, nor-adrenaline and dopamine concentrations (P < 0.05) than non-stimulated animals. Electrical stimulation resulted in a significantly (P < 0.05) more rapid pH fall in muscle during the first 12 h after slaughter. Muscles from electrically-stimulated carcasses had significantly (P < 0.05) lower pH values, longer sarcomere length, lower shear force value, higher expressed juice, MFI and lighter L than those from non-stimulated ones. The muscle samples from the transported sheep had significantly (P < 0.05) smaller and lower proportion of Types I and IIA fibers than those from the non-transported group. This study indicated that pre-slaughter transport at high ambient temperatures can cause noticeable changes in physiological and muscle metabolism responses in sheep. Electrical stimulation improved meat quality characteristics, which indicate that meat quality of transported sheep can be improved by electrical stimulation.     

Prevalence and severity of foot lesions in Danish Holstein heifers through first lactation

Several studies have shown that foot lesions and clinical lameness occur before first calving and develop further during the lactation period. Lameness may cause production losses, but the relationship between foot lesions, particularly in the claw horn, and lameness in heifers is unclear. The objectives of this study were to describe the development of and evaluate the relationship between lameness and foot lesions in Danish Holsteins before and after first calving. In a longitudinal study, 147 heifers were examined for lameness and foot lesions 2–5 times over an 18-month period. Lameness was assessed by means of a visual locomotion score and foot lesion severity was recorded.The prevalence of a locomotion score 3 was 25% before calving, and 90% at approximately 250 days in milk (DIM). Prevalence of moderate to severe sole haemorrhage (SH) was 27% before calving and 56% at 250 DIM, and that of moderate to severe white line lesion (WLL) 44% before calving with a peak of 70% at 200 DIM. There was one case of white line abscess but SH was seen throughout the entire study period. Digital dermatitis (DD) was prevalent prior to first calving (15%) and peaked at 39% at 0–100 DIM. Heel horn erosion (HHE) occurred in almost all cows (93–100%) and was strongly correlated with DD (r = 0.51). The correlation coefficient between SH and WLL was also high (0.42). The relatively high correlations between WLL and both DD and HHE were more surprising (0.38 and 0.35, respectively), those between SH and both DD and HHE were moderate (around 0.18). Interdigital dermatitis was significantly correlated with both HHE and DD, but completely unrelated to SH and WLL.The overall average locomotion score increased by about one-half of a score unit from 1 month prior to calving until 250 DIM, with a large difference between herds, although this was unsurprising as cows may alter their locomotion pattern with management factors (e.g. floor properties). DD and WLL were both associated with a locomotion score 3 but of the cows with severe WLL there was no clear association between a locomotion score 3 and DD. The highest locomotion scores occurred among cows with DD but without WLL.     

Effect of castration on productive performance, carcass characteristics and meat quality of Iberian pig females reared under intensive management systems

Sixty crossbred (Iberian dam × Duroc sire) females, 80 days of age (17.6 ± 0.13 kg body weight, BW), was used to investigate the effect of castration on productive performance, carcass and meat quality and fatty acid profile of backfat (BF). There were 2 treatments (intact females, IF; castrated females, CF) and 5 replicates of 6 pigs per treatment. Pigs were reared indoor under an intensive production system, ovariectomized at 92 days of age (26.1 ± 0.19 kg BW) and slaughtered at 267 days of age (143.6 ± 6.49 kg BW). Meat samples were taken at longissimus dorsi muscle at the level of the last rib and BF samples were taken at the tail insertion. For the entire experiment (18 to 144 kg BW), IF ate less feed and were more efficient than CF (P < 0.05). Also, IF had less carcass yield (P < 0.01) and fat thickness at the gluteus medius muscle (P < 0.05) and tended to have lower backfat depth (P < 0.10) than CF. However, IF had higher shoulder yield at 2 and at 24 h post mortem (P < 0.05) and after trimmed (P < 0.10) than CF. The pH₂₄ of the semimembranosus muscle tended to be lower for IF than for CF. Also, IF had more moisture (710 vs. 691 g/kg) and less fat (66.4 vs. 91.2 g/kg) in the longissimus dorsi muscle than CF (P < 0.05). Meat from IF was more lightness (higher L value; P < 0.01), redder (higher a value; P < 0.001) and had more intensive color (higher c value; P < 0.001) than meat from CF. Backfat was more saturated in CF than in IF (P < 0.05), mostly because of the higher palmitic acid (P < 0.05) and the lower linolenic acid (P < 0.05) content. We conclude that intact females have better productive performance and shoulder yield but less carcass yield than castrated females and that castration does not improve meat quality. Therefore, when animal welfare, cost of castration, productive performance and carcass and meat quality traits are considered, the use of intact females rather than castrated females is recommended for the production of Iberian pigs reared under intensive management systems.