Detection of potential markers for systemic disease in saliva of pigs by proteomics: A pilot study

Animals with different health status have been studied in order to extend the knowledge about protein composition of porcine saliva samples and to discover potential salivary markers for systemic disease in porcine production. Clinical examination of animals was performed at farm level where 10 healthy pigs and 10 animals with evident clinical signs of disease were randomly selected. Saliva and blood samples were obtained and afterwards animals were humanely sacrificed to perform a complete necropsy. Levels of two acute phase proteins, haptoglobin and C-reactive protein, were used to identify possible active infections of the animals. Moreover, serological analysis, to the main porcine infectious diseases in the area, was performed. Salivary proteins were separated by two-dimensional gel electrophoresis followed by mass spectrometry for the identification of specific proteins. A total of 58 spots out of 75 were successfully identified by MS, which correspond to 20 unique proteins. Two different approaches were used to perform a statistical comparison of saliva protein patterns from healthy and diseased animals using the relative spot volume (% spot volume/total volume of all spot in the gel, approach “A”) or taking also into account the total protein content of each saliva sample (μg of spot/mL of saliva, approach “B”). Both analyses showed three proteins in common that are differentially regulated between states. However, approach B was selected for biomarker searching since it gave an estimation of protein concentration and showed differential expression of proteins between both health states in a total of 10 proteins, which were up-regulated in disease. Mass spectrometric analysis identified those proteins as salivary lipocalin, lipocalin 1, double headed protease inhibitor protein, adenosine deaminase, haptoglobin, albumin fragments, S100-A8, S100-A9, S100-A12 and pancreatic alpha amylase. These proteins could be considered as potential salivary markers of disease.     

Changes in pig salivary cortisol in response to transport simulation, food and water deprivation, and mixing

Cortisol concentrations in the saliva of two groups (N = 5/group) of prepubertal pigs were measured by radioimmunoassay. Samples were collected in the home pen under normal husbandry conditions and after a 24 h period when food and water were withheld. The pigs were then transferred to a transport simulator, which was left stationary (control) or set in motion (experimental), and further samples were taken 1 h later before the animals returned to their pens. In the following week, the two groups of pigs were mixed and saliva was collected over a 3 h period. Samples were also taken 2 days later after the pigs had been injected with a maximally stimulating dose of adrenocorticotrophin (ACTH). Transport simulation in deprived animals, and mixing, produced salivary cortisol levels similar to those seen after ACTH. Food and water deprivation alone also increased cortisol secretion whereas transport simulation in non-deprived animals had no effect. These results indicate that salivary cortisol estimation offers a non-invasive means of measuring stress responses in unrestrained pigs.     

Physiological indicators of stress and meat and carcass characteristics in tail bitten slaughter pigs

Background

Tail biting is a common welfare problem in pig production and in addition to being a sign of underlying welfare problems, tail biting reduces welfare in itself. The aim of this study was to evaluate the effects of tail biting on different pre and post mortem indicators of stress in slaughter pigs and on carcass and meat characteristics. A total of 12 tail bitten (TB) and 13 control (C) pigs from a farm with a long-term tail biting problem were selected for salivary cortisol analyses before and after transport to the slaughterhouse. After stunning, samples were taken for the analysis of serum cortisol, blood lactate, intestinal heat shock protein 70 (HSP70), and meat quality characteristics. In addition, body temperature immediately after and muscle temperature 35 min after stunning were measured, as well as lean meat percentage and carcass weight.

Results

TB pigs showed a lower cortisol response to the transport-induced stress than C pigs and also had a lower serum cortisol concentration after stunning. HSP70 content in the small intestine was higher in the TB pigs than in C pigs. TB pigs had a considerably lower carcass weight therefore produced a lower total amount of lean meat per carcass than C pigs.

Conclusions

This study suggests that prolonged or repeated stress in the form of tail biting causes a blunted stress response, possibly a sign of hypocortisolism. In addition, it underlines the importance of reducing tail biting, both from an animal welfare and an economic point-of-view.     

Effects of Butafosfan on salivary cortisol and behavioral response to social stress in piglets

We assessed the efficacy of Butafosfan, a component of Catosal, in the metaphylactic treatment of stress in pigs. Four 6-week-old female littermates were taken from 12 litters. They were confronted with a pig from a different litter for 2 h. There were 24 pairs, each consisting of confronting two unfamiliar pigs in a new pen. This housing of unfamiliar pigs provides a good, but simple, model of the psychosocial stress that pigs experience when housed in large groups on pig farms. Immediately before being housed with an unfamiliar pig, 12 pairs of pigs were injected subcutaneously with Catosal at a dose equivalent to 20 mg Butafosfan per kg body weight; the other 12 pairs received the control solution containing all ingredients of Catosal except Butafosfan. The frequency and duration of aggressive behavior and the salivary cortisol response were measured during the first 2 h of the encounter. No adverse effects associated with Catosal were observed. Subcutaneous injection of Catosal reduced the stress-induced salivary cortisol response and the frequency of aggressive behavior evoked by the social stress of housing two unfamiliar pigs together.     

Slaughter value, meat quality, creatine kinase activity and cortisol levels in the blood serum of growing-finishing pigs slaughtered immediately after transport and after a rest period

The experimental materials comprised 44 hybrid [female (Polish Large White x Polish Landrace) x male Duroc] growing-finishing pigs. The animals were randomly divided into two groups: 24 pigs were slaughtered immediately after transport and 20 pigs were slaughtered after a 24-hour rest period in the lairage. The meat content of pork carcasses, carcass dressing percentage, the proximate chemical composition, physicochemical and sensory properties of meat and shear force values were determined. Serum creatine kinase activity and cortisol levels were determined in blood samples collected before transport and during carcass bleeding. Pigs slaughtered immediately after transport, compared with those slaughtered after a 24-hour rest period, were characterized by a higher meat content of the carcass and a higher carcass dressing percentage. Pre-slaughter handling had no effect on pork quality. The incidence of normal-quality meat, partially PSE (pale, soft, exudative) meat and PSE meat was similar in both groups. Chemical analysis showed that the content of dry matter, total protein, fat and minerals in meat was comparable in both groups. As regards the functional properties of the pork, samples from the carcasses of pigs that had rested before slaughter had a higher contribution of the red color component. Meat from pigs slaughtered immediately after transport had more desirable sensory properties. Pre-slaughter resting had a significant effect on those analyzed physiological parameters which were found to be good indicators of pre-slaughter stress. Serum creatine kinase activity and cortisol levels were higher in blood samples collected after transport (during carcass bleeding) than in samples collected before transport, pointing to a strong stress response of animals to pre-slaughter treatment. The decrease in serum cortisol levels in blood samples collected during bleeding from the carcasses of pigs slaughtered after a 24-hour rest period, compared with samples collected from animals slaughtered immediately after transport, suggests that rest before slaughter alleviated stress induced by pre-slaughter handling operations.     

Intestinal translocation of Streptococcus suis type 2 EF+ in pigs

Sepsis with subsequent multisystem organ failure after translocation of bacteria from the gut is a serious risk associated with stress situations. We showed that intestinal bacterial translocation could be one of the pathways for pathogenic Streptococcus suis infections in the pig. In 24 piglets weighing 10-14 kg, free of the extracellular factor (EF+) producing phenotype of S. suis serotype 2, a silicon canula was placed in the proximal jejunum to enable intestinal inoculation and bypassing the upper alimentary tract. The pigs were individually housed. After stress induction in 18 pigs by means of a truck drive in individual cages for 1h, pigs were inoculated through the intestinal canula either with S. suis type 2 EF+ or with growth medium only, and put back in their original housing. The six not transported pigs were also inoculated with the same strain. To prevent oral self-infection, faeces were collected in a bag that was glued around the anus. Clinical and behavioral symptoms were recorded for 72 h post inoculation, and then the animals were sacrificed for pathological and bacteriological examination. In three animals, the inoculation strain was re-isolated from mesenterial lymph nodes and typically affected organs. No S. suis type 2 EF+ was detected by specific polymerase chain reaction (PCR) in any of the tonsil-swabs and -homogenates. We concluded that infection of the organs had taken place after bacterial translocation out of the gut and that the intestinal tract can be a porte d'entree for S. suis type 2 EF+.     

Effect of prenatal stress on subsequent response to mixing stress and a lipopolysaccharide challenge in pigs

Sows subjected to prenatal stress have been found to produce offspring that have altered responses to stress. Our objective was to determine if exposing a sow to stress would alter the response of the offspring to lipopolysaccharide (LPS) at 2 mo of age or their response to mixing stress at 4 mo of age. Sow treatments consisted of intravenous injections of ACTH (1 IU/kg of BW), exposure to rough handling for a 10-min duration (rough), or no treatment (control) once per week from d 42 to 77 of gestation. At 2 mo of age, pigs from each treatment, 1 per litter (n = 21, 17, and 15 for the ACTH, rough, and control treatments, respectively), were challenged with 2 μg of LPS/kg of BW or saline, or served as a noninjected control. Their behavioral response to a human approach test and salivary cortisol were measured. At 4 mo of age, 1 pig from each treatment (n = 14, 14, and 15 for the ACTH, rough, and control treatments, respectively) was taken from its home pen and placed in a pen of unfamiliar pigs. At this time, a punch biopsy wound (6 × 6 mm) was created to measure the ability of the pig to heal the wound. At this same time, each pig received a 1-mL intramuscular injection of 20% ovine red blood cells (oRBC), and then a second injection of oRBC at 21 d postmixing. Blood samples were collected 3 times per week for 2 wk and then once a week for 4 more weeks. Blood samples were analyzed for cortisol, porcine corticosteroid-binding globulin, antibody response to oRBC, and nitric oxide production by macrophages. Behavior was recorded during the first 5 d after mixing. All pigs in the LPS challenge responded with characteristic sickness behavior; however, pigs in the rough treatment showed less sickness behavior than those in the other 2 treatments (P < 0.05). Maternal stress treatment did not affect (P < 0.43) salivary cortisol. Pigs from all treatments responded similarly to mixing stress with regard to cortisol, porcine corticosteroid-binding globulin, antibody titers, nitric oxide production, and hematology measures, and all pigs experienced the same amount of aggression in response to mixing. Without altering peripheral measures of stress responsivity, prenatal stress enhanced the ability of pigs to cope with a simulated immune challenge, which could prove to be an adaptation to challenging environments.     

Changes in Eye Temperature and Stress Assessment in Horses During Show Jumping Competitions

High stress levels in horses during sporting events can influence the animal’s performance in competitions as well as their welfare. This study investigated the use of infrared thermography (IRT) measurement as a rapid, noninvasive, and accurate method for stress assessment in horses during sport competitions and compared IRT with a commonly used physiological method used for stress assessment and salivary cortisol. IRT and salivary cortisol responses were measured 3 hours before the competition, immediately following the competition, and 3 hours after the competition, each day during the 3 days of a show jumping event. Salivary cortisol values ranged from −0.11 ± 4.51 ng/mL in 5-year-old animals to 1.10 ± 2.20 ng/mL in 4-year-old animals, whereas eye temperature values ranged from 35.84 ± 1.16°C in 6-year-old animals to 36.08 ± 1.11°C in 5-year-old animals. The results obtained suggested that IRT measurements constitute an effective method for detecting stress in horses subjected to the acute events of show jumping.     

Effect of ascorbic acid administration on erythrocyte osmotic fragility of pigs transported by road during the hot-dry season

The experiments were performed in order to determine the effect of 8-h road transportation of pigs on erythrocytes osmotic fragility during the hot-dry season, and the ameliorative role of ascorbic acid. Twenty-three adult pigs comprising of both sexes served as subjects for the study. Thirteen pigs administered ascorbic acid (AA) per os 30 min before transportation, at a dose rate of 100 mg/kg served as experimental animals, while ten pigs administered with distilled water per os served as control, and were transported for 8 h during the hot-dry season. EDTA blood samples collected a day before (pre-transportation), immediately after 8-h transportation and 7 days post-transportation were used to determine erythrocyte osmotic fragility. The ambient temperature (AT) and relative humidity (RH) measured within the vehicle ranged between 30.5 – 39.0°C and 40.0 – 71.0% respectively. These values were outside the thermoneutral zone for the pig, indicating that the season was thermally stressful. Results obtained showed a significant difference (p < 0.05) in percent haemolysis recorded at NaCl concentrations of 0.4% and 0.6% immediately after transportation in experimental pigs and at 0.5, 0.6, 0.8 and 0.9% NaCl concentrations in experimental pigs 7 days post-transportation. In conclusion, result from the present study indicated that 8-h road transportation during the hot-dry season could induce stress resulting in haemolysis of erythrocytes and AA administration ameliorated the stress.     

Preweaning housing effects on behavior and physiological measures in pigs during the suckling and fattening periods

The effect of the preweaning housing system on the stress response of pigs before weaning and during fattening was studied in 33 litters of domestic pigs. Three preweaning housing systems were compared: barren crate (standard farrowing crate without straw), enriched crate (20% larger crate, with straw), and as a control, a farrowing pen (pen, 60% larger than the barren crate, with straw). At 25 d of age, pigs were tested with an isolation test and 1 d later with a human approach test (HumanT). Pigs were weaned at 28 d of age. At 3 and 6 mo of age, pigs were tested with an isolation-human approach test. The latency and frequency of squeal calls and locomotor activity were analyzed for all 3 tests, whereas physical contact with the human was also analyzed for the HumanT and isolation-human approach test. At 6 mo of age, the pigs were transported to a slaughterhouse. One day before transport, immediately after transport, and 1 h after transport, saliva samples were taken for cortisol analysis. The pH of the LM was also measured 45 min after slaughter. Preweaning housing system affected (P < 0.05) the probability of squeal vocalizations, the latency of locomotion, and the duration of locomotion during the HumanT. Pigs from the enriched pens vocalized less, had a longer latency to move, and performed less overall locomotion than pigs from the barren crates. Preweaning housing system did not affect behavior of fattening pigs. Cortisol concentrations before and after transport were not affected by preweaning housing system. An interaction of cortisol concentrations and housing systems was observed between the control sample and the sample taken immediately after transport in pigs from the barren crates (P < 0.05) compared with pigs from the enriched housing systems. Meat from pigs reared in the barren crate tended to have lower pH (P < 0.10) and that of pigs reared in enriched crates had lower pH (P < 0.05) than meat of pigs reared in enriched pens. No differences were observed between pigs from barren or enriched crates. Our results suggest that enrichment of the preweaning environment through enlarged space, provision of straw, and free movement for the sow had a positive effect on the coping behavior of pigs before weaning and prevented an increase in salivary cortisol concentration immediately after transport and a decrease in meat pH 45 min postmortem at the age of 6 mo. Minimal enrichment of the commercial farrowing crate did not affect behavior and physiological measures in pigs before and after weaning.     

Salivary cortisol in pigs following adrenocorticotrophic hormone stimulation: comparison with plasma levels

Four prepubertal pigs were prepared with venous catheters and housed in metabolism cages. Plasma and saliva samples were taken at 15-min intervals over a 105-min period and analysed by radioimmunoassay for total (i.e. free and bound) cortisol content. Adrenocorticotrophic hormone (ACTH) was given i.v. at three different doses (0.5, 1.0 and 2.0 mg) after the second sample and the cortisol responses were compared with pretreatment values and levels observed after saline vehicle administration. Basal levels of salivary cortisol were approximately 10% of those in plasma. ACTH induced significant increases in plasma and salivary cortisol but in no case was a dose/response relationship detected. Plasma cortisol showed a maximum increase of approximately 230% whereas salivary cortisol increased only by about 130%, indicating that salivary cortisol is a less sensitive indicator of adrenal activity than plasma cortisol in this species. Estimation of salivary cortisol concentrations may offer practical advantages for the assessment of stress responses in intensively housed pigs.     

Salivary testosterone measurements in growing pigs: validation of an automated chemiluminescent immunoassay and its possible use as an acute stress marker

This study aimed to validate the use of an automated chemiluminescent immunoassay analyser for salivary testosterone measurements in growing pigs and study how circadian pattern during daytime and stress can influence its values. The test method had intra- and inter-assay coefficient of variation lower than 10%. The method showed good linearity and recovery, and detection limits were low enough to detect salivary testosterone levels. No significant differences were observed in testosterone concentrations at different sampling time, and age and gender did not influence circadian pattern. In addition, this assay was used to quantify testosterone in two models of acute stress and, in both cases, significant increases (P < 0.01) in salivary testosterone were detected. Therefore, the automated assay system tested for porcine testosterone determinations would be suitable for its use in saliva samples and, furthermore, salivary testosterone levels could be used as a possible marker of acute stress in pigs.     

Different stressors elicit different responses in the salivary biomarkers cortisol,haptoglobin, and chromogranin A in pigs

Most commonly, salivary cortisol is used in pig stress assessment, alternative salivary biomarkers are scarcely studied. Here, salivary cortisol and two alternative salivary biomarkers, haptoglobin and chromogranin A were measured in a pig stress study. Treatment pigs (n = 24) were exposed to mixing and feed deprivation, in two trials, and compared to untreated controls (n = 24). Haptoglobin differed for feed deprivation vs control. Other differences were only found within treatment. Treatment pigs had higher salivary cortisol concentrations on the mixing day (P < 0.05). Chromogranin A concentrations were increased on the day of refeeding (P < 0.05). Haptoglobin showed a similar pattern to chromogranin A. Overall correlations between the salivary biomarkers were positive. Cortisol and chromogranin A were moderately correlated (r = 0.49, P < 0.0001), correlations between other markers were weaker. The present results indicate that different types of stressors elicited different physiological stress responses in the pigs, and therefore including various salivary biomarkers in stress evaluation seems useful.     

Proteomic analysis of porcine saliva

Saliva contains a number of proteins that may be useful as biomarkers of health and disease and can be easily obtained from large numbers of animals in a non-invasive, stress-free way. The objective of this study was to explore the protein composition of porcine saliva from 10 specific pathogen free pigs using first one-dimensional SDS–PAGE and then two-dimensional electrophoresis and mass spectrometry. A reference proteome pattern for porcine saliva was established with the identification of 13 different, mainly saliva-specific, proteins. These reference data will facilitate the investigation of salivary proteins potentially altered in disease and could serve as novel diagnostic biomarkers.     

Dynamics of serum antibodies to and load of porcine circovirus type 2 (PCV2) in pigs in three finishing herds,affected or not by postweaning multisystemic wasting syndrome

Background

Despite that PMWS commonly affects pigs aged eight to sixteen weeks; most studies of PMWS have been conducted during the period before transfer to finishing herds. This study focused on PCV2 load and antibody dynamics in finishing herds with different PMWS status.

Methods

Sequentially collected blood samples from 40 pigs in each of two Swedish (A and B) and one Norwegian (C) finishing herds were analysed for serum PCV2-load and -antibodies and saliva cortisol. The two Swedish herds differed in PMWS status, despite receiving animals from the same sow pool (multi-site production). However, the PMWS-deemed herd (A) had previously also received pigs from the spot market. ResultsThe initial serum PCV2 load was similar in the two Swedish herds. In herd A, it peaked after two weeks in the finishing herd and a high number of the pigs had serum PCV2 levels above 10⁷ per ml. The antibody titres increased continually with exception for the pigs that developed PMWS, that had initially low and then declining antibody levels. Pigs in the healthy herd B also expressed high titres of antibodies to PCV2 on arrival but remained at that level throughout the study whereas the viral load steadily decreased. No PCV2 antibodies and only low amounts of PCV2 DNA were detected in serum collected during the first five weeks in the PMWS-free herd C. Thereafter a peak in serum PCV2 load accompanied by an antibody response was recorded. PCV2 from the two Swedish herds grouped into genotype PCV2b whereas the Norwegian isolate grouped into PCV2a. Cortisol levels were lower in herd C than in herds A and B.

Conclusions

The most obvious difference between the Swedish finishing herds and the Norwegian herd was the time of infection with PCV2 in relation to the time of allocation, as well as the genotype of PCV2. Clinical PMWS was preceded by low levels of serum antibodies and a high load of PCV2 but did not develop in all such animals. It is notable that herd A became affected by PMWS after errors in management routine, emphasising the importance of proper hygiene and general disease-preventing measures.     

水牛发情期唾液生殖激素和结晶与卵泡发育的关系

本研究旨在探明水牛唾液生殖激素、唾液结晶和卵泡发育变化的规律。应用ELISA试剂盒测定水牛唾液和血清中的生殖激素,制作唾液结晶,并分析生殖激素和唾液结晶与卵泡变化的关系。运用唾液结晶法、直肠检查法、唾液结晶+B超法、直肠检查+B超法4种方法在生产中进行验证。结果表明,水牛发情当天唾液雌激素（E₂）的水平为233.51 pg/mL,达到一个峰值,随后开始缓慢降低。水牛唾液中孕激素（P₄）水平在发情前2 d达到16.17 ng/mL,发情当天降到8.91 ng/mL。促卵泡素（FSH）在水牛发情前2 d逐渐下降,从37.91 ng/mL降到34.64 ng/mL,但在发情2 d后逐渐升高,达到61.20 ng/mL。促黄体素（LH）在发情前1 d逐渐下降,从5.15 ng/mL降至发情当天4.12 ng/mL,但在发情1 d后升至5.77 ng/mL。B超监测卵巢卵泡从发情前2 d迅速生长,到发情当天达到20 mm大小,直到破裂排卵卵泡的大小变化不显著。排卵后形成黄体,黄体期一直维持到下次发情前4 d左右,期间在发情后14~17 d的黄体最大,与水牛唾液中LH分泌峰一致。唾液结晶+B超鉴定方法判定的发情率最高（86.67%）,与其他几种方法比较差异显著（P<0.05）;妊娠诊断结果也表明,唾液结晶+B超鉴定方法判定的妊娠率（61.53%）显著高于直肠检查法、唾液结晶法和直肠检查+B超法3种方法鉴定的结果（P<0.05）。综上所述,卵巢上卵泡的发育与唾液中生殖激素的浓度显著相关,发情当天唾液结晶呈现典型的树枝状,唾液结晶+B超法的发情鉴定准确率最高。     

Effects of supplemental L-tryptophan on serotonin, cortisol, intestinal integrity, and behavior in weanling piglets

Stress occurs in intensive pig farming when piglets are weaned and mixed. In this study, we investigated whether this stress might be reduced with elevated dietary levels of Trp. The effects of supplemental dietary Trp (5 g/kg of feed, as-fed basis) were tested on the neuroendocrine system, intestinal integrity, behavior, and growth performance in nursery pigs, both before and after mixing. Mixing occurred 5 d after weaning and diet introduction. On d 4, 5, and 6, Trp-fed pigs vs. control pigs showed approximately a 2-fold elevation in plasma Trp concentrations (68 +/- 7 vs. 32 +/- 2 micromol/L; P < 0.001), a 38% increase in hypothalamic serotonin turnover as measured by 5-hydroxyindoleacetic acid:5-hydroxytryptamine (P < 0.001), and an 11 to 18% increase (P < 0.05) in the intestinal villus height:crypt depth. Before (d 4) and at (d 5) mixing, saliva but not plasma cortisol concentrations were reduced (P < 0.02) by approximately 2-fold in Trp-fed pigs vs. control pigs. Intestinal paracellular (horseradish peroxidase) and transcellular (fluorescein isothiocyanate) transport of macromolecules were not affected by dietary treatment, but mixing induced a 2-fold reduction (P < 0.05) in transcellular transport. Behavioral responses (lying and standing) at mixing were not affected by dietary treatment, except on d 10 after diet introduction when Trp supplementation induced more lying and less standing (P < 0.02). Average daily gain and ADFI were not different among dietary groups (P > 0.10). In conclusion, supplemental dietary Trp (5 g/kg) to piglets increased hypothalamic serotonergic activity, reduced the salivary cortisol response to mixing, improved intestinal morphology, and reduced physical activity 10 d after diet introduction. Consequently, diets containing high Trp levels improved neuroendocrine components of stress and increased gastrointestinal robustness but did not affect behavioral reactivity in nursery pigs during weaning and mixing.     

Porcine acute phase protein concentrations in different diseases in field conditions

Five acute phase proteins (APPs) [C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), pig-MAP and albumin] were measured in pigs with naturally occurring infections by porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus (ADV), porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae, and in animals with tail and ear bites, arthritis and other acute inflammatory processes. Healthy specific pathogen-free (SPF) pigs were used as controls. In PRRSV-infected pigs, all APPs with the exception of pig-MAP exhibited significant changes compared with controls. In animals affected with ADV only Hp presented changes of statistical significance, whereas pigs with PCV2 showed marked modifications in all APPs tested. Animals affected with Mycoplasmosis showed concentrations of all positive APPs significantly above levels obtained in SPF pigs, though albumin concentrations did not differ from controls. Finally, all APPs studied showed substantial changes in pigs with acute inflammation. The results indicated that an acute phase response was developed in the different diseases studied, this response being higher in animals with clinical signs and concurrent bacterial processes. Haptoglobin would be the APP that better reflects pathological states; however, to get more complete and valuable information it might be advisable to perform APPs profiles including another APP, such as CRP or SAA.     

Synthesis of heat stress proteins in lymphocytes from livestock

Cultured bovine, equine, ovine and chicken lymphocytes responded to heat stress by the increased synthesis of a specific set of proteins known as heat stress proteins (HSP). Proteins with molecular weights of 70 and 90 kDa were synthesized in all species. Additional proteins were found in bovine, ovine and chicken lymphocytes. A time course of induction showed an increased synthesis of some of these proteins with only 30 min of heat stress and of several proteins with 60 min of heat stress. A specific monoclonal antibody was used to identify HSP70 as one of the stress proteins in bovine lymphocytes. Cells from these animals are capable of responding to heat stress by the production of heat stress proteins. These may be of physiological importance.     

CO2-stunning of slaughter pigs: effects on EEG, catecholamines and clinical reflexes

Investigations were carried out on the response of slaughter pigs to CO2 stunning with two different gas concentrations (80 vol%, 90 vol%, 73 s) under practical conditions in a one gondola-dip-lift system. EEG measurements were performed and blood constituents such as catecholamines (adrenaline, nor-adrenaline) and lactate as well as clinical reactions (nasal septum and corneal reflex, heart beats) investigated. Special EEG-electrodes were adapted for the measurement on pigs and a mobile data logger was prepared for the use in the lairage of the slaughter house and in the stunning unit. The CO2 concentrations were measured continuously close to the head of the pigs when transported up and down in the stunning gondola. The results show that the technique is suited to monitor the effects of different CO2 gas concentrations on the EEG of the pigs under practical conditions. There is strong evidence that CO2 concentrations of 80 vol% applied over 70 s as required by law are not sufficient to stun pigs properly. A large part of the animals still showed typical reflexes when leaving the stunning pit. When an atmosphere of 90 vol% CO2 is applied, most animals are already dead before bleeding commences. This may create problems in respect to meat hygiene. The blood analysis revealed very high concentrations of catecholamines after stunning. The values for adrenaline and nor-adrenaline in the sticking blood rose by a factor of about 1000 as compared to the concentrations in blood samples taken in the lairage before stunning. It seems necessary to revise the current legislation on gas stunning and to look in greater detail in the effects of CO2 stunning on the welfare of slaughter pigs.