Changes in fibre type composition of gluteus medius and semitendinosus muscles of Dutch Warmblood foals and the effect of exercise during the first year postpartum

In order to obtain broader insights into the equine musculoskeletal system, we studied the fibre type composition of 2 locomotory muscles in biopsies from Dutch Warmblood foals taken at 3 different ages in the first year postpartum. The muscle fibre types were determined histochemically as well as immunohistochemically. ATPase-characterised IIB fibres appear to express either IId or type lIa plus IId myosin heavy chain (MHC). A high percentage of fibres classified as IIA with ATPase expressed both fast types of MHC. The type I classification by the 2 methods matched almost completely. There was an increase with age of fibres expressing I and IIa MHC in the gluteus medius. At the same time, there was a decrease of fibres expressing IId MHC and fibres co-expressing MHC IIa and IId. MHC expression of the semitendinosus muscle did not change over time at first, but from age 22-48 weeks there was a decrease in the percentage of type IId fibres. In general, the gluteus medius contained more type I fibres but fewer type IId fibres compared to the semitendinosus. At most ages the fibre type compositions of both muscles correlated with one another. To examine the effect of exercise, one-third of the foals were given box rest, one-third received training and one-third kept at pasture during the first 22 weeks of life. The 3 exercise groups differed in their fibre type composition; however, these differences could not be attributed to the effect of exercise.     

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.     

The effect of diet fed to lambs on subsequent development of Trichostrongylus colubriformis larvae in vitro and on pasture

Contrasting herbage diets were fed to lambs to evaluate their effect on subsequent development of Trichostrongylus colubriformis larvae in faeces and on pasture. The diets had either no condensed tannin (CT), lucerne (Medicago sativa cv. Otaio), white clover (Trifolium repens cv. Tahora), or had moderate to high concentrations of CT, sulla (Hedysarum coronarium cv. Grassland Aokau), Lotus corniculatus (cv. Grasslands Goldie), L. pedunculatus (cv. Grassland Maku), Dorycnium pentophyllum, and Dorycnium rectum. Trials were carried out in summer (warm) and in autumn (cool and moist). In summer, egg viability was evaluated in vitro with egg hatch and larval development assays. In both seasons faeces were placed on pasture to compare recovery of eggs and larvae from faeces and larvae from herbage on the high and low fertility farmlets on the AgResearch Ballantrae Hill Country Research Station. D. rectum and D. pentophyllum diets decreased (P<0.01) egg hatching and larval development in laboratory assays relative to other diets. In summer, the number of larvae recovered from faeces placed on pasture was far greater (P<0.001) if the lambs had been fed lucerne than any other diet, whereas recovery was always lowest from faeces of sheep fed D. rectum and D. pentophyllum. Although dietary differences were lower in autumn than in summer, larval recoveries were lower (P<0.05) from faeces of lambs fed D. rectum and L. corniculatus than from white clover, lucerne and sulla diets. This study indicates that the diet of the host can have a significant impact on egg hatching and the subsequent development of T. colubriformis larvae in the laboratory and in the field. In particular, D. rectum consistently reduced T. colubriformis development. Effects measured in vitro generally under-estimated effects measured under field conditions.     

Welfare assessment of broiler breeders that are food restricted after peak rate of lay

1. Broiler breeder females were reared on one of three growth curves (ad libitum, conventional or modified restriction) and given rations containing a high or low concentration of crude protein. After the peak rate of lay they were fed ad libitum or a decreasing quantity of food in response to declining egg production. The welfare of the birds was assessed by determining the changes in indices of welfare at 36, 48 and 60 weeks of age. 2. Body weight increased rapidly in restricted birds fed ad libitum post-peak and water intake declined. 3. Post-peak food restriction was associated with a decrease in resting and increased drinking and spot-pecking activities. Birds that were food restricted during rearing spent more time foraging and spot-pecking at 36 and 48 but not 60 weeks of age. 4. Immune function increased with age but was not affected by the experimental treatments. The heterophil-lymphocyte ratio in birds fed ad libitum during rearing was numerically lower at 36 and higher at 48 and 60 weeks of age compared with restricted birds. 5. There was no effect of treatment on plasma corticosterone concentration. Creatine kinase activity was high at 60 weeks in treatments that were characterised by poor reproductive status and the activities of other enzymes reflected differences in reproductive status and mortality. 6. There was no long-term welfare or production advantage from feeding low protein rations or more generous feeding during the rearing period compared with conventional food restriction programmes.     

Variation in clinical and parasitological traits in Pietrain and Meishan pigs infected with Sarcocystis miescheriana

Future prophylaxis needs new concepts, including natural disease resistance of hosts against infectious agents. Genomic approaches to detect and improve disease resistance in farm animals and the molecular mechanisms involved in host-parasite interactions depend to a high degree on the trait differences between founder breeds, i.e. on the animal model. The present study evaluates differences in susceptibility/resistance against Sarcocystis miescheriana in the European Pietrain (PI) and the Chinese Meishan (ME) pig breeds, based on 25 individuals, infected orally with 5x10(4) sporocysts of S. miescheriana. Significant differences appeared in clinical, serological, haematological and parasitological findings. The major discriminating period post infection (p.i.) was between days 42 and 45. Severity of signs was negatively correlated with specific immunoglobulin titres during the first 3 weeks p.i. and positively with the load of bradyzoites in muscle tissues of the pigs. Loads of bradyzoites in muscle tissues were 20 times higher in PI than in ME. Sarcocystis-specific differences between the two breeds were in the range of 1-2 standard deviations. The study lays the foundation for further experiments to analyse chromosomal regions, candidate genes, and thus the molecular basis of Sarcocystis susceptibility/resistance as a model for host-parasite interaction in protozoan infectious disease.     

Effects of restricted feeding on physiological stress parameters in growing broiler breeders

1. In previous studies, a lack of agreement in measurements of plasma corticosterone concentrations and heterophil:lymphocyte (H/L) ratio as physiological indices of stress, caused by hunger and frustration in restricted-fed broiler breeders, was observed. It could be suggested that the differences between previous studies were caused by differences in duration of restriction and time of the day of the measurements. Therefore, in the present study the plasma corticosterone concentration and the H/L ratio were again determined in restricted- and ad libitum-fed growing broiler breeders, taking possible causes of disagreement between previous studies into account. In addition, we measured the daily rhythm in body temperature and heart rate, and the corticosterone responses to an acute stressor as physiological indices of stress. 2. Female broiler breeders (64 per treatment, housed in groups of 4 birds) were used in the experiment. Behaviour, baseline plasma corticosterone concentrations and H/L ratio were determined at 21 d of age (immediately after the start of food restriction), and at 42 and 63 d of age. Body temperature, heart rate and activity were measured by radiotelemetry for 36 h at 49 and 70 d of age. In addition, the plasma corticosterone response to acute stress (5 min manual restraint) was measured at 77 or 78 d of age. 3. Restricted broiler breeders had higher plasma corticosterone concentrations at 42 and 63 d of age, but no differences in H/L. ratio were found between restricted birds and unrestricted control birds. Restricted broiler breeders had a higher corticosterone response to 5 min manual restraint than unrestricted birds. Restricted birds displayed a clear day-night rhythm in body temperature, heart rate and activity whereas such a rhythm was blunted in ad libitum-fed birds. 4. It is discussed that some physiological differences (plasma corticosterone concentrations, body temperature and heart rate) between ad libitum-fed and restricted broiler breeders may have been caused by differences in metabolic rate as well as by differences in the level of stress. It is concluded that a combination of behavioural measurements and a wide range of physiological parameters should be used for the assessment of stress in growing broiler breeders.     

Monoclonal antibodies specific for Campylobacter fetus lipopolysaccharides

Four monoclonal antibodies (mAbs) (M1357, M1360, M1823 and M1825) which reacted with Campylobacter fetus lipopolysaccharide (LPS) core region epitopes were produced and characterized. Reactivity of these mAbs with C. fetus core LPS epitopes was determined by enzyme-linked immunosorbent assay (ELISA) with whole cell proteinase K digests and phenol-water extracted LPS, and by immunoblotting with proteinase K digests. The specificities of the four mAbs were evaluated using an indirect ELISA. One of the mAbs reacted with 42 and three of the mAbs reacted with 41 of the 42 C. fetus strains examined. No reaction was observed between the four mAbs and 32 non-C. fetus bacteria tested, with the exception of one mAb with one organism. The four mAbs reacted with serotype A and B strains indicating the presence of shared epitopes in C. fetus LPS core oligosaccharides. The specificities of three mAbs previously produced to C. fetus LPS O-antigens (M1177, M1183 and M1194) were also evaluated and no reaction was observed with these mAbs and the 32 non-C. fetus bacteria tested. Strong immunofluorescence reactions were observed with the anti-O chain mAbs and selected C. fetus strains of the homologous serotype. These anti-LPS core oligosaccharide and anti-LPS O chain mAbs are highly specific for C. fetus and are potentially useful as immunodiagnostic reagents for detection, identification and characterization of C. fetus.     

Serological responses against the pathogenic dimorphic fungus Mucor amphibiorum in populations of platypus (Ornithorhynchus anatinus) with and without ulcerative mycotic dermatitis

Mucor amphibiorum, a dimorphic fungus, causes ulcerative dermatitis and systemic infections in the platypus Ornithorhynchus anatinus in some river systems in Tasmania but apparently not in other regions of Australia. As yet there are no suitable tests for population surveys, nor for detection of internal lesions in live animals. Consequently, immunoglobulins were purified from the serum of platypuses and anti-immunoglobulin antisera were prepared in rabbits in order to develop an enzyme-linked immunosorbent assay (ELISA) for anti-M. amphibiorum antibodies. Antigens from plate-grown cultures resulted in greater signal-to-noise ratios in indirect ELISA than those from broth-grown cultures. Platypuses with clinical ulcerative dermatitis had elevated anti-Mucor antibody levels compared to apparently unaffected individuals. Seroconversion was observed in one animal coincident with the development of cutaneous ulcers. The results suggested that platypuses in affected rivers were exposed to M. amphibiorum at a higher frequency than the occurrence of clinical disease. Some platypuses from New South Wales had elevated antibody levels but these increased significantly with age suggesting exposure to cross-reactive antigens, although exposure to M. amphibiorum cannot be excluded. Further studies are warranted to determine factors that result in progression from infection to disease, the occurrence of the fungus in areas where disease has not been observed and the specificity of antigen used in ELISA.     

Cloning of bovine CD69

CD69 is rapidly inducible on various hematopoietic cells upon stimulation and is detectable as an early activation antigen. Although CD69 is well characterized in human and mouse, no information is available on bovine CD69. We report here that, bovine CD69 was cloned from a cDNA expression library prepared from activated peripheral blood lymphocytes. The full-length cDNA contained an 80bp 5' untranslated region, followed by a 600bp coding region and AU-rich motifs in a 3' untranslated region (GenBank accession number AF272828). Comparison of the bovine CD69 coding sequence reveals 69.4 and 78.2% nucleotide sequence identities with mouse and human CD69, respectively. The predicted amino acid sequence of bovine CD69 shares 56.3 and 62.3% sequence identity when compared with mouse and human CD69, respectively. Bovine CD69 has the highly conserved amino acid sequences found in the C-type lectin family, suggesting that the conserved residues may be important for conformation and binding to the, as yet unidentified ligand. In addition, the cytoplasmic tail of bovine CD69 has two casein kinase-2 (CK-2) phosphorylation sites. These data suggest that bovine CD69 plays an important role in the activation of lymphocytes.     

Probability of and risk factors for introduction of infectious diseases into Dutch SPF dairy farms: a cohort study

A 2-year cohort study was conducted to investigate the probability of disease introduction into Dutch dairy farms. The farms were tested regularly for diseases and were visited biannually to collect management data. Ninety-five specific pathogen-free (SPF) dairy farms were selected from a database of bovine herpesvirus type 1 (BHV1)-free farms to study the probability of, and risk factors for, introduction of BHV1, bovine viral diarrhoea virus (BVDV), Salmonella enterica subsp. enterica serotype Dublin (S. dublin), and Leptospira interrogans serovar hardjo (L. hardjo).Although most of the 95 SPF farms had a low risk on introduction of infectious diseases, one disease was introduced into 12 farms and two diseases were introduced into one farm. Three farms experienced an outbreak of BHV1, one farm an outbreak of L. hardjo, two farms BVDV, six farms S. dublin, and one farm both BHV1 and S. dublin. The total incidence rate was 0.09 (0.06-0.12) per herd-year at risk. The results suggest that the "non-outbreak" farms were significantly more closed than the "outbreak" farms. Direct animal contacts with other cattle should be avoided and professional visitors should be instructed to wear protective clothing before handling cattle.