Failure of a recombinant Schistosoma bovis-derived glutathione S-transferase to protect cattle against experimental Fasciola hepatica infection

The potential of a recombinant Schistosoma bovis 28-kDa glutathione S-transferase (rSb28GST) to protect cattle against Fasciola hepatica was tested in a vaccination trial. Thirty two calves were randomly divided into four groups of eight animals. Calves of the three vaccine groups received two intramuscular injections at 3 weeks interval, of 0.250mg rSb28GST in either aluminium hydroxide (Al(OH)(3)), Quil A, or PBS emulsified in an equal volume of Freund's complete adjuvant (FCA).Animals of the control group received injections of Al(OH)(3)/PBS only. All animals were challenged orally with a total of 360 metacercariae of F. hepatica, spread over 6 weeks.All groups of vaccinated animals produced measurable IgG antibody titers to rSb28GST after vaccination. Animals immunised with FCA adjuvanted vaccine had the highest and more durable antibody titers and only sera from this group recognised an approximately 24kDa protein band from F. hepatica, that is thought to be a F. hepatica GST. Despite a good antibody response differences in cumulative faecal egg output between the groups were not statistically significant. In addition, no significant difference was found between groups in terms of total worm numbers or percentage of immature flukes recovered at necropsy. In conclusion, the recombinant S. bovis 28kDa GST was not found to adequately protect cattle against experimental F. hepatica challenge, using either aluminium hydroxide, Quil A or FCA as adjuvant.     

Morphometry and histochemistry of the semitendinosus muscle of tufted capuchin monkeys (Cebus apella Linnaeus, 1758)

Considering monkeys are animals closely related to the human, and semitendinosus muscle has been used in many postural research experiments, we have decided to study its histochemical characteristics. Samples were removed from the proximal, middle and distal regions of the semitendinosus muscles of five adult male tufted capuchin monkeys and observed for reaction with m-ATPase (with alkaline and acid pre-incubation), nicotinamide adenine dinucleotide tetrazolium reductase, and haematoxylin and eosin. The muscle fibres were classified as fast glycolytic (FG), fast oxidative glycolytic (FOG) and slow oxidative (SO) and quantified in terms of frequency and area. The three regions of the semitendinosus muscle showed no significant differences in frequency or area of the respective fibre types, and therefore the muscle can be considered histoenzymologically homogeneous. FG fibres presented higher frequencies and larger areas. The sum of FOG and SO fibres was 57.5%, suggesting that the semitendinosus muscle of the tufted capuchin monkey is adapted to an action involved in posture maintenance and in long arboreal dislocation. The present data agrees with the notion of differentiated quadrupedalism in some primates, which support a lower percentage of their weight on the fore limbs.     

Extrinsic cardiac nerve segments in the domestic dog (Canis familiaris- Linnaeus, 1758). Comparative study in young and adult dogs

In this paper, important connections between the two main contingents of the autonomic nervous system, intrinsic and extrinsic visceral plexus were analysed. Concerning heart innervation, the territories of extrinsic innervation are very important in the treatment of congenital or acquired cardiopathy, thoracic neoplasia and aortic arch persistence, among others. This research compared young and adult extrinsic cardiac innervation and described the surgical anatomic nerve segments. Animals were perfused with a 10% formaldehyde solution in PBS (0.1 m) (pH 7.4) and submitted to macro- and meso-scopic dissection immersed in 60% acetic acid alcoholic solution and 20% hydrogen peroxide aqueous solution. The nerve segments were assigned as: right vagus nerve segment, left vagus nerve segment, right middle cervical ganglion segment, left middle cervical ganglion segment, right caudal laryngeal nerve segment, left caudal laryngeal nerve segment, right phrenic nerve segment and left phrenic nerve segment.     

A rapid and simple method for the separation of pure lymphocytes from horse blood

A method for the separation of pure and viable lymphocytes and granulocytes from the same blood sample in horses was reported. By centrifuging equine heparinized blood at 100 xg for 10 min at room temperature (r.t.), the resulting supernatant plasma was an almost pure (97.71 +/- 0.30%; n = 15) suspension of highly viable (98.72 +/- 0.28%) lymphocytes. When sodium citrate was used as an anticoagulant, lymphocyte suspensions collected in the same manner showed lower purity (87.89 +/- 1.59%; n = 9) and higher yields (56.56 +/- 3.89%, n = 9 versus 36.11 +/- 2.23%, n = 15). Where needed, a further centrifugation at 250 xg for 3 min (r.t.) of heparinized lymphocyte preparations removed an average of 87.39% (n = 15) contaminating platelets. A suspension of 85.96 +/- 2.20% pure granulocytes (93.23 +/- 1.74% neutrophils; n = 14) with minimal contamination by erythrocytes and high viability (93.11 +/- 1.26%) was obtained by performing a flash red blood cell lysis on the white-greyish layer resulting from the centrifugation of the heparinized blood samples. Among the several methods available, the procedure described herein is easy, rapid, cheap and reproducible.     

Small intestinal morphology and disaccharidase activities in early-weaned piglets fed a diet containing spray-dried porcine plasma

The purpose of this study was to test whether dietary spray-dried porcine plasma (SDPP) in early-weaned piglets prevents small intestinal villus atrophy by trophic or protective activity. Fifty-four weaned, 18-day-old piglets were used to determine the effect of dietary SDPP on small intestinal villus length, crypt depth, enterocyt mitotic activity and brush border enzyme activities during the first week after weaning. The piglets were offered a diet containing either 8% SDPP or 8% casein. At 2 and 7 days after weaning, piglets were anaesthetized to provide samples of the small intestinal wall and killed immediately afterwards. There were no differences in daily gain and daily feed intake between the two dietary treatments. At day 2 after weaning, all piglets showed a marked reduction in villus height when compared with baseline values. In all piglets, small intestinal enterocyte mitotic activity had decreased by day 2 and was increased again on day 7. There were no significant effects of dietary SDPP on small intestinal villus length, crypt depth and enterocyt mitotic activity. This indicates that SDPP has no trophic effect on the small intestinal mucosa and that it does not protect against the damaging effect on the small intestinal villi that is associated with the process of weaning. There was no effect of SDPP on lactase-, sucrase- or maltase-specific activities that are a measure of the digestive function of the small intestine. It can be concluded that SDPP versus casein has no effect on small intestinal morphology and disaccharidase activities in early weaned piglets kept under low infection pressure.     

Differential serological testing by simultaneous indirect immunofluorescent antibody test in canine leishmaniosis and ehrlichiosis

A mixed indirect fluorescence antibody test (IFAT), based on cultured promastigotes Leishmania infantum and formol-inactivated suspension of cells infected with the bacteria Ehrlichia canis, was applied to make a differential diagnosis between canine ehrlichiosis and leishmaniosis. A titre greater than 80 was considered positive for antibodies to E. canis and suggestive of antibodies to L. infantum. Positive sera were titrated subsequently by serial dilutions to confirm antibodies positive to Leishmania and establishing the antibody titre of both pathogens. Fluorescence was absent with negative control sera and background staining was minimal. No serological cross-reactions between positive sera for L. infantum or E. canis were detected. Results obtained by mixed IFAT did not differ when the same serum IFAT standard was compared. The test showed equivalent sensitivity (100%). The specifities were 100% for L. infantum and 98.5% for E. canis. The equivalence in sensitivity was confirmed by calculating the correlation coefficient between IFAT standards and mixed IFAT (r>or=0.99 for both pathogens). The results of our investigations demonstrated that mixed IFAT is a specific means of establishing serological differential diagnosis of canine leishmaniosis and ehrlichiosis.     

The status of Mycobacterium bovis infection in UK wild mammals: a review

Bovine tuberculosis caused by Mycobacterium bovis is a zoonotic infection with a wide range of mammalian hosts. In parts of the UK M. bovis infection in cattle is a persistent problem. The European badger (Meles meles) is implicated in the transmission of M. bovis to cattle, and is widely believed to constitute the most important reservoir of infection in UK wildlife. However, few studies have been carried out on the status of M. bovis infection in other UK mammals. In this review we present information on the incidence and pathology of M. bovis infection in UK wild mammals from both published and previously unpublished sources. Although the evidence does not support the existence of a significant self-maintaining reservoir of infection in any wild mammal other than the badger, there is a clear lack of sufficient data to rule out the involvement of other species. In the light of this and the dynamic nature of epidemiological patterns, further surveillance for M. bovis infection in UK wild mammals, using modern methods of diagnosis, is essential.     

Immune function of bovine leukocytes after in vitro exposure to selected heavy metals

OBJECTIVE: To study effects of in vitro exposure of bovine leukocytes to mercury, cadmium, and lead on phagocytosis, natural killer cell activity, and lymphocyte proliferation. SAMPLE POPULATION: Leukocytes from 6 nonpregnant Holstein heifers. PROCEDURE: Leukocytes were exposed in vitro to the aforementioned metals, and leukocyte functions were assessed. RESULTS: Phagocytosis was suppressed by 10(-5) to 10(-7) M CdCl2 and by 10(-5) and 10(-6) M HgCl2, but not 10(-7) M HgCl2 nor 10(-4) to 10(-6) M PbCl2. Spontaneous and concanavalin A- or phytohemagglutinin-stimulated proliferation of metal-treated bovine blood mononuclear cells was not significantly different from that of nontreated control cells, except for enhanced spontaneous proliferation in response to 10(-5) M HgCl2. When proliferation was expressed as a stimulation index, a dose-dependent increase of spontaneous proliferation was observed in response to exposure to HgCl2 and PbCl2. Compared with response to 10(-6) or 10(-7) M CdCl2, reduction of mitogen-induced and spontaneous proliferation was observed on exposure to 10(-5) M CdCl2. Natural killer cell activity against YAC-1 target cells, evaluated by flow cytometry, was decreased only in cells exposed to 10 M HgCl2. CONCLUSION AND CLINICAL RELEVANCE: Bovine leukocytes are susceptible to the immunomodulatory effects of in vitro exposure to heavy metals at concentrations equal to or higher than those at which similar effects are seen for leukocytes from most other animal species for which data are available for comparison. Exception is phagocytosis, which is severely affected by low concentrations of CdCl2 and HgCl2 in cattle. Reduction of defense mechanisms on exposure to metals could lead to increased susceptibility to potential pathogens.     

Significance of infectious bursal disease serology in an integrated quality control program under European epidemiologic conditions

In this study performed between 1993 and 1997, infectious bursal disease virus (IBDV) antibody titers and performance data were recorded in a vertically integrated monitoring scheme in order to make a follow-up from day-old parents down to the broilers at slaughter. All measured data were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. It appeared that high and/or uniform antibody titers in the parents were correlated with increased daily weight gain and decreased mortality and slaughterhouse condemnation in the broilers. Antibody titers and their CVs were negatively correlated in broiler parents and their offspring at day-old and even at slaughter. Results indicate that high and uniform antibody titers against IBDV in broiler parents are important for good performance of the broiler offspring, at least under the epidemiologic conditions of this study, which included the presence of very virulent IBDV strains in the field and the sole use of live intermediate vaccines in broilers as well as broiler parents.     

Effects of a porcine reproductive and respiratory syndrome virus infection on the development of the immune response against pseudorabies virus

The aim of this study was to investigate the effects of a porcine reproductive and respiratory syndrome virus (PRRSV) infection on the development of the immune response after pseudorabies virus (PRV) vaccination in pigs. Pigs were intranasally inoculated with the European PRRSV strain, Lelystad virus ter Huurne, and were vaccinated intramuscularly with PRV 2 weeks later (LV-PRV group). Control pigs were vaccinated with PRV only (PRV group). Eight weeks after PRV vaccination, pigs from both groups were challenged intranasally with wild-type PRV. We measured the lymphoproliferative, and the cytolytic responses to PRV of peripheral blood mononuclear cells (PBMC), isolated from blood samples. In addition, serum samples were examined for antibodies against PRV and LV. One week after PRV vaccination, PBMC proliferated abundantly to PRV in both groups. However, in the LV-PRV group the lymphoproliferative response declined after 1 week, whereas, in the PRV group, the lymphoproliferative response was high for 3 weeks and declined thereafter (P<0.05). After challenge, the lymphoproliferative response was 1 week earlier and was consistently and significantly higher in the PRV group than in the LV-PRV group. The PRV-specific killing was higher at 3 weeks after PRV vaccination and 5 weeks after PRV challenge 19+/-3 and 24+/-6%, respectively, in the PRV group, compared to 7+/-4 and 6+/-9%, respectively, in the LV-PRV group (P<0.05). However, later after vaccination and challenge the cytolytic response was identical in both groups. The antibody titre against PRV developed equally in both groups. After challenge, no PRV virus was isolated from both groups. From these results we conclude that, although PRRSV infection did cause changes in the time course of the T-lymphocyte response after PRV vaccination, PRRSV infection did not inhibit the development of vaccine-induced protection after PRV.