Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke

The use of cysteine proteinases from Fasciola hepatica adult flukes for the serodiagnosis of caprine fasciolosis by means of an indirect ELISA test was studied. Two proteolytic fractions from adult fluke homogenates, with apparent molecular weights of 28 and 34 kDa (P28 and P34 respectively), were characterised as cysteine proteinases using azocasein assays and gelatin gel analysis. Both P28 and P34 fractions were electroluted and used as antigens in two different indirect ELISA tests. Serum IgG levels against P28 and P34 in goats given an experimental primary infection with 200 metacercariae or in goats given two experimental infections with 200 metacercariae were determined and compared with those observed in an uninfected control group. ELISA tests using both cysteine proteases showed a rapid and consistent detection of specific IgG in all experimentally infected goats. The IgG response to P28 was the first to be detected as early as 2-3 weeks post-infection and remained elevated throughout the experiment. The response to P34 was detected later (4-6 wpi) and disappeared in some animals at 18 wpi, while flukes were still present in the bile ducts. No significant differences were observed between the anti-P28 and anti-P34 IgG responses between animals receiving a primary or a challenge infection. The results of our study, although preliminary, are promising since the P28 ELISA described here may be a reliable method for the immunodiagnosis of F. hepatica infection in goats.     

Evaluation of the local analgesic effect of ketamine in the palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses

OBJECTIVE: To evaluate the local analgesic effect of ketamine in a palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses. ANIMALS: 36 mature healthy Andalusian horses. PROCEDURE: Horses were randomly assigned to 4 groups of 9 horses each and received an abaxial sesamoid block in a randomly chosen forelimb with 1 of the following: saline (0.9% NaCl) solution, 1% ketamine solution, 2% ketamine solution, or 3% ketamine solution. To determine analgesia, the radiant heat lamp-hoof withdrawal model was used as a noxious thermal stimulus. Before each nerve block, baseline hoof withdrawal reflex latency (HWRL, time between lamp illumination and withdrawal of the hoof) was determined; after the nerve block, local analgesic effects were determined by measuring HWRL at 2 and 5 minutes after injection and then every 5 minutes for a total period of 1 hour. RESULTS: Significant differences in HWRL were found between baseline values and values at 2 to 15 minutes following a nerve block with ketamine. Significant differences were found between HWRL values at every time point from 2 to 10 minutes following a nerve block with saline solution, compared with 1 or 2% ketamine solution. Similarly, significant differences were found between HWRL values at every time point from 2 to 15 minutes following a nerve block with saline solution, compared with 3% ketamine solution. CONCLUSIONS AND CLINICAL RELEVANCE: Abaxial sesamoid block with ketamine ensures adequate analgesia in horses with an onset of action of 2 minutes and a maximal duration of action of 15 minutes.     

A bulk tank milk survey of Ostertagia ostertagi antibodies in dairy herds in Prince Edward Island and their relationship with herd management factors and milk yield

The objective of this study was to quantify the relationship of the levels of antibodies to Ostertagia ostertagi in bulk-tank milk samples from Prince Edward Island (PEI) dairy farms to milk production and to herd-management practices potentially related to gastrointestinal nematode infections. The milk samples were obtained from 289 to 322 dairy farms during 2000; production and management data were available from 197 and 200 farms, respectively. Cow exposure to pasture and whole-herd anthelmintic treatment were the only herd management variables significantly associated with antibody levels in the fall of 2000. An increase in antibody levels from the observed 25th percentile to the 75th percentile (interquartile range) was associated with a drop in milk production of 1.2 kg/cow/day. The results of this study indicate that the enzyme-linked immunosorbent assay for O. ostertagi antibody is a potentially useful technique to measure parasite exposure in adult dairy cows and that parasite burdens in lactating cattle in PEI have an important impact on milk production.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

A longitudinal study of gastrointestinal parasites in Canadian dairy farms. The value of an indirect Ostertagia ostertagi ELISA as a monitoring tool

The general objective of this study was to evaluate a crude Ostertagia ostertagi antigen enzyme-linked immunosorbent assay (ELISA) for monitoring gastrointestinal parasites in lactating dairy cattle. A longitudinal study of gastrointestinal parasites in lactating dairy cows was carried out in 38 herds in four provinces of Canada (Prince Edward Island, Quebec, Ontario and Saskatchewan) from September 1999 to October 2000. Bulk tank milk, cow milk, serum and fecal samples were collected monthly or quarterly from all these farms. Information on herd management factors was collected by a standard questionnaire and individual cow production data were obtained from an electronic database. The overall mean optical density ratio (ODR) was 0.30 and ranged from -0.05 to 1.55. Although a clear seasonal pattern was not observed, the ODR values tended to decrease during the housing period and start increasing in the spring before the cows went out to pasture. The second and third or greater lactation cows had significantly higher ODR values compared with first lactation animals. The individual cow ODR had a very low correlation with individual squared root fecal egg counts but showed a reasonably high correlation when herd averages values were computed (r=0.73). A moderate correlation (r approximately 0.50) between the bulk tank and herd average ODR was observed. Milk yield was negatively associated with individual cow milk ODR and a quadratic effect on ODR was observed for days in milk. Twenty-eight of the herds participated in a clinical trial of eprinomectin (Ivomec Eprinex) treatment at calving. The cow level ODR values determined late in the previous lactation had a marginally significant effect (P=0.07) on treatment response, suggesting that high OD cows responded better to the anthelmintic treatment. However, because of the small sample size available in this model, more research is needed to better understand this relationship. In conclusion, the indirect ELISA using an O. ostertagi crude antigen appears useful as a technique for monitoring gastrointestinal parasite burdens in adult dairy cows and holds promise as a potential predictor of response to anthelmintic treatment.     

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8 μg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256 μg/ml (62 strains) and 512 μg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.     

Responses of citrus fine roots to localized soil drying: a comparison of seedlings with adult fruiting trees

We studied the responses of citrus (Citrus volkameriana Tan. & Pasq.) roots to 15 weeks of soil drying. A comparison was made between the fine roots of 1-year-old seedling root systems (seedling) and the fine roots of woody laterals of 6-year-old grafted trees (adult). Each seedling and woody lateral root system was established in a pair of vertically separated and independently irrigated soil compartments located in field root chambers excavated adjacent to the trees to which the woody laterals were attached. Root + soil respiration and fine root survival of seedlings and adults were similar for the first 5 weeks. However, eight weeks after termination of irrigation to the upper soil compartments, mortality of fine roots was high in adults but not seedlings. Fine roots of adults exposed to dry soil for 5, 8 and 15 weeks exhibited 2, 26 and 33% mortality, respectively, whereas the corresponding values for fine roots of seedlings were 2, 6 and 8%. Although root + soil respiration rates of adults and seedlings were similar before the soil drying treatment, rates for adults were only 25% of those for seedlings after 15 weeks of soil drying. We conclude that, although fine roots of adults and seedlings are similar in form, they respond differently to soil drying.     

Shoot development and dieback in progenies of Nothofagus obliqua

Shoot growth and dieback were compared among progenies of nursery-grown seedlings of Nothofagus obliqua belonging to seven progenies of the same provenance (Quila-Quina, Argentina). First-year shoots consisted of one growth unit (GU) and second-year shoots of one or two GUs. The probability of development of two GU was similar for all progenies. Progenies were different in terms of shoot size, terminal bud abscission, the extent of shoot dieback after shoot extension and the node of origin of the relay shoot on the first shoot. Plants with a second-year shoot consisting of two GUs had a thicker stem and more nodes than those with single-GU shoots. The selection of N. obliqua seed trees based on architectural traits suitable for forestry development at specific sites must contemplate variability among progenies and their probabilities of successful development under different conditions.