Characterisation of the 33kDa piroplasm surface antigen of Theileria orientalis/sergenti/buffeli isolates from West Java, Indonesia.

The immunodominant 33/35kDa antigen of a Theileria isolate from West Java, Indonesia, was characterised and immuno-affinity purified by use of a monoclonal antibody, KUL-a4, and was shown to be representative of the T. orientalis/sergenti/buffeli group. The aminoterminal sequence of the purified 35kDa peptide (20 residues) was determined by automated Edman degradation and found to correspond to the predicted amino acid sequence of a prospective p33 gene previously sequenced from the same isolate. The cleavage site of a putative signal peptide was identified and conforms the (-3, -1) rule for signal peptidases. The existence of dimeric and trimeric forms of the p33/35 antigen is hypothesised from Western blot profiles. KUL-a4 appeared specific for the T. orientalis/sergenti/buffeli group. It did not recognise in indirect fluorescence antibody test (IFAT), intraerythrocytic bodies of Anaplasma marginale or piroplasms and schizonts of T. mutans, T. parva and T. annulata, whereas cattle antisera raised to these species showed cross-reactivity in IFAT. It however, appeared weakly cross-reactive in Western blot and ELISA, with the 34kDa piroplasm antigen of one T. annulata (Gharb) isolate. The present study indicates that the isolated antigen belongs to the p33/34 antigen family described within the T. sergenti/orientalis/buffeli group, and documents the group-specificity of one of its epitopes.     

Assessment of intake inadequacy and food sources of zinc of people in China

OBJECTIVES: To assess the intake inadequacy and food sources of zinc of people in China. DESIGN AND SUBJECTS: Diets of 68 962 subjects aged 2-101 years (urban 21 103, rural 47,859) in the 2002 China National Nutrition and Health Survey were analysed. Dietary intake was assessed using 24-hour recall for three consecutive days. Zinc intake inadequacy was calculated based on values suggested by the World Health Organization. RESULTS: The median zinc intake ranged from 4.9 mg day- 1 (urban girls, 2-3 years) to 11.9 mg day-1 (rural males, 19+ years). The zinc density of urban residents (2-3 to 19+ years) was 5.0-5.3 mg day-1 (1000 kcal)-1, significantly higher than that of their rural counterparts (4.7-4.8 mg day-1 (1000 kcal)-1). Differences in food sources of zinc from cereal grains (27.4-45.1 vs. 51.6-63.2%) and animal foods (28.4-54.8 vs. 16.8-30.6%) were found between urban and rural residents. Zinc from vegetables and fruits (8.2-13.8 vs. 9.7-12.4%) and legumes (1.3-3.3 vs. 2.5-3.4%) was comparable between urban and rural residents. The proportion of zinc intake inadequacy ranged between 2.8% (urban females, 19+ years) and 29.4% (rural lactating women). Rural residents had higher proportions of zinc intake inadequacy than their urban counterparts. Significantly higher proportions of zinc inadequacy were found in the category of phytate/zinc molar ratio >15 for both rural and urban residents. CONCLUSIONS: About 20% of rural children are at risk of inadequate zinc intake, with phytate as a potential important inhibitor. Moreover, lactating women are also considered a vulnerable group.     

Competitive enzyme-linked immunosorbent assay for heartwater using monoclonal antibodies to a Cowdria ruminantium-specific 32-kilodalton protein

Hybridomas producing monoclonal antibodies (mAb) to Cowdria ruminantium were raised. Four mAbs of the IgG isotype reacted in western blots with a 32-kilodalton Cowdria protein (Cr32), which had previously been shown to be conserved and immunodominant. A fifth mAb of the IgM isotype recognized a 40-kDa Cowdria protein. The latter mAb was negative in an indirect fluorescent antibody test (IFA), whereas the other four were positive. mAb No. 4F10B4 showed the strongest signal in western blots using three different stocks of Cowdria. Immuno-gold labeling of Cowdria organisms in vitro using 4F10B4 showed that Cr32 has surface-exposed antigenic determinants. Using mAb 4F10B4, a competitive ELISA was developed which detected specific Cowdria antibodies in goat, sheep and cattle sera. Antibodies in animal sera competed with binding of mAb 4F10B4 to a crude sonicated Cowdria antigen obtained from infected endothelial cell cultures. The competition ELISA (CELISA) detected antibodies in 55 out of 70 (79%) goats experimentally infected with one of eight different Cowdria stocks. Fourteen out of the 15 sera which were shown negative in the CELISA were also negative in the IFA. Nevertheless, all 15 sera recognized some epitopes of the immunodominant Cowdria-specific 32 kDa protein as judged from their reaction with this protein in western blots. Overall, there was 89% agreement between CELISA and IFA considering all 70 goat sera. Moreover, antibodies were detected in nine out of nine sheep infected with one of three different stocks of Cowdria and in sera from calves experimentally infected by two different strains of heartwater. There were no cross-reactions with Ehrlichia phagocytophila antibodies in goat sera, nor with Anaplasma marginale antibodies in bovine sera. Lack of cross-reactivity and detection of antibodies to eight geographically widely distributed stocks of Cowdria, makes the competition ELISA a promising test for use in heartwater endemic areas.     

Observations on mouse-infective stocks of Cowdria ruminantium: microscopical demonstration of the Kwanyanga stock in mouse tissue and the carrier-status of the Senegal stock in mice

The behaviour of two different stocks of Cowdria ruminantium was investigated in mice. The mouse-pathogenic Kwanyanga stock of C ruminantium was microscopically demonstrated in mice in capillary endothelial cells of the lung, spleen, kidney, liver and brain. Mice of the ninth passage of the Senegal stock, which is infective but not pathogenic to mice, were kept alive for a year. Their blood and homogenised spleens, inoculated intravenously, caused fatal heartwater in a goat. However, the Senegal stock could not be demonstrated microscopically in mice. These results indicate the possible role of rodents in the epidemiology of heartwater.     

The association of hoof lesions at claw trimming with test-day milk yield in Danish Holsteins

In a cross-sectional study, performed between October 2002 and April 2003 on 55 Danish dairy herds with 6161 predominantly Holstein Friesian cows the prevalence of 9 hoof lesions was determined.

All test-day yields (TDY) of kg energy corrected milk (ECM) in the lactation of diagnosis were recorded. For the purpose of including hoof lesions in a decision support model an attempt was made to aggregate the lesions into digital dermatitis (DD), other interdigital diseases (OID, infectious diseases other than DD) and hoof horn diseases (HHD, related to metabolic disorders and trauma). A division was made based on the stage of lactation in which the lesions were diagnosed; during the first 100 days in milk, between days 101 and 200 and between days 201 and 305. Associations between the presence of hoof lesions at trimming and milk production were analyzed by linear mixed modeling at the cow level, clustered within herd. The data of primiparous cows was analyzed separately from the multiparous cows.

Based on the associations between TDY and the individual lesions aggregated into HHD, this aggregation as initially planned could not be justified and was therefore not maintained. For OID the value of early diagnosis was apparent; an early diagnosis and early treatment (<101 DIM) was associated with either a positive value or a value less negative compared to a diagnosis and treatment in late lactation (201–305 DIM). This pattern was not as clearly present for the other lesions.     

Quantitative correlation of parasitological and serological techniques for the diagnosis of Trypanosoma congolense infection in cattle

A comparison was made between serological and parasitological techniques for the diagnosis of bovine trypanosomiasis in Zambia. Overall sero-prevalence rates as determined by IFAT and ELISA were respectively 2.7-fold and 2.9-fold greater then the percentage of samples found positive with the dark ground/phase contrast buffy coat technique (DG). The results obtained by the two serological techniques were found to be closely correlated (94.2%) agreement) and titres obtained by ELISA tended to be slightly higher than those obtained by IFAT. Linear regression analysis of the results obtained by the IFAT and DG techniques revealed a highly significant correlation. This finding would permit the use of only one of the techniques in an epidemiological survey and to extrapolate the results from the regression line.     

The phenology and population dynamics of Culicoides spp. in different ecosystems in The Netherlands

The Netherlands has enjoyed a relatively free state of vector-borne diseases of economic importance for more than one century. Emerging infectious diseases may change this situation, threatening the health of humans, domestic livestock and wildlife. In order to be prepared for the potential outbreak of vector-borne diseases, a study was undertaken to investigate the distribution and seasonal dynamics of candidate vectors of infectious diseases with emphasis on bluetongue vectors (Culicoides spp.). The study focused primarily on the relationship between characteristic ecosystems suitable for bluetongue vectors and climate, as well as on the phenology and population dynamics of these vectors. Twelve locations were selected, distributed over four distinct habitats: a wetland area, three riverine systems, four peat land areas and four livestock farms. Culicoides populations were sampled continuously using CO(2)-baited counterflow traps from July 2005 until August 2006, with an interruption from November 2005 to March 2006. All vectors were identified to species level. Meteorological and environmental data were collected at each location. Culicoides species were found in all four different habitat types studied. Wetland areas and peat bogs were rich in Culicoides spp. The taxonomic groups Culicoides obsoletus (Meigen) and Culicoides pulicaris (Linnaeus) were strongly associated with farms. Eighty-eight percent of all Culicoides consisted of the taxon C. obsoletus/Culicoides scoticus. On the livestock farms, 3% of Culicoides existed of the alleged bluetongue vector Culicoides dewulfi Goetghebuer. Culicoides impunctatus Goetghebuer was strongly associated with wetland and peat bog. Many Culicoides species were found until late in the phenological season and their activity was strongly associated with climate throughout the year. High annual variations in population dynamics were observed within the same study areas, which were probably caused by annual variations in environmental conditions. The study demonstrates that candidate vectors of bluetongue virus are present in natural and livestock-farm habitats in the Netherlands, distributed widely across the country. Under favourable climatic conditions, following virus introduction, bluetongue can spread among livestock (cattle, sheep and goats), depending on the nature of the viral serotype. The question now arises whether the virus can survive the winter conditions in north-western Europe and whether measures can be taken that effectively halt further spread of the disease.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.