Update on the diagnosis of Haemophilus parasuis infection in pigs and novel genotyping methods

Haemophilus parasuis causes Gl?sser's disease as well as a number of other diseases in pigs. The diagnosis of H. parasuis-associated disease is usually established by clinical signs, pathological findings and bacterial isolation but diagnosis is complicated by the existence of non-virulent strains and the early colonisation of the upper respiratory tract of healthy piglets. Moreover, several strains can be found on a farm and even within a single animal so it is important to determine the specific strain that is causing the clinical outbreak. Recently, genotyping methods have been developed with the goal of correlating genotype with the degree of virulence of H. parasuis strains. The association between genotype and virulence in H. parasuis is challenging due to the lack of knowledge of the complete genomic sequence and virulence factors of this bacterium.     

Morphometric analysis of the corneal endothelium of Yacare caiman (Caiman yacare) using scanning electron microscopy

The objective of this study was to examine the endothelial surface morphology and to perform morphometric analysis of the corneal endothelial cells of Yacare caiman (Caiman yacare) using scanning electron microscopy. Morphometric analysis with regard to polygonality, mean cell area, cell density and coefficient of variation of mean cell area was performed. Cell areas were measured using image analysis software. The normal corneal endothelium of Yacare caiman consisted of polygonal cells of uniform size and shape with interdigitations of the cell borders. Microvilli appeared as protrusions on the cellular surface. The average cell area was 270 +/- 24 microm(2) and the endothelial cell density was 3704 +/- 324 cells/mm(2). The coefficient of variation of cell area was 0.22. This study demonstrates that the Yacare caiman corneal endothelium is similar to those described in other vertebrates.     

Identification of bluetongue virus serotype 2 (Corsican strain) by reverse-transcriptase PCR reaction analysis of segment 2 of the genome

In October 2000, bluetongue virus was detected on the French island of Corsica. The disease was also reported in Sardinia, Calabria, Sicily and on the Spanish islands of Majorca and Minorca. This paper describes the use of molecular techniques for a rapid identification and serotype determination of serotype 2 of the virus. The nucleotide sequences of segments 2 and 7 of the genome of the Corsican strain were determined and its phylogenetic relationships are described.     

Molecular phylogenetic analysis of Onchocerca lupi and its Wolbachia endosymbiont

The morphology of Onchocerca lupi, responsible for canine ocular onchocercosis, is unique within the genus. Earlier analyses of the 5S ribosomal RNA gene spacer region sequence of the parasite and the 16S ribosomal RNA gene sequence of its Wolbachia endosymbiotic bacteria (Rickettsiales) supported the morphological and biological arguments that O. lupi is a distinct species. However, the exact phylogenetic position of O. lupi and its endosymbiont could not be unambiguously determined. Herein we report analyses based on the mitochondrial cytochrome oxidase I (COI) gene of the filarial species and the Wolbachia surface protein (wsp) and the bacterial cell-cycle ftsZ genes of their wolbachiae. Our results indicate that O. lupi separated from other Onchocerca spp. early in evolution. This is in line with the previous morphological analysis demonstrating that O. lupi is an atypical Onchocerca species showing both primitive and evolved characters. The phylogenetic trees generated for the COI sequences of filariae and the wsp and ftsZ sequences of their wolbachiae were congruent with each other, which supports the hypothesis that nematodes and their Wolbachia endobacteria share a long co-evolutionary history.     

Sandwich enzyme-linked immunosorbent assay (ELISA) for measuring the concentration of, and detection of antibodies to, Aujeszky's disease virus

A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for measuring Aujeszky's disease virus (ADV) antigen concentration and an inhibition technique based on the former was developed for detection of antibodies to ADV. The results were checked by determining the cytopathic and serum neutralization titres. The correlation was satisfactory in both cases, with correlation coefficients above 0.8. When measuring ADV antigen concentration, the lower limit of detection was 10(3) TCID 50/0.2 ml. The sensitivity of ELISA in detecting antibodies to ADV was found to be superior to that of the serum neutralization test and, thus, enabled the testing of rabbit and guinea-pig sera.     

The influence of physiological state and dietary nitrogen supply on digestion in the dairy cow

The effects of N supply on digestion were compared in cows in late pregnancy vs early lactation. Two groups of four and one group of three dairy cows received, during a digestion trial, corn silage-concentrate diets (65:35) differing in N supply. Concentrates were formulated so that diets were either insufficient (Diet 1) or sufficient (Diets 2 and 3) in ruminally fermented N and either insufficient (Diets 1 and 2) or sufficient (Diet 3) in protein digestible in the intestines. Experimental periods were 3 wk before and 3 wk after parturition. Organic matter digestibilities were 69.8, 73.1 and 72.5% in late pregnancy vs 64.9, 69.8 and 70.8% in early lactation for Diets 1, 2 and 3, respectively. Digestibility was higher (P less than .05) in late pregnancy than in early lactation. Differences between physiological states were attributed to differences in ruminal digestibility and in fiber digestibility. These differences were not explained by a reduction in large particle retention time, but in situ DM disappearance was reduced in early lactation. Ruminal protozoa concentration and the acetate: propionate ratio decreased between pregnancy and lactation. The duodenal non-ammonia N:N intake ratio was higher for Diet 1 than for Diets 2 and 3, 1.20, .97 and .94, respectively, but it did not vary between physiological states. In conclusion, some of the negative consequences of a shortage in degraded N are more dramatic in early lactation than in late pregnancy.     

Intake and digestibility in sheep and chemical composition during different seasons of some West African browse species

Foliage of Afzelia africana, Pterocarpus erinaceus and Khaya senegalensis, from 10 trees per species, was collected every two weeks during the late dry, rainy and cool season to determine the seasonal effects on chemical composition. Fifteen rams of the Djallonké breed, weighing on average 20.0 kg, were used to evaluate the voluntary intake and digestibility of hay of A. gayanus, foliage of A. africana (as a sole feed), and A. africana, P. erinaceus and K. senegalensis offered with 30% of the diet as A. gayanus hay. The crude protein (CP) content of A. africana, and P. erinaceus decreased significantly from the late dry season to the cool season when that of K. senegalensis tended to increase. The mean CP of A. africana, P. erinaceus and K. senegalensis differed significantly (173 g, 139 g and 114 g/kg DM, respectively). The DM intake of A. africana offered with hay (571 g/d) or as a sole feed (598 g/d) were not significantly different, but was higher than that of P. erinaceus (428 g/d) and K. senegalensis (298 g/d). The digestibility calculated by difference of DM and CP of A. africana (582 g/kg DM and 795 g/kg CP, respectively) did not differ significantly from A. africana as a sole feed, but were higher than for the other species. The nutritive value of A. africana seems to justify the high preference of herders for this species.     

Detection of bovine herpesvirus 4 in CD11b+ leukocytes of experimentally infected rabbits

The presence and numbers of bovine herpesvirus 4 (BoHV-4) infected CD11b+ leukocytes were investigated during experimental infections of New Zealand White rabbits by Fluorescence Activated Cell Sorter (FACS) analysis. Peripheral blood leukocytes (PBL) were collected every second day, and the cells were stained with phycoerythrin-labelled CD11b-specific mouse monoclonal antibody and fluorescein-conjugated bovine herpesvirus 4-specific mouse monoclonal antibody. The numbers of double-stained cells from PBLs of the control and inoculated groups were measured and compared in FACSTREK analyser. Double-stained cells were detected in the virus-inoculated group on postinoculation days (PID) 2-5 and 9-12. The results indicated that CD11b+ PBLs were permissive for BoHV-4 infection, and are probably the main reservoir of the virus during the latent period. The data did not indicate production of infectious viral particles, but virus-specific proteins were expressed on the surface of CD11b+ cells. The two waves of double-stained cells gave similar results to the PCR assays from serum samples, which showed the presence of viral DNA in the serum on the same days when virus-infected CD11b cells were also present. Productive BoHV-4 infection of mast cells or undifferentiated leukocytes in the bone marrow and the antiviral immune response might be responsible for this periodic appearance of the virus in CD11b+ PBLs and in the serum. The paper provides evidence that CD11b+ PBLs are the main target cell populations in the blood for BoHV-4.     

Theileria mutans in Nigeria.

Two strains of bovine Theileria from northern Nigeria were shown to be identical to Theileria mutans in the indirect immunofluorescent antibody test. One of the strains was transmitted by the tick Amblyomma variegatum; large macroschizonts, typical for T mutans, could be demonstrated in infected cattle. It is concluded from these experiments and from the literature that there is reliable evidence so far for the occurrence in Nigeria of only two bovine Tehileria species, T mutans and T velifera.