Expression and location of IGF binding proteins-2, -4, and -5 in developing fetal tissues.

Insulin-like growth factors are associated with myogenesis in vivo, and their actions are mediated by IGF binding proteins (IGFBP). Sites of IGFBP production and their location during early development are not clear. The objective of this research was to examine the developmental expression and location of IGFBP-2, -4, and -5 mRNA and peptides in developing porcine skeletal muscle and liver. Pregnant pigs were euthanatized at various times postconception (pc). Developmental expression of IGFBP was evaluated using total RNA extracted from skeletal muscle and liver of 30-, 44-, 59-, 68-, 75-, 89-, and 109-d pc fetuses and from adult and neonatal pigs. Localization of IGFBP-2, -4, and -5 mRNA and peptides was examined by in situ hybridization and immunocytochemistry of muscle samples from contralateral pelvic limbs of each pig. Overall muscle IGFBP gene expression decreased (P < .05) with increasing age. Moreover, expression of liver IGFBP-2 and -5, but not of IGFBP-4, was greater (P < .05) during prenatal than during postnatal periods. The majority of immunoreactive IGFBP was located in developing muscle cells, with little localized to connective tissue, except at later stages of development. These data show that IGFBP-2, -4, and -5 expression is time- and tissue-dependent in fetal liver and muscle.     

Effects of glutamine supplementation of an amino acid-based purified diet on intestinal mucosal integrity in cats with methotrexate-induced enteritis.

OBJECTIVE: To determine effects of glutamine-supplemented and glutamine-free amino acid-based purified diets, compared with a dry expanded diet, on intestinal structure and function in a model that used cats with methotrexate-induced enteritis. ANIMALS: 18 adult specific-pathogen-free cats. PROCEDURE: 12 cats were given intragastric feedings of an amino acid-based purified diet supplemented with glutamine (7% [wt:wt]) or an isonitrogenous amount of glycine and alanine; 6 cats consumed a dry expanded diet. After 21 days, cats received methotrexate (MTX; 11 mg/kg of body weight, IV). Intestinal permeability testing was performed immediately before and 66 hours after MTX administration. Celiotomy was performed 72 hours after MTX administration for aseptic removal of mesenteric lymph nodes, collection of full-thickness intestinal biopsy specimens, determination of intestinal cellular proliferation, and collection of aortic and portal venous blood samples for determination of arteriovenous amino acid concentrations across the intestine. RESULTS: Administration of MTX was associated with severe enterotoxicosis manifested as diarrhea (8/12 cats), vomiting (12/12), and positive results for bacterial culture of mesenteric lymph nodes (12/12) in cats receiving the purified diets, independent of glutamine supplementation. Diet did not affect villus tip length and villus surface area in the small intestine or cellular proliferation. Administration of MTX was associated with significantly increased intestinal permeability, which was not attenuated by glutamine supplementation. CONCLUSIONS: Feeding of a glutamine-supplemented amino acid-based purified diet was unable to preserve intestinal function in cats with MTX-induced enteritis. CLINICAL RELEVANCE: Intestinal morphologic alterations correlate poorly with intestinal function as measured by means of bacterial translocation and intestinal permeability.     

Mitotane treatment in a dog with a recurring adrenocortical carcinoma--a case report]

In a 7 year old female poodle an adrenocortical tumor was diagnosed on basis of laboratory and ultrasonographic examinations. One year after adrenalectomy, a relapse was diagnosed, at that time the suspicion of metastases in the liver arose for the first time. By treatment with Mitotane in a dose aiming at completely destroying the adrenal cortex, a complete disappearance of the tumor as well as a dramatic reduction of the size of the metastases could be achieved. 12 months after the begin of the chemotherapy, the dog is in good general condition.     

Evaluation of scratches as an essential element in the development of avian cellulitis in broiler chickens.

Currently, the published cellulitis models do not adequately address the actual pathogenesis as seen in the commercial broiler industry. In this model, small dermal scratches were made on the skin of broiler chickens, which were then placed on litter seeded with avian cellulitis-associated Escherichia coli. The research confirms scratches are required for the induction of avian cellulitis. The research also confirms that "type I" cellulitis lesions or those previously thought to be due to hatchery-borne infections can be induced with scratches. The described methods provide a realistic model for cellulitis development that will improve the reliability of prophylactic and therapeutic-regimen efficacy testing data, thereby providing information more directly useful to the commercial broiler industry.     

A PCR based method for the identification of equine influenza virus from clinical samples.

In this paper we describe the development of a nested RT-PCR assay for the rapid diagnosis and characterisation of influenza virus directly from clinical specimens. Viral RNA is extracted from nasal swabs by the guanidine thiocyanate extraction method, and subsequently reverse transcribed. The complementary DNA is then used as template in a nested PCR reaction. Primers designed for use in this assay are specific for three templates; (1) the nucleoprotein (NP) gene, (2) the haemagglutinin gene of the H7N7 equine influenza virus (A1), and (3) the haemagglutinin gene of the H3N8 equine influenza virus (A2). We show that the assays are specific for the target genes chosen, and display sensitivity similar to virus isolation. The NP assay detects a variety of different influenza subtypes, whereas A1 and A2 assays are specific for influenza subtypes H7N7 and H3N8, respectively. Sequencing of amplicons obtained in the A2 assay yields information on antigenic regions of the haemagglutinin molecule, and use of this procedure in the routine surveillance of equine influenza will enable tentative characterisation of circulating viruses despite difficulties in isolating field strains of the H3N8 subtype. The A1 assay will be useful in ascertaining whether viruses of the H7N7 subtype still circulate amongst horses, or whether these are extinct.     

Mycobacterium paratuberculosis detection in bovine feces is improved by coupling agar culture enrichment to an IS900-specific polymerase chain reaction assay.

The feasibility of coupling an agar culture enrichment step with gene amplification (ACE-PCR) as a means to improve turnaround time and detect Mycobacterium paratuberculosis (Mpt) in the presence of contaminants was investigated. Fecal samples from 463 Pennsylvania dairy cows were cultured in duplicate sets. One replicate from each set was processed and interpreted according to standard culture (SC) protocol, whereas cultures from the second replicate were harvested at 6 weeks postinoculation; DNA extracts from the harvested material were evaluated by a polymerase chain reaction (PCR) test for the Mpt-specific IS900 gene. One hundred seventy-six of 463 culture sets were positive by either method. One hundred sixty-five of these (94%) were ACE-PCR positive, and 151 (86%) were positive by SC. Eleven SC-positive samples were ACE-PCR negative, and 9 ACE-PCR-positive samples were negative by SC; these discrepancies could be a consequence of a low organism burden (< or = 5 organisms/g) or slow growth rate of Mpt in cultures of these samples. One hundred thirty-nine of 463 culture sets (30%) were reported as inconclusive because of culture contamination according to SC protocol; 16 of these (11.5%) were ACE-PCR positive. Seventy-four ACE-PCR-positive sets (42% of all positives) were negative or inconclusive by SC at 6 weeks postinoculation. Agar culture enrichment prior to IS900 PCR testing significantly improves Mpt culture turnaround time and sensitivity.     

Microsatellite variation between an African and five European taurine breeds results in a geographical phylogenetic tree with a bison outgroup

Genetic variation is being used extensively for individual identification and linkage analysis, and may be useful for interpopulation studies. Previously, blood groups and biochemical variants in blood cell and serum proteins have been used to study (evolutionary) relationships in mammals. But genetic divergence and gene flow among closely related populations are difficult to measure with these classical markers because their mutation rate is so low that new mutations have not had sufficient time to appear and become fixed. So they have a small number of alleles and a relatively low level of heterozygosity. These markers are now replaced by DNA markers, mostly microsatellites. These microsatellite loci are useful genetic markers at which alleles differ in length due to differences in the number of short sequence motifs arranged adjacent to one another. They are abundantly distributed throughout the mammalian genome. They have a large number of alleles, a high level of heterozygosity and are inherited in true Mendelian fashion. These characteristics make them valuable for parentage control, linkage analysis, genome mapping and phylogenetic studies. In terrestrial vertebrates with limited mobility, genetic differentiation often increases with the distance between populations or corresponds to the extent of geographic and habitat barriers (R oy et al. 1994). Investigations of short tandem repeats yield a considerable volume of genetic data regarding the similarities and divergence times of different cattle populations. Microsatellite markers are suitable for the estimation of these parameters as they are not generally subject to direct selection and environmental influences. Computation of genetic distances based on data from several loci can be used to evaluate the taxonomic relationship between populations. The aim of this study was to estimate the relative genetic variability between Belgian cattle breeds and to reconstruct the evolutionary relationship among them, also using two small genetically isolated cattle-like populations.     

Some Pharmacokinetic Parameters of Oxfendazole in Sheep

Soraci, A.L., Mestorino, N. and Errecalde, J.O., 1997. Some pharmacokinetic parameters of oxfendazole in sheep. Veterinary Research Communications, 21 (4), 283-287