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A 15-year-old female goat suddenly developed right-sided head tilting with anorexia and depression. Post-mortem examination of the brain revealed a large, unilateral, well-demarcated, intraventricular neoplasm which was diagnosed as a choroid plexus carcinoma. The neoplasm, which occupied about 75% of the left lateral ventricle, led to unilateral obstructive hydrocephalus and invaded the white and grey matter of the left piriform lobe, with focal subarachnoid spread and meningeal implantation. Histopathological examination revealed loss of branching papillary architecture, invasive growth, a high mitotic index and marked necrosis in the undifferentiated areas of the tumour. Neoplastic cells expressed vimentin and, multifocally, broad spectrum cytokeratins, but were negative for GFAP, NSE and Sl00 antigen. This is the first report of a choroid plexus carcinoma in a goat.

Bovine leucosis virus (BLV), circulating in the Ukrainian territory, was characterized through the definition of its subspecies affiliation. The pro-viral BLV DNA was isolated from peripheral-blood lymphocytes of naturally-HIV-infected black-variegate animals taken from leucosis-affected farms in the Kharkov Region. The env-gene fragment of pro-viral DNA was amplified, sequenced and analyzed after the amplicon had been treated by three restriction enzymes, i.e. BamH I, Bcl I and Pvu II. According to the analysis of restriction-fragments' length polymorphism, the Ukrainian BLV isolate can be classified as belonging to the Australian subspecies, i.e. to one of the 3 known subspecies. Multiple alignment and phylogenetic analysis of the env-gene fragment of BLV isolates from the EMBL database showed that evolutionally the Ukrainian isolate is distantly located from the isolates' clusters of the Belgian, Japanese and Australian subspecie and has the biggest quantity (4) of non-coinciding nucleotides for the analyzed highly conservative locus of the BLV env-gene with a length of 444 pair of nucleotides.

A first attempt for the investigation of molecular epidemiology of BLV was carried out. PCR amplicons of a part of the env gene of BLV isolated from 309 cattle of different geographical origin were compared with known BLV env sequences. Using RFLPA most of the PCR products can be assigned to the Australian, the Japanese or the Belgian subgroup. A phylogenetic tree resulting from the comparison of the sequences of these env fragments demonstrates the relations and differences between and within the subgroups.

The genomic structure of the murine Bv8 gene was determined in 129/SvJ mouse, and the chromosomal localization was identified. Bv8 has first been characterized from skin secretion of the yellow-bellied toad, Bombina variegata. When injected into rat brain, this polypetide causes hyperalgesia. The murine Bv8 gene was shown to consist of four exons and was localized on chromosome 6 between the microsatellite markers D6Mit66 and D6Mit36 near the gene mem1, whereas the human counterpart was assigned to the non-syntenic region 3p21.1. Furthermore, the primary Bv8 transcript appeared to be alternatively spliced. The first variant contained all four exons yielding a product with a stretch highly enriched in basic amino acids in its central part. This domain is absent in the peptides from frog as well as in a splice variant expressed in mouse testis. A third variant gives rise to a truncated polypeptide.

Enzootic bovine leukosis is caused by the bovine leukemia virus (BLV) and has a world wide distribution in cattle. Due to the program for eradication of BLV-infections in Germany the BLV incidence in cattle declined and only few new cases seem to occur per year. On the other hand, BLV-infected cattle with low, transient or without BLV-antibody titers are difficult to identify as BLV-infected. These animals may be sources for new infections. It was the aim of this study to compare the suitability of agargel-immunodiffusion (AGID), enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) for diagnosis of BLV-infected cattle. We investigated a herd with 10 cows, where after a long period when the herd was negative suddenly a positive serological reaction appeared. In addition 64 animals from 6 federal states of different herds with doubtful serological reactions found in previous tests were included. In the herd with 10 cows we were able to detect BLV-infection in one animal 8 weeks earlier with PCR than with ELISA. Investigation of 56 adult cattle and 3 calves from different herds with both PCR and ELISA showed that 51 animals were positive in ELISA and 55 in PCR. Seven animal were positive in PCR and negative in ELISA. Three calves yielded negative results in PCR and positive results in ELISA. One cow which was positive in previous serological tests was negative in ELISA, AGID and PCR. Restriction fragment length polymorphism analysis demonstrated that the majority of the cattle was infected with the same BLV provirus variant. The four PCR variants used in this study yielded a similar sensitivity for BLV provirus detection. In conclusion, compared to the serological tests, PCR detects BLV-infection earlier in naturally infected cattle. The method is also a useful tool to exclude or confirm BLV-infection in cattle with doubtful serological results. PCR may be used to complement the serological tests in the diagnosis of BLV-infection.

A typical infection with bovine leukemia virus (BLV) induces a permanent antibody (Ab) response with high titers against BLV-antigens. In the last few years atypical courses of infection with low or transient BLV-Ab-titers or even lack of any detectable BLV-Ab-titers in animals with BLV-provirus integration have been described. This makes it difficult to eliminate BLV infection from herds using serological assays only. Whether or not polymerase chain reaction (PCR) is a useful tool to complement serological Ab-assays in BLV-eradication in herds was clarified in three ways: (i) different DNA-quick-preparations of blood were examined in nested PCR, (ii) cows of a BLV infected herd that was involved in a national eradication program were investigated for 6 months und (iii) BLV-provirus-variants occurring in this herd were differentiated. The results show, that even by using PCR it was not possible to detect all infected animals all the time and that eradication of BLV from this herd was not completed in this short time. The PCR is useful for the investigation of herds and more sensitive than ELISA. PCR using LTR-primers (34 positive cattle) was more sensitive than PCR with env-primers (30 positive cattle). Using PCR 34 BLV infected cattle were detected of which only 21 reacted in ELISA. Restriction enzyme analysis or sequence analysis of PCR-amplificates allowed the detection of virus variants and conclusions about the way of infection. PCR should be used for BLV-eradication in cattle herds with low BLV-incidence, for the investigation of new outbreaks or tumor cases in long term BLV free herds and for investigation of breeding cattle.

BACKGROUND & AIMS: Pathophysiological conditions such as sepsis and hepatitis are mediated by inflammatory cytokines and frequently are associated with cholestasis. The aim of this study was to determine the effect of endotoxin (lipopolysaccharide [LPS]) and cytokine administration on hepatocellular transporters involved in bile salt transport. METHODS: LPS and cytokines were administered to Sprague- Dawley rats or C57BL/6 mice, and the expression and function of hepatocyte transporters involved in bile salt secretion were examined. RESULTS: LPS caused gene expression of the hepatocyte basolateral sodium-dependent taurocholate cotransporter (Ntcp) to decrease by more than 90%. Tumor necrosis factor alpha (TNF-alpha) or interleukin (IL) 1beta also produced a time-dependent decrease in Ntcp messenger RNA levels, whereas IL-6 had no effect. LPS administration resulted in a concordant 90% reduction of basolateral protein expression of the hepatocyte sodium taurocholate cotransporter and markedly diminished sodium-dependent taurocholate uptake. Activity of the hepatocyte basolateral Na+,K+-adenosine triphosphatase (ATPase) was also decreased by 50% in a posttranslational manner after endotoxin treatment. CONCLUSIONS: Endotoxin inhibits hepatocellular sodium-dependent bile salt uptake by decreasing both expression of Ntcp and activity of the Na+,K-ATPase. The effects on Ntcp are mediated via TNF-alpha and IL- 1beta. Alterations of these transporters may contribute to the cholestasis of sepsis and inflammation. (Gastroenterology 1996 Jul;111(1):193-8)

The potential effects of cytokines on hepatocellular transport functions remain undefined. Interleukin-6 (IL-6) is a cytokine that is produced in sepsis, hepatitis, and other inflammatory conditions often associated with cholestasis. Using cultured rat hepatocytes, we have investigated the effects of IL-6 on hepatocellular bile salt uptake. Because hepatocyte Na(+)-K(+)-adenosinetriphosphatase (ATPase) produces the electrochemical gradient that drives sodium-dependent bile salt contransport, we also examined the effects of IL-6 on Na(+)-K(+)-ATPase activity. Hepatocytes cultured for 20 h in media containing IL-6 exhibited a dose-dependent noncompetitive inhibition of [3H]taurocholate uptake, which was maximal at an IL-6 dose of 100 U/ml. IL-6 treatment had no effect on hepatocyte sodium-independent taurocholate uptake. Northern blotting of RNA from cultured hepatocytes revealed that IL-6 had no effect on steady-state RNA levels of the Na(+)-taurocholate transporter (Ntcp). Hepatocytes incubated with IL-6 for 20 h, however, exhibited a 55% decrease in hepatocyte Na(+)-K(+)-ATPase activity. This effect also was dose dependent, with maximal inhibition occurring at an IL-6 dose of 100 U/ml. Similar treatment with IL-6 did not influence hepatocyte Mg(2+)-ATPase activity. The inhibition of Na(+)-K(+)-ATPase activity induced by IL-6 provides a putative mechanism for the observed inhibition of sodium-dependent taurocholate uptake. Since modulation of bile salt transport and Na(+)-K(+)-ATPase activity occurred at IL-6 concentrations comparable to the serum levels observed in patients with severe inflammatory states, these findings have potential pathophysiological relevance for the cholestasis of sepsis and other inflammatory disorders.

During the hunting season from 1991/1992 blood samples were collected from wild boar shot in the Federal States of Sachsen-Anhalt (482 samples) and Brandenburg (177 samples) which corresponds to 2.1 and 0.4% of the total hunting bag. All sera were screened in a complex trapping blocking (CTB) ELISA for antibodies against hog cholera virus (HCV) and in an indirect ELISA for antibodies against Aujeszky's disease virus (ADV). Additionally the sera were tested for neutralizing antibodies against HCV strain ALFORT/187, bovine viral diarrhoea virus (BVDV) strains NADL and 1138/69, and against an ADV field isolate. In case of questionable results sera were tested against HCV strain "BERGEN", HCV vaccine strain "RIEMS" and three HCV field isolates from wild boar. The serological testing for antibodies against "porcine reproductive and respiratory syndrome virus" (PRRSV) was carried out in indirect immunoperoxidase monolayer assay (IPMA). Four sera (Sachsen-Anhalt) reacted positive in CTB-ELISA. Seven sera yielded neutralizing antibodies against HCV but only one of the "non-negative" samples scored positive in both techniques, ELISA and VNT. Two sera (Brandenburg) had low neutralizing antibody titres against Alfort/197 but scored negative in CTB-ELISA. Screening for antibodies against ADV of 640 sera led to 13 positive sera including 5 positive findings in both ELISA and VNT. Antibodies against PRRSV were detected in two sera which were collected Sachsen-Anhalt. Estimations resulted in a prevalence of about 5% for antibodies against HCV.

A highly specific monoclonal antibody (mAb) directed against envelope protein gp51 and effectively bonding the antigen (Ag) on account of its high affinity from an unpurified Ag preparation was chosen for use in a double-sandwich enzyme immuno-assay (EIA) for diagnosis of bovine leukaemia virus (BLV). The epitopes recognised in bovine sera by the gp51-specific antibodies were at the same time properly exposed. Some parameters of major importance to testing were optimised (Ab and Ag quantities, dilution of bovine sera for testing). Preliminary testing of the double-sandwich EIA on selected bovine sera and comparison with both the immunodiffusion test and anti-BLV EIA confirmed its good diagnostic specificity and sensitivity. Hence, this double-sandwich EIA, developed by means of an mAB against gp51, on account of the possibility to use as Ag culture supernatant of the FLC cell line, is a sensitive, low-cost alternative to the anti-BLV EIA Dessau MTP which had so far been used. The double-sandwich EIA is recommended for use in final sanitation for its high analytical and diagnostic sensitivity.

An optimal magnesium supply of dairy cows during pasture period is still a pressing problem in our farms and husbandry. Experiences are described concerning the application of different magnesium concentrated preparations in cases of grass tetany in 1970 to 1975 in the district of Potsdam. In 224 cows with magnesium concentrations values in the serum below 1.8 mg/100 ml a 25% MgSO4-solution, Parevert and Vers. A, a test solution of VEB Tierarzneimittelwerk Leipzig are used as a metaphylactic treatment. Losses by grass tetany could be reduced after application of the named preparations. Also in cases of low dosage a further, at least temporary drop of magnesium values in the serum could be prevented. Metaphylactic measures are recommended for andangered herds.

Two antigenic extracts, prepared by different methods, were tried on 317 serum samples in the microscope-slide agglutination, sedimentation and tube agglutination tests. In comparison with the haematological findings, both antigens and all the tests gave a proportion of non-specific positive results, which made it impossible to obtain specific results. Various ways of reducing the proportion of non-specific reactions were tried, without success.

The use of organophosphorus preparations for the control of ectoparasites and endoparasites of sheep, particularly systemic application, is discussed. Experiments on 13 groups of sheep with five preparations produced in the German Democratic Republic in various formulations and concentrations showed that external application had good contact activity, but little was absorbed. Acetylcholinesterase activity was not inhibited, except by pour-on application of doses several times the normal dose. The preparations were arranged in order of cholinesterase inhibition. In contrast to cattle, diminished cholinesterase activity was unreliable as in indicator of systemic toxicity of organophosphorus preparations in sheep.

Four anticoagulants in current use were compared in blood samples used to prepare dried specimens. The most suitable was a mixture of one part of heparin solution (5000 IU strength) and four parts of distilled water. Blood samples treated in this way must be process at the laboratory within 16 hours. Specimens dried in tubes could be stored for a long time, and this was the best way of dealing with samples from individual or export cattle.

Behaviour of Mg and Ca content in the blood serum of 28 cattle with grass tetany and hypocalcaemia was evaluated at short time intervals after infusion of 500 ml of a solution A (containing 12 g Mg-adipat and 5 g Ca-gluconate/100 ml aqua dest.) and a solution B (containing 12 g Mg-adipate and 12 g Ca-gluconate/100 ml) respectively. After treatment with the last-named solutions there was evident a considerable increase of Mg-values in the blood serum for a longer period (up to 6 h), also in the case of distinct hypomagnesaemia while Ca-values of the circulating blood declined more quickly. Efficacy of treatment was the better the earlier infusion was performed after occurence of clinical signs. Application of solution B for treatment of grass tetany and paresis with tetany like symptoms is recommended for several reasons. Necessity of assurance the Mg and Ca supply after infusion is emphasized.

The anticoagulant Chelaplex III in aqueous solution with two new stabilizers (methenamine and tannin) was tested for the preservation of blood samples, together with another combination composed of oxalic acid and methenamine solution. The eventual aim was to test their suitability for use with blood specimens dried in the tube, but this was not done in the present experiments. It was shown that a combination of 5% oxalic acid with 36% methenamine was at least as good as the conventional Chelaplex III-formaldehyde combination for inhibiting coagulation, and it was relatively cheap.

The use of organophosphorus preparations in the treatment of ectoparasites and endoparasites of pigs is discussed. Treatment of 43 store pigs with preparations produced in the German Democratic Republic by washing and by the pour-on method resulted in considerable inactivation of acetylcholinesterase, which implies good systemic action. Overdosage increased this effect, but without producing signs of toxicity. The oral dosages for pigs cited in the literature were confirmed. Repeated treatment was recommended for the best results, as pigs rapidly break down the preparations. It is necessary to take into account adverse effects of oral administration on palatability of the ration and on weight gain. Addition of a preparation to a batch of food for a group of pigs is likely to be less successful than individual dosing, which is the best method for sows.

International audience; The embryonic lethal phenotype observed when DDK females are crossed with males from other strains results from a deleterious interaction between the egg cytoplasm and the paternal pronucleus soon after fertilization. We have previously mapped the Om locus responsible for this phenotype, called the DDK syndrome, to an approximately 2-cM region of chromosome 11. Here, we report the generation of a physical map of 28 yeast and bacterial artificial chromosome clones encompassing the entire genetic interval containing the Om locus. This contig, spanning approximately 2 Mb, was used to map precisely genes and genetic markers of the region. We determined the maximum physical interval for Om to be 1400 kb. In addition, 11 members of the Scya gene family were found to be organized into two clusters at the borders of the Om region. Two other genes (Rad51l3 and Schlafen 2) and one EST (D11Wsu78e) were also mapped in the Om region. This integrated map provides support for the identification of additional candidate genes for the DDK syndrome.