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Detection of cattle-derived BSE prions using transgenic mice overexpressing bovine PrP(C).

In interspecies transmissions of transmissible spongiform encephalopathies, the agent has to overcome a species barrier that is largely influenced by the rate of homology between the prion proteins (PrP(C)) of the two involved species. Generating transgenic mice expressing PrP(C) of a foreign species is an approach to develop TSE models that are at least partly devoid of a species barrier. The availability of such animals would enable the detection of low doses of infectivity in tissues or bodily fluids derived from other species. We generated transgenic mice that overexpress bovine PrP(C) (Tgbov XV mice) for the development of an improved detection assay for cattle derived BSE prions. These mice succumbed to the disease 250 days after inoculation with a brain homogenate from BSE diseased cattle. Diagnosis of BSE in transgenic mice was confirmed using Western blot, as well as histological and immunohistochemical methods. In contrast, transgenic mice overexpressing murine PrP(C) (tga20 mice) did not display shorter incubation times than nontransgenic RIII mice infected with the same inoculum. The expression of a chimaeric PrP of murine and bovine sequences rendered such mice partly, if not entirely resistant to an infection with the BSE agent.

Animals↗

Optimizing replacement of dairy cows: modeling the effects of diseases.

We modified an existing dairy management decision model by including economically important dairy cattle diseases, and illustrated how their inclusion changed culling recommendations. Nine common diseases having treatment and veterinary costs, and affecting milk yield, fertility and survival, were considered important in the culling decision process. A sequence of stages was established during which diseases were considered significant: mastitis and lameness, any time during lactation; dystocia, milk fever and retained placenta, 0-4 days of lactation; displaced abomasum, 5-30 days; ketosis and metritis, 5-60 days; and cystic ovaries, 61-120 days. Some diseases were risk factors for others. Baseline incidences and disease effects were obtained from the literature. The effects of various disease combinations on milk yield, fertility, survival and economics were estimated. Adding diseases into the model did not increase voluntary or total culling rate. However, diseased animals were recommended for culling much more than healthy cows, regardless of parity or production level. Cows in the highest production level were not recommended for culling even if they contracted a disease. The annuity per cow decreased and herdlife increased when diseases were in the model. Higher replacement cost also increased herdlife and decreased when diseases were in the model. Higher replacement cost also increased herdlife and decreased the annuity and voluntary culling rate.

Animal Husbandry↗

[The role anaerobes in dermatitis digitalis et interdigitalis in cattle].

Altogether 52 tissue samples and swabs from the corium of the claw of diseased cattle were investigated. Microbiological examinations revealed the presence of 11 strictly anaerobic bacterial strains that belonged to the species Peptococcus asacharolyticus, Pc. sacharolyticus, Peptostreptococcus anaerobius, Bacteroides asacharolyticus and B. melaninogenicus. The remaining isolates were classified as Bacteroides, Fusobacterium, Streptococcus and Clostridium. In the above-mentioned strains observations of their biochemical, morphological and enzymatic properties (Tab. I) were carried out. Their resistance to antibiotics was evaluated from the qualitative viewpoint (Tab. II). Capillary isotachophoresis was used to determine the amounts of volatile fatty acids that developed during glucose fermentation (Tab. III). In the single clinical samples the above-mentioned strains occurred as association of microorganism. The anaerobic genera Bacteroides and Fusobacterium were found in 52% of the samples (Bacteroides sp. and Fusobacterium sp. were recorded in 25% and 12% of the samples, respectively, whereas in 15% of the samples both species occurred). According to our results the etiology of dermatitis digitalis et interdigitalis is closely connected with the incidence of anaerobes. Of the latter, those belonging to the genera Bacteroides and Fusobacterium are of major importance.

Animals↗

Increased prevalence of Mycoplasma bovis in the Netherlands.

The epidemiology, therapy, and prevention of M. bovis infections are briefly reviewed. In a survey begun in 1982, M. bovis was found frequently in the respiratory tract [corrected] of veal calves and beef cattle with respiratory problems. In replacement calves infected with respiratory disease in dairy herds, however, the organism has only been detected since 1986. Respiratory tract specimens collected from calves with respiratory disease were submitted for examination for M. bovis from 1986 to 1991 and originated from 83 herds. Mycoplasma bovis was detected in specimens from 59 of the herds, 20% of which were dairy herds and 80% fattening herds. Arthritis caused by M. bovis was observed in 12 herds until July 1991. Since 1976 when the first mastitis outbreak caused by M. bovis was diagnosed, M. bovis has caused 14 more outbreaks. The number of diseased cattle varied from 1 tot 16 per farm, and clinical signs of mastitis varied from mild to severe. In all instances the infection has been eradicated from the herds. Because M. bovis can cause great losses in intensively reared cattle herds, it is advisable to separate purchased veal calves and beef cattle from dairy cattle to prevent further spread of M. bovis.

Animals↗

Development of disabled, replication-defective gene transfer vectors from the Jembrana disease virus, a new infectious agent of cattle.

Jembrana disease virus (JDV) is a newly isolated and characterised bovine lentivirus. It causes an acute disease in Bali cattle (Bos javanicus), which can be readily transmitted to susceptible cattle with 17% mortality. There is as yet no treatment or preventive vaccine. We have developed a gene transfer vector system based on JDV that has three components. The first of the components is a bicistronic transfer vector plasmid that was constructed to contain cis-sequences from the JDV genome, including 5'- and 3'-long terminal repeats (LTRs), 0.4kb of truncated gag and 1.1kb of 3'-env, a multiple cloning site to accommodate the gene(s) of interest for transfer, and an internal ribosome entry site plus the neomycin phosphotransferase (Neo) gene cassette for antibiotic selection. The second element is a packaging plasmid that contains trans-sequences, including gag, pol, vif, tat and rev, but without the env and packaging signals. The third is a plasmid encoding the G glycoprotein of vesicular stomatitis virus (VSV-G) to supply the vector an envelope for pseudotyping. Cotransfection of 293T cells with these three plasmid components produced VSV-G pseudotyped, disabled, replication defective, bicistronic JDV vectors encoding the green fluorescent protein (EGFP) and the Neo resistance selection maker simultaneously with a titre range of (0.4-1.2)x10(6)CFU/ml. Transduction of several replicating primary and transformed cells from cattle, primate and human sources and importantly growth-arrested cells with the JDV vectors showed high efficiency of EGFP gene transfer at 35-75%, which was stable and the expression of EGFP was long term. Furthermore, these JDV vectors were designed to suit the inclusion and expression of genes corresponding to JDV specific proteins, such as gag or env, for the development of vaccines for Jembrana disease. This strategy should also be applicable to other bovine diseases as well. The design and construction of the JDV vector system should facilitate the study of the lentivirology and pathogenesis of the diseases associated with JDV or other bovine virus infections. To our knowledge, this is the first such vector system developed from a cattle virus.

Animals↗

Adenovirus-mediated type I interferon expression delays and reduces disease signs in cattle challenged with foot-and-mouth disease virus.

Foot-and-mouth disease (FMD) is an economically important disease of livestock. Eliminating FMD outbreaks in previously disease-free countries often relies on restriction of animal movement and massive slaughter of infected and in-contact susceptible animals. To develop a more effective and humane FMD control strategy, we explored the possibility of using type I interferon (IFN-alpha/beta) as a novel anti-FMD agent. We have demonstrated previously that swine inoculated with replication-defective human adenovirus type 5 (Ad5) vector expressing porcine IFN-alpha (Ad5-PoIFN-alpha) were completely protected from FMD virus (FMDV) challenge. To extend this approach to bovines, we constructed Ad5 vectors that express bovine IFN-alpha or IFN-beta (Ad5-BoIFN-alpha and Ad5-BoIFN-beta). Cells infected with these viruses produced high levels of biologically active BoIFN-alpha/beta, but despite expression in vitro, no detectable IFN-induced biologic activity was found in cattle inoculated with Ad5-BoIFN-alpha. Because PoIFN-alpha inhibits FMDV replication in bovine cells, we evaluated the potential use of PoIFN-alpha against FMD in cattle. In cattle inoculated with Ad5-PoIFN-alpha, the appearance of vesicles was delayed after challenge with FMDV and disease was less severe than in control animals. One Ad5-PoIFN-alpha-inoculated animal never developed clinical disease. Similarly, although all the Ad5-PoIFN-alpha-inoculated animals developed viremia, it was delayed for 1 day as compared with the control group. These results suggest that in vivo expression of PoIFN-alpha partially protected cattle from FMD.

Adenoviridae↗

Clinical features of experimentally induced rabies in cattle and sheep.

A total of 20 cattle and five sheep out of a larger group of animals that were experimentally challenged with virus as part of the required protocol for a vaccine trial developed clinical signs of rabies. All five sheep and 18 of the cattle tested positive for rabies in a direct fluorescent antibody (FA) test. The remaining two cattle had suspicious FA results. Prospective observations are reported in this study. In the diseased cattle, the average incubation period was 15.1 days and the average morbidity period was 3.7 days. Of those, the naive cattle had significantly shorter incubation and morbidity periods than the test-vaccinated cattle. Major clinical signs included excessive salivation (100%), behavioural change (100%), muzzle tremors (80%), vocalization (bellowing; 70%), aggression, hyperaesthesia and/or hyperexcitability (70%), and pharyngeal paresis/paralysis (60%). The furious form of rabies was seen in 70% of the cattle. In the diseased sheep, the average incubation period was 10.0 days and the average morbidity period was 3.25 days. Major clinical signs included muzzle and/or head tremors (80%), aggressiveness, hyperexcitability, and/or hyperaesthesia (80%), trismus (60%), salivation (60%), vocalization (60%) and recumbency (40%). The furious form of rabies manifested in 80% of the sheep. Current rabies vaccines on the market contain higher effective doses than that utilized for the test vaccine and the results of this study do not reflect in any way on commercially available ruminant rabies vaccines.

Animals↗

Antigenic diversity of bovine viral diarrhea-mucosal disease (BVD-MD) viruses recently isolated from persistently infected cattle and mucosal disease, and serologic survey on bovine sera using antigenically different BVD-MD viruses.

Twenty nine recent isolates of bovine viral diarrhea-mucosal disease (BVD-MD) virus, 17 from persistently infected cattle and 12 from mucosal disease, were compared antigenically with the reference strains by a serum neutralization test. The reference viruses were divided into 2 groups, tentatively designated as N and K, based on the antigenic relationships in the cross-neutralization test. Antigenic properties of recent isolates were considerably different with the sources of virus isolation. Seventeen isolates recovered from persistently infected cattle were divided into 3 groups in the neutralization test using antisera to the reference strains; 12 and 2 were considered as the possible members of groups K and N, respectively, and the others belonged to neither group. On the other hand, 10 of 12 isolates recovered from mucosal disease were considered as the possible members of group N, and the others were classified into neither group. Interestingly, none of BVD-MD viruses isolated from cases of mucosal disease belonged to group K. The results of serologic survey on sera collected from 713 cattle at the Hokkaido provinces in 1974 to 1988 indicated that infections of cattle with BVD-MD viruses other than group K were prominent before 1981. Cattle infected with group K BVD-MD virus were first detected in 1982, and increased in number thereafter. The results obtained in this study suggested that BVD-MD viruses with various antigenic properties spread widely among cattle herds, and also a possibility that clinical manifestations in cattle infected with BVD-MD viruses may differ with their antigenic properties.

Animals↗

Technical and economic evaluation method for use in improving infectious animal disease surveillance networks.

With hope of improving the increasing number of epidemiological surveillance networks for animal diseases set up in recent years, a qualitative and quantitative technical and economic evaluation tool was developed and then applied to three epidemiological surveillance networks: RENESA (a French surveillance network for salmonella and mycoplasma contamination in poultry production units subject to official sanitary controls), the French Foot and Mouth Disease Epidemiovigilance Network and REPIMAT (the epidemiological surveillance network in Chad for major cattle diseases). We identified critical points in epidemiological surveillance networks using a modified version of the hazard analysis: critical control point (HACCP) method. An evaluation grid was then developed and validated by experts who were consulted in accordance with the Delphi method. A questionnaire to collect the information required for the evaluation and a scoring guide were then designed. Our evaluation procedure also included a calculation of the annual operating costs for two of the three networks studied. On the basis of the detailed results of the technical and economic evaluation, we formulated specific suggestions for improving the networks. The cost of implementing these proposals was calculated. We then simulated the effects of implementing each of the proposed improvements and a new global evaluation score was determined for each network. The 'cost per point' of each improvement was then calculated and discussed. This tool for the technical and economic evaluation of epidemiological surveillance networks for animal diseases is proposed so that it may be tested on a far wider scale and eventually be used in improving the functioning of such networks and for risk analysis in international trade.

Animals↗

A space-time cluster investigation of an outbreak of acute respiratory disease in Norwegian cattle herds.

Outbreaks of acute respiratory disease occurred in several cattle herds in the south-east regions of Norway during the winter and spring of 1995. The present study was conducted to investigate the temporal and spatial dynamics of the occurrence of acute respiratory disease between January and April 1995 in the affected cattle herds, in two neighbouring counties in Norway. This was done to assess the possible role of an infectious agent in these outbreaks, as well as to investigate the possibility of transmission between neighbouring herds. During the study period, a total of 190 (19.8%) outbreaks occurred in 955 cattle herds located in the defined study area. Descriptive maps of the spatial and spatio-temporal patterns of the outbreak were generated, and two different statistical tests of space-time interaction (the Knox test and k-nearest neighbour test) and a cluster-detection test (the space-time scan statistic) were used. The results of all the space-time interaction tests strongly suggested that the case herds were clustered in time as well as in space. In addition, the space-time scan statistic defined the most-likely cluster to be located within the second veterinary district (Toten where the disease occurred) during 9-15 weeks in 1995. The relative risk of a herd being infected inside the most-likely cluster compared with the surrounding study area was estimated to be 3.59 (p=0.001). The transmission rate between neighbouring farms therefore appeared to have been higher within this cluster than in the surrounding study area. This study supports the hypothesis that one common source of infection was involved in the outbreaks during the study period.

Acute Disease↗

[Isolation of lymphocyte cultures from leukemic cattle and electron microscopic study of the leukemia virus].

Several methods were employed to obtain lymphocyte cultures from blood samples taken from normal cattle and from cattle affected with enzootic leukosis. Biologic and virologic experiments revealed that the cultures from diseased cattle contained an oncogenic leukosis virus, while those obtained from healthy animals were exempt from such virus. Electron microscopy was applied to study the morphologic aspects of the bovine leukosis virus with a total of 18 lymphocyte cultures. The viral particles noted were shown to have typical configuration and size of type C oncogenic viruses as described in the literature--possessing a central dense nucleotide and double membranes. Seen were also various stages in the development of the virus. Mature virus particles were likewise observed in a stable cell line FLK (percent of the virus) as well as in ultrathin cross sections of the spleen of leukosis-affected animals.

Animals↗

A disease monitoring system for dairy herds.

A system for recording and evaluating disease, intended for both dairy herd health monitoring and research purposes, is described. The system was initiated in 1960 with the objective of providing a research database and it has been developed to respond to the herd health management needs of modern farmers and veterinarians. The system is probably one of the oldest disease recording systems in existence and has contributed to the expansion of epidemiological knowledge of dairy cattle diseases and has helped to improve the health management of dairy herds. The history of the system, the herd reporting form and various analyses and reports generated with the system are presented.

Animals↗

A familial degenerative neuromuscular disease of Gelbvieh cattle.

A degenerative skeletal muscle disease with vascular, neurologic, and renal lesions and a probable familial distribution was identified in 4-20-month-old purebred Gelbvieh cattle. Thirteen affected animals were confirmed from 6 separate beef herds, with a mortality rate of 100%. Clinical signs in affected animals consisted of ataxia, weakness, and terminal recumbency. Gross and histologic muscle lesions were indicative of nutritional myopathy of ruminants, with a lack of myocardial lesions in most cases and only rare myocardial changes in a few animals. Acute to chronic lesions in most large skeletal muscle groups consisted of degeneration, necrosis, regeneration, fibrosis, and atrophy. Fibrinoid necrosis of arterioles was a common feature in multiple tissues. Lesions in the spinal cord white matter and peripheral nerves consisted of degeneration of the dorsal columns and axons, respectively. Changes in the kidneys consisted of chronic interstitial nephritis with fibrosis, hyaline droplet change and tubular epithelial vacuolar change and were most severe in the older calves. Intracytoplasmic myoglobin and iron were demonstrated within the hyaline droplets in degenerate renal cortical tubular epithelial cells. Vitamin E levels were deficient in most (6/7) of the animals tested. Investigation of the pedigree of affected animals revealed a common ancestry for all but 1 of the animals whose parentage could be traced. This investigation suggests that a hereditary metabolic defect, possibly involving antioxidant metabolism, could be responsible for this condition. Renal disease, possibly secondary to myoglobinuria, may be unique to this bovine condition.

Animals↗