Scrapie surveillance in Great Britain: results of an abattoir survey, 1997/98

A randomised sample of 2,809 apparently healthy sheep, 55 per cent of them less than 15 months of age, which were slaughtered for human consumption at abattoirs in Great Britain in 1997/98, was taken to establish the prevalence of scrapie infection. The medulla oblongata of each sheep was examined histopathologically at the level of the obex, and fresh brain tissue was examined for scrapie-associated fibrils (SAF) to establish whether there was evidence of scrapie. In addition, histological sections of the medulla from 500 of the sheep were immunostained with an antiserum to PrP, and the same technique was also applied to any animal found positive or inconclusive by the histological or SAF examinations. Any sheep which was positive by any of these diagnostic methods was also examined by Western immunoblotting, for the detection of the disease-specific protein PrP(Sc). A total of 2,798 sheep (99.6 per cent) were negative by all the methods applied. Ten animals were SAF-positive but negative by all the other methods, and in one animal there was immunohistochemical staining which could not be interpreted unequivocally as disease-specific. A mathematical model was used to estimate the prevalence of scrapie infection in the national slaughtered sheep population which would be consistent with these results. By this model, the absence of unequivocally substantiated cases of scrapie in the sample was consistent with a prevalence of infection in the slaughter population of up to 11 per cent.     

Monte Carlo simulation of surveillance strategies for scrapie in Norwegian sheep

Our aim was to compare the efficiency of different surveillance strategies for detecting scrapie-infected sheep flocks in the Norwegian population using simulation modelling.The dynamic Monte Carlo simulation model has the flock as the unit. The input parameters include properties of the sheep population (number of flocks, flock size, age distribution, reasons for culling, breeds, prion protein-allele distribution); properties of scrapie (genotype-dependent infection rate and incubation periods, and age- and genotype-dependent prevalence of scrapie); properties of the surveillance strategy (selection of sheep for examination, period in which infected sheep are detectable, and properties of the diagnostic tests). For simplification, the prion protein-alleles were grouped into three allele groups: VRQ, ARR, and ARQ' (ARQ' represents ARQ, ARH and AHQ).Through either abattoir surveillance or surveillance of fallen stock, 70% of the detected sheep (compared to 33% in the underlying population). The model output was sensitive to the susceptibility of infection for the genotype ARQ'/ARQ'. The effect was large for abattoir surveillance (increased susceptibility increased the efficiency of abattoir surveillance).     

Transmission of sheep scrapie to elk (Cervus elaphus nelsoni) by intracerebral inoculation: final outcome of the experiment.

This is a final report of an experimental transmission of sheep scrapie agent by intracerebral inoculation to Rocky Mountain elk (Cervus elaphus nelsoni). It documents results obtained in experimental (n = 6) and control (n = 2) elk. During the first 2 years postinoculation (PI), 3 animals died or were euthanized because of infection or injuries other than spongiform encephalopathy (SE). In years 3 and 4 PI, 3 other inoculated elk died after brief terminal neurological episodes. Necropsy of these animals revealed moderate weight loss but no other gross lesions. Microscopically, characteristic lesions of SE were seen throughout the brain and spinal cord, and the tissue was positive for proteinase K-resistant prion protein (PrPres) by immunohistochemistry (IHC) and by Western blot. Scrapie-associated fibrils (SAF) were observed by negative-stain electron microscopy in the brain of elk with neurologic signs. PrPres and SAF were not detected in the 3 inoculated elk necropsied during the first 2 years or in the 2 control animals. Retrospective analysis of the gene-encoding cervid PrP revealed a polymorphism at codon 132. The elk with SE were either homozygous (MM) or heterozygous (LM). These findings confirm that intracerebral inoculation of sheep scrapie agent results in SE with accumulations of PrPres in the central nervous system of elk. Based on morphologic and IHC findings, the experimentally induced SE cannot be distinguished from chronic wasting disease of elk with currently available diagnostic techniques.     

Longitudinal study of hepatitis E virus infection in Spanish farrow-to-finish swine herds

Hepatitis E is a zoonotic disease and is highly prevalent in European swine livestock. There is a need to compare the infection dynamics of hepatitis E virus (HEV) between herds with the same production system and determine the percentage of animals that could arrive infected at slaughter age. Therefore, a longitudinal study was performed in six Spanish farrow-to-finish affected farms. Twenty piglets per farm were monitored from nursery to slaughter. RT-PCR and serology techniques were applied to analyze longitudinally collected sera and/or faecal samples. Liver and bile samples were also taken at the abattoir. Anti-HEV IgM were firstly detected at 7 weeks of age in 5 farms whereas at 13 weeks of age in 1 farm (farm 2). At slaughter age 50–100% of pigs had seroconverted to anti-HEV IgG in the former 5 farms whereas in the other herd only 5% of pigs were IgG seropositive (farm 2). Six out of 96 livers and 5 out of 80 biles analyzed were HEV positive at the abattoir (total percentage of infected animals: 11.5%). All these positive animals had already seroconverted except 2 pigs of farm 2. Hence, pigs can be seronegative at slaughter age being infected during the latest fattening period. Manipulation of HEV-infected livers or other organs from pigs could be considered a possible route of transmission in Spanish abattoirs. This study represents the first longitudinal survey on swine HEV infection dynamics conducted in different herds.     

Diagnosis of preclinical and subclinical scrapie in a naturally infected sheep flock utilizing currently available postmortem diagnostic techniques.

Scrapie is a naturally occurring transmissible encephalopathy of sheep and goats. Currently available methods for diagnosis are the presence of characteristic histopathologic changes and detection of an abnormal form of prion protein (PrPres) in the brains of affected animals. This study documents preclinical and subclinical scrapie in a flock of 16 sheep utilizing histopathology, immunohistochemistry (IHC), western blot, and electron microscopy (for scrapie-associated fibrils) for confirmation of the disease. Prior to necropsy, none of the sheep showed signs of clinical scrapie. Based on the results of histopathology and positive PrPres tests, 3 ewes were found to have subclinical scrapie. An additional ewe, which did not have histopathologic changes in the brain but was positive by IHC and western blot,was considered a preclinical case of scrapie. None of the sheep had amyloid in the brain stem.     

Failure to detect abnormal prion protein and scrapie-associated fibrils 6 wk after intracerebral inoculation of genetically susceptible sheep with scrapie agent

Detection of the scrapie-associated protease-resistant prion protein (PrP^res) in sheep brains in the early phase after intracerebral inoculation of the scrapie agent has not been documented. Fourteen 4-mo-old, genetically susceptible lambs (QQ homozygous at codon 171 of the PrP gene) were obtained for this study. Twelve lambs were inoculated intracerebrally with a brain suspension from sheep naturally affected with scrapie, and 2 served as uninoculated controls. Two inoculated animals were euthanized at each of 6 times postinoculation (1 h to 6 wk), and their brains were collected for histopathological study, for detection of PrP^res by the Western blot technique and an immunohistochemical (IHC) method, and for the detection of scrapie-associated fibrils (SAF) by negatively stained electron microscopy (EM). Microscopic lesions associated with introduction of the inoculum were seen in the brains of inoculated animals at all 6 times. However, both the Western blot and IHC techniques did not detect PrP^res after the initial 3 d postinoculation, nor did EM detect SAF in any of the samples. From these findings, it is presumed that until host amplification has occurred, the concentration of PrP^res in inoculum is insufficient for detection by currently available techniques.     

Salmonella in the mesenteric lymph nodes of healthy sows and hogs

Mesenteric lymph nodes were collected from healthy sow and slaughter hogs that had been kept in holding pens for an average of 10 and 2 days, respectively, before slaughter at an abattoir. Salmonella was isolated from 58.2% (67/115) of the sows and 31.3% (16/51) of the slaughter hogs. This difference in Salmonella prevalence was significant (chi 2 = 10.22, P less than 0.005); therefore, the hypothesis that there would be no significant difference in the infection rates of sows and slaughter hogs was rejected.     

Phenotypic and genotypic traits of Staphylococcus aureus strains isolated from pig carcasses

A total of 142 S. aureus strains isolated from pig carcasses from abattoirs A (n = 98) and B (n = 44) were characterized by phenotypic and genotypic traits. Phenotypically, 96% showed yellow-pigmented colonies, 63% β/δ hemolysis, 85% were egg yolk-positive and 99% were positive for clumping factor/protein A. Only 25% of the strains were resistant to the antimicrobials tested (abattoir A: 19%; abattoir B: 39%), especially to penicillin and ampicillin. None of the strains harbored the mecA gene, which is conserved in methicillin-resistant S. aureus. The biofilm associated genes icaA, icaD and bap were present in 100%, 100% and 0% of the strains. Genes for staphylococcal enterotoxin (SE) were detected in 51% (abattoir A) and 14% of the strains (abattoir B). Among strains harboring SE genes (n = 56), 63%, 31%, 4% and 2% tested positive for seg/sei, seg, sei and sec, respectively. The amplification of the 3′ end of the coagulase gene (coa) yielded amplicons of 400, 436, 602, 682 or 776 bp. Coa restriction profile (CRP) analysis using HaeIII resulted in seven patterns (a–d, e1–e3). CRP (c) was detected most frequently at both abattoirs, whereas CRP (a) was restricted to abattoir A and CRP (e3) to abattoir B. In the slaughter process (abattoir B), (i) two CRPs (b and d) were only found before dehairing/singeing, and (ii) four CRPs (c, e1–e3) were identified throughout the process. The genotyping revealed a remarkable homogeneity in S. aureus strains from the two different abattoirs and the slaughter process stages. These results may be explained by the distribution of a limited number of S. aureus genotypes in the pig population. Moreover, as the predominant CRPs (c, e1–e3) persisted throughout the slaughter process in abattoir B, it may be hypothesized that these types are characterized by colonization advantages.     

Micro ELISA and indirect haemagglutination tests in the diagnosis of Taenia hydatigena metacestode infections in lambs

Of 48 lambs slaughtered at a local abattoir, 60.4% were either found to be infected with Taenia hydatigena cycts, or showed evidence of abortive infections (white spots on the liver) with that parasite. The micro ELISA test and the IHA test did not distinguish between animals showing evidence of infection at slaughter and those which did not. Both tests gave results which correlated significantly with each other. The antigens used in the tests cross-reacted with each other, which may be the reason for the large number of false negative results.     

Some observations on the liver copper status of pigs in the northern part of New Zealand

The copper content of the livers from 347 pigs was analysed. Two hundred and three of these samples were from animals varying in age from full term foetus to five years, which were submitted to the laboratory (laboratory cases) for a variety of diagnostic tests. The remaining 144 liver samples were obtained from pigs slaughtered at an abattoir (abattoir survey) at 75-85 kg liveweight (approximately 20-30 weeks old). Liver copper levels of 12 mg/kg D.M. (Dry Matter) or less, a level consistent with copper deficiency, were found in 6.9% of the laboratory cases and in 9.0% of the abattoir survey pigs. Liver levels less than 20 mg/kg suggesting insufficient dietary copper to produce a growth-promoting effect, were found in 21.6% of the former and 36.1% of the latter pigs. A significantly greater frequency of illthrift and anaemia was found in the laboratory cases in which liver copper levels were less than 20 mg/kg compared with animals with higher levels. Liver copper levels less than 20 mg/kg were also more common in pigs derived from dairy farms or fed garbage than in animals from pig farms or fed on meal. All of 26 samples of commercial pig meal analysed contained sufficient copper to satisfy the daily essential copper requirements of pigs (greater than 5-6 mg/kg) but 14 of these (53.8%) contained less than 125 mg/kg levels of copper and were unlikely to produce a growth-promoting effect.     

Some observations on the liver copper status of pigs in the northern part of New Zealand

The copper content of the livers from 347 pigs was analysed. Two hundred and three of these samples were from animals varying in age from full term foetus to five years, which were submitted to the laboratory (laboratory cases) for a variety of diagnostic tests. The remaining 144 liver samples were obtained from pigs slaughtered at an abattoir (abattoir survey) at 75-S kg liveweight (approximately 20–30 weeks old).

Liver copper levels of 12 mg/kg D.M. (Dry Matter) or less, a level consistent with copper deficiency, were found in 6.9% of the laboratory cases and in 9.0% of the abattoir survey pigs. Liver levels less than 20 mg/kg suggesting insufficient dietary copper to produce a growth-promoting effect, were found in 21.6% of the former and 36.1% of the latter pigs.

A significantly greater frequency of illthrift and anaemia was found in the laboratory cases in which liver copper levels were less than 20 mg/kg compared with animals with higher levels. Liver copper levels less than 20 mg/kg were also more common in pigs derived from dairy farms or fed garbage than in animals from pig farms or fed on meal.

All of 26 samples of commercial pig meal analysed contained sufficient copper to satisfy the daily essential copper requirements of pigs (greater than 5–6 mg/kg) but 14 of these (53.8%) contained less than 125 mg/kg levels of copper and were unlikely to produce a growth-promoting effect.     

Prevalence of Salmonella in Ethiopian cattle and minced beef

To estimate the prevalence and distribution of Salmonella in the chain from cattle to the consumer, faeces, mesenteric lymphnodes and beef cuts from 235 cattle, stool samples from 300 workers of the same Addis Ababa abattoir, and 330 minced beef samples from supermarkets in Addis Ababa were analyzed. Isolated Salmonella strains were serotyped and tested for antibiotic susceptibility. Low prevalence in faeces and lymphnodes, and higher contamination rates of beef cuts (diaphragm, abdominal muscles) indicate severe cross-contamination during slaughter. Animals of poor health status were far more frequently carriers of salmonellae, which stresses the need of intensive ante-mortem inspection on slaughter animals. During transport from slaughterhouse to the supermarkets, production and selling of minced beef, the prevalence of Salmonella did not increase.     

Distribution and genetic characterization of porcine Campylobacter coli isolates

The aim of this study was to evaluate the impact of the slaughter process on the Campylobacter (C.) coli prevalence on pig carcasses and finally pork. To detect C. spp., faecal samples, organ samples and surfaces of slaughter pigs were sampled. Additionally, various abattoir surfaces (n=208) and 227 pork and minced meat samples were included in our study.Whereas a high C. spp. prevalence (64.0%) was detectable in the faeces of slaughter pigs (all isolates were identified as C. coli), low detection rates were observed on pig carcasses after the slaughter process before the chilling period (21.1%).The impact of chilling reduced the detection rate of C spp. on pig carcasses even further to 0.8%. Only C. jejuni strains were isolated after the chilling process. Chilling and the associated drying of the skin are responsible for that massive reduction of C. spp prevalence. Significantly more C. spp. were isolated from livers compared to the corresponding carcasses. Only 5 out of 208 swab samples from different surfaces of the abattoir were C. coli positive. Bacteriological investigation could not detect any C. spp. strains from pork and minced pork meat.The low detection rates at the end of the slaughter and processing line indicate that pork may only play a minor role in the transmission of C. coli infections to humans. By genotyping C. coli-isolates from selected animals we were able to demonstrate three possible ways of contamination of the slaughter carcass surface. Genetically highly related strains were detectable on carcass surfaces of consecutively slaughtered animals. Faecal isolates and isolates from the carcass surface showed occasional high similarities. C. coli-genotypes from tonsils and genotypes from the corresponding slaughter carcasses formed a close cluster.     

The prevalence of leptospirosis and its association with multifocal interstitial nephritis in swine at slaughter.

An abattoir survey was undertaken to determine the prevalence of leptospirosis and its association with lesions of multifocal interstitial nephritis (so-called "white spotted kidneys") in swine at slaughter. Both cross-sectional and case-control study designs were used. Of 197 kidneys from hogs randomly selected at slaughter, 11 (5.6%) had generalized grey-white foci typical of multifocal interstitial nephritis (MFIN). Antibody titers greater than or equal to 1:80 against Leptospira pomona were detected in nine (4.6%) hogs and against L. bratislava in 63 (32%) of these hogs. Leptospira pomona (kennewicki) was detected by immunofluorescence in 5/197 (2.5%) of randomly selected hogs. Leptospires identified as genotype kennewicki were isolated from six (9.8%) of 61 kidneys cultured. Leptospira bratislava was not detected by immunofluorescence or culture. There was a highly significant (p = 0.00) and strong association (odds ratio (OR) = 195) between high L. pomona titer (greater than or equal to 1:80) and the presence of leptospires in the kidneys, as detected by culture. There was also a significant (p = 0.046) and strong (OR = 8.10) association between multifocal interstitial nephritis and the presence of renal leptospires as detected by culture. The association between leptospiral titer and MFIN lesions in the prevalence survey group of animals was statistically significant (p = 0.031), but this association was not significant in the case-control study group (p = 0.071) The failure to identify L. bratislava despite serological evidence of infection suggests that some of these seropositive animals may have been transiently infected at an early age, that serological findings were falsely positive, or that immunofluorescence and isolation attempts failed to detect L. bratislava if they were indeed present in the kidneys.(ABSTRACT TRUNCATED AT 250 WORDS)     

Samonellosis and meat hygiene: red meat.

The association between salmonellosis in man and the infection in food animals has been clearly established. There is, moreover, little doubt that abattoir by-products, effluent and solid waste may allow the recycling of infection in animals. The potential hazard posed by salmonellosis to human and animal health will be reduced only by a greater control over the slaughter of infected farm livestock, improved isolation and casualty slaughter accommodation, a stricter control of slaughterhouse hygiene and the provision and full utilisation of adequate laboratory facilities for the bacteriological examination of meat and the abattoir environment.     

Adaptation and evaluation of a rapid test for the diagnosis of sheep scrapie in samples of rectal mucosa.

In recent publications, it was shown that disease-associated prion protein (PrP(d)) accumulates in the lymphoid tissue of the rectal mucosa of a high proportion of scrapie-infected sheep at clinical and preclinical stages, regardless of several host factors; PrP(d) can also be detected in biopsy specimens of rectal mucosa, with an increased probability proportional to age or incubation period and with an efficiency almost identical to that of tonsil biopsies. Rectal biopsies have the advantages of providing higher numbers of lymphoid follicles and of being simpler to perform, which makes them suitable for scrapie screening in the field. In biopsy samples, PrP(d) could be demonstrated by immunohistochemical (IHC) and Western immunoblotting methods, and the purpose of the present study was to optimize and evaluate a "rapid test" for the diagnosis of scrapie in rectal biopsy samples. The HerdChek CWD (chronic wasting disease) antigen EIA (enzyme immunoassay) test was chosen and, once optimized, provided specificity and sensitivity figures of 99.2% and 93.5%, respectively, compared with IHC results in the same samples obtained at a postmortem. The sensitivity of the assay increased from 82.1%, when a single rectal mucosa sample was tested to 99.4% for those sheep in which 3 or more samples were analyzed. Similarly, sensitivity values of the HerdChek CWD antigen EIA test on biopsy samples increased from 95% to 100% for sheep subjected to 1 or 2 sequential biopsies 4 months apart, respectively. Thus, a preclinical diagnosis of scrapie in live sheep can be achieved by a combination of a simple sampling procedure, which can be repeated several times with no detrimental effect for the animals, and a rapid and efficient laboratory method.     

Genital pathology of feral male goats

Genitalia from 1000 feral male goats derived from western Queensland and New South Wales were examined after slaughter at an abattoir and the prevalence of abnormalities determined. Ulcerative balanoposthitis, considered due to caprine herpesvirus infection, was observed in 11 animals (1.1%); acidophilic intranuclear inclusions were found in 7 of these. Other conditions included focal hypoplasia of seminiferous tubules in 2 bucks (0.2%), segmental aplasia of the epididymis (one buck, 0.1%), bulbourethral gland cysts with contained aggregations (33 bucks, 3.3%) and haemangiosarcoma of the bulbourethral gland in one animal. The low prevalence of several conditions such as spermatic granuloma, cryptorchidism, and testicular hypoplasia, was attributed largely to the fact that the bucks examined were horned so that the recognised association between genital abnormalities and polledness did not apply.     

Prevalence of Fascioloides magna in cattle and deer in Michigan

A survey among 1.12 million cattle slaughtered in 357 packing plants in Michigan during 1977 to 1981 was conducted to determine the prevalence of liver fluke infection. The condemnation rate of liver fluke-infected livers was 0.41, 3.7, and 13.9% in the southern, northern-lower, and upper regions of Michigan, respectively. The same trend in infection rates was observed in white-tailed deer that had been examined in diagnostic laboratories in the state. A serologic survey among 50 randomly selected dairy farms detected Fascioloides antibody-carrying cattle in 3 farms. The data on prevalence in slaughter cattle were slightly biased, because many slaughter cattle originated from other states in which F magna may be less common.     

Second passage of sheep scrapie and transmissible mink encephalopathy (TME) agents in raccoons (Procyon lotor)

To determine the transmissibility and pathogenicity of sheep scrapie and transmissible mink encephalopathy (TME) agents derived from raccoons (first passage), raccoon kits were inoculated intracerebrally with either TME (one source) or scrapie (two sources-each in separate groups of raccoons). Two uninoculated raccoon kits served as controls. All animals in the TME-inoculated group developed clinical signs of neurologic dysfunction and were euthanatized between postinoculation month (PIM) 6 and 8. Raccoons in the two scrapie-inoculated groups manifested similar clinical signs of disease, but such signs were observed much later and the animals were euthanized between PIM 12 and 18. Necropsy revealed no gross lesions in any of the raccoons. Spongiform encephalopathy was observed by use of light microscopy, and the presence of protease-resistant prion protein (PrPres) was detected by use of immunohistochemical (IHC) and Western blot analytic techniques. Results of IHC analysis indicated a distinct pattern of anatomic distribution of PrPres in the TME- and scrapie-inoculated raccoons. These findings confirm that TME and sheep scrapie are experimentally transmissible to raccoons and that the incubation periods and IHC distribution for both agents are distinct. Therefore, it may be possible to use raccoons for differentiating unknown transmissible spongiform encephalopathy (TSE) agents. Further studies, with regard to the incubation period and the pattern of PrPres deposition by use of IHC analysis in bovine spongiform encephalopathy and for other isolates of scrapie, chronic wasting disease, and TME in raccoons are needed before the model can be further characterized for differentiation of TSE agents.     

Comparison of two immunochromatographic assays and the indirect immunofluorscence antibody test for diagnosis of Trypanosoma cruzi infection in dogs in south central Louisiana

Two rapid tests evaluated in dogs considered to be of high risk of infection with the Chagas parasite Trypanosoma cruzi using two immunochromatographic assays: Trypanosoma Detect™ for canine, InBios, Seattle, WA and CHAGAS STAT-PAK™ assay, Chembio Diagnostic Systems, Medford, NY, in south central Louisiana. For this purpose a serological survey was carried out in a total of 122 dogs and a serum bank was created. These 122 animals were first tested by IFAT that was used as the standard test. From the serum bank 50 samples were tested using the two rapid Chagas assays and results compared to the standard test IFAT. The serological survey using IFAT showed a prevalence of T. cruzi infection in 22.1% of the tested dogs. In the immunochromatographic assays, 13 and 11 animals were positive on rapid assay: Trypanosoma Detect™ for canine, InBios and CHAGAS STAT-PAK™, Chembio Diagnostic Systems, respectively compared to 11 positive by IFAT. These two immunochromatographic tests have shown high susceptibility and specificity compared to our standard method IFAT. The rapid, easy and accurate screening assays used in conjunction with confirmatory tests, would be an excellent tool for veterinarians to diagnose T. cruzi infection. Early detection of T. cruzi infection may prevent complications through an effective treatment. Greater awareness by veterinarians of the risk, clinical findings, history along with diagnostic methods will contribute greatly to an understanding of the true prevalence of Chagas disease in dogs in Louisiana.