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.     

Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra S. Lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite

Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained.The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue.Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.     

Composted biosolids as a source of iron for hybrid poplars (<Emphasis Type="Italic">Populus</Emphasis> sp.) grown in northwest New Mexico

Composted sewage sludge (biosolids) supply plant available Fe and may represent a sustainable alternative to more costly chelated Fe fertilizers currently used to supplement nutrition in hybrid poplar test plots of elevated soil pH. To test the response of poplars, field plots were amended with composted biosolids at two agricultural rates: 22.75 and 44.5 Mg ha⁻¹. Iron EDDHA served as a fertilizer check and control plots received no amendment. The hybrid poplar OP-367 (Populus deltoides × P. nigra) was planted on a 3.6 m grid spacing. Significant amounts of P and Fe originating from the sewage treatment process were detected in soils 13 months after amending. Chlorosis evaluated with a SPAD-502 meter, showed that poplars amended with biosolids remained the least chlorotic and had greater tree growth when compared to Fe EDDHA and control plots during two growing seasons. Biosolids show promise as a cost effective alternative for the remediation of Fe chlorosis in hybrid poplar agroforestry plantations and present new opportunities in northwestern New Mexico for municipalities seeking solid waste land disposal options.     

Core terrestrial habitat for conservation of local populations of salamanders and wood frogs in agricultural landscapes

Pond-breeding amphibians require aquatic and terrestrial habitats to complete their lifecycles, and preservation of both habitats is necessary for maintaining local populations. Current wetland regulations focus primarily on aquatic habitats, and criteria to define critical upland habitats and regulations to protect them are often ambiguous or lacking. We examined the association between the presence of seven pond-breeding amphibian species and the landscape composition surrounding 54 wetlands located within the Till Plains and the Glaciated Plateau ecoregions of Ohio, USA. We quantified landscape composition within 200 m of the wetland (“core terrestrial zone”) and the area extending from 200 m to 1 km from the wetland (“broader landscape context zone”). We constructed binary logistic regression models for each species, and evaluated them using Akaike Information Criterion. Presence of spotted salamanders (Ambystoma maculatum), Jefferson's salamander complex (A. jeffersonianum) and smallmouth salamanders (A. texanum) was positively associated with the amount of forest within the core zone. Presence of wood frogs (Rana sylvatica) was positively associated with the amount of forest within the core zone and the amount of forest within the broader landscape context zone. Presence of tiger salamanders (A. tigrinum tigrinum) was negatively associated with the cumulative length of paved roads within 1 km of the site, and presence of red-spotted newts (Notophthalmus v. viridescens) was negatively associated with the average linear distance to the five nearest wetlands. Overall salamander diversity was positively associated with the amount of forest within the core zone, and negatively associated with the presence of predatory fish and cumulative length of paved roads within 1 km of the site. Our results confirm the strong association between the structure of surrounding upland areas and amphibian diversity at breeding ponds, and stress the importance of preserving core terrestrial habitat around wetlands for maintaining amphibian diversity.     

Tim50 maintains the permeability barrier of the mitochondrial inner membrane

Transport of metabolites across the mitochondrial inner membrane is highly selective, thereby maintaining the electrochemical proton gradient that functions as the main driving force for cellular adenosine triphosphate synthesis. Mitochondria import many preproteins via the presequence translocase of the inner membrane. However, the reconstituted Tim23 protein constitutes a pore remaining mainly in its open form, a state that would be deleterious in organello. We found that the intermembrane space domain of Tim50 induced the Tim23 channel to close. Presequences overcame this effect and activated the channel for translocation. Thus, the hydrophilic cis domain of Tim50 maintains the permeability barrier of mitochondria by closing the translocation pore in a presequence-regulated manner.     

The postnatal development of the mucosal immune system and mucosal tolerance in domestic animals

The mucosal immune system is exposed to a range of antigens associated with pathogens, to which it must mount active immune responses. However, it is also exposed to a large number of harmless antigens associated with food and with commensal microbial flora, to which expression of active, inflammatory immune responses to these antigens is undesirable. The mucosal immune system must contain machinery capable of evaluating the antigens to which it is exposed and mounting appropriate effector or regulatory responses. Since the immune system is likely to have evolved initially in mucosal tissues, the requirement to prevent damaging allergic responses must be at least as old as the adaptive immune system, and studies of the mechanisms should include a range of non-mammalian species. Despite the importance for rational design of vaccines and for control of allergic reactions, the mechanisms involved are still largely unclear. It is not clear that the classical experimental protocol of "oral tolerance" is, in fact, measuring a biologically important phenomenon, nor is it clear whether tolerance is regulated in the evolutionarily recent organised lymphoid tissue (the lymph nodes) or the more ancient, diffuse architecture in the intestine. The capacity of the immune system to discriminate between "dangerous" and "harmless" antigens appears to develop with age and exposure to microbial flora. Thus, the ability of an individual or a group of animals to correctly regulate mucosal immune responses will depend on age, genetics and on their microbial environment and history. Attempts to manipulate the mucosal immune system towards active immune responses by oral vaccines, or towards oral tolerance, are likely to be confounded by environmentally-induced variability between individuals and between groups of animals.     

Sequence differences in the diagnostic region of the cysteine protease 8 gene of Tritrichomonas foetus parasites of cats and cattle

In order to investigate the genetic variation between Tritrichomonas foetus from bovine and feline origins, cysteine protease 8 (CP8) coding sequence was selected as the polymorphic DNA marker. Direct sequencing of CP8 coding sequence of T. foetus from four feline isolates and two bovine isolates with polymerase chain reaction successfully revealed conserved nucleotide polymorphisms between feline and bovine isolates. These results provide useful information for CP8-based molecular differentiation of T. foetus genotypes.     

Improving the specificity of immunodiagnosis for porcine brucellosis

This report describes the use of cell mediated immunity to improve specificity of current diagnosis for Brucella suis. Diagnosis is problematic due to cross reactions that lead to false positive serological reactions (FPSR) in the standard diagnostic tests. A common cause of this cross reactivity is infection with the organism Yersinia enterocolitica O:9. Gottingen™ mini-pigs were experimentally infected with B. suis biovar I field strain or Y. enterocolitica serotype O:9 biotype 3. Infection was followed for 70 days. During this time whole blood stimulation assays were set up using Brucella specific antigen. IFNγ was measured in the supernatants (SN) from these assays by ELISA. Concurrent standard serological tests were carried out. The results indicate that the IFNγ assay is specifically able to distinguish Y. enterocolitica O:9 infection from a B. suis infection in experimentally infected mini-pigs. These results represent an improvement in diagnostic specificity compared to currently used serological tests. Thus suggesting that in a surveillance setting this test could be applied as a confirmatory test in the face of FPSR. The authors are British Civil Servants and as such their work is subject to British Crown Copyright. This means the exclusive copyright for the article cannot be transferred.