A method for extracting genomic DNA from individual elaphostrongyline (Nematoda: Protostrongylidae) larvae and differentiation of Elaphostrongylus spp. from Parelaphostrongylus spp. by PCR assay.

This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L1) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L1s of all 6 species of elaphostrongylines, based on polymerase chain reaction (PCR) amplification of a partial fragment of the first internal transcribed spacer (ITS-1) of the ribosomal DNA. Differences were detected in the sizes of partial ITS-1 amplicons between the 2 elaphostrongyline genera, Elaphostrongylus and Parelaphostrongylus. The reliability of the ITS-1 PCR assay was tested by using L1s of unknown identity from Newfoundland and Labrador, Canada. The ability to consistently isolate gDNA from individual L1s, together with a simple PCR-based method to distinguish between Parelaphostrongylus and Elaphostrongylus, have important implications for diagnostic testing and for conducting epizootiological studies on these parasites of veterinary importance.     

Intestinal translocation capabilities of wheat allergens using the Caco-2 cell line

Because intestinal absorption of food protein can trigger an allergic reaction, the effect of wheat proteins on intestinal epithelial cell permeability was evaluated and the abilities of these proteins in native or pepsin-hydrolyzed state to cross the epithelial cell monolayer were compared. Enterocytic monolayers were established by culturing Caco-2 cells, a model of enterocytes, on permeable supports that separate the apical and basal compartments. Proteins were added into the apical compartment, and the transepithelial resistance (TER) was measured; proteins that crossed the cell monolayer were detected in the basal medium by ELISA. Wheat proteins did not alter the cell monolayer. TER and Caco-2 cell viability were conserved, and the passage of dextran was prevented. Native and pepsin-hydrolyzed forms of omega5-gliadin and lipid transfer proteins were detected in the basal medium. The results suggest that these two major allergens in food allergy to wheat were able to cross the cell monolayer by the transcellular route.     

Effect of humic acid on copper adsorption by Kaolin

Varying amounts of humic acid (Na-salt; 0 to 10 ppm) were first equilibrated with a dilute suspension of H⁺ -Kaolin and then with spiked (high activity radioactive ⁶⁴Cu) water. After filtration, through a 0.45 µm Millipore membrane, the amount of radioactivity on the filter and the filtrate was measured separately using a Gamma ray well counter. Adsorption of Cu (expressed as distribution coefficient — K _d ) was plotted against humic acid concentration. K _d values decreased sharply upto 1 ppm humic acid concentration and thereafter remained constant up to 10 ppm.     

Biodegradability Improvement of Sulfamethazine Solutions by Means of an electro-Fenton Process

The main objective of this study was to examine the effect of an electro-Fenton pretreatment on the biodegradability of sulfamethazine-polluted solutions. The aim of the pretreatment was only to degrade this molecule in order to increase the biodegradability of the effluent and therefore allow a subsequent biological treatment. Preliminary tests showed the absence of biodegradability of the target compound. The degradation of sulfamethazine by electro-Fenton process was then examined using a carbon felt cathode and a platinum anode in an electrochemical reactor containing 1?L of solution. The influence of some experimental parameters such as initial concentration, temperature and current intensity on the degradation by electro-Fenton step has been investigated. In addition, the biodegradability of the solution after electrochemical pretreatment was examined and showed a Biological Oxygen Demand (BOD₅) on Chemical Oxygen Demand (COD) ratio above the limit of biodegradability, namely 0.4, for several experimental conditions. The feasibility of coupling an electro-Fenton pretreatment with a biological degradation of by-products in order to mineralize polluted solutions of sulfamethazine was confirmed.     

Influence of the allelic variants encoded at the Gli-B1 locus, responsible for a major allergen of wheat, on IgE reactivity for patients suffering from food allergy to wheat

Wheat presents an important genetic diversity that could be useful to look for cultivars with reduced allergencity. omega5-Gliadins have been described as major allergens for wheat allergic patients suffering from wheat-dependent exercise-induced anaphylaxis (WDEIA) and some cases of chronic urticaria (U). Our objective was to study the influence of genetic variability at the Gli-B1 locus encoding for omega5-gliadins on the reactivity of IgE antibodies from these patients. We selected cultivars expressing 13 alleles at Gli-B1 including a wheat/rye translocation and studied the reactivity to gliadins of a rabbit antiserum specific for omega5-gliadins and of IgE from 10 patients. The antiserum and IgE from nine patients with WDEIA and U strongly detected omega5-gliadins expressed by most of the Gli-B1 alleles but showed no or faint responses to the gliadins and secalins extracted from the translocated wheat. The selection of genotypes lacking the Gli-B1 locus may reduce wheat allergenicity.     

Extended surveillance for CBPP in a free country: Challenges and solutions regarding the potential caprine reservoir

Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease of cattle and buffalo caused by Mycoplasma mycoides subsp. mycoides "Small Colony" (MmmSC). The agent of CBPP has been isolated from goats in different countries including CBPP-free areas. Goats can therefore be regarded as a putative MmmSC reservoir. No diagnostic test for CBPP surveillance in goats has been proposed as yet. Furthermore, serological tests could be seriously hampered by a widespread caprine infection due to the subspecies M. mycoides subsp. capri (Mmc), which is antigenically very close to MmmSC and displays high levels of genetic variability. A competition ELISA (cELISA) is currently used to screen for CBPP in cattle at the herd level in infected areas. The aim of this study was to see if the same cELISA would be specific enough to be used to screen goats despite the potential concomitant infection with Mmc. The cELISA titers of goats from Mmc-infected and non-infected herds were comparable and negative using the accepted cutoff for bovine sera. In contrast, seroconversion was observed in goats experimentally inoculated with an Mmc strain that cross-reacted with a monoclonal antibody targeting the same epitope as that used in cELISA. The probability of such false positivity occurring under field conditions is very low since Mmc strains with such an atypical antigenic profile emerge only rarely as a result of random nucleotide variation of the epitope-coding region. In conclusion, the commercially available cELISA can be considered specific enough to be used as a primary test to monitor passage of the CBPP agent in goats, but its sensitivity in goats requires further investigation.     

Piloting a livestock identification and traceability system in the northern Tanzania–Narok–Nairobi trade route

We designed and piloted a livestock identification and traceability system (LITS) along the Northern Tanzania–Narok–Nairobi beef value chain. Animals were randomly selected and identified at the primary markets using uniquely coded ear tags. Data on identification, ownership, source (village), and the site of recruitment (primary market) were collected and posted to an online database. Similar data were collected in all the markets where tagged animals passed through until they got to defined slaughterhouses. Meat samples were collected during slaughter and later analyzed for tetracycline and diminazene residues using high-performance liquid chromatography (HPLC). Follow up surveys were done to assess the pilot system. The database captured a total of 4260 records from 741 cattle. Cattle recruited in the primary markets in Narok (n = 1698) either came from farms (43.8%), local markets (37.7%), or from markets in Tanzania (18.5%). Soit Sambu market was the main source of animals entering the market from Tanzania (54%; n = 370). Most tagged cattle (72%, n = 197) were slaughtered at the Ewaso Ng’iro slaughterhouse in Narok. Lesions observed (5%; n = 192) were related to either hydatidosis or fascioliasis. The mean diminazene aceturate residue level was 320.78 ± 193.48 ppb. We used the traceability system to identify sources of animals with observable high drug residue levels in tissues. Based on the findings from this study, we discuss opportunities for LITS—as a tool for surveillance for both animal health and food safety, and outline challenges of its deployment in a local beef value chain—such as limited incentives for uptake.     

Erythrocyte pyruvate kinase deficiency in three West Highland white terriers in Ireland and the UK

Erythrocyte pyruvate kinase (PK) deficiency is described for the first time in three apparently unrelated West Highland white terriers (WHWT) from Ireland and the UK. All three dogs were diagnosed with markedly regenerative but persistent anaemia and had been treated for presumed immune-mediated haemolytic anaemia (IMHA) before hereditary erythrocyte PK-deficiency was confirmed by breed-specific DNA mutation analysis. This hereditary erythroenzymopathy causes haemolytic anaemia and affects several canine breeds with varying degrees of severity. Although eventually causing osteosclerosis, haemosiderosis and death, PK-deficient dogs can adapt to their anaemia for many years.PK-deficiency should be considered in anaemic WHWTs worldwide particularly in dogs with haemolytic anaemia where evidence for an immune-mediated, infectious or toxic underlying cause is lacking.     

Effects of management regimes and extreme climatic events on plant population viability in Eryngium alpinum

Extreme climatic events like the 2003 summer heatwave and inappropriate land management can threaten the existence of rare plants. We studied the response of Eryngium alpinum, a vulnerable species, to this extreme climatic event and different agricultural practices. A demographic study was conducted in seven field sites between 2001 and 2010. Stage-specific vital rates were used to parameterize matrix population models and perform stochastic projections to calculate population growth rates and estimate extinction probabilities. Among management regimes, spring grazing and land abandonment decreased vital rates and population growth, while autumn grazing and late mowing had positive effects on population viability. The 2003 heatwave reduced fecundity rates and survival rates. Only spring grazed sites presented considerable extinction risk. Stochastic projections showed that an increased frequency of 2003-like events may exacerbate extinction risk, but extinction probability depends mainly on land management regimes. To better conserve E. alpinum populations, we recommend conversion of presently spring grazed and abandoned sites to late mowing or autumn grazing.