Development of an indirect competitive ELISA for flumequine residues in raw milk using chicken egg yolk antibodies

To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA-flumequine) and to cationized ovalbumin (cOVA-flumequine). For the immunization of chickens, cBSA-flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA-flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 microg/kg). No cross-reactivity was noticed with various related quinolones.     

Enzymatic Maceration of Witloof Chicory by the Soft Rot Bacteria Erwinia carotovora Subsp. carotovora: The Effect of Nitrogen and Calcium Treatments of the Plant on Pectic Enzyme Production and Disease Development

Disease incidence of bacterial soft rot caused by Erwinia carotovora subsp. carotovora and activities of bacterial pectolytic enzymes were studied in witloof chicory. Disease incidence after forcing of the chicory heads was enhanced by the nitrogen treatment and reduced by the calcium treatment of the chicory plants prior to forcing. Significant differences in susceptibility to bacterial soft rot were found for the tested chicory cultivars. Disease severity after 96h ranged from 6% in Clause R2 to 100% in Bea and Tabor. Chicory cultivars Rumba and Salsa showed a final average severity of 65–70%. Activity of the pectolytic enzymes, polygalacturonase and pectate lyase increased in artificially inoculated chicory heads of cultivar Rumba and was not affected by calcium and nitrogen treatments of the plants. Polygalacturonase showed highest activity 48h after inoculation while pectate lyase activity increased throughout 72h of incubation. Maceration of the chicory tissue and bacterial growth increased continuously until 96h after inoculation, when more than 60% of the chicory heads was macerated by pectolytic enzymes of the bacteria. Enzyme activity of Erwinia carotovora subsp. carotovora grown on chicory cell wall extracts was influenced by nitrogen and calcium treatments of the chicory plants. The activity of polygalacturonase reached its highest level at 48h after incubation and pectate lyase activity peaked at 24h followed by a continuous decrease until 72h after inoculation.     

Studies on neonatal calf diarrhoea caused by rotavirus: transmission of the disease and attempted vaccination of colostrum-deprived calves

Mild to severe scouring could be produced in colostrum-deprived calves with tissue culture-adapted rotavirus and feacal material from field cases of calf diarrhoea. The feaces of experimentally infected calves contained rotavirus for at least 3 days. Pathogenic bacteria were presented in one calf only and this calf also showed the most severe gastroenteritis. Eight calves were vaccinated with a live rotaviral calf diarrhoea vaccine and subsequently challenged with infective rotavirus. Mild scouring was observed after vaccination, but the calves remained normal after challenge. Rotavirus particles were detectable in the faeces for a few days after vaccination and challenge.     

Production, disinfection and evaluation for aquaculture applications of rotifer resting eggs from Bohai Bay, P.R. of China

The dynamics of resting egg production of the rotifer Brachionus plicatilis originating from a wild population in the Tanggu Saltworks (P.R. China) was investigated. In the natural environment as well as in semi-controlled rearing conditions an increased resting egg production was noticed with declining food availability. Processed resting eggs had a hatching efficiency of 3 x 10 6 rotifers per gramme irrespective of their origin. Hatching started 22 h after the initiation of incubation and was completed after 36 h. Rotifers obtained from resting eggs could be further cultured on Culture Selco® and enriched with Super Selco®. The fatty acid profile of these rotifers were not divergent from reference rotifers (originating from the Laboratory of Aquaculture and Artemia Reference Centre) demonstrating that this strain was not catabolizing essential fatty acids and could be used for enrichment purposes. Storage of resting eggs at 4°C resulted in a 50% lower hatching after 1 year but remained stable during the next 2 years. The resting eggs used for storage could easily be disinfected without affecting their hatching characteristics. These results indicate that this material could be used as inocula for mass cultures of live food for commercial hatcheries.     

Transmission of methicillin resistant Staphylococcus aureus among pigs during transportation from farm to abattoir

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in pigs at abattoirs is higher than in pigs sampled on farms. This study investigated whether MRSA negative pigs can become MRSA positive during transportation from the farm to the abattoir after exposure to other pigs and environmental sources of MRSA. Nasal swabs were collected from four batches of pigs during loading at the farm, on arrival at the abattoir and after stunning. Environmental wipes were taken from lorries after transporting pigs and from lairages after holding pigs. All pigs (n = 117) tested MRSA negative before transportation. On arrival at the abattoir, 12/117 (10.3%) pigs in two batches tested MRSA positive. In lorries that tested positive after transportation, the prevalence of MRSA positive pigs was 21.1%, whereas no MRSA was detected in pigs that had been transported in lorries that tested negative after transportation. At stunning, all batches and 70/117 (59.8%) pigs tested MRSA positive. Pigs can become MRSA positive in the short period of time during transportation from the farm to stunning at the abattoir.     

Soil dynamics of the origination of soil tare during sugar beet lifting

High soil tare of sugar beet on wet clay soil after uprooting with share lifters is usually attributed to the fact that the soil becomes sticky due to mechanical impact during uprooting. Results of field experiments have shown good potential for obtaining low soil tare of sugar beet on wet clay soil by beet extraction with a high average vertical acceleration and a spiral extraction path with a small pitch. The objective of this research was to provide models for soil–beet–lifter systems used in these experiments, and to simulate some of the observed effects of the uprooting method on soil tare and soil adherence. As the rootlet system plays an important role in the origination of soil tare, the rootlet system was also modelled. A constitutive soil model, assuming elastic–perfectly-plastic soil behaviour, was used as a basis for modelling soil–beet–lifter system variants, including beet with and without rootlets, uprooting by extraction, rotation and extrusion, and uprooting with low and high vertical acceleration. The origination of soil tare is presented on the basis of the calculated zone of initial soil failure during extraction. Expected soil adherence is discussed in relation to the stress state of the soil. The simulated initial soil failure and stress state of the soil agreed well with experimental results on soil tare and soil adherence.     

Total CK and CK-BB activity in serum from sheep with scrapie.

Total CK and iso-enzyme CK-BB activity was measured in serum from four sheep with scrapie and in serum from four healthy control sheep. Blood samples were taken weekly for about six months. There was a clear overlap between the total CK and CK-BB activity in serum from sheep with scrapie and that in serum from control sheep. Thus measurement of these enzymes does not aid the clinical diagnosis of scrapie.     

Isolation of Cowdria ruminantium by means of Percoll density gradient centrifugation and detection by an enzyme-linked immunosorbent assay

The isolation of Cowdria ruminantium by means of Percoll density gradient centrifugation permits the recovery of partially purified viable populations of the organism possessing distinctly different densities. These conclusions are based upon results of analyses of density fractions by intravenous inoculation into sheep, protein determination, electron microscopy and enzyme-linked immunosorbent assay. Morphological differences were observed in the density fractions obtained from infected brain tissue and Amblyomma hebraeum nymphae.