A proposal for enhanced EU herbage VCU and DUS testing procedures

Herbage production is regarded as having environment-friendly credentials. However, as the ruminant production it supports is facing major challenges on sustainability, environmental footprint and human health concerns, EU herbage cultivar testing must contribute to the solutions. Before new cultivars can be sold in a member state (MS) and gain EU-wide marketing, they must pass official tests to prove they are both novel (distinct, uniform and stable, DUS) with improved value for cultivation and use (VCU). Herbage species present specific challenges, as their allogamy imposes a wide within-cultivar variation that adds complexity to DUS tests and their “value” is only realized in ruminant produce. Current VCU systems measure production, chemical composition and disease/stress tolerances, often on large numbers of candidate cultivars, but prohibitive labour costs and logistics mean that animal intake, ruminant output or environmental benefits cannot be measured directly. Furthermore, some candidate cultivars with proven superior VCU fail DUS even though the non-distinct comparison is with a significantly lower performing registered cultivar. To resolve these problem cases, a “vmDUS” distinctness tool is proposed, which uses molecular markers but conforms to UPOV-declared principles. A short overview of current grassland research shows smart proxy measures of animal value can easily and quickly be adopted into an integrated pan-European (EU-VCU) test network. The proposed EU-VCU scheme will reallocate test resources to conduct these additional tests by placing MS in data sharing collaborations, while retaining their national listing authority. The benefits to all stakeholders from adopting these new testing procedures are considered.     

Micro-environmental conditions modulate protein secretion and infectivity of the Trichinella spiralis L1 larva

After digestion of infected meat the free L1 of Trichinella spp. penetrate the intestinal mucosa where they moult to the mature adult stage. We have used proteomics to identify changes in protein secretion during in vitro culture of free T. spiralis muscle larvae under different environmental conditions, and to correlate these changes with their infectivity in mice. Muscle larvae were cultured in different media (RPMI-1640, C-199 and HBSS) under conditions of anaerobiosis, microaerobiosis and in 5% CO₂ at 37 °C. Following incubation the larval excretory/secretory proteins were analysed by two-dimensional gel electrophoresis and the larvae were used to orally infect naïve CD1 mice. For all culture media tested, infectivity of the L1 was preserved following incubation in anaerobic conditions. In contrast, the infectivity of worms cultured in nutrient-rich media was almost completely abolished in both microaerobiosis and in the presence of 5% CO₂. Some infectivity was retained in poor or reduced culture media. Comparative analysis of larval infectivity and protein secretion showed that loss of infectivity correlated with the appearance of non-tyvelosylated proteins that in turn may be related to the onset of moulting.     

Architecture of succinate dehydrogenase and reactive oxygen species generation

The structure of Escherichia coli succinate dehydrogenase (SQR), analogous to the mitochondrial respiratory complex II, has been determined, revealing the electron transport pathway from the electron donor, succinate, to the terminal electron acceptor, ubiquinone. It was found that the SQR redox centers are arranged in a manner that aids the prevention of reactive oxygen species (ROS) formation at the flavin adenine dinucleotide. This is likely to be the main reason SQR is expressed during aerobic respiration rather than the related enzyme fumarate reductase, which produces high levels of ROS. Furthermore, symptoms of genetic disorders associated with mitochondrial SQR mutations may be a result of ROS formation resulting from impaired electron transport in the enzyme.     

A COMPARISON OF DIATRIZOATE* AND IOXAGLATE† FOR POSITIVE CONTRAST SHOULDER ARTHROGRAPHY IN DOGS

In six experimental dogs, arthrographic quality and synovial inflammatory response with shoulder arthrography comparing meglumine-sodium diatrizoate (Urovison) and the monoacidic dimer, meglumine-sodium ioxaglate (Hexabrix) was evaluated. In our study initial films were of equally high diagnostic quality for both contrast media, but delayed films significantly favored ioxaglate for diagnostic quality. The rise in white blood cells in synovial fluid samples collected 24 hours after the arthrographic procedure was significantly lower after the use of ioxaglate. Histologic examination performed 14 days after the intra-articular injection revealed no drug related lesions.     

Comparison of methods to detect gastrointestinal parasites in llamas and alpacas

OBJECTIVE: To compare relative sensitivity and overall yields of various methods of fecal examination for gastrointestinal parasites in llamas and alpacas. DESIGN: Prospective study. SAMPLE POPULATION: Fecal samples from 42 alpacas and 62 llamas. PROCEDURES: Fecal samples were analyzed via direct smear, a modified McMaster technique with sucrose solution or saturated saline (approx 36% NaCl) solution, and a centrifugation-flotation procedure. McMaster flotation chambers were examined 15 and 60 minutes after loading. Centrifugation-flotation samples were examined after 10 and 60 minutes of flotation. The proportions of samples with positive results and concentrations of parasites were compared among methods. RESULTS: The centrifugation-flotation technique yielded more positive results than other methods for all parasites except small coccidia. Longer flotation time increased the proportion of positive results and parasite concentrations for all parasites except Nematodirus spp. Longer time in the McMaster chamber made little difference. By use of the modified McMaster technique, sucrose solution yielded more positive results for Trichuris spp, Eimeria macusaniensis, and strongyles, whereas saline solution yielded more positive results for Nematodirus spp and small coccidia. The saline solution McMaster test yielded more positive results for small coccidia than did most other methods, and the sucrose McMaster technique yielded more positive results for Trichuris spp. CONCLUSIONS AND CLINICAL RELEVANCE: The centrifugation-flotation technique appeared to offer clear advantages in detecting infection with E macusaniensis, Trichuris spp, Nematodirus spp, and capillarids. The saline McMaster technique appeared to offer an advantage in detecting small coccidia.     

Calf health from birth to weaning. III. housing and management of calf pneumonia

Calfhood diseases have a major impact on the economic viability of cattle operations. A three part review series has been developed focusing on calf health from birth to weaning. In this paper, the last of the three part series, we review disease prevention and management with particular reference to pneumonia, focusing primarily on the pre-weaned calf. Pneumonia in recently weaned suckler calves is also considered, where the key risk factors are related to the time of weaning. Weaning of the suckler calf is often combined with additional stressors including a change in nutrition, environmental change, transport and painful husbandry procedures (castration, dehorning). The reduction of the cumulative effects of these multiple stressors around the time of weaning together with vaccination programmes (preconditioning) can reduce subsequent morbidity and mortality in the feedlot. In most studies, calves housed individually and calves housed outdoors with shelter, are associated with decreased risk of disease. Even though it poses greater management challenges, successful group housing of calves is possible. Special emphasis should be given to equal age groups and to keeping groups stable once they are formed. The management of pneumonia in calves is reliant on a sound understanding of aetiology, relevant risk factors, and of effective approaches to diagnosis and treatment. Early signs of pneumonia include increased respiratory rate and fever, followed by depression. The single most important factor determining the success of therapy in calves with pneumonia is early onset of treatment, and subsequent adequate duration of treatment. The efficacy and economical viability of vaccination against respiratory disease in calves remains unclear.     

Measurement of serum immunoglobulin concentration in killer whales and sea otters by radial immunodiffusion

Killer whales and sea otters maintained in captivity are the subjects of routine health monitoring programs, and interest in immunologic studies in sea otters has been rising recently in response to potential impacts from infectious disease and environmental pollution on the threatened southern sea otter population. Development of species-specific reagents for immunologic studies in these two marine mammals is currently in its infancy. In this study, killer whale and sea otter immunoglobulin-specific polyclonal antibodies were generated, and used to develop tests for serum Ig concentration in the killer whale (Orcinus orca) and the southern (Enhydra lutris nereis) and northern sea otter (Enhydra lutris lutris). Killer whale serum IgG was purified using caprylic acid/ammonium sulfate precipitation. Sea otter plasma IgG was purified using protein-A-agarose. Polyclonal anti-Ig antisera were produced in rabbits, and specificity confirmed by immunoelectrophoresis. Radial immunodiffusion was used to measure Ig concentration in serum or plasma samples derived from 21 captive killer whales, 18 wild and 4 captive southern sea otters and 15 wild and 4 captive northern sea otters grouped by age. Mean killer whale serum Ig concentration (+/-95% confidence interval) ranged from 15.04 +/- 3.97 g/l for animals aged 0-5 years to 26.65 +/- 9.8 g/l for animals aged >10 years. Mean sea otter serum Ig concentration (+/-95% confidence interval) ranged from 28.39 +/- 11.00 g/l for southern sub-adults to 32.76 +/- 11.58 g/l for southern adults. No significant difference in serum Ig concentration was found between southern and northern sea otters. Serum Ig concentrations in two northern sea otter pups were low compared to those of adult sea otters. The two serum Ig quantitation assays produced were highly specific and reproducible and will be useful additions to the limited number of tests available for immune function in these marine mammal species.     

Chemiluminescent immunoassay in comparison with the indirect ELISA as reference method for detecting Salmonella antibodies in swine meat juice

The efficiency of chemiluminescent immunoassay (CLIA) in detecting Salmonella antibodies in the meat juice of slaughter swine was compared with the indirect ELISA (BgVV method). Based on the screening test results of 987 meat juice samples obtained from different laboratories in Germany, a good level of agreement between the two systems was obtained with a kappa value of 0.824 at 20% cut-off and 0.798 at 40% cut-off. At 20% and 40% cut-off levels, a sensitivity of 96.2% and 97.3%, respectively, and a specificity of 94.6% and 95.1%, respectively, were demonstrated between CLIA and ELISA. The detecting LPS antigen was tested for specificity and a cross-reaction with two E. coli and Yersinia strains was found when tested with ELISA. This reaction was not observed in CLIA, possibly because of the broader measurement spectrum of this test which allows a more distinctive definition of immunologic reactions. The same explanation can be given for the increased number of meat juice samples which were positively detected only in ELISA but not in CLIA. The positively classified samples in screening were further tested for reciprocal titers in both test systems, and a higher correlation between screening and titration results was obtained for CLIA. Towards the end of the study, a preliminary comparison of CLIA with two available commercial ELISA test kits was conducted and the same tendency was observed, namely, wider detection range of CLIA compared to the other tests. Based on the results of this study, CLIA can be used as a reference method in detecting Salmonella antibodies in the meat juice of slaughter pigs.     

The Role of Saprotrophy and Virulence in the Population Dynamics of Botrytis cinerea in Vineyards

ABSTRACT Change in relative frequencies of the three main genetic types of Botrytis cinerea (Group I, Group II vacuma, and Group II transposa) were monitored over time from 1998 to 2000 in three Bordeaux vineyards not treated with fungicides. During 2000, Group I isolates, detected mostly at flowering comprised only 2.5% of the entire population. Within Group II, the complementary frequencies of vacuma and transposa isolates differed significantly depending on grapevine phenological stages and organs. Every year and at all sites, including one noble rot site, transposa isolates dominated at every stage, particularly on overwintering canes and at harvest (greater than 86.7% on berries). The complementary frequency of vacuma isolates reached a maximum on senescing floral caps (between 23.5 and 71.4%) and then decreased significantly until harvest on leaves and berries. In pathogenicity tests on grape berries, transposa isolates were significantly more virulent than were vacuma isolates. Mycelial growth rate was negatively correlated with virulence, notably on leaves in transposa and with double resistance to the fungicides carbendazim and vinclozolin. In vacuma, this double resistance was positively correlated with virulence on leaves. Change in the vacuma and transposa frequencies was most likely caused by differences in saprotrophic and pathogenic fitness. Possible interactions between fungicide resistance profiles and fitness are discussed.