Maximum-likelihood estimation of sensitivity and specificity of ELISAs and faecal culture for diagnosis of paratuberculosis

The accuracy of three diagnostic tests for paratuberculosis was evaluated using maximum-likelihood estimation of sensitivity and specificity. We also explored the variety of estimates that can be obtained if the tests are to be used in populations of different composition with regard to infection and disease states. Two enzyme-linked immunosorbent assays (ELISAs) were evaluated separately with the faecal culture (FC). The study was carried out as a cross-sectional field study to cover all likely states of infection with Mycobacterium avium subsp. paratuberculosis.The three basic assumptions for the maximum-likelihood technique were evaluated to validate the results. Our accuracy estimates for the ELISAs were not very different from those previously published, but those for faecal culture differed if a different cut-off value was chosen for the ELISA. If faecal culture was used for screening in a Danish dairy region where the median ELISA reading was a measure of the general disease situation, the sensitivity of the faecal culture was 20-25%. If faecal culture was used as a confirmatory test on cows with a high ELISA reading (and thus high level of antibodies), the sensitivity of the faecal culture would be in the range 60-70%. These results emphasise the importance of the composition of a target population before selecting a specific diagnostic test for a given purpose. We concluded that faecal culture is useful for confirmation but not for screening purposes.     

Cellulase Supplementation Does Not Improve the Digestibility of a High-Forage Diet in Horses

Six mature Arabian geldings were used in a two-period crossover study to investigate the effects of cellulase supplementation on fiber digestion. Horses were randomly assigned to either a control (CO; n = 3) or a cellulase (CE; n = 3) treatment for the first period and then treatments were switched for period 2. Each period consisted of a 10-day diet adaptation followed by a 3-day total fecal collection. The enzyme mixture contained 40,000 cellulase units/g and was fed at a rate of 3 g/day split evenly between two feedings. During the diet adaptation period, horses had ad libitum access to timothy hay and were also fed 165 g whole oats as a carrier for the supplement. When eating the CO treatment, horses consumed 16% more hay than when on the CE treatment (P = .004). Fecal output also tended to be greater when horses consumed the CO treatment as compared with CE treatment (P = .07). No differences were found between treatments for fecal percent dry matter (DM%), fecal neutral detergent fiber (NDF), fecal acid detergent fiber (ADF), fecal nitrogen (N), or fecal gross energy (GE). There was a trend for horses consuming the CO treatment to digest more NDF than when consuming the CE treatment (34.6% ± 1.5 vs 31% ± 1.5; P = .07). Horses also digested a greater %ADF, %N, and Mcal of energy when consuming the CO treatment than when consuming the CE treatment (P < .05). Cellulase addition to a hay-based horse diet decreased digestion of fiber components.     

DNA‐analysis to monitor fisheries and aquaculture: Too costly?

Evidence from DNA‐analysis is commonplace in human criminal investigations, and while it is increasingly being used in wildlife crime, to date, its application to control and enforcement activities in fisheries and aquaculture has only been sporadic. Contemporary DNA‐analysis tools are capable of addressing a broad range of compliance issues, species identification, mislabelling of fish products, determining the origin of catches and the farm of origin of aquaculture escapees. Such applications have the potential to ensure traceability along the fish product supply chain and to combat consumer fraud and Illegal, Unreported and Unregulated fishing. Nevertheless, DNA‐analysis is not yet used routinely in investigations into compliance with fisheries and aquaculture legislation. One potential reason for this is that DNA‐analysis techniques may have been regarded as too expensive. However, costs have plummeted over the past decade prompting us to objectively assess whether the costs associated with routine use of DNA‐analysis techniques for fisheries and aquaculture control and enforcement activities do constitute an impediment. Based on a number of recent fisheries and aquaculture compliance investigations that incorporated DNA‐analysis, our results indicate that the use of genetic analysis was justified and worthwhile in all cases examined. We therefore conclude that the costs associated with DNA‐analysis do not represent a barrier to the routine adoption of DNA‐analysis techniques in fisheries and aquaculture compliance investigations. Thus, control and enforcement agencies should be encouraged to use such techniques routinely.     

A meta-analysis comparing the effect of vaccines against Mycoplasma hyopneumoniae on daily weight gain in pigs

Our aims were to evaluate the published literature concerning the effect of swine vaccination against Mycoplasma hyopneumoniae on the average daily weight gain (ADWG). This was done by re-evaluating the influence of selected factors on ADWG by a meta-analysis of published studies from 1991 to 1999, fulfilling certain inclusion criteria.With ADWG as the outcome, an analysis of variance was performed for such variables as treatment, vaccination schedule, age during study, housing system and publication quality. Each clinical trial was considered as a random effect and the numbers of pigs in each trial were weightings.Of 63 published studies, 16 describing three commercial vaccines fulfilled the criteria for the meta-analysis. Due to few studies with one of the vaccines (n=3), only two vaccines were included.Vaccinated pigs gained an average of 592g (S.E.=15) with Stellamune and 590g (S.E.=15) with Suvaxyne compared to non-vaccinated pigs that gained an average of 569g (S.E.=14)(P<0.01) when adjusted for age during the study. Vaccine type, vaccination schedule, housing system and publication quality were not significantly associated with ADWG.     

Progression of Mycoplasma hyosynoviae infection in three pig herds. Development of tonsillar carrier state, arthritis and antibodies in serum and synovial fluid in pigs from birth to slaughter

In this investigation, natural infection with Mycoplasma hyosynoviae was followed in groups of individual pigs in three different herds with regard to occurrence of tonsillar carrier state, clinical arthritis and development of antibodies in serum and in synovial fluid. Antibodies were detected by a polyclonal enzyme-linked immunosorbent assay (ELISA) developed for experimental use. The infection with M. hyosynoviae progressed very differently in the three herds investigated. In one herd, the infection was apparently limited to adult pigs. In a second herd, all pigs became tonsillar carriers of M. hyosynoviae, but no mycoplasma-related arthritis nor any serological response was demonstrated within the growing-finishing period. In the third herd investigated, tonsillar infection was detected in all pigs, clinical cases of M. hyosynoviae arthritis followed and a moderate serological response was observed in some, but not all, pigs. In all three herds, M. hyosynoviae infection was carried in the tonsils of the adult pigs, but it was only occasionally transmitted from sows to piglets. Maternal antibodies were transferred to the piglets and persisted for approximately 8-12 weeks. After weaning, some pigs became infected before 20 weeks of age, while others did not. In the majority of cases, the tonsillar infection was established from 11 weeks of age or older. A latent tonsillar infection was present for a period of several weeks within the group of investigated pigs before cases of generalized infection and arthritis were seen. In some cases, generalization of M. hyosynoviae infection in the blood and in joints was observed in spite of the detection of an active serological response a few weeks earlier. The present work suggests that generalization of the infection and development of arthritis may depend on age, immunity, virulence factors and/or infection pressure; in some herds maybe combined with certain triggering mechanisms such as stress and lowered general resistance.     

Protein constituency of unbound fraction of bovine serum applied to anion exchange columns at different pHs and molarities with or without ethylenediamine tetraacetic acid. Application to preparation of bovine IgG1

Ethylenediamine tetraacetic acid disodium salt (EDTA) was found to be an important addition to anion exchange buffers in terms of the profile and amounts of eluted bovine serum proteins. Hydrogen ion concentration and buffer composition were important when different types of anion exchanger (DEAE 52 cellulose and QAE Sephadex A 50) were used for separation of bovine serum proteins as functional groups of the different anion exchangers did not behave similarly and were therefore not interchangeable. These findings applied to the purification of bovine serum IgG1 amounting to 15 to 25% of the total IgG1 was accomplished using QAE Sephadex A 50 anion exchange chromatography. This was followed by absorption of the IgG1 fraction with Staphylococcus aureus containing protein A to remove minor IgG2 contaminants and gel filtration to exclude traces of the third component of complement (C3).     

Gastrointestinal and body growth in colostrum-deprived piglets in response to whey, casein or soy protein diets

Cow's milk stimulates growth in neonates and supplementation with specific milk proteins may benefit immuno-compromized neonates exhibiting poor growth. We investigated the effects of specific milk proteins (casein and whey) and plant protein (hydrolysed soy) on body and organ growth and bone mineralization in colostrum-deprived newborn pigs. Six days after birth, both casein and whey increased body and small intestinal weight relative to soy protein. Villus morphology and epithelial permeability were similar among groups. Casein significantly increased bone mineralization, while whey stimulated soft tissue growth (internal organs, muscle and fat). The differential effects of casein and whey proteins in the early postnatal period show that specific milk proteins rapidly affect whole body and gut development, even in a state of impaired growth induced by artificial feeding of colostrum-deprived pigs.