The role of phagocytic cells in enhanced susceptibility of broilers to colibacillosis after Infectious Bronchitis Virus infection

Colibacillosis results from infection with avian pathogenic Escherichia coli bacteria. Healthy broilers are resistant to inhaled E. coli, but previous infection with vaccine or virulent strains of Infectious Bronchitis Virus (IBV) predisposes birds for severe colibacillosis. We investigated whether IBV affects recruitment and function of phagocytic cells and examined NO production, phagocytic and bactericidal activity, and kinetics of peripheral blood mononuclear cells (PBMC) and splenocytes. Moreover, we measured cytokine mRNA expression in lung and spleen samples. Broilers were inoculated with IBV H120 vaccine or virulent M41 and challenged 5 days later with E. coli 506. A PBS control and E. coli group without previous virus inoculation were also included. Birds were sacrificed at various time points after inoculation (h/dpi). Inoculation with IBV induced extended and more severe colibacillosis than with E. coli alone. At 4dpi, the number of KUL-01(+) PBMC in all E. coli-inoculated groups was significantly higher than in PBS-inoculated birds, which correlated with lesion scores. From 1 to 4dpi, NO production by PBMC from all E. coli-inoculated animals was elevated compared to PBS birds. Bactericidal activity of PBMC in IBV-inoculated animals at 7dpi was lower than in PBS- and E. coli-inoculated birds, but phagocytic capacity and recruitment were not severely impaired. In spleen samples of IBV-infected animals reduced expression of IL-1beta, IL-6, IL-8, IL-10, IL-18 and IFN-gamma mRNA was found 1dpi. Our results suggest that enhanced colibacillosis after IBV infection or vaccination is caused at least by altered innate immunity and less by impairment of phagocytic cell function.     

Urinary aldosterone to creatinine ratio in cats before and after suppression with salt or fludrocortisone acetate

Background: The endocrine diagnosis of primary hyperaldosteronism in cats currently is based on an increased plasma aldosterone to renin ratio, which has several disadvantages for use in veterinary practice. Objectives: To establish a reference range for the urinary aldosterone to creatinine ratio (UACR) and to determine whether oral administration of either sodium chloride or fludrocortisone acetate is effective for use in a suppression test. Animals: Forty‐two healthy cats from an animal shelter and 1 cat with primary hyperaldosteronism from a veterinary teaching hospital. Methods: Morning urine samples for determination of the basal UACR were collected from 42 healthy cats. For the suppression tests, urine samples for the UACR were collected after twice daily oral administration for 4 consecutive days of either sodium chloride, 0.25 g/kg body weight (n = 22) or fludrocortisone acetate, 0.05 mg/kg body weight (n = 15). Results: The median basal UACR was 7.2 × 10^?9 (range, 1.8–52.3 × 10^?9), with a calculated reference range of <46.5 × 10^?9. Administration of sodium chloride resulted in adequate salt loading in 10 of 22 cats, but without significant reduction in the UACR. Administration of fludrocortisone resulted in a significant decrease in the UACR (median, 78%; range, 44–97%; P < .001) in healthy cats. In the cat with an aldosterone‐producing adrenocortical carcinoma, the basal UACR and the UACR after fludrocortisone administration were 32 × 10^?9 and 36 × 10^?9, respectively. Conclusions and Clinical Importance: Using the UACR for an oral fludrocortisone suppression test may be useful for the diagnosis of primary hyperaldosteronism in cats.     

The use of a genetic-counselling program by Dutch breeders for four hereditary health problems in boxer dogs

Our group developed a genetic-counselling program for boxer-dog breeders in The Netherlands, using data for cryptorchidism (uni- and/or bilateral), epilepsy, knee-problems (including ligament rupture, fractured or ruptured meniscus, severe osteo-arthrosis of the knee, or a combination of these disorders), and schisis (including cheiloschisis, palatoschisis, or cheilopalatoschisis). We transformed the estimated breeding values (EBVs) into odds ratios (ORs), to enable the breeder to compare the risk for each of the traits for a certain dam-sire combination with the average population risk (set at 1). The goal of the study was to evaluate the use of our genetic-counselling program by Dutch breeders of boxer dogs. We asked breeders of the Dutch Boxer Club to send in an application form for genetic-counselling from June 1 to December 1, 2000. Breeders indicated on this application form three desirable sires for their dam (sire 1, sire 2, sire 3) in random order. On the basis of this information, a counselling report was produced which included ORs for the four diseases in litters of the dam-sire combinations indicated on the application form. Together with the counselling report, the breeders received an evaluation form. We received 129 application forms from 70 breeders, and collected 125 evaluations. Of these evaluations, 96 were informative about the influence of the counselling report on sire choice. The most-important criteria used by breeders to select sires were: the exterior characteristics (60%) and known progeny (52%). Although it was the first time breeders could make use of genetic-counselling, 32% of the breeders indicated that the genetic-counselling played a major role in their sire selection. Breeders expressed little difference in importance for the four genetic traits, but there was a tendency to consider epilepsy more than the others. Breeders hesitated to put long-term population interest above short-term personal interest. Nevertheless, the general conclusion of this study is that breeders acknowledge the value of counselling, especially to reduce the frequency of a threatening disease such as epilepsy.     

TB diagnosis in non-human primates: comparison of two interferon-gamma assays and the skin test for identification of Mycobacterium tuberculosis infection

To examine the effect of recombinant bovine interferon-gamma (rbIFN-gamma) on cattle persistently infected with bovine leukemia virus (BLV), BLV-infected cattle were inoculated intraperitoneally with IFN-gamma. All cattle were febrile after inoculation with IFN-gamma and then recovered within 48 h. Flow cytometric analysis showed that the numbers of CD4+ and CD8+ T cells were decreased for 2-3 days and then their numbers were recovered. The number of gammadelta T cells increased after the fever. In contrast, the number of IgM+ lymphocytes remained low for about 1 week. Moreover, the numbers of syncytia produced by peripheral blood lymphocytes decreased and remained low compared to that before IFN-gamma administration. These results suggest that IFN-gamma induces the up-regulation of gammadelta T cells, decreases the number of IgM+ lymphocytes and suppresses the growth of BLV in BLV-infected cattle in vivo.     

DNA testing for type III von Willebrand disease in Dutch Kooiker dogs

Von Willebrand disease type III is widespread in Dutch Kooiker dogs. To eradicate von Willebrand disease from the breed, affected dogs and nonsymptomatic carriers must be excluded from breeding. Previous efforts to detect carriers in Kooiker dogs by a von Willebrand factor antigen assay were not satisfactory because of considerable overlap of plasma concentrations in normal dogs and carriers. The aim of this study was to develop and apply a DNA test for the detection of von Willebrand disease carriers in the Kooiker breed. Two mutations in the von Willebrand factor gene in affected Kooiker dogs have been described previously, a splice site mutation at the border of intron 16 and exon 16 and a missense mutation in exon 3. We have developed polymerase chain reaction tests for both mutations in genomic DNA. The missense mutation most likely is a neutral variant and appears to be a polymorphism present in many breeds. The allele-specific oligonucleotide test for the splice site mutation was applied in the selection of animals cleared to breed by the Dutch breeding club. In a few years, the mutation has been eliminated from the breeding stock without apparent increase of inbreeding or preferential sire usage.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Longitudinal development of kinematics in shetland ponies and the influence of feeding and training regimes

Nowadays, ponies are used more and more for competitive equine sports, driving and racing. The locomotor performance of ponies, however, has been evaluated in only a few kinematic and kinetic studies. To compare the longitudinal development of locomotor characteristics in the pony to the same process in the horse, the kinematics of 23 Shetland ponies were recorded at the trot on a treadmill (3 m/s) at ages 4 and 30 months using a modified CODA-3 apparatus. Furthermore, the influence of specific feeding and training regimes on this development was evaluated. It appeared that stride and stance duration increased with age, but the swing duration and pro/retraction range were similar at foal and mature age. Fore- and hindlimb maximal joint flexion at swing was larger in the mature ponies compared to foals. The elbow and shoulder joint were kept in a more extended position and the stifle joint more flexed, while scapula and pelvis were kept in a more vertical position. These effects are similar to those found in horses. The majority of both temporal and angular kinematic variables correlated significantly between foals and mature ponies. Ponies that were restricted in their feeding, resulting in a low body condition score, demonstrated a flatter gait than ponies fed ad lib (4 cm more height, 60 kg more weight), which trotted at the same velocity with significantly more joint flexion. Extra sprint training on top of free paddock exercise did not significantly influence the longitudinal development of kinematics. From these data, it can be concluded that the longitudinal development of locomotor characteristics in ponies is similar to that in horses. Fatter animals show a more animated gait, while extra sprint training added to free paddock exercise has no detectable effect on the development of kinematics. Aside from training or feeding regimes, in the Shetland pony breed, it appears possible to recognise 'good movers' at a young age.     

Heparinised blood ionised calcium concentrations in horses with colic or diarrhoea compared to normal subjects

Our objectives were to 1) establish ionised calcium (ICa), C-terminal PTH and biologically active PTH (intact molecule) concentrations in blood from normal horses, 2) examine the stability of ionised calcium and acid-base values in stored equine heparinised blood and serum and 3) check the applicability of the formulas based on these parameters in certain disease states. Mean +/- s.d. % ionised calcium in heparinised blood of normal Warmbloods was 51 +/- 2.7 (n = 20) of total calcium, range 1.45-1.75 mmol/l (n = 15) at Michigan State University and 1.43-1.69 mmol/l (n = 20) at Utrecht University. Mean +/- s.d. EDTA plasma concentration for intact +/PTH in normal horses measured 0.6 +/- 0.3 pmol/l (n = 11). Both mean serum and the heparinised blood ionised calcium concentrations changed (not significantly) after 102 h storage at room temperature. Six cycles of freezing and thawing did not affect serum ionised calcium concentration significantly. Ionised calcium concentration and pH in heparinised blood of 20 normal Warmbloods were used to calculate the regression equation for the prediction of the adjusted ionised calcium concentration to a pH of 7.4. The linear regression equation found was: adjusted plasma ICa at pH 7.4 mmol/l = -6.4570 + 0.8739 x (measured pH) + 0.9944 x (measured ICa mmol/l). By means of this formula, mean adjusted ionised calcium concentration in heparinised blood calculated was 100% of the actual value given by the analyser in the normal horses. When using this formula in horses with colic or diarrhoea, mean adjusted ionised calcium concentration was underestimated by 0.2 and 0.3%, respectively. Furthermore, to adjust the measured ionised calcium concentration in heparinised blood to a pH of 7.4 in healthy as well as in 2 groups of diseased horses 2 formulas with a good prediction are now available.