Feeding-unrelated factors influencing the plasma leptin level in ruminants

The triglyceride content of lipid depots associated with the current feeding level is the primary determinant of leptin gene expression and the circulating leptin level. In laboratory rodents and primates the plasma leptin is influenced also by the age, gender and physiological status (puberty, pregnancy, lactation, postpartum period), and by the health condition such as sepsis due to Gram-negative (GN) bacteria. Some pathologic conditions with intensive cytokine release evoke an increase in plasma leptin, which is thought to depress the subsequent feed intake. However, the effect of these secondary factors may be species-dependent, with still unknown clinical relevance in ruminants. In our ovine and bovine models plasma leptin increased after castration and dexamethasone treatment, decreased after experimental administration of synthetic androgens in castrated rams, but remained unchanged throughout the ovarian cycle and after ovariectomy. The circulating leptin level increased temporarily during synthetic progestin (fluorogestone) treatment in ewes, but similar changes were not seen in progesterone-supplemented ewes and norgestomet-treated cows. In a second trial on dairy cows we wanted to study whether elevated plasma leptin levels are induced by experimental endotoxin mastitis, or by natural outbreak of GN mastitis and puerperal metritis. Experimental endotoxin mastitis resulted in some-hour elevation in cortisol and insulin, with a simultaneous decrease in IGF-I and thyroid hormones. In the first 14 days of lactation GN mastitis induced the same endocrine alterations as the experimental endotoxin challenge, but in natural cases these changes varied within a wider range, and were more protracted and robust. Cows with puerperal metritis had more obvious catabolic changes in the early weeks of lactation, than their healthy counterparts. However, both mastitis and puerperal metritis failed to increase the circulating leptin level, showing that in cows the plasma leptin is not responsible for the anorexia associated with these inflammatory diseases.     

Effect of environmental enrichment during the rearing phase on subsequent eggshell quality in broiler breeders

This study set out to investigate if access to an enriched environment during the rearing phase influences subsequent production and eggshell quality in broiler breeders. Broiler breeder pullets were reared under standard rearing conditions (control treatment) or under standard conditions with an environmental enrichment of bales of plastic-wrapped wood shavings (enriched treatment). At 19 weeks of age both groups were transported to a laying facility and housed in standard pens according to treatment type. Egg weight, shape, shell thickness, non-destructive deformation, breaking strength, stiffness and ultrastructural quality were compared at beginning of lay (25 weeks old), peak production (31 weeks old), mid-lay (45 weeks old) and end of lay (57 weeks old). Production and hatchery data were also collected throughout lay. Significant treatment-age interactions were detected for non-destructive deformation, breaking strength and stiffness, such that birds which had received prior enrichment went on to maintain a better shell quality with age. The measurements of total shell thickness, mammillary thickness and effective shell thickness all showed a trend for a treatment-age interaction. Significant age effects were found for weight, shape index, shell thickness, breaking strength and stiffness. All production and hatchery data also demonstrated significant changes with age. It is concluded that early enrichment with bales of wood shavings may provide a novel approach to the maintenance of eggshell quality at a time when calcium metabolism begins to reduce in efficiency.     

Parasitemia in an immunocompetent horse experimentally challenged with Sarcocystis neurona sporocysts

Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in Americans. Most cases are attributed to infection of the central nervous system with Sarcocystis neurona. Parasitemia has not been demonstrated in immunocompetent horses, but has been documented in one immunocompromised foal. The objective of this study was to isolate viable S. neurona from the blood of immunocompetent horses. Horses used in this study received orally administered S. neurona sporocysts (strain SN 37-R) daily for 112 days at the following doses: 100/day for 28 days, followed by 500/day for 28 days, followed by 1000/day for 56 days. On day 98 of the study, six yearling colts were selected for attempted culture of S. neurona from blood, two testing positive, two testing suspect and two testing negative for antibodies against S. neurona on day 84 of the study. Two 10 ml tubes with EDTA were filled from each horse by jugular venipuncture and the plasma fraction rich in mononuclear cells was pipetted onto confluent equine dermal cell cultures. The cultures were monitored weekly for parasite growth for 12 weeks. Merozoites grown from cultures were harvested and tested using S. neurona-specific PCR with RFLP to confirm species identity. PCR products were sequenced and compared to known strains of S. neurona. After 38 days of in vitro incubation, one cell culture from a horse testing positive for antibodies against S. neurona was positive for parasite growth while the five remaining cultures remained negative for parasite growth for all 12 weeks. The Sarcocystis isolate recovered from cell culture was confirmed to be S. neurona by PCR with RFLP. Gene sequence analysis revealed that the isolate was identical to the challenge strain SN-37R and differed from two known strains UCD1 and MIH1. To our knowledge this is the first report of parasitemia with S. neurona in an immunocompetent horse.     

Treatment of 10 dogs with discospondylitis by fluoroscopy-guided percutaneous discectomy

Ten dogs suffering from discospondylitis were treated by percutaneous discectomy and local and systemic antibiotic therapy. With fluoroscopic guidance, a cylinder 5 mm in diameter was removed from the centre of the intervertebral space, yielding a fenestration and decompressing the disc without producing any spinal instability. The causative bacteria were identified in nine of the 10 biopsy specimens, but in only three urine cultures and four blood cultures. In two cases, the antibiotics used initially had to be changed owing to the organisms' antibiotic resistance. The clinical signs of the dogs improved markedly after two to nine days (mean 4.2 days) and had resolved completely after five to 14 days (mean 9.1 days). In all the cases the disease could be classified histologically as either acute or chronic, and the disease was classified as chronic in one dog. No side effects were observed.     

Rapid isolation of Brachyspira hyodysenteriae and Brachyspira pilosicoli from pigs

The aim of this study was to compare and evaluate the time required to isolate Brachyspira hyodysenteriae and Brachyspira pilosicoli from porcine faeces. This was done using previously described selective media (spectinomycin) S400, (colistin, vancomycin and spectinomycin) CVS and (spectinomycin, vancomycin, colistin, spiramycin and rifampin with swine faecal extract) BJ, compared with the method based on blood agar modified medium, with spectinomycin and rifampin (BAM-SR), including a pre-treatment step. Fourteen spirochaetal strains were obtained in pure cultures after 5 days (48 h in BAM-SR primary plate and three passages every 24 h in brain heart infusion (BHI) without antibiotics) pre-treating simulated samples in brain heart infusion broth with spectinomycin (400 microg/ml) and rifampin (15 microg/ml), before streaking on the selective BAM-SR medium. Spirochaetes from samples in S400, CVS and BJ, with and without pre-treatment, were obtained in pure cultures only after repeatedly transferring on plates of the same selective medium requiring 15-18 days according to the strain. BAM-SR used after the pre-treatment step showed a detection limit ranging from 3.5 x 10(2) to 6.7 x 10(7) cells/g faeces and was the only method able to support the growth of spirochaetes after 48 h.     

Evaluation of an Electrolyte Analyser for Measurement of Concentrations of Ionized Calcium and Magnesium in Cats

The goal of this study was to evaluate the Nova CRT 8 electrolyte analyser for determination of concentrations of ionized calcium (Ca_i) and magnesium (Mg_i) in cats, todetermine the effects of sample handling and storage and to establish reference ranges. The precision and analytical accuracy of the Nova CRT 8 analyser were good. The concentrations of Ca_i and Mg_i were significantly lower in aerobically handled serum samples than in those handled anaerobically. The concentrations of Ca_i and Mg_i differed significantly among whole blood, plasma and serum. In anaerobically handled serum, the concentration of Ca_i was stable for 8 h at 22°C, for 5 days at 4°C and for 1 week at −20°C. The concentration of Mg_i was stable for 4 h at 22°C but for less than 24 h at 4°C and for less than 1 week at −20°C. In serum from 36 cats, the reference ranges were 1.20–1.35 mmol/L for Ca_i and 0.47–0.59 mmol/L for Mg_i. The Nova CRT 8 electrolyte analyser is suitable for determination of Ca_i and Mg_i concentrations in cats. Anaerobically handled serum samples are recommended and, stored at room temperature, they yield accurate results when analysed within 4 h.     

Neurogenesis in a young dog with epileptic seizures

Epileptic seizures can lead to various reactions in the brain, ranging from neuronal necrosis and glial cell activation to focal structural disorganization. Furthermore, increased hippocampal neurogenesis has been documented in rodent models of acute convulsions. This is a report of hippocampal neurogenesis in a dog with spontaneous epileptic seizures. A 16-week-old epileptic German Shepherd Dog had marked neuronal cell proliferation (up to 5 mitotic figures per high-power field and increased immunohistochemical expression of proliferative cell nuclear antigen) in the dentate gyrus accompanied by microglial and astroglial activation. Some granule cells expressed doublecortin, a marker of immature neurons; mitotically active cells expressed neuronal nuclear antigen. No mitotic figures were found in the brain of age-matched control dogs. Whether increased neurogenesis represents a general reaction pattern of young epileptic dogs should be investigated.     

Response of white leghorn chickens to infection with avian leukosis virus subgroup J and infectious bursal disease virus

The effects of viral-induced immunosuppression on the infectious status (viremia and antibody) and shedding of avian leukosis virus (ALV) were studied. Experimental white leghorn chickens were inoculated with ALV subgroup J (ALV-J) and infectious bursal disease virus (IBDV) at day of hatch with the ALV-J ADOL prototype strain Hcl, the Lukert strain of IBDV, or both. Appropriate groups were exposed a second time with the Lukert strain at 2 wk of age. Serum samples were collected at 2 and 4 wk of age for IBDV antibody detection. Samples for ALV-J viremia, antibody detection, and cloacal shedding were collected at 4, 10, 18, and 30 wk of age. The experiment was terminated at 30 wk of age, and birds were necropsied and examined grossly for tumor development. Neoplasias detected included hemangiomas, bile duct carcinoma, and anaplastic sarcoma of the nerve. Control birds and IBDV-infected birds were negative for ALV-J-induced viremia, antibodies, and cloacal shedding throughout experiment. By 10 wk, ALV-J-infected groups began to develop antibodies to ALV-J. However, at 18 wk the incidence of virus isolation increased in both groups, with a simultaneous decrease in antibody levels. At 30 wk, 97% of birds in the ALV-J group were virus positive and 41% were antibody positive. In the ALV-J/IDBV group, 96% of the birds were virus positive at 30 wk, and 27% had antibodies to ALV-J. In this study, infection with a mild classic strain of IBDV did not influence ALV-J infection or antibody production.     

Disseminated pheochromocytoma in a North American river otter (Lontra canadensis)

A 21-yr-old male North American river otter (Lontra canadensis) with a chronic history of degenerative osteoarthritis was evaluated for acute posterior paralysis. Because no definitive cause was identified and a poor prognosis was expected, the otter was euthanatized. A malignant neoplasm of adrenal gland origin with disseminated metastases to the central nervous system, lymph nodes, diaphragm, pancreas, spleen, and liver was diagnosed on postmortem examination. No clinical signs of disseminated neoplasia had been noted throughout the otter's history. The adrenal neoplasm was composed of nests of epithelial cells surrounded by a fine fibrovascular stroma. Neoplastic cells were immunohistochemically positive for chromogranin A, PGP9.5, metencephalin, and endorphin and negative for melan A and inhibin, confirming a diagnosis of a malignant pheochromocytoma. On the basis of the necropsy finding, metastasis of the pheochromocytoma might have contributed to the observed clinical signs.