Renal handling of calcium and phosphorus in experimental renal hyperparathyroidism in dogs

Twenty-four hour urinary excretion, fractional excretion and the filtered load of calcium and phosphorus were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Thirteen beagles of both sexes aged four and a half months were used. Nine of them were subjected to a renal damaging schedule (neomycine, 60 mg/kg/48 h, IM, 32 weeks) in order to induce chronic renal failure leading to secondary hyperparathyroidism (2HPT group). The remaining four were kept as the control group. The experiment was conducted over 32 weeks. Blood and 24 h urine were collected every four weeks. Calcium, phosphorus and creatinine were analyzed. Plasma parathormone and calcitonin were determined at weeks 0, 12, 24 and 32. The level of renal function in the 2HPT animals was reduced to 25% of that of the controls (endogenous creatinine clearance was 0.45 +/- 0.22 mL/min/kg as opposed to 1.81 +/- 0.54 mL/min/kg). Hyperparathyroidism was confirmed by a progressive increase in the levels of the parathyroid hormone. Calcitonin levels were not modified. A tendency to hypocalcaemia was observed, reaching statistically significant levels from the twenty-eighth week of the study, when hyperphosphataemia also became significant. Daily urinary excretion of calcium and phosphorus remained at values considered normal throughout the experiment with no alteration imputable to the impaired renal function. This is explained by the decrease in the filtered load of these elements (in both cases statistically significant from the 24th week on) being associated with an increase in their fractional excretion. Thus, calcium and phosphorus urinary excretion values could be maintained in a normal range up to the end of the experiment, showing that renal calcium handling in dogs with experimentally induced renal failure seems to differ from that observed in human patients.     

Genital lesions in an outbreak of caprine contagious agalactia caused by Mycoplasma agalactiae and Mycoplasma putrefaciens

This paper reports on the genital lesions observed in adult male and female goats from a commercial flock in the Extremadura region of southwestern Spain, following an outbreak of contagious agalactia syndrome caused by Mycoplasma agalactiae and M. putrefaciens. Although both species were isolated from several organs, M. putrefaciens was the only agent isolated from the genital lesions reported here, characterized by desquamative salpingitis and cystic catarrhal metritis in females and by testicular degeneration in males. Mycoplasma putrefaciens was isolated from the testes of only one of the males examined.     

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Coinfection by Cryptosporidium parvum and porcine circovirus type 2 in weaned pigs

Routine histopathological diagnosis of one representative 3-month-old pig from a group suffering from diarrhoea revealed a massive degree of parasitation by Cryptosporidium parvum, with a concomitant infection by porcine circovirus type 2 (PCV2), that was confirmed by immunohistochemical procedures. The areas of intestine where parasites were more numerous presented abundant PCV2 infected cells in mucosa and submucosa. The concurrence of C. parvum, a rare primary intestinal pathogen in post-weaning and growing pigs, and PCV2 infections suggest an increased susceptibility as a result of an immunosuppression state.     

Prevalence of antibodies to Borrelia burgdorferi sensu lato in southern chamois (Rupicapra pyrenaica) in Spain

Ungulates are involved in the epidemiology of Borreliosis as maintenance hosts for vector tick species. We evaluated the prevalence of antibodies to Borrelia burgdorferi sensu lato in two different populations of southern chamois in Spain. Forty-six sera (40.4%) were positive at titres >/=1 : 64 using an indirect immunofluorescence assay (IFAT).     

Evaluation of jitter by stimulated single-fiber electromyography in normal dogs

Single-fiber electromyography (SFEMG), a technique used to investigate neuromuscular transmission, has been described previously in the pelvic limb of dogs. Because preferential involvement of isolated muscle groups can occur in disorders of neuromuscular transmission, SFEMG was done in the peroneus longus (PL), extensor carpi radialis (ECR), and orbicularis oculi (OO) muscles of 10 adult, clinically normal dogs. Jitter was calculated as the mean absolute value of the consecutive differences in latency of 50 single muscle fiber action potentials after stimulation of intramuscular nerve bundles at the level of the motor point in at least 20 muscle fibers per muscle. Bilateral recordings were performed in 3 dogs. Mean jitter values were determined for each muscle, and differences among muscle groups and among dogs were compared. The upper limits of mean consecutive difference (mean plus 3 standard deviations) for the PL, ECR, and OO muscles were 21.94, 22.53, and 23.39 micros, respectively, and the upper limit of mean consecutive difference for individual muscle fibers in the respective fiber pools was 28.62, 36.39, and 35.68 micros. Jitter values for the ECR and OO were significantly higher than the jitter value for the PL muscle (P < .05). Significant differences among muscles or dogs or between sides were not observed for the ECR. Significant differences among dogs were observed for OO jitter values and were attributed to extremely low jitter values in 1 dog. Significant differences were demonstrated between sides for the PL and were attributed to small sample size. Results of this study provide normative data that can be used in the application of the stimulated SFEMG technique to dogs with suspected disorders of neuromuscular transmission.     

Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

Effect of fast protein liquid chromatography fractionated salivary gland extracts from different ixodid tick species on interleukin-8 binding to its cell receptors

Interleukin-8 plays a critical role in inflammatory processes. Hence generation of molecules with anti-IL-8 activity is likely to be important for successful feeding and for survival of the ticks. Anti-IL-8 activity was studied in saliva of three ixodid tick species--Dermacentor reticulatus (Fabricius, 1794), Rhipicephalus appendiculatus Neumann, 1901, and Amblyomma variegatum (Fabricius, 1794). The greatest activity was shown in saliva prepared from D. reticulatus. The activity was attributed to tick salivary gland molecules that bind to IL-8, preventing binding of the chemokine to its specific receptor, rather than to occupation of the IL-8 cell receptor by the tick molecules. The distribution of anti-IL-8 activity in fast protein liquid chromatography (FPLC) fractions of salivary gland extracts (SGE) derived from adult female D. reticulatus, R. appendiculatus and A. variegatum was compared directly by both ELISA and receptor-binding inhibition assays. The correspondence in results with fractions of SGE from ELISA is consistent with detection of tick molecules that inhibit IL-8 binding to its receptor. As IL-8 is an important chemoattractant and activator of neutrophils, the presence of an anti-IL-8 activity in tick saliva indicates that neutrophils play an important role in the host response to parasitism by ticks.     

Liquid Formulation of the Postharvest Biocontrol Agent Candida sake CPA-1 in Isotonic Solutions

ABSTRACT Viability of the postharvest biocontrol agent Candida sake CPA-1 stored as liquid formulation was evaluated by studying the effect of growth, preservation medium, and temperature. C. sake was grown in molasses medium with unmodified water activity (a(w)) and in the same with a(w) modified to 0.98 with the addition of several solutes. Cells were preserved with isotonic solutions of different substances. Efficacy of liquid formulations stored for different periods was tested against infection by Penicillium expansum on apples. The best growth media were the (unmodified one and those modified to 0.98 a(w) with the addition of glycerol or sorbitol. For all growth media, the best preservation medium was the isotonic solution prepared with trehalose. When the effect of trehalose concentration in the preservation medium was studied, generally, at trehalose concentrations below the isotonic one, C. sake viabilities increased with increased trehalose. However, the best results were obtained when cells were preserved with the trehalose solution which was isotonic with cells. After 7 months of storage at 4 degrees C, cells that were grown in the sorbitol-modified medium and preserved with the isotonic solution of trehalose (0.96 M) maintained their viability and efficacy against P. expansum infection of apples.