Exogenous leptin advances puberty in domestic hen

The present study was undertaken to examine the effect of recombinant chicken leptin administered to fed ad libitum and feed-restricted immature chickens of a layer strain on ovarian development and the timing of sexual maturity. In the first experiment 11-week-old pullets (77 days of age) fed ad libitum were injected daily with leptin at four dose levels (4, 16, 64 and 256 microg/kg body weight) until sexual maturity (lay of the first egg). Leptin treatment at the highest dose significantly (P<0.05) advanced the onset of puberty (day 116.3+/-1.0) in comparison to controls (day 121.3+/-1.2). The rises of luteinizing hormone, estradiol and progesterone in blood plasma were also advanced by leptin treatment. In the second experiment, both full-fed and feed-restricted pullets (79 days of age) were injected daily with leptin (256 microg/kg body weight). In birds fed ad libitum, exogenous leptin again significantly (P<0.05) advanced first ovipostion (day 118.4+/-1.4 versus day 124.4+/-1.7), while abolishing the significant (P<0.05) delay caused by feed restriction (day 131.5+/-1.6) and restoring the normal onset of sexual maturity (day 125.7+/-1.6). Analysis of the ovaries in 106-day-old pullets revealed that leptin injections advanced follicular development, particularly in birds fed ad libitum, and significantly (P<0.01) reduced follicular apoptosis both in full-fed and feed-restricted birds. In conclusion, we have shown that in female chickens exogenous leptin advances the onset of puberty by attenuation of ovarian apoptosis and enhancement of folliculogenesis.     

Cytoarchitecture, morphology, and lumbosacral spinal cord projections of the red nucleus in cattle

OBJECTIVE: To analyze the morphology, cytoarchitecture, and lumbosacral spinal cord projections of the red nucleus (RN) in cattle. ANIMALS: 8 healthy Friesian male calves. PROCEDURES: Anesthetized calves underwent a dorsal laminectomy at L5. Eight bilateral injections (lateral to the midline) of the neuronal retrograde fluorescent tracer fast blue (FB) were administered into the exposed lumbosacral portion of the spinal cord. A postsurgical calf survival time of 38 to 55 days was used. Following euthanasia, the midbrain and the L5-S2 spinal cord segments were removed. Nissl's method of staining was applied on paraffin-embedded and frozen sections of the midbrain. RESULTS: The mean length of the RN from the caudal to cranial end ranged from 6,680 to 8,640 microm. The magnocellular and parvicellular components of the RN were intermixed throughout the nucleus, but the former predominate at the caudal portion of the nucleus and the latter at the cranial portion with a gradual transitional zone. The FB-labeled neurons were found along the entire craniocaudal extension of the nucleus, mainly in its ventrolateral part. The number of FB-labeled neurons was determined in 4 calves, ranging from 191 to 1,469 (mean, 465). The mean cross-sectional area of the FB-labeled neurons was approximately 1,680 microm2. CONCLUSIONS AND CLINICAL RELEVANCE: In cattle, small, medium, and large RN neurons, located along the entire craniocaudal extension of the RN, contribute to the rubrospinal tract reaching the L6-S1 spinal cord segments. Thus, in cattle, as has been shown in cats, the RN parvicellular population also projects to the spinal cord.     

Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations

Three serological methods, indirect fluorescent immunoassay (IFI), enzyme-linked immunosorbent assay (ELISA) and direct agglutination test (DAT) that are commonly employed in the diagnosis of canine visceral leishmaniasis (CVL), have been assessed. A total of 234 domestic dogs, drawn from an area in the municipality of Belo Horizonte, Minas Gerais, Brazil, endemic for visceral leishmaniasis, were submitted to clinical and parasitological examinations and serological assay. Sera collected from confirmed non-infected dogs (n=20), and from dogs with other parasitic diseases including Trypanosoma cruzi (n=7), Leishmania braziliensis (n=5), Toxoplasma gondii (n=5) and Ehrlichia canis (n=3), were also included in the study. IFI presented a lower sensitivity (72%) than ELISA (95%), although the specificities of these assays were low (52 and 64%, respectively) and both exhibited cross-reactivity with sera from dogs infected with T. cruzi, L. braziliensis and E. canis. In contrast, DAT exhibited a high sensitivity (93%) and a high specificity (95%) and cross-reacted with only one serum sample derived from an E. canis-infected dog. The reproducibilities of the ELISA and DAT assays were excellent, whilst that of IFI was considered to be acceptable. The results produced by ELISA and DAT were in complete agreement, those between ELISA and IFI were at an acceptable level of agreement, whilst the concurrence between the IFI and DAT results were either acceptable or poor depending on the clinical conditions of the group of dogs examined. Since there is no readily accessible method for the diagnosis of CVL that offers 100% specificity and sensitivity, the choice of technique employed must depend on the aim of the investigation.     

Evaluation of agreement between numerical rating scales, visual analogue scoring scales, and force plate gait analysis in dogs

Objective— To evaluate the accuracy of numerical rating (NRS) and visual analogue (VAS) scoring scales compared with force plate gait analysis and agreement between observers for each scoring scale.
Study Design— Experimental study.
Animals— Mixed breed dogs (n=21) with a right limb tibial osteotomy repaired with an external fixator.
Methods— Three small-animal veterinarians with orthopedic training scored lameness using NRS and VAS before surgery, and at 4 and 8 weeks after surgery. Peak force and impulse were determined at the same time points using a force plate. Agreement between observers and with force plate data was assessed. Significance was set at P ≤.05.
Results— Agreement was generally low among observers for both NRS and VAS scores. When evaluated at each time point, an acceptable level of agreement was present only at 4 weeks after surgery. Only impulse had a significant relationship with some of the observers' subjective scores. No significant relationships between any observer's scores and force plate data existed if very lame dogs were omitted.
Conclusions— Subjective scoring scales do not replace force plate gait analysis. Agreement is low unless lameness is severe, and each observer uses an individually unique scale. Subjective scoring scales most accurately reflect force plate gait analysis when lameness is severe.
Clinical Relevance— Subjective lameness scoring scales may not accurately reflect lameness and do not replace force plate gait analysis. Observers must stay the same during the duration of a study for accurate analyses.     

A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

NECROTIZING ENCEPHALITIS IN A YORKSHIRE TERRIER: CLINICAL, IMAGING, AND PATHOLOGIC FINDINGS

A 5-year-old, male Yorkshire Terrier had chronic and progressive neurologic signs compatible with lesions in the right brain stem and right forebrain. In magnetic resonance images of the brain there were multifocal lesions at different stages of evolution, consistent with an inflammatory disease. The lesions were located in the white matter of the cerebrum, in the diencephalon and mesencephalon. These lesions were hypointense in T1-weighted images and hyperintense in T2-weighted images with no mass effect and minimal enhancement with gadolinium. Necrotizing encephalitis was confirmed post mortem.     

Canine fertility: The consequences of selection for special traits

Pedigree dogs and cats are bred aiming to conform breed standards with very poor consideration for breeding stock fertility. At the same time, the genetic asset underlining reproductive traits could be effectively analysed like in other species under selection. The definition of selection targets is very important in breeding protocols determination. The aim of the present work is to present an overview of the different correlations between reproduction and genetics, starting from selection procedure and inbreeding coefficient moving to genomic and the application of SNPs and GWAS on population study and identification of genes involved in phenotypical variation of reproductive traits in dogs. Particular relevance has been given to the concept of inbreeding which effects on canine reproduction have been presented. The use of genomic information in inbreeding coefficient calculation can be considered an improved effective procedure in the evaluation of the genetic variability loss in canine population and its negative effects on reproductive traits.     

Participation of Rhipicephalus sanguineus (Acari: Ixodidae) in the epidemiology of canine visceral leishmaniasis

The vectorial competence of the tick Rhipicephalus sanguineus is discussed in relation to the epidemiology of canine visceral leishmaniasis, taking into account its strict association with dogs and the low indices of natural infection presented by its known vector, the phlebotomine sand fly Lutzomyia longipalpis. In order to evaluate natural infection by Leishmania chagasi and the infectivity of these parasites in the tick, 39 specimens (6 females, 11 males and 22 nymphs) of R. sanguineus were removed from 21 dogs showing diverse symptoms of zoonotic visceral leishmaniasis (ZVL). Six ticks (15.4%) gave positive results for the genus Leishmania using the PCR technique. To determine the infectivity of the parasites, 36 hamsters were inoculated orally and peritoneally with macerates of ticks removed from nine dogs symptomatic for visceral leishmaniasis. After 6 months the hamsters were sacrificed and necropsied. Serum was removed for IFAT, as well as spleen and liver fragments to make imprint smears and for PCR. Eight (88.9%) of these dogs presented ticks that were infective for 14 hamsters (41.2%), 12 (85.7%) of them infected peritoneally and two (14.3%) orally. PCR revealed 27 smears (40.9%) to be positive, 20 (62.5%) of them infected peritoneally and seven (20.6%) orally. IFAT showed 14 positive animals (41.2%). Based on these findings, we suggest that the vectorial capacity of R. sanguineus for L. chagasi should be evaluated further, opening new perspectives in the epidemiology of ZVL.     

Determination of tumor necrosis factor-alpha responsiveness in piglets around weaning using an ex vivo whole blood stimulation assay

Ex vivo whole blood stimulation with endotoxin has proved to be a useful method for quantitative evaluations of the pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) response ability in humans. In the present study, a dose- and time-response study was carried out in order to develop an ex vivo whole blood stimulation assay for the quantification of TNF-alpha production in pigs. The TNF-alpha response was enhanced with increasing endotoxin stimulation dose. The maximum TNF-alpha response occurred after 2-8 h of stimulation. Subsequently, the assay was used to evaluate the TNF-alpha response in pigs (n=32) in relation to weaning. The TNF-alpha response was 332 pg ml(-1) (+/-59 pg) plasma 2 days before weaning and 127 pg ml(-1) (+/-23 pg) plasma 2 days after weaning, which was a significant reduction (p<0.001). Total and differential counts of leucocytes were the same before and after weaning. Thus the lower TNF-alpha response may be due to reduced monocyte responsiveness to the endotoxin, rather than caused by a reduction in monocyte numbers. The reduced TNF-alpha response in piglets after weaning may be a factor of importance to the increased disease susceptibility seen in piglets in this period.     

Vitamin D status of wild Ricord's iguanas (Cyclura ricordii) and captive and wild rhinoceros iguanas (Cyclura cornuta cornuta) in the Dominican Republic.

Calcidiol (25-hydroxyvitamin D) values are reported for 22 wild Ricord's iguanas (Cyclura ricordii) and seven wild rhinoceros iguanas (Cyclura cornuta cornuta). Calcitriol (1,25-hydroxyvitamin D) values are reported for 12 wild Ricord's iguanas and seven wild rhinoceros iguanas. These animals were captured as part of a larger health assessment study being conducted on Ricord's iguanas in Isla Cabritos National Park, Dominican Republic. A total of 13 captive rhinoceros iguanas held outdoors at Parque Zoológico Nacional were also sampled for comparison. Mean concentrations of 25-hydroxyvitamin D were 554 nmol/L (222 ng/ml) with a range of 250-1,118 nmol/L (100-448 ng/ml) for wild Ricord's iguanas, 332 nmol/L (133 ng/ml) with a range of 260-369 nmol/L (104-148 ng/ml) for wild rhinoceros iguanas, and 317 nmol/L (127 ng/ml) with a range of 220-519 nmol/L (88-208 ng/ml) for captive rhinoceros iguanas. On the basis of these results, serum concentrations of at least 325 nmol/L (130 ng/ml) for 25-hydroxyvitamin D should be considered normal for healthy Ricord's and rhinoceros iguanas.