Viability assessment of spermatozoa in large falcons (Falco spp.) using various staining protocols

Viability assessment is an important part of semen analysis, and various live/dead staining protocols have been used in semen of avian species. Results of live/dead count differed between dyes, staining protocols and bird species, impeding comparability between studies and requiring species-specific comparisons of viability stains. In raptor semen, similar comparisons are absent. Thus, the aim of the present study was to compare eight conventional viability stains. Eosin blue 2% [EB], eosin blue 2% with the addition of 3% sodium citrate [EB2], eosin blue–nigrosin 5% [EBN5], eosin yellow–nigrosin 5% [EYN5], eosin yellow–nigrosin 10% [EYN10], eosin blue–aniline blue [EBA], eosin yellow–aniline blue [EYA] and bromophenol blue–nigrosin [BBN] were evaluated in comparison with the fluorescence stain SYBR^® Green–propidium iodide [SYBR-PI] in spermatozoa of falcons. The comparison was performed using conventional light microscopy which is applicable in breeding centres, veterinary practices and field studies. Additionally, live/dead stains were correlated to motility values of the same samples to validate sperm viability. Light microscopy using EB and using SYBR-PI enabled an effective and clear differentiation between alive and dead spermatozoa of falcons. Motility values correlated significantly and strongly with EB only (r = .629; p < .001), but not with any other stain used in the study. Therefore, our results suggest EB as the most suitable stain for viability assessment in the semen of large falcons.     

Humoral immune response in hens naturally infected with Salmonella Enteritidis against outer membrane proteins and other surface structural antigens

A simple procedure for obtaining surface exposed antigens of Salmonella Enteritidis is described. A heat treatment of whole bacteria in saline solution induced the release of small membrane vesicles containing outer membrane components as well as surface appendage components, such as fimbriae and flagellin. The characterization of the structural components of this extract, called HE, was established by SDS-PAGE and immunoblotting using polyclonal and monoclonal specific antibodies. Five major groups of proteins were identified: flagellin, porins, OmpA, SEF21 and SEF14 fimbriae. The immunogenicity of these proteins was studied by immunoblotting with serum samples from naturally infected hens. Flagellin, porins, OmpA, SEF14 and SEF21 fimbriae were immunogenic in the S. Enteritidis infected hens (frequency of reactants: 47.3, 97.3, 64.7, 50.0 and 60.8%, respectively); porins also reacted with sera from non infected hens (66.7%). The immunogenicity of these antigens in infected birds provide promise that they may serve as components of an effective subcellular vaccine for poultry salmonellosis.     

Determination of creatinine excretion and evaluation of spot urine sampling in Holstein cattle

Four experiments were carried out to determine urinary creatinine excretion in Holstein growing bulls, lactating cows, and replacement heifers. In addition, we evaluated the use of spot sampling technique to estimate purine derivatives (PD) excretion. In Experiment I, 15 lactating cows were used in a randomized block design to compare creatinine excretion obtained in different time-spans of urine collection (during 6, 9, 12, 15, 18, 21, and 24 h). In Experiment II, four bulls were allocated in a 4 × 4 Latin square to evaluate the effect of diet (levels of cottonseed hulls of 0, 10, 20, and 30% of the DM) on excretion of creatinine. In Experiment III, 15 lactating cows were used to evaluate the effect of milk production (ranging from 3.9 to 36.7 kg/d) on daily creatinine and PD excretions. In Experiment IV, 22 replacement heifers were utilized to evaluate the effect of body weight (BW, ranging from 107 to 545 kg) on daily creatinine and PD excretions. For all experiments, total urine collections were made over 24 h and daily creatinine and PD excretions were determined. Different time-spans for total urine collection had no effect (P = 0.70) on creatinine excretion compared to the 24-h collection period, indicating a constant excretion rate of creatinine. The roughage source did not influence (P = 0.64) creatinine excretion by bulls, averaging 0.248 ± 0.008 mmol/kg BW. Similarly, milk production did not affect (P = 0.82) creatinine excretion in cows, averaging 0.212 ± 0.004 mmol/kg BW. In contrast, the creatinine excretion (mmol/kg BW) decreased linearly (P < 0.001) as BW of heifers increased, suggesting that creatinine excretion might vary with the degree of maturity of growing animals. There were no differences (P > 0.14) between the 24-h total collection and spot sampling technique in estimating daily PD excretion. The spot sampling technique may be used to estimate the daily excretion of urinary PD in Holstein cattle under practical conditions.     

Intramedullary epidermoid cyst in the thoracic spine of a dog

A 5 yr old female spayed mastiff was evaluated for a 3-4 mo history of paraparesis and 3 days of acutely worse paraparesis and incontinence. On magnetic resonance imaging, a spinal cord lesion was present at the ninth thoracic vertebra. The lesion was hyperintense on T2-weighted images (T2-W), and a hyperintense rim was present on T1-weighted postcontrast images. Histologic examination showed a cystic mass lined by squamous epithelial cells. Histopathologic diagnosis was an intramedullary epidermoid spinal cyst. Epidermoid cyst should be a differential diagnosis in young dogs with a myelopathy and an intramedullary spinal cord lesion on magnetic resonance imaging examination.     

The effect of induced hindlimb lameness on thoracolumbar kinematics during treadmill locomotion

REASON FOR PERFORMING STUDY: There are no detailed studies describing a relationship between hindlimb lameness and altered motion of the back. OBJECTIVES: To quantify the effect of induced subtle hindlimb lameness on thoracolumbar kinematics in the horse. METHODS: Kinematics of 6 riding horses were measured during walk and trot on a treadmill before and during application of pressure on the sole of the left hindlimb using a well-established sole pressure model. Reflective markers were located at anatomical landmarks on the limbs, back, head and neck for kinematic recordings. Ground reaction forces (GRF) in individual limbs were calculated from kinematics to detect changes in loading of the limbs. RESULTS: When pressure on the sole of the hindlimb was present, horses were judged as lame (grade 2 on the AAEP scale 1-5) by an experienced clinician. No significant unloading of this limb was found in the group of horses (unloading was observed in 4 animals, but was not detectable in the other 2), but statistically significant effects on back kinematics were detected. The overall flexion-extension (FE) range of motion (ROM) of the vertebral column was increased at walk, especially in the thoracic segments. Axial rotation (AR) ROM of the pelvis was also increased. At trot, the FE ROM was decreased only in the segment L3-L5-S3. During the stance phase of the lame limb, the segment T6-T10-T13 was more flexed and the neck was lowered at both gaits; the thoracolumbar segments were more extended at walk and trot. There were no significant changes in the stride length or protraction-retraction angles in any of the limbs. CONCLUSIONS: Subtle hindlimb lameness provoked slight but detectable changes in thoracolumbar kinematics. The subtle lameness induced in this study resulted in hyperextension and increased ROM of the thoracolumbar back, but also in decreased ROM of the lumbosacral segment and rotational motion changes of the pelvis. POTENTIAL RELEVANCE: Even subtle lameness can result in changes in back kinematics, which emphasises the intricate link between limb function and thoracolumbar motion. It may be surmised that, when chronically present, subtle lameness induces back dysfunction.     

IBR and BVD control: the key to successful herd management

Control of IBR and BVD should be possible in Europe. Effective vaccines and reliable tools for monitoring are available. Systematic approach and strict implementation of control measures are essential. Voluntary or mandatory programs are ongoing on regional or national level in a lot of countries. Successful programs put pressure on surrounding regions/countries to initiate control program as well.     

Balloon Valvuloplasty for Treatment of Pulmonic Stenosis in English Bulldogs with an Aberrant Coronary Artery

Background: English Bulldogs (EB) with pulmonic stenosis (PS) sometimes have an aberrant coronary artery (CA) type R2A encircling the pulmonary artery (PA). Balloon valvuloplasty (BV) is treatment of choice for severe PS, but is considered to be contraindicated in dogs with aberrant CA. Hypothesis: Conservative BV in EB with aberrant CA is safe and improves clinical signs and quality of life. Animals: Four client‐owned EB with severe PS were retrospectively reviewed/analysed. Methods: Retrospective study: Case records, echocardiography, BV, and follow‐up investigations of EB diagnosed with severe PS and treated with BV were reviewed. The ratios of PA to aortic (Ao) velocity time integral (VTI) were calculated to assess progression/improvement of PS. Results: An aberrant CA was confirmed on angiography in all EB. Conservative BV was performed, using a balloon of the size of the PA annulus or smaller (0.6–1 × PA annulus size). All dogs survived the procedure, but only a mild reduction in pressure gradient was achieved. There was an improvement in PA to Ao VTI in 3 of 4 dogs (P≤ .017), which were free of evidence of congestive heart failure (CHF) 5, 10, and 15 months after BV. One dog that had right‐sided CHF when BV was performed died due to progressive right‐sided CHF within 3 months. Conclusions: Conservative BV in EB might be safe and might improve quality and quantity of life.     

A male linkage map constructed for QTL mapping in Spanish Churra sheep.

A male ovine linkage map has been constructed on the basis of 11 half-sib families of a commercial population of Spanish Churra sheep as part of a genome scan for quantitative trait loci mapping. A total of 1421 daughters and their sires were genotyped for 182 microsatellite markers evenly distributed along the ovine autosomes. A total of 259,192 genotypes were obtained, generating an average of 669 informative meioses per marker. An autosomal genome length of 3262 cM was estimated for the Churra population with a mean marker interval of 17.86 cM. Our map represents an approximate 90% coverage of the autosomal ovine genome and constitutes a useful tool for the genetic dissection of complex traits in this breed. General agreement was found between the Churra map and other published maps for sheep, despite certain length discrepancies.     

Serological prevalence of Babesia caballi and Theileria equi in horses of Lara State, Venezuela

The main objective of this study was to demonstrate the occurrence of equine piroplasmosis (EP) in horses of Lara State, Venezuela, and to correlate it with the factors host's sex and age in order to know the epidemiology of this disease at the Venezuelan Centroccidental Region. Antibody levels to Babesia caballi and Theileria equi were assessed in 360 equine serum samples, collected from 9 municipalities of Lara State, using an ELISA technique with recombinant antigens and monoclonal antibodies (Mabs). Antibodies to B. caballi were found in 254 horses (70.6%), whereas 181 animals (50.3%) were detected as seropositives to T. equi. In addition, 128 samples (35.56%) were seropositives to both hemoparasites. There were no significant differences between the seropositivity to B. caballi and T. equi with the factors sex and age of the horses. These results show that Lara State is an enzootic area for equine piroplasmosis, and are a contribution to a partial knowledge of the dynamic of this disease in Venezuela.     

3-Phosphoinositide-dependent protein kinase-1/Akt signalling and inhibition in a canine prostate carcinoma cell line

Deregulation of the 3‐phosphoinositide‐dependent protein kinase‐1 (PDK‐1)/Akt signalling pathway is associated with prostate cancer development and progression. Inhibition of PDK‐1/Akt signalling can be achieved using structurally optimized celecoxib derivatives such as OSU‐03012. In this study, we treated the novel canine prostate cancer cell line, Ace‐1, with OSU‐03012 or dimethyl sulphoxide in vitro. We found that Akt was constitutively phosphorylated in the canine prostate cancer cell line Ace‐1 and that there was a dose‐dependent decrease in cell viability, and Akt and glycogen synthase kinase‐3β phosphorylation, in response to OSU‐03012 treatment. This was accompanied by a dose‐dependent increase in apoptosis. These data suggest that Akt signalling pathway inhibition is a potential strategy for the treatment of dogs with prostate cancer and that canine prostate cancer is a relevant large animal model for evaluating Akt pathway inhibitors such as OSU‐03012 for use in people.