The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques

In order to develop procedures to label the main bovine leucocyte populations in paraffin embedded sections, the immunoreactivity of 25 monoclonal antibodies (mAbs) to different leucocyte antigens was assessed with formal dichromate (FD5) and 10% formalin fixation, a battery of antigen retrieval (AR) methods, and the biotin-tyramide amplification system. All the leucocyte populations investigated (CD2+, CD4+, CD8+, WC1+ T lymphocytes, B cells and macrophages) were strongly and specifically detectable under an appropriate combination of mAb, AR method and signal amplification system. CD4 and CD8 required the most stringent conditions and could only be demonstrated in FD5 fixed sections. For detection of CD2, WC1+ T lymphocytes, B cells and macrophages, all the mAbs produced immunoreactivity in FD5 or formalin fixed tissues. The need to check a range of different AR methods is stressed, as the method of choice varied for each individual mAb. The incorporation of the signal amplification system was necessary to observe a strong signal and the complete distribution of CD4, CD8 and B cells. Fixation by FD5 proved to be better than formalin for the preservation of surface antigens but it was inferior for the detection of markers which were found to show cytoplasmic immunoreactivity, such as the macrophage marker MAC387 or the B cell markers BAQ155 or IL-A59.     

Pharmacokinetics and renal toxicity of three once-a-day doses of amikacin in cows

Pharmacokinetic variables of amikacin in cows were determined after administration of amikacin sulphate either intravenously (IV) or intramuscularly (IM) at a dose of 25 mg/kg per day for three days. Amikacin concentrations at time zero and maximum serum concentrations were 240.8 microg/mL and 122.53 microg/mL, respectively. The elimination half-life remained unchanged during the three days of administration (T1/2beta = 1.33 +/- 0.029 h for the IV route and T1/2beta = 2.75 +/- 0.38 h for the IM route). Apparent volumes of distribution suggest limited distribution out of the central compartment (VdAUC = 0.154 +/- 0.005 L/kg; Vdc = 36.50 +/- 2.35 L; Vdss = 0.092 +/- 0.004 L/kg). Bioavailability after IM administration was 95%. Serum profiles of urea, creatinine, albumin, electrolytes and pH after 5-day treatment with amikacin at a dose of 25 mg/kg per day IM revealed no changes. Assessment of diffusion of amikacin to milk by a commercially available screening method to detect antibiotic residues revealed that amikacin could not be detected by the fifth milking period after the last treatment. These results suggest that it would be rational to use a large single-daily dose of amikacin for future clinical trials in cows.     

GROSS AND HISTOPATHOLOGIC CORRELATION OF LOW‐FIELD MAGNETIC RESONANCE IMAGING FINDINGS IN THE STIFLE OF ASYMPTOMATIC HORSES

With the recent introduction of a 0.25T rotating MRI system, clinical evaluation of the equine stifle joint is now possible in the average equine athlete. A recent publication described common abnormalities of horses with stifle lameness detected with a low‐field MRI system; however, postmortem corroboration of the lesions detected was not possible. Therefore, our objective was to compare postmortem findings with low‐field MRI findings in equine cadaver stifle joints. Ten fresh cadaver stifle joints from horses without clinical signs of stifle disease were evaluated using low‐field MRI, gross dissection, and histopathology. In eight stifles, either the lateral or medial cranial meniscotibial ligament had an irregular shape, fiber separation, or moderate abnormal signal intensity (SI) on all sequences. In five stifles, the medial femoral condyle had articular cartilage fibrillation with or without an osteochondral defect over the weight bearing surface of the medial femoral condyle. All stifles had abnormal SI on all sequences within the patellar ligaments that corresponded with adipose tissue infiltrating between the collagen bundles. Other abnormalities identified included articular cartilage fibrillation of the tibial condyles in three stifles, and articular cartilage fibrillation with chondral defects in the patella in three stifles. All abnormalities detected with low‐field MRI were corroborated by gross dissection. Findings from the current study supported the use of low‐field MRI for detection of stifle joint lesions in horses and demonstrated that some stifle joint pathologies may be subclinical in horses.     

Ocular lesions associated with Trypanosoma evansi in experimentally infected goats

Ocular lesions associated with Trypanosoma spp. infection have been described in man and many animal species. However, loss of vision has not been demonstrated in humans presenting Chagas disease or in animals affected by different trypanosome species. In order to assess the possible ocular disorders caused by Trypanosoma evansi infection, six goats were inoculated with 1 x 10(5) T. evansi and maintained for 12 months and four goats were used as control. The inoculated animals became positive at serological and parasitological tests at 1-month post-inoculation and showed a subclinical course of the disease. Unilateral superficial corneal ulceration and retinochoroiditis were observed in two inoculated animals. Data from ocular neurologic examination and electroretinography showed no significant differences between inoculated and non-inoculated goats. It could be concluded that Trypanosoma evansi can produce ocular lesion but without apparent loss of vision in goats.     

Genetic and phenotypic aspects of early reproductive performance in Raeini Cashmere goats

This study used pedigree information and data collected from 1979 to 2012 at the Raeini Cashmere goat breeding station, located in Baft City in Kerman Province in southeastern Iran. Genetic and phenotypic parameters for early reproductive traits of breeding does, including total numbers of kids born at first kidding (LSB1), total numbers of kids weaned at first kidding (LSW1), total birth weight of all kids born at first kidding (LWB1), total weaning weight of all kids weaned at first kidding (LWW1), and age at first kidding (AFK), were estimated using a Bayesian approach via Gibbs sampling. Posterior means for heritability estimates of LSB1, LSW1, LWB1, LWW1, and AFK were statistically significant, with values of 0.12, 0.23, 0.17, 0.15, and 0.46, respectively. Low-to-moderate additive genetic variation was present for the studied reproductive traits. Estimated genetic correlations among LSB1, LSW1, LWB1, and LWW1 were statistically significant and ranged from 0.12 between LWB1 and LWW1 to 0.72 between LSB1 and LSW1. Corresponding phenotypic correlation estimates were also statistically significant and ranged from 0.04 between LWB1 and LWW1 to 0.55 between LSB1 and LSW1. Posterior means of genetic and phenotypic correlations between AFK and other studied traits were statistically significant only for LSB1 and LWB1. For LSB1, LSW1, LWB1, and LWW1, we conclude that genetic and phenotypic improvement in any of these traits in Raeini Cashmere does would favorably influence all of the other traits. However, does that first kidded at younger ages have smaller litters at birth and lower litter birth weights at their first parity.

     

A PROPOSED RADIOGRAPHIC CLASSIFICATION SCHEME FOR CONGENITAL THORACIC VERTEBRAL MALFORMATIONS IN BRACHYCEPHALIC “SCREW‐TAILED” DOG BREEDS

Congenital vertebral malformations are common in brachycephalic “screw‐tailed” dog breeds such as French bulldogs, English bulldogs, Boston terriers, and pugs. The aim of this retrospective study was to determine whether a radiographic classification scheme developed for use in humans would be feasible for use in these dog breeds. Inclusion criteria were hospital admission between September 2009 and April 2013, neurologic examination findings available, diagnostic quality lateral and ventro‐dorsal digital radiographs of the thoracic vertebral column, and at least one congenital vertebral malformation. Radiographs were retrieved and interpreted by two observers who were unaware of neurologic status. Vertebral malformations were classified based on a classification scheme modified from a previous human study and a consensus of both observers. Twenty‐eight dogs met inclusion criteria (12 with neurologic deficits, 16 with no neurologic deficits). Congenital vertebral malformations affected 85/362 (23.5%) of thoracic vertebrae. Vertebral body formation defects were the most common (butterfly vertebrae 6.6%, ventral wedge‐shaped vertebrae 5.5%, dorsal hemivertebrae 0.8%, and dorso‐lateral hemivertebrae 0.5%). No lateral hemivertebrae or lateral wedge‐shaped vertebrae were identified. The T7 vertebra was the most commonly affected (11/28 dogs), followed by T8 (8/28 dogs) and T12 (8/28 dogs). The number and type of vertebral malformations differed between groups (P = 0.01). Based on MRI, dorsal, and dorso‐lateral hemivertebrae were the cause of spinal cord compression in 5/12 (41.6%) of dogs with neurologic deficits. Findings indicated that a modified human radiographic classification system of vertebral malformations is feasible for use in future studies of brachycephalic “screw‐tailed” dogs.     

Plasma concentrations of leptin, insulin-like growth factor-I, and insulin in relation to changes in body condition score in heifers

The objective of this study was to determine the relationships among plasma concentrations of leptin, insulin, and IGF-I with dynamic changes in body condition scores (BCS) in heifers. Nineteen Zebu-Brown Swiss crossbred heifers, 24 to 30 mo old, weighing 322 +/- 9 kg, and with an initial BCS of 2.6 +/- 0.11 (range = 1 to 9) were used. Heifers were fed 60% of their maintenance requirements until they reached a BCS of < or = 2. Heifers were then maintained at that level for 25 d, after which they were fed to gain 1 kg of body weight daily until a BCS of 6 was reached. Heifers were weighed weekly and BCS was measured every 2 wk. Plasma samples were collected twice weekly, and leptin and insulin were determined by RIA. An immunoradiometric assay was used to measure IGF-I from one sample every 2 wk. Plasma concentrations of leptin were positively correlated during nutritional restriction (NR) and weight gain (WG) periods with BCS (r = 0.47 for NR, and r = 0.83 for WG; P < 0.01) and body weight (r = 0.40 for NR, and r = 0.78 for WG; P < 0.01). Plasma concentrations of leptin decreased during nutritional restriction (P < 0.01) as BCS decreased. During weight gain, leptin concentration increased at BCS 3 and thereafter for each integer change in the BCS. Regression analysis showed that changes in body weight affect leptin concentrations within a given BCS. There was a decrease in IGF-I as BCS declined (P < 0.01). During weight gain, by contrast, IGF-I increased significantly (P < 0.01) with every unit change in body condition up to BCS of 4 and plateaued thereafter. Insulin concentrations did not change during nutritional restriction when BCS decreased from 3 to 1. However, once the diet was improved, there was a large increase in insulin concentrations in heifers with BCS 1 (P < 0.01). Among heifers of BCS 2 and 3, insulin did not differ and was lower than in heifers of BCS 1 (P < 0.01). Insulin increased (P < 0.01) among heifers at BCS 4 to 6. Leptin was positively correlated (P < 0.01) with both IGF-I (r = 0.34 for NR, and r = 0.36 for WG) and insulin (r = 0.18 for WG). Insulin was correlated with IGF-I (r = 0.60; P < 0.01). During nutritional restriction, insulin did not correlate with leptin (r = -0.05), BCS (r = -0.03), or IGF-I (r = 0.07). It was concluded that leptin serves as a dynamic indicator of body condition in heifers, as well as an indicator of nutritional status.