OC1Effects of Cryopreservation on the Expression of Glut‐3, Glut‐5 and As‐A Proteins in Iberian Boar Sperm Membranes

In order to determine the injure produced in boar spermatozoa through cryopreservation process, we analyzed the expression of the hexose transporters Glut‐3 and Glut‐5 and the zona pellucida binding protein As‐A (P68) in three different steps of the freezing‐thawed protocol: at 17°C (fresh BTS‐diluted semen, 1 : 2 v/v, step 1), at 5°C (after glycerol addition; step 2), and post‐thawing (step 3). All sperm analyses were carried out with immunogold techniques under electronic microscopy. For this study eight healthy post‐pubertal Iberian boars were submitted to a collection of twice per week through 3 months, evaluating two ejaculates from each boar. Glut‐3 maintains the expression in the acrosome region post‐thawing but not along the tail where is reduced. The expression of Glut‐5 and As‐A is majority located at the post‐acrosome region of the spermatozoa at step 1, but in step 2 and step 3 this expression is relocated to sperm tail area. In conclusion, while cryopreservation affects the localization and the expression of Glut‐3 and Glut‐5, its fertilizing capacity is not significantly reduced. The stabilization of boar semen at 5°C was found to be the most crucial step for sperm survival.     

In vitro Capacitation and Acrosome Reaction of Dog Spermatozoa can be Feasibly Attained in a Defined Medium Without Glucose

Incubation of dog spermatozoa in a medium without glucose and in the presence of lactate and pyruvate (l-CCM) for 4 h at 38.5 degrees C in a 5% CO(2) atmosphere induced in vitro capacitation of these cells. This was verified after the combined specific capacitation-like changes in percentages of viability and altered acrosomes, motility characteristics, sperm location of reactivity against Pisum sativum, Arachis hypogaea and Helix pomatia lectins and the tyrosine phosphorylation pattern. Furthermore, a feasible acrosome reaction (AR) was induced when spermatozoa incubated in l-CCM for 4 h were further co-incubated for 1 h with canine oocytes. This was demonstrated by AR-like changes in percentages of viability, altered acrosomes, motility characteristics and sperm location of reactivity against P. sativum, A. hypogaea and H. pomatia lectins. All these results clearly indicate that in vitro capacitation, and subsequent AR, can be feasibly achieved without the presence of sugars. This ability can be related to the specific characteristics of energy-metabolism regulation reported in dog spermatozoa.     

Neuronal Responses in the Brainstem and Hypothalamic Nuclei Following Insulin Treatment During the Late Follicular Phase in the Ewe

The aim of this study was to determine the neuronal responses following insulin administration during the late follicular phase. Intact ewes were given either saline or insulin (5 IU/kg, i.v.) at 35 h after progesterone withdrawal and killed 3 h later. There was a marked increase in the number of Fos‐positive noradrenergic neurones in the caudal brainstem of insulin‐treated ewes. In the hypothalamic paraventricular nucleus, insulin treatment increased the presence of Fos‐positive corticotrophin‐releasing hormone neurones (from 2% to 98%) and Fos‐positive arginine vasopressin parvocellular neurones (from 2% to 46%). Interestingly, after insulin treatment, despite a general increase in Fos‐positive neurones in the arcuate nucleus (ARC), there was a marked reduction (from 47% to 1%) in Fos‐positive β‐endorphin neurones. Similarly, colocalized Fos and oestradiol receptor (ER) α‐positive neurones decreased in the ARC after insulin (from 7% to 3%). Conversely, in the ventromedial nucleus, ERα‐positive neurones with Fos increased (from 7% to 22%) alongside a general increase in Fos‐positive neurones. Overall, a complex system of neurones in brainstem and hypothalamus is activated following insulin administration during the late follicular phase.     

Inhalant anesthetic requirement in cats is animal-dependent

Minimum alveolar concentration (MAC) of an inhalant is an indicator of its anesthetic potency. Individuals vary in their sensitivity to anesthetic agents as demonstrated by different individual MAC values. We hypothesized that individual animal sensitivity would be maintained with different inhalant anesthetics. As part of separate studies, six female DSH cats, aged 24 ± 2.5 (mean ± SD) months and weighing 3.5 ± 0.3 kg, were studied similarly on three separate occasions over a 12‐month period to determine the MAC of isoflurane (ISO), sevoflurane (SEVO), and desflurane (DES), respectively. In each study, chamber induction was followed by orotracheal intubation, and anesthesia was maintained via a nonrebreathing circuit. ECG, pulse oximetry, Doppler systolic blood pressure, end‐tidal gases, and esophageal temperature were monitored. End‐tidal gases were hand‐sampled from a catheter whose tip lay level with the distal end of the ET tube. Gases were analyzed by Raman spectrometry and, for each agent, the analyzer was calibrated with at least three gas standards. MAC was determined in triplicate using standard tail‐clamp technique. Data were analyzed by two‐way anova followed by Tukey's test and significant differences were found. Average MACs (%) for ISO, SEVO, and DES were 1.90 ± 0.18, 3.41 ± 0.65, and 10.27 ± 1.06, respectively. Body temperatures, Doppler systolic blood pressure, and SpO₂ were recorded at the time of MAC determinations for ISO, SEVO, and DES were 38.3 ± 0.3, 38.6 ± 0.1, 38.3 ± 0.35 °C; 71 ± 8, 75 ± 16, 88 ± 12 mm Hg; 99 ± 1, 99 ± 1, 99 ± 1%, respectively. Both the anesthetic agent and the individual cat had significant effects on MAC (p = 0.0001 and 0.0185, respectively). MAC varied between individuals and cats were consistent in their order of sensitivity to inhalant anesthetics across the three agents. Within this group of cats, the relationship of individual MAC to the group MAC for each of the three inhalant agents was maintained. This suggests that any individual may be consistently more or less sensitive to a variety of inhalant agents.     

Concentrations of buparvaquone in milk and tissue of dairy cows

AIM: To determine the concentration of the anti-theilerial drug buparvaquone in the milk and tissue of dairy cattle following treatment with two different formulations, and to assess the effect of clinical theileriosis on the concentration of buparvaquone in milk.

METHODS: Healthy lactating dairy cows (n=25) were injected once (Day 0) I/M with 2.5?mg/kg of one of two formulations of buparvaquone (Butalex; n=12 or Bupaject; n=13). Milk samples were collected from all cows daily until Day 35. Five cows were slaughtered on each of Days 56, 119, 147, 203 and 328, and samples of liver, muscle and injection site tissue collected. Milk samples were also collected from cows (n=14) clinically affected with theileriosis for up to 21 days after treatment with buparvaquone. Milk and tissue samples were analysed by liquid chromatography-mass spectrometry; limits of detection (LOD) were 0.00018?mg/kg for muscle and 0.00023?mg/L for milk. Concentrations of buparvaquone in milk and tissues were log₁₀-transformed for analysis using multivariate models.

RESULTS: In healthy cows, concentrations of buparvaquone in milk declined with time post-treatment (p<0.001), but were above the LOD in 11 of 25 cows at Day 35. Concentration in milk was higher one day after treatment in cows treated with Butalex than in cows treated with Bupaject, but not different thereafter (p=0.007). Concentrations of buparvaquone in muscle were below the LOD for four of five animals at Day 119 and for all animals by Day 147, but were above the LOD at the injection site of one cow, and in the liver of three cows at Day 328. Tissue concentrations did not differ with formulation nor was there a formulation by time interaction (p>0.3).

Concentrations of buparvaquone in the milk of clinically affected animals were not different from those of healthy animals at 1 and 21 days post-treatment (p=0.72). Between 21 and 25 days post-treatment concentrations were below the LOD in 9/14 milk samples from clinically affected cows.

CONCLUSIONS: Detectable concentrations of buparvaquone were found in the milk of some cows for at least 35 days and in the liver and injection site of some cows until at least 328 days after injection. There were no biologically meaningful differences in milk or tissue concentrations between the formulations, or in the milk concentrations for cows that were clinically affected compared with those that were healthy at the time of treatment.     

Efficacy of doxorubicin as an induction agent for cats with lymphosarcoma

OBJECTIVE: To determine the efficacy of doxorubicin when used alone in inducing remission in cats with lymphosarcoma. DESIGN: Prospective multi-institutional study of naturally occurring disease. METHODS: Cases were accrued from veterinary institutions in Australia and New Zealand after obtaining consent from informed owners. Cats were treated with doxorubicin every 3 weeks for three treatments. If there was no response to the first dose of doxorubicin or if the cat relapsed during the doxorubicin regimen, the cat was withdrawn from the trial and either euthanased or treated with other agents. Age, breed, gender and anatomic site of the lymphosarcoma (multicentric, alimentary, mediastinal, extranodal) were recorded for each cat. Clinical remission was assessed before each treatment by physical examination, radiography, ultrasonography and computed tomography where appropriate. Complete remission was defined as the disappearance of all clinical signs and clinically detectable tumour. RESULTS: Twenty-one cases were accrued over a 2-year-period but only 19 were available for data analysis. Young Siamese cats were over-represented and all cats with mediastinal tumours were young Siamese. There was a significant difference between the mean ages of cats with mediastinal or multicentric lymphosarcoma (mean +/- SD: 3.5 +/- 3.0 and 4.3 +/- 2.6 years, respectively) and cats with alimentary or extranodal LSA (11.4 +/- 0.9 and 11.0 +/- 0.9 years, respectively). Of 19 cats treated with doxorubicin alone, 6 (32%) had complete remission, 6 (32%) had partial remission and 7 (36%) did not respond. CONCLUSIONS: The results suggest that doxorubicin cannot be recommended as a single agent for treatment of feline lymphosarcoma because of the rather poor remission rate achieved.     

Associations between hoof shape and the position of the frontal plane ground reaction force vector in walking horses

AIMS: To determine the frontal plane position of the ground reaction force vector at its centre of pressure under the hoof of walking horses, and its projection through the distal limb joints, and to relate this to hoof geometric measurements.

METHODS: Reflective markers were glued to the forelimb hooves and skin of 26 horses, over palpable landmarks representing centres of the coffin, fetlock and carpal joints, and the dorsal toe at its most distal point. A 4-camera kinematic system recorded the position of these markers as the horse walked in hand across a force platform, to generate a frontal plane representation of the ground reaction force vector passing between the markers at the joints. The position of the vector was calculated as the relative distance between the lateral (0%) and medial (100%) markers at each joint. Digital photos were taken of the hoof in frontal and sagittal views to determine hoof geometric measurements. Associations between these and the position of the force vector at each joint were examined using Pearson correlation coefficients.

RESULTS: Mean vector position for both forelimbs at the toe, coffin, fetlock and carpal joint was 50.1 (SD 8.9), 53.0 (SD 9.2), 54.6 (SD 11.4) and 50.5 (SD17.3)%, respectively, of the distance between the lateral and medial sides of the joint in the frontal plane. Across all four joints, the vector position was slightly more medial (2–4%) for the right than left limb (p>0.05). Medial hoof wall angle was correlated (p<0.05) with force vector position at the fetlock (r=?0.402) and carpal (r=?0.317) joints; lateral hoof wall angle with vector position at the toe (r=0.288) and carpal (r=?0.34) joint, and medial hoof wall height with vector position at the fetlock (r=?0.306) and carpal (r=?0.303) joints.

CONCLUSION: The position of the two-dimensional frontal plane ground reaction force vector at the toe, and at the fetlock and carpal joints was associated with hoof shape. Mediolateral hoof balance has been shown in vitro to affect articular forces, which may be a factor in development of joint disease. The effect of hoof shape needs to be evaluated at faster gaits to determine the potential for joint injury in the presence of larger forces.     

Isolation and characterization of a 6/85-like Mycoplasma gallisepticum from commercial laying hens

Eighty-three-week-old table egg layers with swollen sinuses were presented with a history of increased mortality. Serology revealed positive titers to Mycoplasma gallisepticum (MG). The birds were part of a flock in which some birds had been vaccinated with 6/85 live MG vaccine at 18 wk of age. Tracheal cultures were obtained from both vaccinated and unvaccinated birds within the flock. The cultures were indistinguishable from 6/85 vaccine by both random amplified polymorphic DNA analysis and DNA sequence analysis. Challenge studies were performed to compare the field isolates with 6/85 vaccine and the R strain of MG. The field isolates produced a greater antibody response by serum plate agglutination than did the 6/85 vaccine. The isolates effectively colonized the trachea without increasing the tracheal mucosal thickness; however, they did not extensively colonize the air sacs or cause airsacculitis in the experimental birds.     

Efficacy of grain protectants against four psocid species on maize,rice and wheat

BACKGROUND: Psocids are emerging pests in stored products, particularly in amylaceous commodities such as grains. Currently, their control is based on the use of fumigants and contact insecticides; however, newer data indicate that psocids are tolerant to insecticides used to control other stored‐grain species. This study evaluated the insecticides registered in the USA for use on stored maize, rice and wheat for control of the psocid species Lepinotus reticulatus, Liposcelis entomophila, L. bostrychophila and L. paeta. Mortality of exposed adult females was recorded after 7 and 14 days of exposure, while progeny production was assessed after 30 days of exposure. RESULTS: On wheat and rice, chlorpyriphos‐methyl + deltamethrin was generally more effective against exposed parental adults than spinosad or pyrethrin, while pirimiphos‐methyl was more effective on maize than spinosad or pyrethrin. In most cases, progeny production was suppressed in the treated grains. Progeny production was consistently lowest on wheat and rice treated with chlorpyriphos‐methyl + deltamethrin and maize treated with pirimiphos‐methyl. CONCLUSIONS: Chlorpyriphos‐methyl + deltamethrin and pirimiphos‐methyl were the most effective insecticides for all species and commodities. Conversely, efficacy of spinosad or pyrethrum was highly dependent on the psocid species and commodity. Published in 2009 by John Wiley & Sons, Ltd.     

Efficacy of layer treatment with methoprene for control of Rhyzopertha dominica (Coleoptera: Bostrychidae) on wheat, rice and maize

BACKGROUND: Insect growth regulators are promising alternatives to traditional pesticides in stored grain. The efficacy of the juvenile hormone analogue methoprene was evaluated as a layer treatment in a laboratory experiment for control of Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) in wheat, rice and maize. RESULTS: Adults of R. dominica were placed in vials containing 33, 26 and 29 g (to a depth of 6.5 cm) of wheat, rice and maize, respectively, that was entirely or partially treated with 1, 5 or 10 mg kg^?1 methoprene. In wheat and rice, the layer treatments were not as effective as the whole‐grain treatment, but there was decreased progeny production as the application rate increased. However, on maize the partial treatments were as effective as the whole‐grain treatment at 5 and 10 mg kg^?1. CONCLUSIONS: The results suggest that partial layer treatments with methoprene can be used to control R. dominica on maize but may not be effective for control of this species on wheat and rice. Copyright © 2011 Society of Chemical Industry