Breeding Soundness Evaluation and Semen Analysis for Predicting Bull Fertility

Bull fertility is influenced by numerous factors. Although 20–40% of bulls in an unselected population may have reduced fertility, few are completely sterile. Breeding soundness refers to a bull's ability to get cows pregnant. A standard breeding soundness evaluation identifies bulls with substantial deficits in fertility, but does not consistently identify sub-fertile bulls. In this regard, the use of frozen-thawed semen (from bulls in commercial AI centres) that meets minimum quality standards can result in pregnancy rates that differ by 20–25 percentage points. Although no single diagnostic test can accurately predict variations in fertility among bulls that are producing apparently normal semen, recent studies suggested that a combination of laboratory tests were predictive of fertility. This review is focused on recent developments in prediction of bull fertility, based on assessments at the molecular, cellular and whole-animal levels.     

Cell Cycle Stage Analysis of Rabbit Foetal Fibroblasts and Cumulus Cells

Synchronization of the cell cycle stages in G0/G1 phase is one of the key factors determining the success of nuclear transplantation. Serum deprivation, contact inhibition and chemical inhibitors are widely used methods for this purpose. In this study, cell cycle stages of foetal fibroblasts and cumulus cells were determined using flow cytometry [fluorescence-activated cell scan (FACS)]. Foetal fibroblasts (in vitro cultured for 72-120 h) and fresh cumulus cells were analysed in Experiment 1. Fifty to 55% proliferating fibroblasts remained in G0/G1 phase compared with 78% in confluent culture (p <0.05). In contrast to foetal fibroblasts, fresh cumulus cells maintained 90% of the population in the G0/G1 stage. When serum was retrieved from the proliferating fibroblasts from day 1 to day 5 (Experiment 2), proportions of G0/G1 cells increased from the initial ratio of 53 to 87% at day 4 of starvation, which was significantly higher than the non-starved proliferating cells (p <0.05). In Experiment 3, fibroblasts were treated with aphidicolin (0.1 microg/ml, 6 h), demicolcine (0.5 microg/ml, 10 h), or a combination of these two chemicals to synchronize the cell cycle stages. Surprisingly, no differences or significantly lower in the proportions of G0/G1)phase cells were detected (25-50%) compared with the uncontrolled growing cells (53%). These results suggested that fresh cumulus cells rest their cell cycle in G0/G1 stage. Serum deprivation became effective in the first 24 h and reached the highest proportions during days 4-5 after deprivation. Chemical synchronization of the cell cycle stage of rabbit foetal fibroblasts to G0/G1 phase appeared less effective compared to serum deprivation.     

Experimental Transmission of Corynebacterium pseudotuberculosis Biovar equi in Horses by House Flies

Background

The route of Corynebacterium pseudotuberculosis infection in horses remains undetermined, but transmission by insects is suspected.

Objectives

To investigate house flies (Musca domestica L.) as vectors of C. pseudotuberculosis transmission in horses.

Animals

Eight healthy, adult ponies.

Methods

Randomized, controlled, blinded prospective study. Ten wounds were created in the pectoral region where cages for flies were attached. Three ponies were directly inoculated with C. pseudotuberculosis. Four ponies were exposed for 24 hours to 20 hours C. pseudotuberculosis‐inoculated flies. One negative control pony was exposed to noninoculated flies. Ponies were examined daily for swelling, heat, pain, and drainage at the inoculation site. Blood was collected weekly for CBC and biochemical analysis, and twice weekly for synergistic hemolysis inhibition titers.

Results

Clinical signs of local infection and positive cultures were observed in 7/7 exposed ponies and were absent in the negative control. In exposed ponies, peak serologic titers (1 : 512 to 1 : 2,048) were obtained between days 17 and 21. Seroconversion was not observed in the negative control. Neutrophil counts were higher in the positive and fly‐exposed groups than in the negative control (P = .002 and P = .005) on day 3 postinoculation. Serum amyloid A concentrations were higher in the positive control than in the negative control and fly‐exposed ponies on days 3 (P < .0001) and 7 (P = .0004 and P = .0001). No differences were detected for other biochemical variables.

Conclusions and Clinical Importance

House flies can serve as mechanical vectors of C. pseudotuberculosis and can transmit the bacterium to ponies.     

Keratitis and periocular lesions associated with equine herpesvirus‐3 in a 3‐month‐old filly

A 3‐month‐old Quarter Horse filly presented with corneal ulceration in the right eye with extensive coalescing periocular ulcerations, erosions, and cutaneous crusts. Similar periocular lesions were present around the left eye, on the gingival mucosa, and on the cutaneous and mucosal surfaces of the lips. Based on the severity of the filly's corneal lesions, expense and duration of treatment, euthanasia was elected. Histological post mortem examination revealed numerous hyperplastic and/or dysplastic epithelial cells adjacent to areas of ulceration and erosion with intranuclear viral inclusion bodies. Equine herpesvirus‐3 (EHV‐3) was identified by polymerase chain reaction from the right cornea and lip. The virus was isolated from the right cornea, right eyelid and lip. The dam presented with multifocal to coalescing perineal vesicles. EHV‐3 was confirmed by polymerase chain reaction from the vulvar lesions and the mare recovered spontaneously. This is the first case of EHV‐3 corneal infection reported in horses and emphasises that EHV‐3 should be included as a differential diagnosis for vesicular lesions involving the equine periocular and oronasal epithelium.     

Age-related differences in collagen crimp patterns in the superficial digital flexor tendon core region of untrained horses

Objective To measure collagen fibril crimp angles and lengths as well as collagen fibril mass-average diameters in central and peripheral regions of the superficial digital flexor tendon of wild horses, to ascertain any age-related changes in either region in the absence of imposed galloping exercise.
Design Measurements from a random cull of wild horses.
Sample population Superficial digital flexor tendon samples were taken from 23 wild horses ranging in age from two to ten years.
Procedure Horses were divided into 'young' (< 5 years, n = 10), 'middle-aged' (5 to < 10 years, n = 9) and 'old' groups (10+ years, n = 4) and the mean crimp angle, mean crimp length and collagen fibril mass-average diameter calculated for the central region, and for the peripheral region of each group. Differences between groups and regions were analysed using two-tailed t tests.
Results The crimp angle for the central region was found to decrease with age, so that in old horses it was smaller than that for the tendon periphery (P < 0.05). The crimp angle for the latter region did not alter significantly with age. Crimp period lengths and collagen fibril mass-average diameters did not show significant changes with age in either region.
Conclusion Reduction of the crimp angle in the core of the superficial digital flexor tendon occurs normally with age, as tendons of older animals would have undergone a higher number of loading cycles. It is possible that athletic training increases the frequency and/or the magnitude of high loading cycles experienced by the tendon, and may accelerate and worsen the normal load-related ageing process in the superficial digital flexor tendons of young performance horses, particularly in the central regions where lesions usually occur.