Apoptosis in the Antral Follicles of Swamp Buffalo and Cattle Ovary: TUNEL and Caspase-3 Histochemistry

The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process.     

Cell Proliferation in the Atretic Follicles of Buffalo and Cattle Ovary

The present study was carried out to describe the proliferative activity of granulosa and theca cells in healthy antral and atretic follicles of Philippine buffaloes (BU) and Holstein-Friesian (HF) cows. Paraffin sections of ovary were immunostained with mouse monoclonal antibody to proliferating cell nuclear antigen (PCNA). Then the follicles were classified into healthy and various stages of atretic follicles. The granulosa layer of healthy follicles had a significantly higher frequency of PCNA-positive cells than the early and advanced atretic follicles in both breeds. In the theca interna, significantly reduced populations of the PCNA-positive cells were found in both breeds as atresia progressed. Moreover, HF had significantly higher PCNA-positive cells in the theca interna of healthy, early atretic and advanced atretic follicles than BU. A reduction of PCNA-positive cells during atresia was also noted in the theca externa in both animals although differences were not significant. The results of the present work suggest that the proliferative activity of granulosa and theca cells decreases in association with follicular atresia in the BU similar to HF. Furthermore, a significantly deficient cell proliferative activity of theca interna was found in BU compared with HF.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post‐thaw viability of Piau‐breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800  g for 10 min and 2400  g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour – freezing 1 and automated cooling using a programmed freezing machine – freezing 2) were tested. Therefore, the treatments were divided into M3 – centrifugation at 2400  g for 3 min and freezing 2; M10 – centrifugation at 800  g for 10 min and freezing 2; R3 – centrifugation at 2400  g for 3 min and freezing 1; and R10 – centrifugation at 800  g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post‐thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo‐osmotic test (HO), sperm–egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post‐thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury.     

Financial incentive to control paratuberculosis (Johne's disease) on dairy farms in the United Kingdom

This paper estimates the financial incentive to control paratuberculosis on dairy farms by establishing the level of expenditure that would minimise the total cost of the disease (output losses plus control expenditure). Given the late onset of the clinical signs and the lack of treatments, control was focused on minimising the financial impact of paratuberculosis by adjusting the dairy cow replacement policy. The optimum replacement policies for disease-free herds and infected herds were compared by using dynamic programming. At the standard settings, the disease justified adjusting the culling policy; under constant bioeconomic assumptions, it reduced the expected annuity from milk production under the optimal replacement policy by about 10 per cent (27 pounds sterling per cow annually), a considerably lower figure than for other major endemic diseases that affect dairy cows in the uk. The effect was even less at lower milk prices, suggesting that there is at present little incentive for dairy farmers to put more resources into controlling the disease. However, the incentive could be increased if more information were available about how best to manage the disease under specific farm circumstances. Any effect that paratuberculosis may have on the future demand for milk and hence on milk prices would also be an important consideration.     

Optimal risk management versus willingness to pay for BVDV control options

Previous work has shown that the least-cost BVD-control option for cow-calf herds in Scotland was not necessarily the risk-minimising option. Thus, assessing BVDV prevention measures must account for risk reduction as well as decision-makers' attitude towards risks. We therefore describe a method to do this using a hypothetical example. Data for this analysis were generated using a simulation model of BVDV transmission in a typical Scottish cow-calf herd over a 10-year period. Herd infection/re-infection was adjusted to reflect the expected risk of infection/re-infection and the use of either biosecurity or vaccination strategies at various levels of effectiveness. The level of risk-free financial return that maximises farmers' utility of wealth was significantly affected by the assumed effectiveness of the control strategy. More importantly, it was observed that utility maximisation of wealth as a business objective is not an optimal solution in terms of animal welfare when dealing with a BVDV outbreak.     

Assessing economic and social pressure for the control of bovine viral diarrhoea virus

The objective of this paper is to present a preliminary assessment of variation in the economic impact of bovine viral diarrhoea virus (BVDV) at dairy farm level between a sample of nations within the EU and hence assess differences in pressure to respond to this disease that may be impeding progress in control and hence restricting collective benefits from healthier livestock. We used a questionnaire to obtain national average values of key epidemiological and economic parameters for a typical dairy farm from BVDV experts in the countries concerned. These parameters were converted into assessments of economic impact using a computer simulation model. Uncontrolled output losses for a BVDV-na?ve herd with virus introduced in year 1 of a 10-year epidemic represented 22, 7, 8, 5, 8 and 20% of the BVDV-free annuity for the UK, Northern Portugal, Holland, Norway, Italy and Germany, respectively. Differences between countries will be widened by differences in the risk of acquiring BVDV. These will be much reduced in countries, such as Norway that have a national BVDV eradication programme. Farmers in such countries can therefore justify spending much less on maintaining BVDV-free status than BVDV-free farms in other countries. This result illustrates the paradox that in countries where BVDV prevalence is high, farmers have least to gain from unilateral BVDV eradication because of the high cost of maintaining freedom from the disease. We discuss this issue in the light of increasing recognition at international level of the importance of BVDV control.