Apoptosis-independent Poor Morphology of Bovine Embryos Produced by Multiple Ovulation

In multiple ovulation and embryo transfer (MOET) programmes in cattle, a considerable number of morphologically poor-quality embryos continue to be produced; this is one of the limiting factors of the technique. Apoptosis has often been implicated in developmental arrest and fragmentation; these are regarded as poor traits of embryonic quality in mammalian pre-implantation embryos. In the present study, apoptosis was assessed in morphologically poor-quality embryos in comparison with good-quality embryos that were recovered from a MOET programme. Retarded embryos (two to 16 cell stage), morulae with severe fragmentation and morphologically good-quality morulae recovered from superstimulated cows at day 7 post-insemination were subjected to TdT-mediated dUTP nick-end labelling (TUNEL) and Hoechst staining. Cell nuclei that showed both TUNEL staining and apoptotic morphology were considered to be apoptotic. Apoptotic index (AI) was calculated as the percentage of apoptotic cells per embryo. Fifteen of 17 retarded embryos and 10 of 15 morphologically poor-quality morulae did not show signs of apoptosis. The mean AIs in the morphologically poor-quality embryos (two to 16 cell stage, 2.2%; poor morulae, 1.3%) were as low as that in the good-quality embryos (2.9%). These results suggest that another mode of developmental arrest and/or fragmentation that is independent of apoptosis occurs in morphologically poor-quality embryos recovered from MOET programmes.     

High dose rate interstitial 192-Ir brachytherapy for the treatment of a recurrent dermal vascular hamartoma in a horse

The successful application of an interstitial brachytherapy course is reported in a 14-month-old male Anglo European Studbook (AES) foal for the treatment of a recurrent dermal vascular hamartoma at the dorsal aspect of the pastern on the right hindlimb. The lesion was irradiated four times over 4 days to a total dose of 28 Gy. Progressive tumour shrinkage and necrosis were observed over 2 months. No significant acute side effect was reported at the end of the treatment, with excellent and long-lasting cosmetic results observed. The only reported late effect was permanent leukotrichia within the treated area, which remained otherwise tumour-free 1224 days after treatment. Results of this report suggest the potential use of radiation therapy (and specifically brachytherapy techniques), as an effective and safe treatment modality for vascular malformations in horses.     

Zero-inflated models for identifying disease risk factors when case detection is imperfect: Application to highly pathogenic avian influenza H5N1 in Thailand

Logistic regression models integrating disease presence/absence data are widely used to identify risk factors for a given disease. However, when data arise from imperfect surveillance systems, the interpretation of results is confusing since explanatory variables can be related either to the occurrence of the disease or to the efficiency of the surveillance system. As an alternative, we present spatial and non-spatial zero-inflated Poisson (ZIP) regressions for modelling the number of highly pathogenic avian influenza (HPAI) H5N1 outbreaks that were reported at subdistrict level in Thailand during the second epidemic wave (July 3rd 2004 to May 5th 2005). The spatial ZIP model fitted the data more effectively than its non-spatial version. This model clarified the role of the different variables: for example, results suggested that human population density was not associated with the disease occurrence but was rather associated with the number of reported outbreaks given disease occurrence. In addition, these models allowed estimating that 902 (95% CI 881–922) subdistricts suffered at least one HPAI H5N1 outbreak in Thailand although only 779 were reported to veterinary authorities, leading to a general surveillance sensitivity of 86.4% (95% CI 84.5–88.4). Finally, the outputs of the spatial ZIP model revealed the spatial distribution of the probability that a subdistrict could have been a false negative. The methodology presented here can easily be adapted to other animal health contexts.     

Influence of Temperament Score and Handling Facility on Stress,Reproductive Hormone Concentrations,and Fixed Time AI Pregnancy Rates in Beef Heifers

The objectives were (i) to evaluate the effect of temperament, determined by modified 2‐point chute exit and gait score, on artificial insemination (AI) pregnancy rates in beef heifers following fixed time AI and (ii) to determine the effect of temperament on cortisol, substance‐P, prolactin and progesterone at initiation of synchronization and at the time of AI. Angus beef heifers (n = 967) at eight locations were included in this study. At the initiation of synchronization (Day 0 = initiation of synchronization), all heifers received a body condition score (BCS), and temperament score (0 = calm; slow exit and walk or 1 = excitable; fast exit or jump or trot or run). Blood samples were collected from a sub‐population of heifers (n = 86) at both synchronization initiation and the time of AI to determine the differences in serum progesterone, cortisol, prolactin and substance‐P concentrations between temperament groups. Heifers were synchronized with 5‐day CO‐Synch+ controlled internal drug release (CIDR) protocol and were inseminated at 56 h after CIDR removal. Heifers were examined for pregnancy by ultrasound 70 days after AI to determine AI pregnancy. Controlling for synchronization treatment (p = 0.03), facility design (p = 0.05), and cattle handling facility design by temperament score interaction (p = 0.02), the AI pregnancy differed between heifers with excitable and calm temperament (51.9% vs 60.3%; p = 0.01). The alley‐way with acute bends and turns, and long straight alley‐way had lower AI pregnancy rate than did the semicircular alley‐way (53.5%, 56.3% and 67.0% respectively; p = 0.05). The serum hormone concentrations differed significantly between different types of cattle handling facility (p < 0.05). The cattle handling facility design by temperament group interactions significantly influenced progesterone (p = 0.01), cortisol (p = 0.01), prolactin (p = 0.02) and substance‐P (p = 0.04) both at the initiation of synchronization and at the time of AI. Inter‐ and intra‐rater agreement for temperament scoring were moderate and good (Kappa = 0.596 ± 0.07 and 0.797 ± 0.11) respectively. The predictive value for calm and pregnant to AI was 0.87, and excited and non‐pregnant to AI was 0.76. In conclusion, the modified 2‐point temperament scoring method can be used to identify heifers with excitable temperament. Heifers with excitable temperament had lower AI pregnancy. Further, cattle handling facility design influenced the temperament and AI pregnancy.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

European oak powdery mildew: impact on trees,effects of environmental factors,and potential effects of climate change

? Context

Powdery mildew is one of the most common diseases of oaks in Europe. After alarming reports in the beginning of the twentieth century following the presumed introduction of the invasive fungus, the disease has become familiar to foresters. However, its impact may vary greatly according to intrinsic and extrinsic factors.

? Aims

We aimed at providing updated and synthesised information on the impact of powdery mildew on oak and on the effects of environment on disease.

? Methods

A comprehensive literature review was performed, including old reports of the early epidemics to more recent data.

? Results

Tree growth patterns are of critical importance to explain the severity of the disease and the differences between juvenile and mature trees. A critical element, especially for infection of mature trees, is the availability of spores during the production of the first leaf flush. High disease impact is often related to modified growth patterns, either by environmental factors (insects or frost) or silvicultural practices (e.g., coppicing).

? Conclusion

Powdery mildew can have important impacts in natural oak regenerations and a significant role in decline of mature trees. Climate change might influence the disease severity mainly by altering the host pathogen phenological synchrony. Process-based models are required for reliable predictions.