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The present study was undertaken to examine cell cycle characteristics of endangered Goral (CITES Appendix I) adult skin fibroblasts. Seven experiments were performed, each with a one-way completely randomized design involving three replicates. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused on the effects of cycling, serum-starved, and fully confluent stages of Goral cells. In Experiments II and III, the effects of different antioxidants like beta-mercaptoethanol (beta-ME, 10 microM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 24 h and 4 h, respectively. In Experiments IV and V, three protease inhibitors, namely 6-dimethylaminopurine (6-DMAP, 2 mM), cycloheximide (7.5 microg/ml) and cytochalasin B (7.5 microg/ml), were used as in Experiment II. In Experiments VI and VII, the effect of different levels of dimethylsulphoxide (DMSO) at 0%, 0.5%, 1.0% and 2.5% were tested by flow cytometry (FACS). In Experiment I, 68.7% of Goral skin fibroblasts reached the G(0)/G(1) stage (2C DNA content) when subjected to the serum-starved medium, which was more than the cycling (64.9%) and fully confluent groups (61.0%) (P > 0.05). Among the chemically treated group, beta-ME, cysteine and DMSO showed better results for synchronization of G(0) + G(1) phases than cycling, serum-starved and fully confluent groups. It can thus be concluded that beta-ME, cysteine and DMSO at certain concentrations can synchronize the cell cycle effectively, which could have a positive impact on somatic cell nuclear transfer in the goral.

The present study was conducted to examine the effect of cell culture conditions, antioxidants, protease inhibitors (PI), and different levels of dimethylsulfoxide (DMSO) for the promotion of synchronization of different cell cycles of Siberian tiger skin fibroblasts. We also compared the ability of somatic cell nuclei of the Siberian tiger in pig cytoplasts and to support early development after reconstruction. Cell cycle synchronization between nuclear donor and recipient cells is considered to be one of the most crucial factors for successful cloning. Five experiments were performed each with a one-way completely randomized design involving three replicates of all treatments. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused in the effects of cycling, serum starved and fully confluent stages of Siberian tiger cells on different cell cycles. In Experiment II, the effects of different antioxidants like b-Mercaptoethanol (b-ME, 10 mM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 4 hr. In Experiment III, three PI, namely 6-dimethylaminopurine (6-DMAP, 2 mM), cycloheximide (7.5 mg/ml) and cytochalasin B (7.5 mg/ ml) were used in the sane manner as in Experiment II. In Experiment IV, different levels of DMSO at 0%, 0.5%, 1.0%, and 2.5% were tested on different cell cycle stages of Siberian tiger examined by Flowcytometry (FACS). In Experiment I, 67.2% of the Siberian tiger skin fibroblasts reached the G 0 /G 1 stage (2C DNA content) in fully confluent conditions which was more than the cycling (49.8%) and serum starved (SS) medium (65.5%; P < 0.05). Among the chemically treated group, glutathione (72.6%) and cycloheximide (71.3%) had little bit better results for the synchronization of G 0 þ G 1 phases than serum starved and fully confluent. After nuclear transfer we did not see any significant differences on the development of tigerporcine reconstructed embryos at cycling, SS and fully confluent. Data indicate that prolonged culture of cells in the absence of serum as well as using different chemicals for this experiment does not imply a shift in the percentage of cells that enter G 0 /G 1 and that confluency is sufficient to induce quiescence. This finding can be beneficial in nuclear transfer programs in Siberian tiger, because there are negative effects, such as apoptosis associated with serum starvation. Mol. Reprod. Dev. 74: 403-411, 2007. ß

Poultry is an integral part of the rural livelihoods in Cambodia, with more than half of the households keeping poultry in their small-scale, traditional, and extensive backyards. More than 20 highly pathogenic avian influenza (HPAI) outbreaks have been reported since 2004 with deaths of over 21,000 birds. During the HPAI outbreaks, some of the flocks in the rural areas were culled without compensation and producers were not allowed to sell outside of the community. Heifer International worked with 2,000 rural families through local project partners in the target communities to develop an effective intervention mechanism to mitigate the impact of the HPAI crisis. Heifer International provided training, public education, and networking as well as promoting model farms based on improved scavenging poultry management. Each community selected one farm family to serve as a model farm. They were trained in Heifer's working approach and committed to practicing integrated farming systems based on scavenging poultry management. One Village Animal Health Worker (VAHW) in each community participated during the project implementation, playing a key role in the information exchange and the interaction between the communities and the avian influenza experts. Formal and informal trainings were conducted for all project partners and project recipients through experts and VAHWs, respectively. There have been no outbreaks reported in the communities in the project areas. Farmers have started using appropriate techniques to maintain biosecurity. They are passing on the knowledge and the skills to the surrounding communities. This participatory approach in educating rural farmers can serve as a model to mitigate HPAI in the developing countries around the world.

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