Epigenetic modifiers during in vitro maturation as a strategy to increase oocyte competence in bovine.

Oocyte in vitro maturation (IVM) is a key procedure for the in vitro production (IVP) of bovine embryos; however, the efficiency of this step is limited by the intrinsic developmental competence of oocytes. This study aimed to investigate possible epigenetic changes resulting from using of histone deacetylase (HDAC) inhibitors in the maturation of bovine oocytes and subsequent embryo development. Initially, we investigated the meiotic progression of bovine oocytes matured in vitro in the presence of trichostatin A (TSA). We then evaluated histone H3k9 acetylation levels in oocytes exposed to different TSA concentrations, and the relative expression of genes linked to oocyte competence. Finally, we studied pre-implantation embryonic development by analyzing the cleavage, blastocysts, and hatching rates. Acetylation levels of H3k9 increased (p < 0.05) when oocytes were exposed to 50 nM or 100 nM TSA during IVM, but there were no significant changes in the relative expression of the evaluated genes p34cdc2, cyclin B1, MAPK, GDF9, G6PDH, and HSP70. We found that 5 nM TSA promoted the attenuation of meiotic progression and positively affected pre-implantation embryo development in bovine species, allowing a 10% increase in the blastocyst rate. We concluded that TSA treatment during IVM was efficient in promoting changes in H3k9 acetylation levels from 50 nM and promoted attenuated meiotic progression in bovine oocytes at all concentrations evaluated, with a positive impact on pre-implantation development when used at low concentrations.

Isoflurane MAC determination in dogs using three intensities of constant-current electrical stimulation.

OBJECTIVES: To compare isoflurane minimum alveolar concentrations (MACs) in dogs determined using three intensities of constant-current electrical stimulation applied at the tail, and thoracic and pelvic limbs, and to compare isoflurane MACs obtained with all combinations of electrical stimulation and anatomic site with those obtained using the tail clamp as the noxious stimulus. STUDY DESIGN: Randomized trial. ANIMALS: Six mixed-breed, adult female dogs aged 1-2 years and weighing 11.1 ± 4.4 kg. METHODS: In each dog, MAC was determined by the bracketing method with the tail clamp (MACTAILCLAMP ), and three electrical currents (10 mA, 30 mA, 50 mA) at three anatomic sites (thoracic limb, pelvic limb, tail). Each MAC achieved with electrical stimulation was compared with MACTAILCLAMP using a mixed-model anova and Dunnett's procedure for multiple comparisons. The effects of current intensity and anatomic site on isoflurane MAC were tested using a mixed-model anova followed by Tukey's test for multiple comparisons (p < 0.05). RESULTS: Mean MACTAILCLAMP was 1.69%. MACs achieved with currents of 30 mA and 50 mA did not differ independently of anatomic site. When currents of 10 mA were applied to the tail and thoracic limb, resulting MACs were lower than those obtained using currents of 30 mA and 50 mA. Currents of 30 mA and 50 mA provided MACs that did not differ from those of MACTAILCLAMP , whereas a current of 10 mA achieved the same result only for the pelvic limb. CONCLUSIONS AND CLINICAL RELEVANCE: Isoflurane MAC is affected by current intensity and anatomic site. Current intensities of 30 mA and 50 mA provided consistent results when applied to the tail, and thoracic and pelvic limbs that did not differ from those obtained using the tail clamp. Consequently, they can be used in place of the tail clamp in MAC studies in dogs.

Natural Plasmodium infection in monkeys in the state of Rondônia (Brazilian Western Amazon).

BACKGROUND: Simian malaria is still an open question concerning the species of Plasmodium parasites and species of New World monkeys susceptible to the parasites. In addition, the lingering question as to whether these animals are reservoirs for human malaria might become important especially in a scenario of eradication of the disease. To aid in the answers to these questions, monkeys were surveyed for malaria parasite natural infection in the Amazonian state of Rondônia, Brazil, a state with intense environmental alterations due to human activities, which facilitated sampling of the animals. METHODS: Parasites were detected and identified in DNA from blood of monkeys, by PCR with primers for the 18S rRNA, CSP and MSP1 genes and sequencing of the amplified fragments. Multiplex PCR primers for the 18S rRNA genes were designed for the parasite species Plasmodium falciparum and Plasmodium vivax, Plasmodium malariae/Plasmodium brasilianum and Plasmodium simium. RESULTS: An overall infection rate of 10.9% was observed or 20 out 184 monkey specimens surveyed, mostly by P. brasilianum. However, four specimens of monkeys were found infected with P. falciparum, two of them doubly infected with P. brasilianum and P. falciparum. In addition, a species of monkey of the family Aotidae, Aotus nigriceps, is firstly reported here naturally infected with P. brasilianum. None of the monkeys surveyed was found infected with P. simium/P. vivax. CONCLUSION: The rate of natural Plasmodium infection in monkeys in the Brazilian state of Rondônia is in line with previous surveys of simian malaria in the Amazon region. The fact that a monkey species was found that had not previously been described to harbour malaria parasites indicates that the list of monkey species susceptible to Plasmodium infection is yet to be completed. Furthermore, finding monkeys in the region infected with P. falciparum clearly indicates parasite transfer from humans to the animals. Whether this parasite can be transferred back to humans and how persistent the parasite is in monkeys in the wild so to be efficient reservoirs of the disease, is yet to be evaluated. Finding different species of monkeys infected with this parasite species suggests indeed that these animals can act as reservoirs of human malaria.