Organogenic responses of Pinus pinea cotyledons to hormonal treatments: BA metabolism and cytokinin content

Isolated cotyledons from mature Pinus pinea L. embryos were cultured in vitro in a factorial combination of 4.4, 10 and 44.4 microM N6-benzyladenine (BA) for 2, 4, 8, 16 and 35 days to optimize shoot regeneration. Incubation of explants in 44.4 microM BA for 4 days, in place of the standard incubation in 4.4 microM BA for 35 days, reduced the entire culture period to 4 weeks. Shortening the culture period had no significant effect on the caulogenic response or the number of buds formed per cotyledon. To establish the relationship between key moments in the caulogenic process induced by 4.4 microM BA and the endogenous concentrations of the active forms of BA and other isoprenoid-type cytokinins (CKs), we examined uptake, metabolism and amount of BA, as well as the amounts of zeatin, dihydrozeatin and their ribosides in P. pinea cotyledons after 1, 2, 6, 12 and 24 h, and 2, 4, 8, 16 and 35 days of exposure to 8-[14C]BA. Uptake and release of BA were associated with water movement between explants and the medium during the first 8 days of culture. The interconvertible forms of BA were the main metabolites formed in the tissues. Inactivation of BA as a result of conjugation or oxidation was insignificant. The endogenous concentration of BA + N6-benzyladenosine was 20-fold higher than the exogenously applied BA during the competence acquisition phase (Days 0-3). The concentration of isoprenoid-type CKs also increased 16-fold and then decreased during this time. Induction of shoot buds (Days 4-8) was characterized by a second peak of BA uptake by explants that triggered the synthesis of N6-benzyladenosine-5 -monophosphate and by the maintenance of isoprenoid-type CKs. Reestablishment of CK homeostasis marked the shift from the induction phase to the shoot development phase in this organogenic process (Days 8-12).     

Risk factors to incidental leptospirosis and its role on the reproduction of ewes and goats of Espírito Santo state,Brazil

Ovine and caprine stockbreeding have been gaining attention in developing countries as an attractive investment. On these animals, infectious diseases of the reproductive tract, such as leptospirosis, can compromise the production leading to economic losses. The present study aimed to determine the risk factors associated with incidental leptospirosis and its influence on the reproductive parameters of ewes and goats of Espírito Santo state, Brazil. A total of 737 animals distributed on 24 herds/flocks were studied, and an overall prevalence of 10.9 % seroreactive animals was observed. Serogroup Icterohaemorrhagiae was the most frequent in goats (97.0 %) as well as in ewes (78.3 %). Regarding risk factors related to leptospirosis, the presence of waterholes and the semi-intensive breeding system were the most important associated to seroreactivity. Besides, there was an observed association between seroreactivity and reproductive failures (P?<?0.05). Moreover, seroreactive ewes (relative risk (RR)?=?1.3) and goats (RR?=?1.9) presented more chances to have abortions than seronegative animals. Furthermore, seroreactive ewes presented 11.6 more chances of having premature births when compared to the seronegative ones. It can be concluded that Leptospira infection, mainly those caused incidental strains (such as Icterohaemorrhagiae serogroup), is a significant factor to reduce the productivity of small ruminants’ herds/flocks in the studied region, and environmental measures must be considered on control programs.