Variations in sex ratio, feeding, and fecundity of Triatoma dimidiata (Hemiptera: Reduviidae) among habitats in the Yucatan Peninsula, Mexico.

Chagas' disease is a major public health concern in most Latin American countries and its prevention is based on insect vector control. Previous work showed that in the Yucatan peninsula of Mexico, houses are transiently infested by adult Triatoma dimidiata, which then fail to establish sustained colonies. The present study was designed to evaluate the seasonality and possible causes of the dispersal of sylvatic T. dimidiata toward the houses and the subsequent failure of colonization. Dispersal was highly seasonal and correlated with temperature, pressure, and wind speed. Analysis of sex ratio, feeding status, and fecundity of sylvatic populations of T. dimidiata indicated a rather low feeding status and low potential fecundity, suggesting that seasonal dispersal may be associated with foraging for better conditions. Also, feeding status and potential fecundity tended to improve in the domestic habitat but remained largely suboptimal, suggesting that these factors may contribute to the ineffective colonization of this habitat.

Construction of avian adenovirus CELO recombinants in cosmids.

The avian adenovirus CELO is a promising vector for gene transfer applications. In order to study this potentiality, we developed an improved method for construction of adenovirus vectors in cosmids that was used to engineer the CELO genome. For all the recombinant viruses constructed by this method, the ability to produce infectious particles and the stability of the genome were evaluated in a chicken hepatocarcinoma cell line (LMH cell line). Our aim was to develop a replication-competent vector for vaccination of chickens, so we first generated knockout point mutations into 16 of the 22 unassigned CELO open reading frames (ORFs) to determine if they were essential for virus replication. As the 16 independent mutant viruses replicated in our cellular system, we constructed CELO genomes with various deletions in the regions of these nonessential ORFs. An expression cassette coding for the enhanced green fluorescent protein (eGFP) was inserted in place of these deletions to easily follow expression of the transgene and propagation of the vector in cell monolayers. Height-distinct GFP-expressing CELO vectors were produced that were all replication competent in our system. We then retained the vector backbone with the largest deletion (i.e., 3.6 kb) for the construction of vectors carrying cDNA encoding infectious bursal disease virus proteins. These CELO vectors could be useful for vaccination in the chicken species.

Transcriptional organization of the avian adenovirus CELO.

A detailed map of the transcriptional organization of the CELO virus genome was produced. Recent computer analysis of CELO virus has indicated the presence of 38 putative open reading frames (ORFs). This study, based on analysis of the transcriptional products of CELO in vitro, confirmed the presence of RNAs for 26 of these 38 ORFs. All of the results were obtained by cDNA isolation or specific reverse transcriptase PCR. Observation of ORF transcription kinetics postinfection revealed the existence of early and late expression, with the earliest starting at 2 h postinfection. The 5' untranslated regions of some RNAs were also studied, and this revealed the existence of a bipartite leader sequence, comparable in structure to the tripartite leader of mastadenovirus. The leader most probably involved in transcriptional activity was observed in most of the structural protein genes of the CELO virus genome. This suggests some homology in transcriptional organization between the avian adenovirus CELO and known mastadenoviruses such as human adenovirus.