Influence of Leishmania (Viannia) braziliensis infection on the attractiveness of BALB/c mice to Nyssomyia neivai (Diptera: Psychodidae).

BACKGROUND: Phlebotomine sand flies are vectors for several pathogens, with Leishmania being the most important. In Brazil, the main aetiological agent of American cutaneous leishmaniasis (ACL) is Leishmania (Viannia) braziliensis, and Nyssomyia neivai is one of its main vectors in São Paulo state and other areas of South America. Similar to other haematophagous insects, sand flies use volatile compounds called kairomones to locate their hosts for blood meals. A possible increase in the attractiveness of hosts infected with Leishmania infantum to their vectors has been demonstrated. In the present study, we aimed to investigate whether L. braziliensis-infected hosts present higher attractiveness to Ny. neivai and to identify differences in the volatile compounds released by infected and uninfected mice. RESULTS: Behavioural experiments in which sand fly females directly fed on infected or uninfected mice showed no significant differences in the attractiveness of the mice or the blood volume ingested. Y-tube olfactometer bioassays also revealed no significant differences in the attractiveness of these hosts to Ny. neivai. No differences were observed in the profiles of the volatile compounds released by the two groups of mice. However, PCA and cluster analysis were able to classify the 31 identified compounds into three clusters according to their abundances. This classification showed a possible role for individual variation in the absence of differences in volatile profiles and attractiveness between infected and uninfected mice. CONCLUSION: In this first cross-sectional study with an aetiological agent of ACL, there were no statistically significant differences in the attractiveness of infected hosts to their vector.

Breeding protocol for the sand fly Nyssomyia neivai (Diptera: Psychodidae) in laboratory conditions.

The information in this protocol covers from the basic steps and material necessary to start a sand fly colony up to the specific details which are important to the success of a Nyssomyia neivai colony. The greatest problems in our colony of Ny. neivai were solved with specific care, for instance, using vermiculite and an adequate number of adults in oviposition containers; the control of fungus with the exact amount of diet for the larvae and humidity control; a second blood meal for females and control of the number of times animals are used for blood meals. Currently, our colony is at F22 generation.

Solid phase microextraction, sand flies, oviposition pheromones, plaster of Paris and siloxanes-What is in common?

Sand flies are natural hosts of various microorganisms. Due to their epidemiological importance, sand fly colonies are kept in laboratories to be studied in terms of their biology and vector/host/parasite interactions. In order to investigate the presence of oviposition pheromones in Nyssomyia neivai, experiments using Solid Phase Microextraction (SPME) were performed. However, siloxanes which is an external class of contamination, present in breeding containers made by plaster used to maintain sand flies in colonies, may be hindered the experiments.

A Coxiella mutualist symbiont is essential to the development of Rhipicephalus microplus.

The cattle tick Rhipicephalus microplus is a hematophagous ectoparasite that causes important economic losses in livestock. Different species of ticks harbor a symbiont bacterium of the genus Coxiella. It was showed that a Coxiella endosymbiont from R. microplus (CERM) is a vertically transmitted mutualist symbiont, comprising 98% of the 16S rRNA sequences in both eggs and larvae. Sequencing of the bacterial genome revealed genes for biosynthetic pathways for several vitamins and key metabolic cofactors that may provide a nutritional complement to the tick host. The CERM was abundant in ovary and Malpighian tubule of fully engorged female. Tetracycline treatment of either the tick or the vertebrate host reduced levels of bacteria in progeny in 74% for eggs and 90% for larvae without major impact neither on the reproductive fitness of the adult female or on embryo development. However, CERM proved to be essential for the tick to reach the adult life stage, as under antibiotic treatment no tick was able to progress beyond the metanymph stage. Data presented here suggest that interference in the symbiotic CERM-R. microplus relationship may be useful to the development of alternative control methods, highlighting the interdependence between ticks and their endosymbionts.