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.
Nanomaterials composed of natural matrices associated with biopesticides have promising applications in sustainable agriculture. In this study, the biopesticide neem Powered by Editorial Manager® and ProduXion Manager® from Aries Systems CorporationAnswer: Thank you for your comment. We have inserted the requested information throughout the manuscript. Please verify the revised version.Reviewer: Figure 5. I do not agree with the analysis. I will suggest to apply repeated measures ANOVA or survival analysis and their curves. Answer: Thank you for your comment. We would like to explain that in Figure 5, there is no need to perform repeated measurements ANOVA because our data do not represent measurements over time of the same sample. In fact, different leaves were collected on different days after the application of the treatments to perform the test, and not a single leaf was collected and analyzed 1, 6 and 12 days latter. We have modified the figure caption for better understanding. Please verify the revised version of the manuscript.Reviewer: Line 591-801. Firstly, discussion section seems like a review of literature and many of the sentenses are awkwardly placed. I will suggest to rewrite the whole sentense. Secondly, hard to get the idea due to linguistic and syntax errors. Answer: Thank you for your comment. We have abbreviated the discussion and some cited studies were deleted. Please verify the revised version of the manuscript.Reviewer: Line 592-600. The opening paragraph of the discussion section is not appropriately written. I will suggest to rewrite this section. Answer: Thank you for your comment. We have rewritten the discussion. Please verify the revised version of the manuscript.Reviewer: Line 553-554. The sentense is awkwardly placed. Answer: Thank you for your comment. We have deleted the indicated sentence.Reviewer: Line 801. I will suggest the authors to provide a comprehensive conclusion of the study. Answer: Thank you for your comment. We have rewritten the conclusion. Please verify the revised version of the manuscript. Reviewer #2: Reviewer: I have read the manuscript entitled " Nanobiopesticide based on zein nanoparticles and neem oil: a study using target and nontarget organisms". The manuscript presents a well-written and appropriately analyzed series of experiments to determine the pesticidal and biological activity of neem oil-loaded zein nanoparticles against three pests (Acanthoscelides obtectus, Bemisia tabaci, and Tetranychus urticae), in addition to the phytotoxic effects of these nanoparticles using Phaseolus vulgaris. Overall, the manuscript is generally clear and concise report of a well-executed study. The objectives are clear; the experiments are pertinent and follow a logical reasoning; the main findings of the study are convincing and the conclusion is appropriate. The paper is clearly organized and the contribution is interesting and falls within the scope of the journal. The work is generally well written, except for certain parts of the manuscript, where a good technical ...
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.
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.
Leishmaniasis encompasses a spectrum of diseases caused by a protozoan belonging to the genus Leishmania. The parasite is transmitted by the bite of sand flies, which inoculate the promastigote forms into the host’s skin while acquiring a blood meal. Nyssomyia neivai is one of the main vectors of tegumentary leishmaniasis (TL) in Brazil. Southeastern Brazil is an endemic region for TL but also overlaps with an endemic focus for pemphigus foliaceus (PF), also known as Fogo Selvagem. Salivary proteins of sand flies, specifically maxadilan and LJM11, have been related to pemphigus etiopathogenesis in the New World, being proposed as an environmental trigger for autoimmunity. We present a comprehensive description of the salivary transcriptome of the N. neivai, using deep sequencing achieved by the Illumina protocol. In addition, we highlight the abundances of several N. neivai salivary proteins and use phylogenetic analysis to compare with Old- and New-World sand fly salivary proteins. The collection of protein sequences associated with the salivary glands of N. neivai can be useful for monitoring vector control strategies as biomarkers of N. neivai, as well as driving vector-vaccine design for leishmaniasis. Additionally, this catalog will serve as reference to screen for possible antigenic peptide candidates triggering anti-Desmoglein-1 autoantibodies.
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