Attractiveness of golden hamster infected with Leishmania amazonensis (Kinetoplastida: Trypanosomatidae) to laboratory-reared Lutzomyia longipalpis (Diptera: Psychodidae).

Lutzomyia longipalpis is the primary vector of Leishmania infantum in the Americas and a permissive vector for Leishmania amazonensis. Previous studies showed that Leishmania infantum-infected hosts can release different volatile organic compounds (VOCs) compared with uninfected hosts, presenting a higher attractiveness to vectors. In this study, we aimed to evaluate a possible effect of L. amazonensis infection of golden hamsters in three parameters: attractiveness to Lu. longipalpis females; blood volume ingested by sand fly females; and VOCs released by the animals.. Attractiveness was measured indirectly by the number of Lu. longipalpis females that blood fed in each L. (L.) amazonensis-infected and uninfected animal. For VOCs extraction, solid phase micro extraction fibers were used, which were analyzed by gas chromatography-mass spectrometry. Behavioral trials did not show any effect of L. amazonensis infection on the attraction of sand flies nor difference on blood meal rates of Lu. longipalpis fed in both goups of hamsters. Additionally, there was no difference between the VOCs profiles of L. (L.) amazonensis-infected or uninfected hamsters.

Semiochemical delivery systems based on natural polymers to attract sand flies (Diptera: Psychodidae).

BACKGROUND: The successful use of semiochemicals to attract insects to traps is based on research on the most suitable compounds and their release profiles over time. Based on the group's promising results, matrices with a more adequate release profile and more eco-friendly properties for the release of 1-hexanol were developed. To use a more suitable prototype in the field, the most promising systems were added to a capsule and evaluated in a wind tunnel. Behavioral experiments were performed using the sand fly species, Lutzomyia longipalpis, to evaluate the efficacy of the proposed system. METHODS: Different delivery systems were developed by varying the polymer (gellan gum and pectin) ratio, crosslinker (aluminum chloride) concentration, and glutaraldehyde removal.The delivery systems were loaded with 1-hexanol, and their release profiles were evaluated using gravimetric analysis under ambient and high-humidity conditions. When the matrix system was placed inside a plastic container, modulations in the active release profile were observed and the system could be reused. Actid attraction behaviors of the sand fly species, Lu. longipalpis, were evaluated in a wind tunnel when exposed to 1-hexanol-loaded release systems at different times. RESULTS: Among the four formulations evaluated, System 2 (gellan gum and pectin in a 1:1 ratio with 5% aluminum chloride) exhibited the most promising release profile, with greater uniformity and longer compound release time. The maximum 1-hexanol release uniformity was achieved over a longer time, mainly every 24 h, under both ambient and high-humidity conditions. System 2 can be reused at least once with the same structure. The wind tunnel trials exhibited efficient activation and attraction of Lu. longipalpis to 1-hexanol after 24, 48, and 72 h in System 2 placed inside the capsules. CONCLUSIONS: The polymeric matrix supplemented with 1-hexanol and introduced in plastic capsules showed promising results in attracting sand flies. This system can be used as a solution for other attractive compounds as well as in other applications where their release needs to be controlled or prolonged.

Light-emitting diodes as visual attractants to phlebotomine sand flies (Diptera: Psychodidae): A mini-review.

Phlebotomine sand flies are responsible for the transmission of different pathogens, including flagellate protozoan of the genus Leishmania. To locate a vertebrate host, females use chemical cues called kairomones, which are also used by males and females to locate carbohydrate sources of food. In addition to these chemical cues, visual cues (such as light and color) can play a key role in determining host location. In this way, incandescent white light is widely used to capture these insects in the field, being more recently replaced by light-emitting diode (LEDs) technology, which can be used with different colors. In this review, we summarize the main results of the use of LEDs as attractants for sand flies. Considering the different LEDs colors, blue and green are responsible for catching the highest number of sand fly species, especially for field trials conducted in New World countries. The intensity of light colors is an important factor to be considered, but so far little explored. In general, the use of LEDs can be considered as a feasible tool for the improvement of light traps. Future research is required to assert the contribution of different colors individually, in different combinations and intensities, as well as the possible synergistic effect of LEDs in association with attractive compounds in the behavior of sand flies.

Phlebotominae Fauna (Diptera: Psychodidae) and the Spatial Distribution of Species in Sergipe, Brazil.

Some sand fly species are the vectors responsible for the transmission of Leishmania spp. (Kinetoplastida: Trypanosomatidae), the etiological agent of leishmaniasis, and in the state of Sergipe, the two main forms of the disease (visceral and cutaneous) are recorded. Few works show information about the species that form the Phlebotominae fauna in Sergipe. This study aimed to update and determine how they are distributed throughout the state. The study used data from surveys about phlebotomines in Sergipe, from the Program of Surveillance and Control of Visceral Leishmaniasis, carried out by the Central Public Health Laboratory of Sergipe, from 2008 to 2018, along with review and original data from 2022. The commentary on this information was developed with focus on the species that can be vectors for the disease. Sergipe has, up to now, 27 registered species of phlebotomines from 12 genera, 15 of them are of sanitary relevance. Twenty two of these species are in the mesoregion East, 19 in the Agreste, and 11 in the Sertão of Sergipe. The species with the greatest distribution was Evandromyia lenti (Mangabeira, 1938), present in 74.6% of the municipalities of Sergipe, followed by Lutzomyia longipalpis (Lutz & Neiva, 1912), the main vector of the etiological agent of visceral leishmaniasis in Brazil, present in 68% of the municipalities studied. This study expands the number and distribution of species recorded in the state. This information can contribute to disease containment plans and support health education actions aimed at the control of leishmaniasis in Sergipe.

Behavioral responses of sand fly Nyssomyia neivai (Psychodidae: Phlebotominae) to 1-hexanol and light.

BACKGROUND: The search for attractive baits that may facilitate the capture of haematophagous insects has been epidemiologically relevant. Sand flies use chemical cues in different phases of their life cycles to find carbohydrate meals, mates, blood meals and oviposition sites. Few studies have related the behaviours of sand flies with volatile compounds that can influence their life cycles. Previous studies in our laboratory have shown that 1-hexanol released on filter paper is a good attractant for the sand fly Nyssomyia neivai, which is suspected in the transmission of the aetiologic agent of American cutaneous leishmaniasis. METHODS: In this study, we developed two release systems to modulated 1-hexanol release: system 1 contained gellan gum and pectin (4:1 ratio), 3% aluminium chloride and 1% glutaraldehyde; system 2 contained: gellan gum and pectin (4:1 ratio) and 3% aluminium chloride. After addition of 1-hexanol to each release system, trials were performed in a wind tunnel with Ny. neivai males and females (unfed, blood-fed and gravid) to evaluate activation and attraction responses. RESULTS: Males and unfed females showed the same response pattern to the systems. For both systems, the males and unfed females of the sand flies showed an activation response up to 24 h. The number of responsive gravid females was lower than unfed females for both systems. The blood-fed females showed no responses in any of the release systems. CONCLUSIONS: Our findings indicate that the state of the females (unfed, fed and gravid) can interfere with the sand fly responses to volatile compounds. Additionally, both systems evaluated with the compound showed effectiveness for sand fly attraction.

Immature instars of three species of Rhodnius Stål, 1859 (Hemiptera, Reduviidae, Triatominae): morphology, morphometry, and taxonomic implications.

BACKGROUND: Among the 18 genera of the Triatominae subfamily, three stand out for their diversity and epidemiological importance: Triatoma, Panstrongylus, and Rhodnius. Rhodnius includes 21 species that can transmit Trypanosoma cruzi (the etiological agent of Chagas disease, also known as American trypanosomiasis) and Trypanosoma rangeli. The Rhodnius prolixus complex comprises seven species, including Rhodnius marabaensis, Rhodnius prolixus, and Rhodnius robustus, which occur in the northern region of Brazil. Since both adults and immatures can carry T. cruzi, in this study the five nymphal instars of the three species mentioned were dorsally characterized. METHODS: Using microscopy, morphometrics, and geometric morphometrics, the present work measures and describes the morphological characters of the five nymphal instars of R. marabaensis, R. prolixus, and R. robustus. RESULTS: The study enabled the characterization of all five nymphal instars, as well as the distinction between the three species in each of their instars. CONCLUSIONS: The morphological, morphometrics of the head, thorax, and abdomen and geometric morphometrics studies of the head enabled the specific distinction of these three species in all five instars.

Nanotechnology as a tool for detection and treatment of arbovirus infections.

Arboviruses are medically important viruses that cause high rates of infection all over the world. In addition, the severity of the symptoms and the inadequate diagnostic methods represent a challenge far beyond eradicating the vector. The lack of specific treatments for arbovirus infections reflects the imminent need for new research for safe and efficient medicines to treat these infections. Nanotechnology is an innovative approach currently used as a platform for developing new treatments, thus improving the biopharmaceutical properties of drugs. It can also be applied to the development of diagnostic devices, improving their detection capacity. The purpose of this paper is to review recent research on the use of nanotechnology for developing new treatments and detection devices for arbovirus infections. Interestingly, it was found that only a few studies report on the use of nanotechnology to treat arbovirus infections and that most of these reports focus on the fabrication of diagnostic tools. Also, some papers report on the use of nanotechnology for the development of vaccines, which in association with mosquito eradication programs could effectively reduce the high rates of infections by these viruses.

Botanical insecticide-based nanosystems for the control of Aedes (Stegomyia) aegypti larvae.

Aedes (Stegomyia) aegypti is a cosmopolitan species that transmits arbovirus of medical importance as dengue, Zika, and chikungunya. The main strategy employed for the control of this mosquito is the use of larvicidal agents. However, the overuse of synthetic chemical larvicides has led to an increase in resistant insects, making management difficult. Therefore, the use of botanical insecticide-based nanosystems as an alternative to the use of synthetic agents for the control of Ae. aegypti has gained more considerable attention in the last years, mainly due to the advantages of nanostructured delivery systems, such as (a) controlled release; (b) greater surface area; (c) improvement of biological activity; (d) protection of natural bioactive agents from the environment and thus achieving stability; and (e) lipophilic drugs are easier dispersed even in aqueous vehicles. This review summarizes the current knowledge about botanical insecticide-based nanosystems as larvicidal against Ae. aegypti larvae. The majority of papers used metallic nanoparticles (NPs) as larvicidal agents, mainly silver nanoparticles (AgNPs), showing potential for their use as an alternative, followed by nanoemulsions containing vegetable oils, most essential oils, nanosystems that allow the dispersion of this high hydrophobic product in water, the environment of larval development. The final section describes scientific findings about the mode of action of these NPs, showing the gap about this subject in literature.

Bovine fasciolisis in São Paulo state, Brazil.

Fasciolosis is a zoonotic disease with a worldwide distribution caused by Fasciola hepatica, which leads to severe economic losses in cattle such as reducing meat and milk production, livers condemnation, growth retardation and increase in mortality. From October 2008 to April 2011, condemned bovine livers in slaughterhouses of different municipalities from São Paulo state, Brazil were evaluated for the presence of Fasciola hepatica. Out of 20,635 analyzed livers, 1422 were infected with F. hepatica. These cattle came from 33 municipalities, out of which 16 showed infected animals and where 7 municipalities did not show statistical difference between each month throughout the year: Tuiuti - 276/1408 (19,6%), Atibaia - 44/257 (17,1%), Joanópolis - 116/738 (15,7%), Bragança Paulista - 318/2316 (13,3%), Piracaia - 182/1442 (12,6%), Santo Antonio de Posse - 118/1005 (11,7%), Amparo 131/2003 (6,5%). The other nine municipalities, Monte Alegre do Sul, Descalvado, Campinas, Morungaba, Pedreira, Socorro, Munhoz, Jaguariúna and Itatiba showed a positive percentage varying from 5.08% to 1.46%. Our results demonstrated the presence of F. hepatica in this region was higher than official data, bringing the need for control measures. There is also an apparent increase in fasciolosis two to three months after low to medium precipitation, however high precipitation causes a decrease in fasciolosis prevalence.

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.

Three new records of the genus Lutzomyia of the subgenus Helcocyrtomyia (Diptera: Psychodidae: Phlebotominae) from Southwestern Brazilian Amazonia.

The sand fly phlebotomine fauna of the Brazilian municipality of Assis Brasil, Acre was sampled between April 2013 and March 2014 using modified CDC light traps. A total of 22,334 sand flies were collected, 41 of them belonging to three species: Lutzomyia gonzaloi, Lu. kirigetiensis and Lu. munaypata, of the subgenus Helcocyrtomyia, which have previously only been recorded in Peru. This paper presents their first reported collection in Brazil.

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.

First Record of Lutzomyia (Lutzomyia) longipalpis (Diptera: Psychodidae: Phlebotominae) on the Trinational Frontier (Brazil-Peru-Bolivia) of South-Western Amazonia.

In South America, the main sand fly species involved in the transmission of Leishmania infantum chagasi (Cunha & Chagas, 1937), etiological agent of the visceral leishmaniasis (VL), is Lutzomyia longipalpis (Lutz & Neiva, 1912). The species has been recorded in Colombia, Venezuela, Bolivia, Argentina, Paraguay, Uruguay, and Brazil, where it is recorded in 24 of the 27 Brazilian states, except Acre, Amazonas, and Santa Catarina. Collections were carried out for one year (April 2013 to March 2014) using modified CDC light traps in different environments in Assis Brasil municipality, state of Acre. Two males of Lu. longipalpis were found in peridomiciliary location in a peri-urban area. This is the first record of the species in Acre. This finding may be considered by the health agencies located in the trinational frontier, and new collections are needed to evaluate the real distribution of the species.

Na América do Sul, a principal espécie de flebotomíneo envolvido na transmissão de Leishmania infantum chagasi , agente etiológico da Leishmaniose visceral (VL), é Lutzomyia longipalpis (Lutz & Neiva, 1912). Esta espécie já foi registrada na Colômbia, Venezuela, Bolívia, Argentina, Paraguai, Uruguai e Brasil, onde foram registradas em 24 dos 27 estados brasileiros, com exceção do Acre, Amazonas e Santa Catarina. As coletas foram realizadas por um ano (abril de 2013 a março de 2014) utilizando-se armadilhas de luz tipo de CDC em diferentes ambientes no município de Assis Brasil, estado do Acre. Dois machos de Lu. longipalpis foram encontrados no peridomicílio de uma residência localizada na área periurbana, sendo o primeiro registro desta espécie no estado. Esse achado deve ser considerado pelas agências de saúde localizadas na fronteira trinacional. Novas capturas serão necessárias para avaliar a distribuição real da espécie.

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.

Techniques to improve the maintenance of a laboratory colony of Nyssomyia neivai (Diptera: Psychodidae).

BACKGROUND: The most critical phase in sand fly colonization is the high mortality in the larval instars. In this study, we sought out strategies for improving the colonization of Nyssomyia neivai, one of the vectors of cutaneous leishmaniasis agent in South America. METHODS: A colony of Ny. neivai was established in the laboratory from a field population, and the productivity of adults was evaluated considering carrying capacity, diet for larvae and surface for oviposition. RESULTS: The highest emergency rate of adults was achieved with the fewest couples inside 150 mL rearing chambers on a sterilized diet made of rabbit feces, rabbit food, soil and fish food and with vermiculite as a substrate for oviposition and the development of larvae. CONCLUSION: Our data on Ny. neivai colonization showed that the best adult productivities were achieved with fewer couples inside the rearing chambers; smaller rearing containers of 150 mL (due to less fungi growth); sterilized diet made of rabbit feces, rabbit food, soil and fish food; and vermiculite as the substrate for oviposition and development of larvae.

Attraction of the sand fly Nyssomyia neivai (Diptera: Psychodidae) to chemical compounds in a wind tunnel.

BACKGROUND: Similar to other hematophagous insects, male and female sand flies must feed on plants to obtain sugar and, subsequently, energy to complete their life cycles. A large number of compounds emitted by plants may act as volatile signals to these insects. Primary alcohols have been detected in some plants, but in small amounts. In a previous report, the attractiveness of saturated primary alcohols with 7 to 9 carbons was evaluated for Lutzomyia longipalpis, the vector of American visceral leishmaniasis, with positive results. METHODS: In the present study, a wide range of primary alcohols, 3 to 10 carbons, were tested to investigate their attractiveness to another sand fly species, Nyssomyia neivai, a putative vector of American cutaneous leishmaniasis. The mixture of compounds that induced the best sand fly response was also evaluated. RESULTS: Of the eight compounds evaluated, hexanol and octanol elicited the best attractive responses for sand fly females. CONCLUSION: Phytochemicals may be an interesting source of search for new sand fly attractants.

Molecular detection of the blood meal source of sand flies (Diptera: Psychodidae) in a transmission area of American cutaneous leishmaniasis, Paraná State, Brazil.

The feeding behavior of sand flies provides valuable information about the vector/host interactions and elucidates the epidemiological patterns of American cutaneous leishmaniasis (ACL) transmission. The aim of this study was to identify the blood meal sources of sand flies in endemic areas of leishmaniasis in Paraná State through polymerase chain reaction (PCR) amplification of a prepronociceptin (PNOC) gene fragment and its subsequent DNA sequencing. Moreover, molecular assays were conducted to evaluate the sensitivity and reproducibility of the PNOC gene amplification. Besides that, a time-course digestion test of the blood using sand flies that fed artificially on BALB/c mice was performed. Of 1263 female sand flies collected in the field, 93 (3.6%) specimens were engorged and 27 allowed efficient amplification of the PNOC gene. These flies had fed on equine (Equus caballus), porcine (Sus scrofa) and canine (Canis lupus familiaris) species. The results also showed that the identification of the blood meal sources of the sand flies using the molecular method was directly linked to the level of digestion of the blood (time-course) and not to the amount of blood that had been ingested or to the presence of inhibitors in the blood.

A laboratory evaluation of alcohols as attractants for the sandfly Lutzomyia longipalpis (Diptera:Psychodidae).

BACKGROUND: The potential attraction from 1-octen-3-ol for sandflies has been documented; however, studies using other primary alcohols are limited. FINDINGS: We used a wind tunnel to compare the activation and attractive behaviors in male and female Lutzomyia longipalpis using 1-octen-3-ol and three additional alcohols, 1-octanol, 1-heptanol and 1-nonanol at three different concentrations: neat (100%) and diluted in hexane (10% and 50%). The compounds 1-octen-3-ol and 1-nonanol induced a clear concentration-dependent activation and attraction response in females. In males, 1-octen-3-ol, 1-nonanol and 1-heptanol yielded the same results. CONCLUSIONS: L. longipalpis is attracted to 1-octen-3-ol, 1-nonanol and 1-heptanol, which are found in many plant volatiles.

Study of the external female genitalia of 14 Rhodnius species (Hemiptera, Reduviidae, Triatominae) using scanning electron microscopy.

BACKGROUND: Among the vectors of Chagas disease (Hemiptera: Reduviidae:Triatominae), there are eighteen Rhodnius species described and some are difficult to identify. The aim of this article is to contribute to the specific identification of fourteen Rhodnius spp. through morphological characters of the external female genitalia. METHODS: Female abdomens were cut transversely. The specimens were then prepared for examination by using scanning electron microscopy. RESULTS: The careful examination of the dorsal, posterior and ventral sides revealed characteristics that allowed the identification of each of the fourteen species. CONCLUSION: The use of external female genitalia as characteristics are proposed as a tool for specifically identifying Rhodnius species, and an identification key for these species is presented.