Toxicity of Trixis vauthieri DC essential oil on Lutzomyia longipalpis (Diptera, Psychodidae, Phlebotominae), a proven vector of visceral leishmaniasis in Brazil.

BACKGROUND OBJECTIVES: In Brazil, one of the visceral leishmaniasis control measures in urban environments is the elimination of Lutzomyia longipalpis, which occurs through the application of pyrethroid insecticides with residual action in homes and outbuildings. Due to the loss of sensitivity of this vector to these insecticides, the search for more efficient insecticide compounds against L. longipalpis has been intensified. The objective of this work was to evaluate the toxicity of Trixis vauthieri essential oil on adult sandflies of the species L. longipalpis, and identify the phytochemical composition of these essential oils. METHODS: Essential oils from leaves collected from T. vauthieri at different times were obtained at concentrations of 5, 10 and 20 mg/mL. Twenty sandflies were exposed to the essential oils and the mortality was evaluated after 1, 2, 4, 16, 24, 48 and 72 h. The chemical constituents of the essential oil were also identified. RESULTS: The essential oils of T. vauthieri at a concentration of 20 mg/mL were the most toxic to sandflies, reaching a mortality rate of 98.33% and 95%, respectively, after 72 h of exposure. The analysis of chemical constituents revealed the presence of triterpenes and/or steroids, tannins, flavonoids, alkaloids, saponins and coumarins. INTERPRETATION CONCLUSION: The results obtained suggest that T. vauthieri essential oil is fairly promising as an insecticidal potential against L. longipalpis. A more detailed analysis of the oil's phytochemical composition is necessary to identify active and pure compounds that can be used in vector control of visceral leishmaniasis.

Survey of parasites in sea turtles rescued off the coast of Santa Catarina and Paraná, Brazil (2020-2022) and their relationship with marine pollution.

The identification of parasite fauna can contribute to the assessment of the health status of sea turtles, and thus make it possible to develop effective management and treatment strategies aimed at conserving these animals. The present study aimed to investigate the parasite fauna in sea turtles rescued in six sections of the coast of Santa Catarina and Paraná (Brazil) from January 2020 to December 2022 through the recording of exam reports available in the Information System of Aquatic Biota Monitoring. In total, records of 334 sea turtles were analyzed during this period, of which 219 (65.6 %) were infected with at least one parasite. Chelonia mydas was the most frequent species with 86.5 % of rescues and presented the highest species richness. At least 43 species belonging to 36 different genera have been recorded parasitizing sea turtles. The most common helminths were Cricocephalus albus, Metacetabulum invaginatum, Pronocephalus obliquus, with 12.3 %, 8.7 % and 8.4 %, respectively, while protozoa of the genus Entamoeba were the most prevalent in the turtles analyzed (8.1 %). An interesting finding was the unprecedented finding of Hymenolepis sp. eggs in faeces of some turtle species. The results showed that the parasite fauna was quite diverse, with several species known to be capable of harming the physical health and well-being of sea turtles. In view of this, it is necessary to adopt strategies for monitoring the health of the different rescued species with a view to conserving sea turtles in Brazilian territory.

The Program for the Control of Visceral Leishmaniasis in Brazil: The Effect of the Systematic Euthanasia of Seropositive Dogs as a Single Control Action in Porteirinha, a Brazilian City with an Intense Transmission of Visceral Leishmaniasis.

BACKGROUND: Porteirinha is endemic for visceral leishmaniasis (VL), with intense disease transmission of the disease. We evaluated the impact of canine euthanasia as a single control measure on the incidence of VL in humans and canines. METHODS: A prospective observational cohort study was carried out over four years (1998-2002) in 8 of the 12 neighborhoods of the city. The dynamics of canine visceral leishmaniasis (CVL) transmission were evaluated for 2 years, before beginning the screening-culling intervention. The comparative morbidity index (CMI) was used to stratify areas with the greatest risk of CVL, and the spatial distribution of human and canine VL cases was compared using univariate and bivariate K-functions. RESULTS: Human cases conglomerated in three neighborhoods. Spatial clusters were detected for CVL in 1998, 2000, and 2001, but not in 1999, when greater spatial dispersion occurred. The screening and culling intervention reduced the number of human VL cases and decreased the incidence of CVL, mainly in neighborhoods with a high CMI. CONCLUSIONS: The systematic euthanasia of seropositive dogs was shown to be an effective control action of the Program for Control of Visceral Leishmaniasis (PCLV) in Brazil. The fundamental role of domestic dogs in the epidemiological chain of VL was reaffirmed.

Sandflies (Diptera: Psychodidae) from caves of the quartzite Espinhaço Range, Minas Gerais, Brazil.

This study investigated the sandfly fauna of two quartzite caves in the Espinhaço Mountain Range, located in the municipality of Diamantina, state Minas Gerais. From August 2010-July 2011, entomological sampling was performed in the caves of Salitre and Monte Cristo with two HP light traps exposed in the photic and aphotic zones of each cave. The sandfly fauna consisted of 17 species, among which Lutzomyia cipoensis was predominant (54.76%). The male/female ratio in the total captures was 1:2.5. The aphotic zone showed the highest frequency of specimens captured (65%). A greater density of sandflies occurred during the summer (January-February), coinciding with the period of higher temperatures, humidity and rainfall. The presence of Lutzomyia longipalpis, Lutzomyia whitmani and Lutzomyia pessoai, proven or suspected vectors of leishmaniasis, is of concern because the area is visited by many tourists.

Lutzomyia diamantinensis sp. nov., a new phlebotomine species (Diptera: Psychodidae) from a quartzite cave in Diamantina, state of Minas Gerais, Brazil.

A new species of Brazilian phlebotomine sandfly found in Brazil, municipality of Diamantina, state of Minas Gerais, is described based on males and females collected in a quartzite cave. The body of spermathecae is continuous to the individual duct, lanky and tapering at the end, with conical shaped, not striated and presenting the head with dense setae. The male presents gonostyle with four spines and a small subterminal seta and gonocoxite with one group of persistent setae. The paramere is simple with a group of small setae on the dorsal apex. The morphological features of this new species permit its inclusion in the migonei group.

Evaluation of the vectorial capacity of Rhipicephalus sanguineus (Acari: Ixodidae) in the transmission of canine visceral leishmaniasis.

The vectorial capacity of Rhipicephalus sanguineus in the transmission of canine visceral leishmaniasis has been evaluated through a laboratory-controlled experiment. One healthy Leishmania-free dog and two dogs naturally infected with Leishmania were infested with R. sanguineus in various stages of development. Engorged larvae, unfed nymphs, engorged nymphs, unfed adults, engorged female adults and fed male adults were collected from the experimental animals and examined for Leishmania infection by optical microscopy, polymerase chain reaction (PCR) and parasite culture. Leishmania forms were not detected in any of the 433 smears prepared from engorged colonies nor in any of the 118 smears prepared from unfed colonies. However, one flagellate structure was identified in one of the smears. All pools of R. sanguineus that had fed on the infected dogs tested PCR-positive for Leishmania DNA, with the single exception of the pool of engorged larvae. In contrast, all pools of ticks that had fed on the Leishmania-free dog were PCR-negative. Leishmania growth was not observed in any of the tick colonies following incubation on culture medium. Considering that no Leishmania forms were identified in any of the meticulously analysed smears derived from engorged colonies of R. sanguineus, it appears somewhat unlikely that the maintenance and multiplication of Leishmania occurs within the tick.

Association of Lutzomyia longipalpis (Diptera: Psychodidae) population density with climate variables in Montes Claros, an area of American visceral leishmaniasis transmission in the state of Minas Gerais, Brazil.

In the present paper, we evaluate the relationship between climate variables and population density of Lutzomyia longipalpis in Montes Claros, an area of active transmission of American visceral leishmaniasis (AVL) in Brazil. Entomological captures were performed in 10 selected districts of the city, between September 2002-August 2003. A total of 773 specimens of L. longipalpiswere captured in the period and the population density could be associated with local climate variables (cumulative rainfall, average temperature and relative humidity) through a mathematical linear model with a determination coefficient (Rsqr) of 0.752. Although based on an oversimplified statistical analysis, as far as the vector is concerned, this approach showed to be potentially useful as a starting point to guide control measures for AVL in Montes Claros.

Phlebotominae distribution in Janaúba, an area of transmission for visceral leishmaniasis in Brazil.

In Brazil, visceral leishmaniasis (VL) is caused by Leishmania chagasi parasites that are transmitted to man through the bites of infected females of Lutzomyia longipalpis sand flies. In order to evaluate transmission risk and to clarify the epidemiology of this tropical disease, studies focused on the vector and favorable environmental conditions are of fundamental importance. In this work, we surveyed the phlebotomine sand fly fauna in Janaúba, a Brazilian municipality that is endemic for VL. During a two-year period, entomological captures were performed monthly in 15 districts with high, moderate and low profiles of VL transmission. A total of 14,591 phlebotomine sand flies were captured (92% L. longipalpis), with a predominance of males. Most specimens were captured in the peri-domicile setting, although the number of specimens captured in the intra-domicile setting emphasises the anthropophilic behaviour of this insect. The population density of L. longipalpis was modulated by climate variations, particularly with clear increases immediately after the rainy season. However, the pattern of distribution did not coincide with the occurrence of human or canine cases of VL. This suggests that the eco-epidemiology of VL is particular to each area of transmission and must be taken into account during the design of public health control actions.

Infectivity of seropositive dogs, showing different clinical forms of leishmaniasis, to Lutzomyia longipalpis phlebotomine sand flies.

Visceral leishmaniasis (VL) is a growing zoonosis with an increasing number of new cases and a rapid geographical spreading of the disease. In the present study, a canine survey was carried out in the city of Montes Claros (320,000 inhabitants), an endemic area of American visceral leishmaniasis in the state of Minas Gerais, Brazil. A total number of 4795 dogs were examined by serology, which showed a rate of seropositivity of 5%. Isoenzymatic analysis confirmed Leishmania infantum chagasi as the local aetiological agent of CVL. Canine tissues were assayed for the presence of Leishmania parasite DNA using different techniques. The infectivity of asymptomatic, oligosymptomatic and symptomatic seropositive dogs was tested by xenodiagnosis using laboratory reared Lutzomyia longipalpis. Rates of infection of 5.4%, 5.1% and 28.4% were found for the phlebotomine sand flies that fed in asymptomatic, oligosymptomatic and symptomatic dogs, respectively. Our results indicate that, under experimental conditions, symptomatic dogs are about four times more infective to VL vectors than oligosymptomatic or asymptomatic animals. The lower infectivity rates of dogs displaying any of the last two clinical forms of leishmaniasis, however, must be taken into account in the epidemiology of CVL.

[Sandflies (Diptera: Psychodidae) in an outbreak of cutaneous leishmaniasis in the State of Minas Gerais]. / Flebotomíneos (Diptera: Psychodidae) de um foco de leishmaniose tegumentar no Estado de Minas Gerais.

Between January and December 2000, systematic monthly capture of phlebotomine sandflies was undertaken by means of light traps of CDC type in Brejo do Mutambal, an endemic area for American cutaneous leishmaniasis located in the municipality of Varzelândia, State of Minas Gerais. Nineteen phlebotomine species were captured, totaling 6,756 specimens. The species correlated with ACL transmission were captured in low numbers, as follows: Lutzomyia intermedia (5.1%), L. migonei (0.4%) and L. whitmani (0.1%). Lutzomyia longipalpis was the predominant species captured (34.8%), thus also suggesting a risk of visceral leishmaniasis transmission. The proportion of sandflies captured in areas surrounding homes was 91.7 % while 8.3% were captured inside homes. The interference of climatic factors (temperature, relative air humidity and rainfall) on phlebotomine population dynamics was evaluated.

DETECTION OF INTESTINAL PARASITES IN THE ENVIRONMENTS OF A PUBLIC SCHOOL IN THE TOWN OF DIAMANTINA , MINAS GERAIS STATE, BRAZIL.

Intestinal parasites are a major public health problem in developing countries, most prevalent in areas where sanitation is poor and the population's hygiene is inadequate. They affect people of all ages, although school-age children are the most susceptible. In this study, we investigated the presence of intestinal parasites in the shared environments of a public school in the town of Diamantina, Minas Gerais State. From December 2012 to February 2013, samples were collected for three months (once a month) by using the Graham method (1941), in duplicate, by affixing a 6 x 5 cm clear tape, six times in each collection site, in a space of about 30 cm2. Then, each tape was positioned longitudinally on a microscope slide and the identification of the biological forms of the parasites was performed with the aid of a 40X objective from an optical microscope. Eleven sites were selected for sampling. Cysts of Entamoeba coli were the most frequently found in this study (50%), followed by Hymenolepis diminuta eggs (27.6%), Iodamoeba butschllii cysts (5.6%), Ascaris lumbricoides eggs (5.6%), Taenia species eggs (5.6%) and hookworm eggs (5.6%). The highest positivity rates were found in the samples drawn from the cafeteria's eating table. The results have indicated the need to improve the cleaning in the school environments, as well as the development of educational practices that may help in the preservation of public health.

[Aspects of the ecology and behaviour of phlebotomines in endemic area for visceral leishmaniasis in State of Minas Gerais]. / Aspectos da ecologia e do comportamento de flebotomíneos em área endêmica de leishmaniose visceral, Minas Gerais.

Studies on the behavioral and feeding habits of some species of phlebotominae sand flies have contributed to the comprehension of the epidemiology of leishmaniasis. In the present work, systematic captures were performed monthly in the municipality of Porteirinha (MG) using 28 light traps (CDC) from January to December 2002. Fourteen different species of phlebotomine were captured in a total of 1,408 specimens. The highest percentage of individuals (53.3%) was collected in the peridomicile against 46.7% in the intradomicile. Lutzomyia longipalpis was the predominant species in that region. The blood feeding of 38 females of this species from the field was analyzed by precipitin reaction. The results indicated that Lutzomyia longipalpis is an opportunist (65.1%) species that feeds on a wide variety of vertebrates in nature.

[Visceral leishmaniasis: a study on phlebotomine sand flies and canine infection in Montes Claros, State of Minas Gerais]. / Leishmaniose visceral: estudo de flebotomíneos e infecção canina em Montes Claros, Minas Gerais.

Visceral leishmaniasis in Brazil was initially associated with rural areas. However, due to several environmental modifications such as deforestation, urbanization and intense migratory processes, there has been an expansion of endemic areas, leading to urbanization of the disease, mainly in the central and northeastern regions of Brazil. In the municipality of Montes Claros, located in the north of the state of Minas Gerais, an epidemiological survey on VL was carried out. A canine serological inquiry was carried out in 2002 and an entomological survey, using luminous CDC traps, was performed from September 2002 to August 2003. Canine VL prevalence showed an average infection rate of approximately 5%. An estimated 16 species comprised the phlebotomine sand fly fauna, based on a total of 1043 specimens. The predominant species was Lutzomyia longipalpis with a rate of 74%, suggesting its participation in the transmission of VL in the municipality of Montes Claros.

IDENTIFICATION OF SANDFLIES (Diptera: Psychodidae: Phlebotominae) BLOOD MEALS IN AN ENDEMIC LEISHMANIASIS AREA IN BRAZIL.

The aim of this study was to identify blood meals of female sandflies captured in the municipality of Governador Valadares, an endemic area of visceral and cutaneous leishmaniasis, in the State of Minas Gerais, Brazil. From May 2011 to January 2012, captures were performed using HP light traps in four districts. There were 2,614 specimens (2,090 males and 524 females) captured; 97 engorged females were identified belonging to the species Lutzomyia longipalpis(82.1%) and Lutzomyia cortelezzii(17.9%). Considering simple and mixed feeding, the enzyme-linked immunosorbent assay revealed a predominance of chicken blood (43.6%) in Lutzomyia longipalpis, showing the important role that chickens exert around the residential areas of Governador Valadares. This finding increases the chances of sandflies contact with other vertebrates and consequently the risk of leishmaniasis transmission.

Purification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lini.

An alkaline serineprotease, capable of hydrolyzing Nalpha-benzoyl- dl arginine p-nitroanilide, was secreted by Fusarium oxysporum var. lini grown in the presence of gelatin as the sole nitrogen and carbon source. The protease was purified 65-fold to electrophoretic homogenity from the culture supernatant in a three-step procedure comprising QSepharose chromatography, affinity chromatography, and FPLC on a MonoQ column. SDS-PAGE analysis of the purified protein indicated an estimated molecular mass of 41 kDa. The protease had optimum activity at a reaction temperature of 45 degrees C and showed a rapid decrease of activity at 48 degrees C. The optimum pH was around 8.0. Characterization of the protease showed that Ca2+ and Mg2+ cations increased the activity, which was not inhibited by EDTA or 1,10-phenanthroline. The enzyme activity on Nalpha-benzoyl-DL arginine p-nitroanilide was inhibited by 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride, p-aminobenzamidine dihydrochloride, aprotinin, 3-4 dichloroisocoumarin, and N-tosyl-L-lysine chloromethyl ketone. The enzyme is also inhibited by substrate concentrations higher than 2.5 x 10(-4)M. The protease had a Michaelis-Menten constant of 0.16 mM and a V(max) of 0.60 mumol released product.min(-1).mg(-1) enzyme when assayed in a non-inhibiting substrate concentration. The activity on Nalpha-benzoyl- dl arginine p-nitroanilide was competitively inhibited by p-aminobenzamidine dihydrochoride. A K(i) value of 0.04 mM was obtained.

Rattus norvegicus (Rodentia: Muridae) Infected by Leishmania (Leishmania) infantum (syn. Le. chagasi) in Brazil.

In the present study we surveyed the fauna of phlebotomine sand flies and small mammals in peridomestic areas from a Brazilian municipality where the American cutaneous leishmaniasis (ACL) is endemic. A total of 608 female phlebotomine sand flies were captured during nine months in 2009 and 2010. Seven different species were represented with 60% of them being Lutzomyia intermedia and Lu. whitmani, both incriminated vectors of ACL. Lu. longipalpis, a proven vector of visceral leishmaniasis (VL) was also captured at high proportion (12.8%). Genomic DNA analysis of 136 species-specific pools of female sand flies followed by molecular genotyping showed the presence of Leishmania infantum DNA in two pools of Lu. longipalpis. The same Leishmania species was found in one blood sample from Rattus norvegicus among 119 blood and tissue samples analysed. This is the first report of Le. infantum in R. norvegicus in the Americas and suggests a possible role for this rodent species in the zoonotic cycle of VL. Our study coincided with the reemergence of VL in Governador Valadares.

Ecological aspects of the Phlebotominae fauna (Diptera: Psychodidae) in the Xakriabá Indigenous Reserve, Brazil.

BACKGROUND: Sand fly collections were performed to study ecological aspects of the Phlebotominae fauna of the Xakriabá Indigenous Reserve, an area with endemic cutaneous leishmaniasis, located in the state of Minas Gerais, Brazil. METHODS: The collections were performed in peridomicile areas and along trails previously selected for the study of wild and synanthropic Leishmania hosts. Differences in the distribution patterns of the sand fly species as well as in species richness and abundance between the different ecotopes were investigated during both rainy and dry seasons over the course of the study period. RESULTS: A total of 8,046 sand flies belonging to 11 genera and 28 species were collected. Lutzomyia longipalpis and Nyssomyia intermedia were the most abundant species in peridomicile areas, whereas Martinsmyia minasensis and Lutzomyia cavernicola were the most abundant species among the different trail ecotopes. CONCLUSION: The different composition of the sand fly fauna observed in the peridomicile areas and in the trails during the study, reinforces the importance of sampled different areas in a phlebotomine fauna survey. The presence of Lutzomyia longipalpis and Ny. Intermedia most abundant in peridomicile can be important to Leishmania infantum and Leishmania braziliensis transmission in the Imbaúbas native village.

Epidemiology of visceral leishmaniasis in a reemerging focus of intense transmission in Minas Gerais State, Brazil.

This study was developed in the urban area of Governador Valadares, a reemerging focus of intense transmission of visceral leishmaniasis (VL) in Brazil, presenting 86 human cases of VL from 2008 to 2011. The disease prevailed in males (73.2%) with most patients between 0 and 9 years (44.1%) and a lethality rate of 16.2%. A canine survey was carried out on 16,529 domestic dogs in 35 districts in the area and it showed that 30.2% of them (4,992 dogs) were positive for VL by serum assays. Prevalence ratios for canine VL varied between 13.6% and 53.4%. The clinical exam of 343 seropositive dogs showed that 49.9% of them were considered symptomatic, with larger prevalence of canine VL being in short-furred animals (90%). The entomological survey was performed in eight districts, where 2,539 phlebotomines were captured, preferentially in the peridomicile (84.5%). Lutzomyia longipalpis was the predominant species (90%) suggesting its participation in the VL transmission in the area. The correlation between canine prevalence and L. longipalpis density was evaluated.

Distribution of the assemblage of phlebotomine sandflies (Diptera: Psychodidae) along an environmental gradient / Distribuição da assembléia de flebotomíneos (Diptera: Psychodidae) em um gradiente ambiental

Abstract The geographical distribution of leishmaniasis has undoubtedly expanded, and is now being reported to be in areas that were previously non-endemic. This study therefore aimed to investigate whether there is any variation in the structure and in the composition of phlebotomine communities along an environmental gradient - from a mesic to a xeric environment, across different climatic periods (rainy and dry seasons). Furthermore, we test whether there were differences in sex ratio of sand flies between wet and dry environments. A total of 535 specimens of sand flies belonging to 18 species were recorded. Richness, abundance, and composition of species were significatively higher in the xeric environment (semidecidual Forest) compared to the mesic environment (Riparina Forest). We unveiled site specific differences in sand fly collections since the community composition changes with the geographic distance among the sampling points. These results led to the conclusion that for sand flies species, the environmental conditions available for each species could be different within the habitat taking into account the degree of humidity in the forest as well as the climatic season (dry or wet). The present study gives a valuable contribution to the knowledge of sand flies, as a group of insects, from the Atlantic Forest biome.

Resumo A distribuição geográfica das leishmanioses, sem dúvida, tem se expandido, e agora está sendo relatada em áreas previamente consideradas como não endêmicas. Assim, este estudo teve como objetivo investigar se existe alguma variação na estrutura e na composição das comunidades de flebotomíneos ao longo de um gradiente ambiental - em habitats classificados como mesicos e xericos, em diferentes períodos climáticos (chuva e seca). Além disso, testamos se havia diferenças na razão sexual entre ambientes mais umidos e mais secos. Um total de 535 espécimes de flebotomíneos pertencentes a 18 espécies foram registradas. Riqueza, abundância e composição de espécies foram significativamente maiores nos habitats xericos (Floresta semidecidual) em comparação com os habitats mesicos (Floresta Ciliar). Revelamos diferenças específicas entre os habitats, nas assembleias de flebotomíneos, uma vez que a composição da comunidade muda com a distância geográfica entre os pontos de amostragem. Estes resultados levaram à conclusão de que as condições ambientais disponíveis para cada espécie de fletomonineo varia dentro do habitat, tendo em conta o grau de umidade na floresta assim como a estação climatica (seca-chuva). O presente estudo dá uma contribuição valiosa para o conhecimento da fauna de flebotomíneos, no bioma Mata Atlântica.

Assessment of sand fly (Diptera, Psychodidae) control using cypermethrin in an endemic area for visceral leishmaniasis, Montes Claros, Minas Gerais State, Brazil.

Montes Claros in Minas Gerais State, Brazil, was considered an intense transmission area for visceral leishmaniasis. This study evaluated sand fly fauna after insecticide application. Captures were performed in 10 districts from September 2005 to August 2006 with CDC light traps inside and outside each residence. Cypermethrin was sprayed in two cycles during November/2005 and May/2006. The 636 specimens collected, belonging to 10 species, were predominantly Lutzomyia longipalpis (79%), and most frequently males (70%). The highest percentage of specimens were captured in areas surrounding domiciles (85.8%). The main species were observed to be sensitive to treatment with the insecticide. The results showed a reduction in the number of sand flies collected after use of cypermethrin in homes and annexes, and with residual effect lasting from two to four months.