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The Rhipicephalus sanguineus group is an assembly of species morphologically and phylogenetically related to Rhipicephalus sanguineus sensu stricto. The taxonomy and systematics of this species group have remained obscure for a long time, but extensive research conducted during the past two decades has closed many knowledge gaps. These research advancements culminated in the redescription of R. sanguineus sensu stricto, with subsequent revalidation of former synonyms (Rhipicephalus linnaei, Rhipicephalus rutilus, and Rhipicephalus secundus) and even the description of new species (Rhipicephalus afranicus and Rhipicephalus hibericus). With a much clearer picture of the taxonomy of these species, we present an updated list of species belonging to the R. sanguineus group, along with a review of their geographic distribution and vector role for various pathogens of animals and humans. We also identify knowledge gaps to be bridged in future studies. [ABSTRACT FROM AUTHOR]

Rodents are the primary reservoir hosts for zoonotic cutaneous leishmaniasis (ZCL) caused by Leishmania major. Knowing reservoir hosts is crucial for leishmaniasis surveillance and control programs in endemic areas. In this study, we examined an archived spleen of Rattus norvegicus obtained during a pest control program in 2000 in Tehran, the capital of Iran. The sample was analyzed using polymerase chain reaction (PCR) and sequencing to determine the presence of Trypanosomatidae based on the internal transcribed spacer (ITS) 1 gene. Amplification and sequencing of the discriminative region of the ITS1 gene followed by BLAST analysis showed the highest similarity with L. major isolates. Also, the phylogenetic analysis revealed that our sample was grouped with L. major isolates retrieved from the GenBank database. This finding might support the claim that R. norvegicus acts as a potential reservoir host for L. major. Further studies, including a survey on more rodent samples as well as studying sandflies in the area, might uncover the possible presence of such pathobiological conditions in ZCL transmission in urban and suburban settings. [ABSTRACT FROM AUTHOR]

Entomological investigations were conducted for the first time in urban forest remnants of Porto Velho, state of Rondônia, Brazil, to explore the transmission dynamics of Leishmania. Sand fly collections were carried out at ten sites, encompassing both canopy and ground strata, from October to December 2021. A total of 1,671 sand flies were collected, representing 42 species within 12 genera. Nyssomyia Antunesi (n = 384) and Psychodopygus davisi (n = 111) were the most abundant species. Molecular analyses targeting the V7V8 region (18S gene) unveiled the presence of sequences 100% identical to Leishmania infantum in females of Bichromomyia flaviscutellata (1), Nyssomyia Antunesi complex (6), Nyssomyia umbratilis (1), Nyssomyia sp. (1), Psychodopygus ayrozai (1), Ps. davisi (3), Psychodopygus paraensis (1), and Sciopemyia sordellii (1). Sequences 100% similar to Trypanosoma minasense were found in two samples of the Nyssomyia Antunesi complex, and two samples of Sc. sordellii presented 100% identity to a Trypanosoma sp. strain, previously identified in this same sand fly in Rondônia. Sequencing of Cytb fragment suggested Homo sapiens, Dasypus novemcinctus and Tamandua tetradactyla as the blood source for distinct sand flies. The identification of sequences similar to L. infantum in sand flies collected in urban forest fragments is noteworthy, correlating with the recent local and regional occurrence of autochthonous cases of human visceral leishmaniasis. However, further studies are imperative to ascertain the presence of hosts/reservoirs and evaluate the risk of L. infantum transmission to humans. [ABSTRACT FROM AUTHOR]

Phlebotomine sand flies are crepuscular and nocturnal small dipteran insects in the family Psychodidae. Several disease agents, including Leishmania parasites, are transmitted to humans and other vertebrate hosts by the bite of an infected female sand fly. As part of leishmaniasis surveillance programs, light traps have been routinely used in sand fly collections. In this context, new trapping devices are always being required to improve vector monitoring. Here, the efficiency of a new suction light trap, named Silva suction trap or SS trap, was field evaluated in collecting sand flies. Two SS traps, one with green (520 nm, 15,000 mcd) and the other with white (wide spectrum, 18,000 mcd) LEDs, and one CDC-type trap were deployed in a rural forested environment. A total of 4686 phlebotomine sand flies were captured. The most frequent species were females of the Ps. Chagasi series (77.8%) followed by males of Ps. wellcomei (11.6%), Nyssomyia whitmani (3.3%), and Bichromomyia flaviscutellata (2.4%). The CDC-type light trap collected 101.9 ± 20.89 sand flies and 14 species, followed by the white-baited SS trap (87.78 ± 16.36, 14), and the green-baited SS trap (70.61 ± 14.75, 15), but there were no statistically significant differences among traps. A discussion on the considerable advantages of the use of SS traps over CDC traps is included. In this study, the Silva suction trap proved to be efficient and can be an alternative to CDC traps for monitoring adult phlebotomine sand fly populations. [ABSTRACT FROM AUTHOR]

Trichophoromyia auraensis (Mangabeira, 1942) (Diptera, Psychodidae, Phlebotominae) has a wide geographic distribution in the western region of the Amazon biome, where it is a putative Leishmania vector. Here, we reported for the first time a population of this species in the Brazilian state of Maranhão, in the eastern Amazon, from which we DNA-barcoded and compared with previously processed specimens from Acre State, in the western Amazon. For this, we analyzed the DNA barcoding fragment (658 bp) of the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer 2 (ITS2) of Trichophoromyia species using phylogenetic gene trees, and species delimitation algorithms. The analyses of COI barcodes showed high values of genetic distance (mean K2P = 5.17) and well-supported clades/MOTUs for the eastern and western populations of T. auraensis, which may indicate a possible complex of cryptic species. The western population of this taxon merged with the close-related sand fly Trichophoromyia velezbernali Posada-López, Galvis and Galati, 2018 from Colombia, which may be associated with the recent speciation history and introgression between these populations. These evidences should be evaluated with a more comprehensive sampling in terms of analyzed populations and molecular markers. [ABSTRACT FROM AUTHOR]

Light traps have been widely used for monitoring malaria vectors, although drawbacks remain. In this context, new tools and attractants are always becoming available to perform monitoring tasks, like the Silva trap, a passive and low-cost LED-light trap for host-seeking anopheline mosquitoes. In this work, the effectiveness of the Silva trap by using UV-LED and at different heights as well as a comparison with the conventional CDC-type (HP) light trap was studied. A total of 9009 mosquitoes and nine species were caught, Anopheles triannulatus, An. argyritarsis, and An. goeldii being the most frequent species. The green (520 nm) and blue (470 nm) LEDs attracted almost equal numbers of anopheline mosquitoes, but UV LEDs (395 nm) attracted a significantly lower number of individuals (Kruskal–Wallis = 19.68, P = 0.0001). Even with the predominance of mosquitoes trapped at the height of 1.5 m, no significant statistical difference was found among the four heights tested (0.5 m; 1.0 m; 1.5 m; 2.0 m). Green-baited Silva traps collected significantly more individuals than incandescent-baited CDC-type traps (U = 60.5; P = 0.0303). LEDs have been useful as light sources for attracting insect vectors and together with a low-cost trap, as the Silva trap, a feasible alternative to conventional trap-based monitoring Anopheles mosquitoes that can be implemented in the field. [ABSTRACT FROM AUTHOR]

Millipedes are well-known for their limited dispersal abilities with species restricted to mountains, islands, and patches of forest being important models for formulating and testing biogeographic hypotheses. The order Spirostreptida is composed of nearly 1300 species distributed across the Afrotropical, Oriental, and Neotropical regions. The order is divided into the suborders Cambalidea and Spirostreptidea with the families Pseudonannolenidae (Cambalidea) and Spirostreptidae (Spirostreptidea) occurring in Brazil. To date, there have been no studies on the biogeography of Neotropical Spirostreptida. We employed a multi-approach analysis with Parsimony Analysis of Endemicity, Endemicity Analysis, and Infomap, to detect areas of endemism and patterns of distribution of the order in Brazil. Six areas of endemism are proposed for the 133 Brazilian species: Northern Serra Geral, Southeastern mountain ranges, Cerrado and Atlantic Forest zone, Eastern Cerrado and Serra do Espinhaço, Pantanal, and Southern Amazon and Cerrado zone. Most endemic areas fall within the Atlantic Forest, which has been previously shown to be an important area of endemism for many invertebrate taxa. The richest biomes are the Atlantic Forest with 75 species and the Cerrado with 55 species, while the least rich biomes are the Caatinga with six species and the Pampa with three species. Although the southeastern region of Brazil has the highest species richness, it also has the highest concentration of threatened species. Based on International Union for Conservation of Nature (IUCN) criteria, most Brazilian Spirostreptida are either endangered or critically endangered, with the highest concentration of endangered taxa occurring in the Atlantic Forest biome. [ABSTRACT FROM AUTHOR]

The present study is the second and last part of the study that investigated the fauna and behavior of sand flies in areas prone to cutaneous leishmaniasis outbreaks, in the State of Rio de Janeiro, in the municipality of Paraty. To collect the sand flies, CDC and Shannon light traps were used, installed in the peridomiciliary and forest areas, and manual suction tubes on the walls of the home and in the animal shelters. A total of 102,937 sand flies, belonging to nine genera and 23 species were captured from October 2009 to September 2012. Regarding the monthly frequency of sand flies, the period of highest density was from November to March, with a maximum peak in January. The lowest density was observed in June and July. In the studied area, the species of epidemiological importance, Nyssomyia intermedia, Pintomyia fischeri, Migonemyia migonei and Nyssomyia whitmani, were found in all months of the year, a period in which residents may be in contact with these vectors of the etiological agent of cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

The Oligocene and Miocene tectonic and biogeographical alterations of the peri-Mediterranean area could strongly impact the speciation processes and migrations of Mediterranean sandfly species. To understand the possible former role of this palaeobiogeographic factor on ancestral sandfly species, the potential suitability values of five Larroussius, two Paraphlebotomus and one Phlebotomus species were modelled from the Rupelian to the Tortonian stage in the Central Paratethys. The bioclimatic extrema of this sandfly species and the Coexistence Approach-based palaeoclimatic reconstructions made the basis of hypothesis testing. In the case of the Tortonian stage, a georeferenced climatic model was created. The models indicate that the suitability values could notably vary by species and periods. The monsoon-influenced humid subtropical climate of the Oligocene epoch could be less suitable for the ancestors of Mediterranean sandfly taxa than the later, drier humid subtropical climates-characterised Miocene stages. The Central Paratethys area could be less suitable for ancestors of the extant Paraphlebotomus, and Phlebotomus in the Miocene epoch compared to Larroussius species. It may indicate that the Central Paratethys formed a barrier against the east-to-west migration of the ancestors of Phlebotomus and Paraphlebotomus species. In contrast, Larroussius species could prefer the Miocene climate of the Central Paratethys. In the Tortonian stage, mainly the coastal areas of the sea could be colonised by sandflies. These results indicate that the coasts of the Central Paratethys should be not only considered as a potential former migration route but could be a part of the speciation area of Larroussius sandflies. [ABSTRACT FROM AUTHOR]

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Sand flies are hematophagous dipterans and are considered vectors of the parasites belonging to the genus Leishmania, which can infect mammals, including humans, and subsequently cause leishmaniasis. Rio Grande do Sul State (RS), southern Brazil, was considered free of this disease for the last few decades. However, several autochthonous cases of visceral leishmaniasis (VL) have been recorded in different municipalities in recent years. Since 2017, cases of canine visceral leishmaniasis (CVL) have been reported in the municipality of Santa Maria, located in the central region of RS. In 2021, two cases of human VL were confirmed in the city, resulting in one death. To capture the possible vectors involved in the disease transmission cycle, Centers for Disease Control (CDC) modified light traps were installed in the peridomestic residential areas. These points were situated in two city districts where cases of CVL had been reported from January 2021 to January 2022. In the 41 sample collections of sand flies, 9 specimens of Lutzomyia longipalpis were identified, confirming the municipality as an area of VL transmission, according to Brazilian sanitary standards. The spread of Leishmania spp. has been occurring efficiently, resulting in a negative impact on the One Health. This study reports the first detection of L. longipalpis in the central region of RS. [ABSTRACT FROM AUTHOR]

Indoor Residual Spraying (IRS) is one of the interventions to control the vectors of Visceral Leishmaniasis (VL). Different insecticides are used in affected countries, also in the Regional Initiative for the Elimination of VL in South-East Asia. This systematic review assesses all available studies analysing the effectiveness of IRS on the key vectors of VL. The systematic review followed PRISMA guidelines, with a broad search strategy, applied to seven key databases. Inclusion criteria were studies focusing on 1) Visceral leishmaniasis 2) Indoor Residual Spraying (IRS) or synonyms, and 3) all primary research methods. 21 studies were included, five cluster randomised controlled trials (cRCTs), one randomised controlled trial (RCT), 11 intervention studies, also included were three modelling studies and one survey. 19 out of 21 included studies were published between 2009 and 2020. 18 of the studies were conducted in the context of the Regional Initiative. Effects of IRS on vector populations are positive, confirmed in terms of effectiveness and by the availability of studies. Deltamethrin and alpha-Cypermethrin reduce total sandfly counts, and/or Phlebotomus argentipes counts by up to 95% with an effect of a minimum of one month. Prolonged effects are not regularly seen. DDT has been used in India only: whereas in the 1990s a good effect could be measured, this effect waned over time. Two intervention studies, embedded in larger programmes in 2019 and 2020, replaced DDT with alpha-Cypermethrin throughout the study. Combinations of different interventions are not systematically researched, however showing some promising results, for example for the combination of IRS and Temephos. Constant monitoring of insecticide resistancies and quality delivery of IRS are confirmed as key issues for programmes. No human transmission data are available to directly relate an effect of IRS–although modelling studies confirm the effect of IRS on human transmission. Concluding, IRS continues to be an effective intervention for Phlebotomus argentipes control. Delivery requires constant monitoring and quality assurance. Further studies need to assess IRS in different geographical areas affected by VL and combinations of interventions. Author summary: Visceral leishmaniasis (VL) is a vector borne disease endemic in Southeast Asia (SEA) and present in other regions of the world. VL is transmitted by different species of sandflies and key strategies against VL rely on the control of the vectors. This systematic review assesses the efficacy and community effectiveness of indoor residual spraying (IRS) in controlling sandflies and VL occurrence among humans. After a revision of 8822 initial hits, 21 studies were included in this systematic review. Effects of IRS on vector populations are positive, Deltamethrin and alpha-Cypermethrin reduce total sandfly counts by up to 95% for at least one month. DDT has been used in India only: whereas in the 1990s a good effect could be measured, this effect waned over time. Combinations of different interventions are not systematically researched, however showing some promising results, for example for the combination of IRS and Temephos. Constant monitoring of insecticide resistancies and quality delivery of IRS are confirmed as key issues for control programmes. No human transmission data are available to directly relate an effect of IRS on human transmission,–although by modelling studies an effect of IRS on human transmission can be demonstrated. [ABSTRACT FROM AUTHOR]

In this work, graphene oxide (GO) was synthesized by the modified Hummers method. The nanomaterial was characterized by FTIR and Raman spectroscopy, SEM, and pH at the point of zero charge. GO exhibited typical characteristics of graphene-based materials, indicating that graphite oxidation and exfoliation occurred successfully. Cd (II) and Pb (II) adsorption onto GO was carried out in batch systems, in which the effect of adsorbent dosage, contact time, and initial adsorbate concentration were evaluated. Langmuir, Freundlich, and Sips isotherm models, as well as pseudo order models and Elovich kinetic equation were applied to adsorption experimental data. Results indicated that increasing adsorbent mass, the removal efficiency of Cd (II) and Pb (II) increased. Freundlich isotherm better described Pb (II) adsorption (R2 = 0.96), while Cd (II) isotherm showed linear behavior. From the Akaike's AIC parameter, kinetic data were satisfactorily described by pseudo-first order (Cd (II)) and pseudo-n order (Pb (II)) models. GO was successfully subjected to five regeneration cycles, maintaining high efficiency (> 90%) in all cycles. GO showed high potential for the adsorption of Cd (II) and Pb (II) from aqueous solution, due to its high adsorption capacity, rapid Cd (II) and Pb (II) intakes, and great regeneration performance. [ABSTRACT FROM AUTHOR]

The present study aimed to check the sand flies' fauna on the municipality of Lassance, Minas Gerais, Brazil and detect the presence of Leishmania DNA on the female captured and determine the risk areas of the municipality. Sand flies were collected monthly from May 2018 to April 2019 using automatic light traps for 3 consecutive nights. Eight houses were selected as sample points due its previous reports of tegumentary leishmaniasis and/or canine leishmaniasis. The sand fly's fauna found on the present study it's represented by several medical importance species and the most abundant species found were Lutzomyia longipalpis (77.09%) and Nyssomyia intermedia (10.06%). Leishmania infantum DNA was detected in a pool of Lu. longipalpis resulting on a 2.81% of infection rate. By the frequency of the two most abundant species on this study, we developed a risk area map and it draws attention to sample point 6 due to disparate abundance of sand flies at this site (81.81%). Statistical overview shows Lu. longipalpis as dominant species and, still, Non-Metric Multidimensional Scaling analysis reveal high similarity on fauna's diversity on the study area. Our findings suggest that the diversity of sand flies from the municipality of Lassance may promote the circulation of Leishmania infantum parasites putting in risk the habitants and other mammal's species. Still, our study reinforces the necessity of specific studies focused on breed sites of phlebotomine and its' ecology to expand the knowledge about the behaviour of this group of insects applying directly to leishmaniases' epidemiology. [ABSTRACT FROM AUTHOR]

Cobalt-grafted-mesoporous silica nanoparticles with the Si/Co molar ratio of 70 were synthesized by using sol–gel method (denoted as Co-MSN). Characterization of the Co-MSN was carried out by X-ray diffraction analysis (XRD), Fourier-transform infrared spectroscopy (FTIR), energy dispersive X-ray analysis (EDX), scanning electron microscopy (SEM), and N2 adsorption/desorption analysis. Photocatalytic activity of the Co-MSN sample was studied for removal of neutral red as one of the most widely used dye pollutants, in presence of H2O2 and UV irradiation. In this regard, effect of various influencing parameters such as pH, Co content, dye concentration, amount of oxidant, and presence/absence of UV irradiation were investigated. It was found that incorporation of the Co2+ ions into the mesoporous silica nanostructure significantly improves the photocatalytic degradation of neutral red. Under the optimized conditions including 0.3 mmol of H2O2 (as an oxidant), catalyst dosage of 0.075 mg, initial dye concentration of 0.08 mM, and UV irradiation (32 W, 254 nm), up to 81% of neutral red was degraded at room temperature for 220 min. [ABSTRACT FROM AUTHOR]

Background: The municipality of Caratinga is an important endemic area for American Tegumentary Leishmaniasis (ATL) and no epidemiological studies were performed during the past two decades. Here, we analyzed the epidemiological situation and the geographical distribution of ATL cases in the municipality of Caratinga from 2007 to 2018 using geographic information systems (GIS) Also, we evaluated the impact of several demographic parameters in ATL distribution and the sand flies incriminated in its transmission. Methods: All demographic information (gender, age, educational level, clinical form, diagnostic criteria and case evolution) used in this study was retrieved from the public health archives and confirmed in the State Health Services databases. All cases were analyzed using GIS software based on ATL distribution. Also, non-systematic sand fly collections and molecular detection of Leishmania were performed in the hotspots. Results and conclusions: During the period, ATL cases continued and increased especially in the past years (2016–2018). Hotspots included urban Caratinga areas and the districts of Patrocínio de Caratinga and Sapucaia. The species Nyssomyia whitmani, Nyssomyia intermedia, Migonemyia migonei and Evandromyia cortelezzii complex were captured. However, ITS1-PCR did not detect Leishmania DNA in those insects. Based on our analyses, urbanization of ATL in Caratinga has occurred in the past years. The increase in the number of cases and vectors presence, it is recommended that health authorities focus on control measures in the most affected areas (Patrocínio of Caratinga and Sapucaia districts and urban Caratinga). Author summary: Leishmaniasis is an important health problem in Latin American countries and worldwide. In some places, notification is limited and underreported hindering correct assessment of existing data. In Brazil, ATL is mainly caused by Leishmania (Viannia) braziliensis. It is transmitted by sand fly vectors Nyssomyia whitmani and Nyssomyia intermedia. ATL comprises different clinical manifestations including cutaneous leishmaniasis, muco-cutaneous leishmaniasis and atypical leishmaniasis. The objective is to present the spatial distribution of cases of ATL notified by the Public Health System of Caratinga between the period of 2007 and 2018. This city is one of the most important endemic areas in the State of Minas Gerais and compose the panel of knowledge on epidemiological aspects of cutaneous leishmaniasis caused by L. braziliensis. This information will be important for developing strategies for ATL control in the affected areas. There is strong evidence that supports ATL urbanization in the city of Caratinga and increase of cases in two rural districts (Sapucaia and Patrocínio de Caratinga). [ABSTRACT FROM AUTHOR]

Genus and species of phlebotomine sand flies have been recorded and described in caves in Brazil, but no study has provided the food source used by sand flies in these environments. Herein, we identified the blood source used by sand fly species in caves located at "Quadrilátero Ferrífero" (QF), Minas Gerais state. Specimens were manually collected near or on anurans inside ferruginous caves in Serra do Gandarela National Park and Serra do Rola Moça State Park. Males and females were placed in vials with 70% alcohol and stored at −10°C. Females engorged, after specific identification, had DNA extracted and followed for PCR amplification using specific primers. Sequencing was analyzed in the GenBank and Barcode of Life. A total of 198 specimens were collected (107 females and 91 males), all of them belonging to species Sciopemyia aff. microps (88.89%), Sciopemyia sordellii (10.61%), or Martinsmyia oliveirai (0.50%). When it comes to the females, 89 were S. aff. microps and 18 S. sordellii. Nineteen engorged females of S. aff. microps were analyzed and most of them (n=18) presented blood from Bokermannohyla martinsi and one contained blood from Scinax fuscovarius. The blood present in engorged females of S. sordellii (n=4) was from B. martinsi. Sciopemyia genus specimens are commonly found in collections carried out inside natural caves, but this was the first study to prove that females of this genus feed on cold-blooded animals in nature. Highlights: • Here we proved that sand flies feed in cold-blooded animals in in Brazilian caves. • Females of the Sciopemyia genus were for the first time found feeding in natural habitats. • Anurans of the family Hylidae were identified as source by molecular analyzes. • Insect bloodmeal identification can help assessing the fauna in several biomes. • This is the first record of S. aff. microps in caves of Brazil. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is endemic in 70 countries and has been reported in 12 countries of Latin America, with over 90% of the cases reported in Brazil, where epidemics have occurred since 1980. The objective of this review is to describe the factors associated with the occurrence of VL epidemics in humans in urban areas. A systematic review was conducted according to the PRISMA-P guidelines. The databases PubMed (by Medline), Cochrane Library, Embase, Amed, LILACS and grey literature [Google Scholar and handsearch of the database of the Information System for Notifiable Diseases (SINAN) of Brazil's Unified Health System] were used. The protocol was registered under PROSPERO (CRD42019128998). Climatic, environmental factors and indicators of urban social structure were described as influencing the outbreaks in the North and Northeast regions. Gender and age characteristics were related to a greater chance of developing VL in the Central-West, Northeast and Southeast regions. Vector indicators showed a positive correlation with the incidence of VL in studies in the Northeast region. In the Southeast and Northeast regions, studies revealed the presence of dogs with positive correlation with VL. Knowledge gaps remain regarding the contribution to the increase in the risk factors described in ecological approaches, as no analysis was performed at the individual level, and it is still necessary to discuss the influence of other associated elements in epidemic episodes in the spread of VL. [ABSTRACT FROM AUTHOR]

Globally visceral leishmaniasis (VL) causes thousands of human deaths every year. In South America, the etiologic agent, Leishmania infantum, is transmitted from an infected canine reservoir to human hosts by the bite of the sand fly vector; predominantly Lutzomyia longipalpis. Previous evidence from model rodent systems have suggested that the odour of infected hosts is altered by the parasite making them more attractive to the vector leading to an increased biting rate and improved transmission prospects for the pathogen. However, there has been no assessment of the effect of Le infantum infection on the attractiveness of dogs, which are the natural reservoirs for human infection. Hair collected from infected and uninfected dogs residing in a VL endemic city in Brazil was entrained to collect the volatile chemical odours present in the headspace. Female and male Lu. longipalpis sand flies were offered a choice of odour entrained from infected and uninfected dogs in a series of behavioural experiments. Odour of uninfected dogs was equally attractive to male or female Lu. longipalpis when compared to a solvent control. Female Lu. longipalpis were significantly more attracted to infected dog odour than uninfected dog odour in all 15 experimental replicates (average 45.7±0.87 females attracted to infected odour; 23.9±0.82 to uninfected odour; paired T-test, P = 0.000). Male Lu. longipalpis did not significantly prefer either infected or uninfected odour (average 36.1±0.4 males to infected odour; 35.7±0.6 to uninfected odour; paired T-test, P = 0.722). A significantly greater proportion of females chose the infected dog odour compared to the males (paired T-test, P = 0.000). The results showed that the odour of dogs infected with Le. infantum was significantly more attractive to blood-seeking female sand flies than it was to male sand flies. This is strong evidence for parasite manipulation of the host odour in a natural transmission system and indicates that infected dogs may have a disproportionate significance in maintaining infection in the canine and human population. Author summary: Visceral leishmaniasis (VL) is a disease caused by the Protist parasite Leishmania infantum. In Brazil and other South and Central American countries, the parasite is transmitted by the bite of infected female Lutzomyia longipalpis sand flies. The disease leads to thousands of human deaths every year. Domestic dogs are the reservoir of infection therefore understanding how the infection affects the attractiveness of dogs to the insect vector is important in understanding the epidemiology of the disease. Golden Hamsters infected with Le. infantum are more attractive than uninfected hamsters to Lu. longipalpis however, it is not known if the attractiveness of infected dogs is similarly altered. In this study we showed that the odour of infected dogs is significantly more attractive to female sand flies, that can transmit the pathogen, than it is to male sand flies which can not. This clear-cut difference in attraction of female and males suggests that the females are preferentially attracted by parasite infected hosts and may lead to enhanced infection and transmission opportunities for the parasite. [ABSTRACT FROM AUTHOR]

Mosquito diversity and disease transmission are influenced by landscape modifications, i.e., vectors and pathogens previously found only in forests are now found close to human environments due to anthropic changes. This study determined the diversity and distribution of mosquitoes in forest environments in order to analyze the potential vectors of Amazonian forest arboviruses. Mosquitoes were collected by 1) vertical stratification from forest canopy and ground areas using Hooper Pugedo (HP) light traps and human attraction and 2) horizontal stratification using HP light traps in peridomicile, forest edge, and forest environments near the Rio Pardo rural settlement, Amazonas, Brazil. A total of 3,750 mosquitoes were collected, representing 46 species. 3,139 individuals representing 46 species were sampled by vertical stratification. Both the Shannon-Weaver diversity index (H') and equitability (J') were higher in the canopy than on the ground. 611 individuals representing 13 species were sampled by horizontal stratification. H' decreased in the following order: forest edge > forest > peridomicile, and J' was greater at the forest edge and smaller in the peridomicile environment. Moreover, H' was higher for the human attraction collection method than the HP traps. A total of 671 pools were analyzed by RT-qPCR; three species were positive for Oropouche-like viruses (Ochlerotatus serratus, Psorophora cingulata, and Haemagogus tropicalis) and the minimum infection rate was 0.8%. The composition of mosquito species did not differ significantly between anthropic and forest environments in Rio Pardo. Some mosquito species, due to their abundance, dispersion in the three environments, and record of natural infection, were hypothesized to participate in the arbovirus transmission cycle in this Amazonian rural settlement. [ABSTRACT FROM AUTHOR]

Background: The rising incidence of visceral leishmaniasis due to Leishmania infantum requires novel methods to control transmission by the sand fly vector. Indoor residual spraying of insecticide (IRS) against these largely exophilic / exophagic vectors may not be the most effective method. A synthetic copy of the male sex-aggregation pheromone of the key vector species Lutzomyia longipalpis in the Americas, was co-located with residual pyrethroid insecticide, and tested for its effects on vector abundance, hence potential transmission, in a Brazilian community study. Methods: Houses within eight defined semi-urban blocks in an endemic municipality in Brazil were randomised to synthetic pheromone + insecticide or to placebo treatments. A similar number of houses located >100m from each block were placebo treated and considered as "True Controls" (thus, analysed as three trial arms). Insecticide was sprayed on a 2.6m2 surface area of the property boundary or outbuilding wall, co-located within one metre of 50mg synthetic pheromone in controlled-release dispensers. Vector numbers captured in nearby CDC light traps were recorded at monthly intervals over 3 months post intervention. Recruited sentinel houses under True Control and pheromone + insecticide treatments were similarly monitored at 7–9 day intervals. The intervention effects were estimated by mixed effects negative binomial models compared to the True Control group. Results: Dose-response field assays using 50mg of the synthetic pheromone captured a mean 4.8 (95% C.L.: 3.91, 5.80) to 6.3 (95% C.L.: 3.24, 12.11) times more vectors (female Lu. longipalpis) than using 10mg of synthetic pheromone. The intervention reduced household female vector abundance by 59% (C.L.: 48.7, 66.7%) (IRR = 0.41) estimated by the cross-sectional community study, and by 70% (C.L.: 56.7%, 78.8%) estimated by the longitudinal sentinel study. Similar reductions in male Lu. longipalpis were observed. Beneficial spill-over intervention effects were also observed at nearby untreated households with a mean reduction of 24% (95% C.L.: 0.050%, 39.8%) in female vectors. The spill-over effect in untreated houses was 44% (95% C.L.: 29.7%, 56.1%) as effective as the intervention in pheromone-treated houses. Ownership of chickens increased the intervention effects in both treated and untreated houses, attributed to the suspected synergistic attraction of the synthetic pheromone and chicken kairomones. The variation in IRR between study blocks was not associated with inter-household distances, household densities, or coverage (proportion of total households treated). Conclusions: The study confirms the entomological efficacy of the lure-and-kill method to reduce the abundance of this important sand fly vector in treated and untreated homesteads. The outcomes were achieved by low coverage and using only 1–2% of the quantity of insecticide as normally required for IRS, indicating the potential cost-effectiveness of this method. Implications for programmatic deployment of this vector control method are discussed. Author summary: The predominant sand fly vector of the intracellular parasite Leishmania infantum, that causes human and canine visceral leishmaniasis in the Americas, is Lutzomyia longipalpis. Vector control tools to reduce transmission are needed. A sex-aggregation pheromone released by male Lu. longipalpis attracts female conspecifics which facilitates blood-feeding and transmission. This study, conducted in Brazil, quantifies the effects of community deployment of a synthetic version of the sex-aggregation pheromone, in a controlled-release dispenser, and co-located with lethal insecticide applied to a small area of the household compound or outbuilding wall. 50mg synthetic pheromone dispensers were used since they attracted substantially more vectors than 10mg dispensers. Deploying this novel lure-and-kill method to houses in eight replicate study blocks in two suburban endemic areas, demonstrated that it reduces vector numbers at both pheromone + insecticide treated houses, and neighbouring untreated houses. The presence of chickens (a known additional attraction to blood-seeking female Lu. longipalpis) increased the beneficial effects of the intervention. The method used only 1–2% of the quantity of insecticide necessary for IRS for an average sized house. The results demonstrate the efficacy and potential cost-effectiveness of this novel lure-and-kill control method. [ABSTRACT FROM AUTHOR]

The present study aimed to evaluate the distribution of sand fly species in iron ore caves in the State of Pará, Brazil and to associate the richness and abundance of these insects with the capacity of leishmaniasis transmission. Entomological captures were carried out in the years 2010, 2013, 2014 and 2015, throughout active samples with brushes, along the entire caves' extension, in dry and rainy periods. A total of 9,807 sand flies were counted during the 532 samplings events, being 4,340 in the dry period and 5,467 in the rainy period. A random sample of 802 morphologically identified specimens consisted of 8 genera and 17 species, being 369 males (46%) and 433 females (54%). The predominant species was Sciopemyia sordellii with 60.6% of the total of sand flies collected. Differences in composition and richness were observed between caves located inside of forest and anthropized areas. The mean richness and abundance were different between the wet and rainy periods, with a greater abundance of these insects in the rainy period. The phlebotomine fauna proved to be rich and abundant in the sampled caves, however, environmental degradation seems to be the main factor determining changes in the composition and richness, reinforces the importance of these places as a shelter for sand flies in degraded areas. [ABSTRACT FROM AUTHOR]

Background: Leishmania infantum is the etiological agent of visceral leishmaniasis (VL) in the New World, where the sand fly Lutzomyia longipalpis and domestic dogs are considered the main vector and host reservoirs, respectively. Systemic insecticides have been studied as an alternative to control vector-borne diseases, including VL. Fluralaner, an isoxazoline class compound, is a systemic insecticide used in dogs, with proven efficiency against different species of phlebotomine sand flies. However, to date no studies have demonstrated the efficacy of fluralaner on Lu. longipalpis. The aim of this study was to evaluate the insecticidal effect of fluralaner (Bravecto®) on the sand fly Lu. longipalpis after blood meal in treated dogs. Methods: Healthy mongrel dogs (n = 8) were recruited from the Zoonoses Control Center in the city of Natal, Rio Grande do Norte, Brazil, and randomized into two groups: fluralaner treated (n = 4) and non-treated control (n = 4). Colony-reared female specimens of Lu. longipalpis (n = 20) were allowed to feed on all dogs for 40 min before treatment (for fluralaner-treated dogs), at day 1 after treatment and then monthly until 1 year post-treatment. Results: In the treatment group, there was 100% mortality of Lu. longipalpis for up to 5 months after treatment initiation, decreasing to 72.5% at 6 months post-treatment initiation. The efficacy of fluralaner ranged from 100% at day 1 (P = 0.0002) to 68% (P = 0.0015) at 6 months, decreasing to 1.4% at 1 year post-treatment. Sand fly mortality carried out blood meal in non-treated control dogs remained constant at ≤ 15%. Conclusions: Taken together, our results suggest that fluralaner may be used as a control strategy for VL in dogs in VL endemic areas. [ABSTRACT FROM AUTHOR]

Background: We assessed the impact of two sand fly insecticide interventions (insecticide spraying and insecticide-impregnated dog collars) on the peridomestic abundance and distribution of mosquitoes (Culicidae) and biting midges (Ceratopogonidae) in western São Paulo (Brazil) in a long-term (42-month) evaluation. Both of these dipteran groups are vectors of diseases of medical and veterinary relevance to humans and domestic animals in Brazil. Methods: The interventions in the 3-arm stratified randomised control trial were: pheromone + insecticide (PI) (chicken roosts were sprayed with microencapsulated lambda-cyhalothrin; pheromone lure has no effect on the Diptera pests studied here); dog-collars (DC) (dogs fitted with deltamethrin-impregnated collars); and control (C) (unexposed to pyrethroids) were extended by 12 months. During that time, adult mosquitoes and midges were sampled along 280 households at three household locations (inside human dwellings, dog sleeping sites and chicken roosts). Results: We collected 3145 culicids (9 genera, 87.6% Culex spp.) distributed relatively uniformly across all 3 arms: 41.9% at chicken roosts; 37.7% inside houses; and 20.3% at dog sleeping sites. We collected 11,464 Culicoides (15 species) found mostly at chicken roosting sites (84.7%) compared with dog sleeping sites (12.9%) or houses (2.4%). Mosquitoes and Culicoides were most abundant during the hot and rainy season. Increased daytime temperature was marginally associated with increased mosquito abundance (Z = 1.97, P = 0.049) and Culicoides abundance (Z = 1.71, P = 0.087). There was no significant association with daily average rainfall for either group. Household-level mosquito and midge numbers were both significantly reduced by the PI intervention 56% [incidence rate ratio, IRR = 0.54 (95% CI: 0.30–0.97), P ≤ 0.05] and 53% [IRR = 0.47 (95% CI: 0.26–0.85), P ≤ 0.05], respectively, compared to the control intervention. The abundance of both dipteran groups at dog sleeping sites was largely unaffected by the PI and DC interventions. The PI intervention significantly reduced abundance of mosquitoes inside houses (41%) and at chicken roosting sites (48%) and reduced midge abundance by 51% in chicken roosting sites. Conclusions: Sprayed insecticide at chicken roosting sites reduced the abundance of mosquitoes and midges at the peridomestic level while dog collars had no effect on numbers for any group. [ABSTRACT FROM AUTHOR]

Background In South America the sand fly Lutzomyia longipalpis is the predominant vector of Leishmania infantum, the parasite that causes canine and human visceral leishmaniasis. Co-location of synthetic male sex-aggregation pheromone with an insecticide provided protection against canine seroconversion, parasite infection, reduced tissue parasite loads, and female sand fly densities at households. Optimising the sex-aggregation pheromone + insecticide intervention requires information on the distance over which female and male Lu. longipalpis would be attracted to the synthetic pheromone in the field. Methodology/Principal findings Wild Lu. longipalpis were collected at two peridomestic study sites in Governador Valadares (Minas Gerais, Brazil). Sand flies were marked with coloured fluorescent powder using an improved protocol and then released into an existing domestic chicken shed at two independent sites, followed by recapture at synthetic-pheromone host-odour baited traps placed up to 30 metres distant from the release point. In total 1704 wild-caught Lu. longipalpis were released into the two chicken sheds. Overall 4.3% of the marked flies were recaptured in the pheromone baited experimental chicken sheds compared to no marked flies recaptured in the control sheds. At the first site, 14 specimens (10.4% of the marked and released specimens) were recaptured at 10m, 36 (14.8%) at 20m, and 15 (3.4%) at 30m. At the second site, lower recapture rates were recorded; 8 marked specimens (1.3%) were recaptured at 5 and 10m and no marked specimens were recaptured at 15m. Approximately 7x more marked males than females were recaptured although males were only 2x as common as females in the released population. 52% of the marked Lu. longipalpis were collected during the first night of sampling, 32% on the second night, and 16% on the third night. Conclusions/Significance The study established that both male and female sand flies can be attracted to the synthetic sex-aggregation pheromone in the presence of host odour over distances up to at least 30m in the field depending on local environmental and meterological conditions. [ABSTRACT FROM AUTHOR]

Anthropogenic environments provide favorable conditions for some species, which is especially true of mosquitoes that present eclecticism at the moment of choice for the site of oviposition. In the present study, the diversity of mosquitoes was assessed by providing plastic containers, bamboo internodes, and tires in a forest, the forest edge, and peridomicile environments in a rural settlement area. Eighteen sampling points were chosen, delimited by a buffer of 200 m, placed in three environments: forest, forest edge, and peridomicile. In each environment, larvitraps were installed, separated by a minimum distance of 7 m and 1 m from the ground. A total of 10,131 immature mosquitoes of 20 species were collected. The most abundant species was Culex urichii (29.5%), followed by Trichoprosopon digitatum (27.1%), and Cx. (Melanoconion) spp. (10.4%). There was a difference in the composition of immature mosquito populations between larvitraps (p < 0.0005), and the plastic container hosted a greater diversity of species, whereas tires presented a greater abundance of individuals. The forest, forest edge, and peridomicile environments were also different with regard to diversity of immature mosquito populations (p < 0.0010). The forest edge was the environment with the greatest diversity of species, followed by the peridomicile and forest environments. In the forest and peridomicile, plastic container larvitraps had the greatest diversity, whereas the forest edge tire presented the largest number of individuals. Further, tire larvitraps collected the largest number of individuals in all environments. Ten species associated with the bamboo internode and tire were identified. The preference of species for artificial larvitraps, such as the plastic container and tire, even in wild environments was noted. These artificial objects may represent a risk factor for the population living in this region, as all vector species found in the study were present in plastic containers and tires. [ABSTRACT FROM AUTHOR]

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This study aimed to describe the sand fly fauna and detect trypanosomatids in these insects from Casa Branca, state of Minas Gerais, Brazil, an endemic area of both visceral (VL) and tegumentary leishmaniasis (TL). Sand flies were collected bimonthly from May 2013 to July 2014, using automatic light traps exposed for three consecutive nights in peridomiciliary areas of nine houses with previous reports of VL and TL. ITS1-PCR and DNA sequencing were performed for trypanosomatids identification. A total of 16,771 sand flies were collected belonging to 23 species. The most abundant species was Nyssomyia whitmani (Antunes & Coutinho, 1939) (70.9%), followed by Lutzomyia longipalpis (Lutz & Neiva, 1912) (15.2%) and Migonemyia migonei (França, 1920) (9.1%). Leishmania amazonensis DNA was detected in Ny. whitmani (four pools) and Le. braziliensis DNA was detected in Psychodopygus lloydi (one pool). In seven pools of Ny. whitmani and in one pool of Lu. longipalpis positive for Leishmania DNA, the parasite species was not determined due to the low quality of the sequences. Moreover, DNA of Herpetomonas spp. was detected in Ny. whitmani (two pools) and Cortelezzii complex (one pool). DNA of Crithidia spp. was detected in Ny. whitmani and Ps. lloydi (both one pool). Our results suggest that Ny. whitmani may be involved in the transmission of Le. amazonensis in the study area. The molecular detection of Le. amazonensis suggests the presence of this species in a sylvatic cycle between vertebrate and invertebrate hosts in the region of Casa Branca. Our data also reveal the occurrence of other non-Leishmania trypanosomatids in sand flies in Casa Branca District. [ABSTRACT FROM AUTHOR]

Leishmaniosis is caused by the protozoa of the genus Leishmania with a wide spectrum of clinical and epidemiological manifestations which are characterized into four clinical groups: cutaneous, mucocutaneous, diffuse cutaneous, and visceral. American visceral leishmaniosis (AVL) or visceral leishmaniosis (VL) has been known as the most severe form of the disease. However, despite the growing number of people exposed to the infection risk and the great effort done by the scientific community worldwide to significantly increase the knowledge about these diseases, there is no vaccine capable of preventing VL in humans. In this short review, we present some of the plasmids used for the expression of recombinant protein by Escherichia coli strains used mainly for the second generation of vaccines for leishmaniosis. It can be emphasized that currently, these vectors and hosts play an important role in developing vaccine strategies against the disease. Indeed, use of the E. coli BL21 (DE) strain is remarkable mainly due to its characteristics for being a stable protein producer as well as the use of histidine tags for antigen purification. Key points: • Plasmid vectors and E. coli will continue being important for studies about leishmaniosis. • Protein purification exploiting histidine tags is a key technique. [ABSTRACT FROM AUTHOR]

Caves are extreme and inhospitable environments that can harbor several species of vertebrate and invertebrate animals. Among these animals are phlebotomine sand flies, vectors of parasites of the genus Leishmania that cause leishmaniasis. This study aimed to evaluate the species composition of sand flies of four caves, a cave located at Moeda Sul (MS) and three at Parque Estadual Serra do Rola Moça (PESRM), in the region of the Quadrilátero Ferrífero in the state of Minas Gerais, southeastern Brazil. Sand flies were collected with automatic light traps. Non-Metric Multidimensional Scaling, using a dissimilarity matrix calculated with the Jaccard index, and Multivariate Permutation Analysis were used to evaluate sand fly species composition among entrance, interior, and the surrounding environments of each sampled cave and to infer biological mechanisms from patterns of distribution among these different cave environments. A total of 375 phlebotomine sand flies representing 14 species and six genera were collected. The most abundant species were Evandromyia tupynambai (54.7%), Brumptomyia troglodytes (25.6%), Evandromyia edwardsi (6.1%), Psathyromyia brasiliensis (4.8%) and Lutzomyia longipalpis (4.3%). Thirty individuals were collected at MS, 16 inside the cave and 14 from its surroundings. At PESRM, five individuals were collected from the surroundings of cave RM38, 190 individuals from cave RM39 (48 in the cave and 142 from its surroundings) and 150 individuals from cave RM40 (42 in the cave and 108 from its surroundings). The results revealed a rich sand fly fauna with similar species compositions among the entrance, interior, and surrounding environments of each sampled cave, suggesting that both caves and their surroundings are important for maintaining sand fly communities. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is a neglected tropical disease, caused by Leishmania (Kinetoplastida, Trypanosomatidae) species. In Brazil, the transmission of this parasite essentially occurs through the bite of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) previously infected with Leishmania infantum. Aiming at preventing VL expansion over the country, integrated control actions have been implemented through a Visceral Leishmaniasis Surveillance and Control Program (VLSCP). Among the actions currently adopted by the program, the screening-culling of seropositive dogs for canine VL (CVL) is particularly polemic. Dogs with negative or divergent serology for CVL remain in their owner's domicile and are monitored by public health agents. In the present study, we determined the prevalence of CVL and analyzed the implementation of the VLSCP screening-culling action, in an area in Brazil where there has been a recent expansion of VL. Canine census surveys were conducted semiannually for two years (Aug/2015 to Feb/2017). Serological diagnosis of CVL was performed in accordance with current VLSCP protocol: immunochromatography (TR-DPP) followed by enzyme-linked immunosorbent assay (ELISA EIE). 6,667 dogs were serologically screened for CVL, of which 567 (8.5%) were positive in both tests and 641 (9.6%) had divergent results. A variable percentage (6.3% to 65.4%) of the dogs in the latter group became positive within nine months from the first result. Xenodiagnosis was conducted in canine samples belonging to any of the three possible serological statuses for CVL–positive, divergent or negative. Leishmania spp. DNA was detected in Lu. longipalpis that fed on 50.0% (5/10) of dogs with positive serology and on 29.4% (5/17) of dogs with divergent serological status for CVL. Therefore, dogs with divergent serology for CVL may be as Leishmania-infective to Lu. longipalpis as seropositive ones. Even with the adoption of euthanasia for seropositive dogs, part of the canine population will continue to serve as a source of Leishmania infection for phlebotomine sand flies. Author summary: Visceral leishmaniasis (VL) is a neglected tropical disease caused by a protozoan of the genus Leishmania. In Brazil, the parasites are mainly transmitted through the bite of female Lutzomyia longipalpis sand flies that have been previously infected with Leishmania infantum. Dogs are the main domestic reservoirs of Leishmania. The Brazilian Ministry of Health has implemented euthanasia of VL-seropositive dogs as a measure to prevent VL expansion in the country, using the Dual-Path Platform Immunochromatographic Rapid Test (TR-DPP) as the screening method and the enzyme-linked immunoassay (ELISA EIE) as the confirmatory test for diagnosis. Dogs with positive results in both tests are considered seropositive and recommended for euthanasia. Dogs with divergent serology between these tests remain at their owners' home and part of them become seropositive within months. Therefore, even adopting euthanasia of seropositive dogs, part of the canine population will continue to serve as a source of Leishmania infection for sand flies, causing new canine and / or human cases of the disease. Difficulties and limitations to implement the screening-culling policy are also discussed. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a emergent disease characterized by different clinical manifestations in both humans and dogs. Predominant clinical features of cutaneous leishmaniasis are ulcerative painless skin lesions. Several data reported that pain is associated with human and dog leishmaniasis, out with areas of painless ulcerative lesions per se. Actually, current medications used for leishmaniasis management are characterized by several side effects and, in addition, some cases of the disease are refractory to the treatment. On this background it is mandatory the identification of new and safe candidates for designing less toxic and low-cost remedies. Therefore, the search for new leishmanicidal compounds is indispensable. Methods: In the present paper we investigated the effect of orally N-acetyl-L-cysteine (NAC) supplementation at dose of 200 mg/Kg for 10 weeks, in subcutaneous Leishmania (L). amazonensis infected BALB/c mice. And evaluating the effect of NAC on inflammatory response such as TNF-α, IL-6, IL-1β levels, and on thermal and mechanical hyperalgesia. Results: In the present paper we showed how NAC supplementation affected parameters of oxidative stress (GSH, MDA, SOD), inflammation such as cytokines levels (IL-1β, IL-6, TNFα) and mast cell activation and consequently on induced pain, during leishmaniosis in BALB\c mice. Conclusions: The findings of our study provided the scientific data demonstrating that L. amazonensis infection induces inflammation and pain in BALB/c mice that are reversed by administration of NAC. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis is a highly lethal zoonosis transmitted by a sandfly. It is caused by a Leishmania protozoan parasite and dogs are the main reservoir. Ceara State is endemic to visceral leishmaniasis and it is considered a high risk transmission area. Temporal and spatial epidemiological studies have been used as tools to analyze the distribution and frequency of human visceral leishmaniasis (HVL). This study aimed to characterize HVL in its epidemiological andtemporal aspects in Ceara State, from 2003 to 2017, as this is a neglected disease and a public health problem. This is an ecological study carried out with HVL confirmed cases in Ceara, using three blocks of years (2003 to 2007, 2008 to 2012 and 2013 to 2017). The disease presented an endemic behavior, affecting mainly male residents in the urban area, especially children under five and young adults between 30 and 49 years old. HVL is recorded in all the municipalities, for more than 10 years, with a growing trend and territorial expansion to the Central and Eastern regions of the State. The results of this study indicated the increase in the incidence and lethality, as well as the expansion of leishmaniasis in Ceara State. [ABSTRACT FROM AUTHOR]

Presence of Leishmania spp. was evaluated in the blood of nine red howler monkeys (Alouatta seniculus) from a specific area of French Guiana, located in the northeast of the Amazon. The molecular detection was performed based on PCR targeting the markers 18S rRNA, kDNA and ITS2 genes, as well as rapid immunomigration tests. Two monkeys were positive for Leishmania infantum and one for Leishmania guyanensis. While L. guyanensis cutaneous leishmaniasis is common, visceral leishmaniasis (human and canine) caused by L. infantum has never been described in this area. The howler monkey proved to be a sentinel and a potential reservoir of a serious zoonosis. These results must be carefully considered by public health officials and veterinarians in the future. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is a zoonosis caused by the protozoan Leishmania infantum and in Brazil is transmitted mainly by the bite of Lutzomuyia longipalpis sand flies. Data about the presence, distribution, natural infection rate, seasonal and monthly dynamics of the vector population are important for optimizing the measures to control VL in endemic areas. This study aimed to identify sand fly fauna in an endemic area for VL to detect the prevalence of L. infantum infection in the Lu. longipalpis population and to elucidate the influence of bioclimatic factors on the monthly fluctuations of this vector. HP light traps were monthly set in the intradomicile and peridomicile of residences located in the central and beachfront areas of Camaçari, a VL endemic area. The sand fly collection was conducted in two periods: i) period 1—between December 2011 and November 2012 and ii) period 2—August 2014 and July 2015. Sand fly species were identified and detection of L. infantum infection by qPCR was performed in pools of female Lu. longipalpis. For the first time, the parasite load of positive pools was correlated with the number of Lu. longipalpis captured per month in both periods. Correlation analyses between the monthly fluctuation of the sand fly population and bioclimatic indices of the municipality in both collection periods were also performed. In both evaluated periods, more than 98% of the collected sand flies were Lu. longipalpis, confirming the predominance of this species in the region. It was captured mostly in the beachfront area in all months evaluated (99%). For the period 1, Leishmania DNA was detected in 81% of tested pools representing a minimal infection rate of 9.6%. In the period 2, 40% of the pools were positive with a minimal infection rate of 10.2%. Infected sand flies were only detected in the beachfront area in both periods. The parasite load was low and did not vary in the evaluated months despite the number of collected sand flies. No correlation was observed for climatic factors in both areas of Camaçari. These findings emphasize the high risk of Leishmania transmission in Camaçari regardless of the season and that other factors, aside from bioclimatic elements, are influencing the sand fly population monthly fluctuation. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL) in Brazil is a neglected, vector-borne, tropical parasitic disease that is responsible for several thousand human deaths every year. The transmission route involves sand flies becoming infected after feeding on infected reservoir host, mainly dogs, and then transmitting the Leishmania infantum parasites while feeding on humans. A major component of the VL control effort is the identification and euthanasia of infected dogs to remove them as a source of infection. A rapid, non-invasive, point-of-care device able to differentiate between the odours of infected and uninfected dogs may contribute towards the accurate diagnosis of canine VL. Methodology/Principal findings: We analysed the headspace volatile chemicals from the hair of two groups of dogs collected in 2017 and 2018 using a bench-top eNose volatile organic chemical analyser. The dogs were categorised as infected or uninfected by PCR analysis of blood samples taken by venepuncture and the number of parasites per ml of blood was calculated for each dog by qPCR analysis. We demonstrated using a robust clustering analysis that the eNose data could be discriminated into infected and uninfected categories with specificity >94% and sensitivity >97%. The eNose device and data analysis were sufficiently sensitive to be able to identify infected dogs even when the Leishmania population in the circulating blood was very low. Conclusions/Significance: The study illustrates the potential of the eNose to rapidly and accurately identify dogs infected with Le. infantum. Future improvements to eNose analyser sensor sensitivity, sampling methodology and portability suggest that this approach could significantly improve the diagnosis of VL infected dogs in Brazil with additional potential for effective diagnosis of VL in humans as well as for the diagnosis of other parasitic diseases. [ABSTRACT FROM AUTHOR]

Canine Visceral Leishmaniasis (CVL) prevalence, spatial distribution and associated factors were assessed in four locations in Iguazú department in 2014 and in Puerto Iguazú city again in 2018. The city areas were divided into a grid of 400x400m cells. All cells were sampled in 2014 and a random subsampling was developed in 2018. In each cell, five dogs clustered in a ‘critical scenario’ (prone to have vectors) were sampled. A rapid immunochromatographic dipstick was used to detect antibodies against Leishmania infantum, confirming by lymph node smears observation and PCR. For Puerto Iguazú, Generalized Linear Models (GLMs) were constructed considering environmental, dog and clinical variables. Pearson's Chi square and Fisher's exact tests were employed to evaluate the association between CVL, dog clinical signs and infestation with other parasites. Cartographic outputs were made and Moran's I indices were calculated as spatial autocorrelation indicators. CVL prevalence rates were 26.18% in 2014 and 17.50% in 2018. No associations were established in environmental models, but dog age and repellent use were significant when running 2014 dog models. Clinical models showed significant associations between seropositive dogs and ophthalmological, dermal signs and onychogryphosis in 2014. In 2018, only adenomegaly was associated. The results of global Moran´s I were not significant but regarding local analysis, six sites in 2014 and one in 2018 presented autocorrelation with neighboring sites. The decrease in CVL prevalence may be associated to transmission stabilization, which could explain the lack of associations with dog-related variables. Further, spatial distribution of CVL is a poor evidence for design of transmission control measures but could be important in case of intensive parasite circulation or when the first autochthonous cases appear. For control success, sensitivity of diagnostic methods, political will and adequate material resources remain critical. Modeling of multiple variables will be required to identify factors that drive disease stabilization/destabilization. [ABSTRACT FROM AUTHOR]

In the present study, we assessed the annual screening coverage (i.e., the percentage of dogs that are screened for anti-Leishmania antibodies annually) in the municipality of Sobral, Ceará state, Brazil. Data on the number of dogs screened during 2008−2017 (except 2010) were obtained from the Centre for Zoonoses Control of Sobral. The annual screening coverage during 2012−2017 was calculated. Data on human visceral leishmaniasis (VL) cases during 2008−2017 were compiled from the National Disease Notification System. Correlation analyses were performed to assess the correlation between canine and human data. During 2008−2017, 73,964 dogs (range, 0 to 13,980 dogs/year) were serologically screened and 2,833 (3.8%) were positive. The annual screening coverage during 2012−2017 ranged from 11.1% to 45.7%. There were no significant correlations between the number of dogs culled and the number of human VL cases, canine positivity and human VL incidence, number of dogs culled and human VL incidence, or between canine positivity and number of human VL cases. An inconsistent and relatively low annual screening coverage was found in the study area, with no dog being screened in 2010 due to the lack of serological tests. Our results highlight that many dogs potentially infected with Leishmania infantum have been virtually overlooked by public health workers in the study area, perhaps with a negative, yet underestimated, impact on the control of canine and human VL. Hence, the failure of the dog culling strategy in controlling human VL in Brazil may be due to the low screening coverage and low percentage of culled dogs, rather than the absence of associations between canine and human infections. [ABSTRACT FROM AUTHOR]

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Background: Leishmaniasis caused by protozoan parasite Leishmania is a neglected disease which is endemic in the northwest of China. Reptiles were considered to be the potential reservoir hosts for mammalian Leishmaniasis, and Leishmania had been detected in lizards from the epidemic area in the northwest of China. To date, few studies are focused on the natural infection of snakes with Leishmania. Methods: In this study, 15 snakes captured from 10 endemic foci in the northwest of China were detected Leishmania spp. on the base of mitochondrial cytochrome b, heat shock protein 70 gene and ribosomal internal transcribed spacer 1 regions, and identified with phylogenetic and network analyses. Result: In total, Leishmania gene was found in 7 snakes. The phylogenetic inference trees and network analysis suggests that the species identification was confirmed as Leishmania donovani, L. turanica and L. (Sauroleishmania) sp. Conclusion: Our work is the first time to investigate the natural Leishmania spp. infection of snakes in the northwest of China. Mammalian Leishmania (L. donovani and L. turanica) was discovered in snakes and the reptilian Leishmania (Sauroleishmania sp.) was closely related to the clinical strains both prompt the importance of snakes in the disease cycle. To indicate the epidemiological involvement of snakes, a wide sample size in epidemic area and the pathogenic features of reptilian Leishmania promastigotes are recommended in the future research. [ABSTRACT FROM AUTHOR]

Leishmaniasis encompasses a group of diverse clinical diseases caused by protozoan parasites of the Leishmania genus. This disease is a major public health problem in the New World affecting people exposed in endemic regions. The city of Governador Valadares (Minas Gerais/Brazil) is a re-emerging area for visceral leishmaniasis, with 191 human cases reported from 2008 to 2017 and a lethality rate of 14.7%. The transmission of the parasite occurs intensely in this region with up to 22% of domestic dogs with positive serology for the visceral form. Lu. longipalpis is one of the most abundant sand fly species in this area. Despite this scenario, so far there is no information regarding the circulating Leishmania species in the insect vector Lutzomyia longipalpis in this focus. We collected 616 female Lutzomyia longipalpis sand flies between January and September 2015 in the Vila Parque Ibituruna neighborhood (Governador Valadares/MG), which is located on a transitional area between the sylvatic and urban environments with residences built near a preserved area. After DNA extraction of individual sand flies, the natural Leishmania infections in Lu. longipalpis were detected by conventional PCR, using primers derived from kDNA sequences, specific for L. (Leishmania) or L. (Viannia) subgenus. The sensitivity of these PCR reactions was 0.1 pg of DNA for each Leishmania subgenus and the total infection rate of 16.2% (100 positive specimens). Species-specific PCR detected the presence of multiple Leishmania species in infected Lu. longipalpis specimens in Governador Valadares, including L. amazonensis (n = 3), L. infantum (n = 28), L. (Viannia) spp. (n = 20), coinfections with L. infantum and L. (Viannia) spp. (n = 5), and L. (Leishmania) spp (n = 44). Our results demonstrate that multiple Leishmania species circulate in Lu. longipalpis in Governador Valadares and reveal a potential increasing risk of transmission of the different circulating parasite species. This information reinforces the need for epidemiological and entomological surveillance in this endemic focus, and the development of effective control strategies against leishmaniasis. [ABSTRACT FROM AUTHOR]

In South America, the Protist parasite that causes visceral leishmaniasis, a potentially fatal human disease, is transmitted by blood-feeding female Lutzomyia longipalpis sand flies. A synthetic copy of the male produced sex-aggregation pheromone offers new opportunities for vector control applications. We have previously shown that the pheromone placed in plastic sachets (lures) can attract both females and males to insecticide treated sites for up to 3 months. To use the pheromone lure in a control program we need to understand how the application of lures in the field can be optimised. In this study we investigated the effect of increasing the number of lures and their proximity to each other on their ability to attract Lu. longipalpis. Also for the first time we applied a Bayesian log-linear model rather than a classic simple (deterministic) log-linear model to fully exploit the field-collected data. We found that sand fly response to pheromone is significantly related to the quantity of pheromone and is not influenced by the proximity of other pheromone sources. Thus sand flies are attracted to the pheromone source at a non-linear rate determined by the amount of pheromone being released. This rate is independent of the proximity of other pheromone releasing traps and indicates the role of the pheromone in aggregation formation. These results have important implications for optimisation of the pheromone as a vector control tool and indicate that multiple lures placed in relatively close proximity to each other (5 m apart) are unlikely to interfere with one another. [ABSTRACT FROM AUTHOR]

To reduce morbidity and mortality caused by visceral leishmaniasis (VL), the Brazilian Visceral Leishmaniasis Control and Surveillance Program promotes the diagnosis and treatment of cases, vector control, euthanasia of seropositive dogs, and health education. Nevertheless, the effectiveness of these measures is questionable as they lead to little reduction in the transmission of the disease. Thus, the effectiveness of strategies such as insecticide-impregnated collars, spot-on insecticides, and immunization of dogs should be assessed. Herein, we evaluated the effectiveness of deltamethrin-impregnated collars on reducing the incidence of Leishmania infantum infection in dogs living in an endemic area of VL. An intervention study was conducted and a total 5,850 dogs were analyzed in baseline. Of these 3,742 seronegative dogs were divided into two groups: collared and uncollared (control). Dogs were followed for 12 months and three interventions were performed. The Cox regression model was used to evaluate the effectiveness of the collar. All analyzes were performed by Intention-to-treat and per-protocol. By intention-to-treat, the incidence rates of L. infantum infection were 7.5 and 7.9 in the collar group, and 6.5 and 13.2 per 1,000 dogs-months in the control group after 6 and 12 months, respectively. In the per-protocol analysis, the incidence rates in the control group were similar to those observed in the intention-to-treat analysis. In the collar group, the incidence rate was 5.1/1,000 dogs-months after 6 and 12 months. The effectiveness by intention-to-treat after adjustment by the multivariate Cox model was 48%. In the analysis per-protocol, the effectiveness increased to 63%. Although collar use was effective when it was evaluated by intention-to-treat, higher effectiveness was found in the per-protocol analysis after one year of follow-up. The data emphasize the importance of the uninterrupted use of deltamethrin-impregnated collars to increase protection against canine VL. [ABSTRACT FROM AUTHOR]

Leishmaniases are a group of infectious diseases transmitted by phlebotomine sand flies, and their distribution depends on the presence of vectors, parasites, reservoirs and susceptible hosts in the same environment. In the last decades, visceral leishmaniasis (VL) has become urbanized and reached economically important cities in countries within the transmission zone. Our study was conducted in one of those cities–Ipatinga–in the state of Minas Gerais, Brazil, where the first autochthonous case of VL dates back to 2011. Since no data regarding the epidemiological triad of VL (etiological agent/vector/domestic reservoir) were available for this city, we characterized the local entomological fauna, identified the presence of specific Leishmania DNA in the captured phlebotomine sand flies, and assessed the incidence of canine and human VL. For the entomological survey, we set twenty light traps in ten districts of the city with reports of human and canine VL. The insect captures were performed monthly, during one year, starting in March 2015. A total of 1501 specimens of phlebotomine sand flies belonging to 16 distinct species were captured, with predominance (61.9%) of Lutzomyia longipalpis. Leishmania infantum DNA was detected in L. longipalpis and in Evandromyia cortelezzii test samples. A total of 9,136 dogs were examined, 1,355 of which (14.8%) were serologically positive for VL. The cases were georeferenced and the data were plotted in thematic maps, along with human cases of VL registered by the local Department of Health, during the study period. Our results confirm that the VL transmission cycle is active in Ipatinga, with the presence of vectors carrying Leishmania DNA, canine and human cases of the disease. Spatial analysis allowed for the observation of a positive relationship between canine and human cases of VL and the identification of areas with high priority for control actions in the city. The mapping of high-risk areas, together with an epidemiological study in urban areas, is fundamental to improve the efficacy of the Program for Surveillance and Control of VL (PSCVL) in Brazil. [ABSTRACT FROM AUTHOR]

Purpose: To develop an infectious keratitis model using caprine (goat) corneas and to investigate the expression of virulence factors during infection. Methods: Goat eyes were surface-sterilized and dissected, and the corneas were placed on an agarose-gelatin solid support (0.5% in phosphate-buffered saline) in a 12-well culture plate containing 10% fetal bovine serum-supplemented culture medium for 3 weeks. Cell viability tests (trypan blue and MTT) were performed on the cultured corneas. Corneas were infected with Pseudomonas aeruginosa and Fusarium solani separately. Infection progression was observed via histological analysis and hematoxylin and eosin (H-E) staining. For Pseudomonas-infected corneas, expression of eight virulence genes (exoS, exoT, exoY, alpR, prpL, lasA, lasB, and algD) was determined via quantitative real-time PCR (qRT-PCR) at 48-h and 72-h time-points. For Fusarium-infected corneas, expression of five proteases (C7Z0E6, C7ZFW9, C7Z7U2, C7ZNV5, and C7YY94) was quantified via qRT-PCR at 2, 4, and 8 days after infection. Protease from infected corneas was detected via gelatin zymography. Results: Goat corneas with a viable epithelium could be maintained for 15 days. Pseudomonas infection progressed rapidly, and complete corneal degradation was observed on day 4 after infection. Fusarium infection progressed more slowly. Histological analysis and H-E staining of Fusarium-infected cornea revealed mycelia penetrating all layers of the cornea. qRT-PCR revealed expression of all eight virulence factors, and statistically significant difference in expression of prpL and alpR in Pseudomonas-infected corneas. Expression of C7ZNV5 was highest in Fusarium-infected corneas. Conclusion: Goat corneas can be used to evaluate the expression of virulence factors involved in Pseudomonas and Fusarium infection. [ABSTRACT FROM AUTHOR]

Background: Many studies have investigated what could attract insects of medical importance and a crucial role has lately been attributed to human skin odors. Most of these researches have been concerned with mosquitoes, e.g., vectors of dengue and malaria. Little is known about volatile organic compounds (VOCs) from human skin odors and their effects on leishmania vectors. Objective: The present study aimed to identify the VOCs from human skin that can be attractive to female anthropophilic phlebotomine sandflies. Results: Forty-two VOCs were identified from skin odors of 33 male volunteers, seven of which were tested in wind tunnel assays employing field-captured phlebotomine sandflies (75.4% identified as Lutzomyia intermedia). Hexane and (E)-oct-3-en-1-ol (octenol) were used as negative and positive controls, respectively. 2-Phenylacetaldehyde (hereafter called phenylacetaldehyde), 6-methylhept-5-en-2-one (also known as sulcatone), nonadecane and icosane were found to activate female phlebotomine sandflies, but only phenylacetaldehyde, 6-methylhepten-5-en-2-one and icosane elicited attraction responses. Conclusions: These results suggest that phenylacetaldehyde, 6-methylhepten-5-en-2-one and icosane may be suitable candidates for attractiveness experimentation in the field which can be an important tool to develop strategies concerning human beings protection against phlebotomine sandflies bites and consequently against leishmaniasis. [ABSTRACT FROM AUTHOR]