Molecular investigation of Leishmania in sandflies (Diptera: Psychodidae) and rodents (Mammalia: Rodentia) in Nahavand, west of Iran.

Cutaneous leishmaniasis caused by different species of Leishmania is transmitted by Phlebotominae sandflies. This disease remains a public health concern in Iran. Therefore, the present study aimed to examine Leishmania infection in sandflies and reservoir rodents in six rural regions of Nahavand, located in western Iran. From May to October 2022, sandflies and rodents were collected and identified at the species level. Additionally, rodents' skin lesions and earlobe specimens were collected separately for microscopic and molecular examination. All specimens were tested for Leishmania DNA by PCRs targeting the parasite's ITS-2 and 18S rRNA gene and positive were Sanger sequenced. A total of 3396 sandflies belonging to seven subgenera and 11 species, i.e., Phlebotomus papatasi (42.7%), P. major (20.6%), P. mascitti (0.3%), P. neglectus (0.2%), P. alexandri (0.2%), P. turanicus (0.3%), Sergentomyia murgabiensis (18.1%), S. dentata (10.5%), S. theodori (5.8%), S. antennata (1.1%), and S. pawlowski (0.1%) were identified. Based on the species population, 29 pools of sandflies were examined for the presence of Leishmania DNA using conventional PCR (cPCR), and individual DNAs were tested when positive. Leishmania major DNA was detected in two P. papatasi and Leishmania sp. in one P. major individual sandfly. This is the first report of Leishmania infection in sandflies from Hamadan province. The captured rodents (n = 61) belonged to four families and seven species, i.e., Arvicola amphibius (37.7%), Mus musculus (29.5%), Microtus socialis (13.1%), Apodemus sylvaticus (11.5%), Talpa davidiana (4.9%), Apodemus witherbyi (1.6%), and Rattus norvegicus (1.6%). Microscopic and molecular examinations of the rodent lesions and earlobes scored negative results. The presence of Leishmania in the Phlebotominae sandflies in Nahavand indicates a potential threat to humans and animals in the region. Regular monitoring and examination of the sandflies' population and timely diagnosis and treatment of new patients are strongly recommended.

Chemical control of medically important arthropods in Panama: A systematic literature review of historical efforts.

Vector-borne diseases are a major source of morbidity in Panama. Herein, we describe historical usage patterns of synthetic insecticides to control arthropod disease vectors in this country. We examine the influence of interventions by vector control programs on the emergence of insecticide resistance. Chemical control has traditionally focused on two mosquito species: Anopheles albimanus, a major regional malaria vector, and Aedes aegypti, a historical vector of yellow fever, and current vector of dengue, chikungunya, and Zika. Countrywide populations of An. albimanus depict hyperirritability to organochlorine insecticides administered by indoor residual spraying, although they appear susceptible to these insecticides in bioassays settings, as well as to organophosphate and carbamate insecticides in field tests. Populations of Ae. aegypti show resistance to pyrethroids, particularly in areas near Panama City, but the spread of resistance remains unknown in Ae. aegypti and Aedes albopictus. A One Health approach is needed in Panama to pinpoint the insecticide resistance mechanisms including the frequency of knockdown mutations and behavioral plasticity in populations of Anopheles and Aedes mosquitoes. This information is necessary to guide the sustainable implementation of chemical control strategies and the use of modern vector control technologies such as genetically modified mosquitoes, and endosymbiont Wolbachia-based biological control.

Immune Reactions of Vector Insects to Parasites and Pathogens.

This overview initially describes insect immune reactions and then brings together present knowledge of the interactions of vector insects with their invading parasites and pathogens. It is a way of introducing this Special Issue with subsequent papers presenting the latest details of these interactions in each particular group of vectors. Hopefully, this paper will fill a void in the literature since brief descriptions of vector immunity have now been brought together in one publication and could form a starting point for those interested and new to this important area. Descriptions are given on the immune reactions of mosquitoes, blackflies, sandflies, tsetse flies, lice, fleas and triatomine bugs. Cellular and humoral defences are described separately but emphasis is made on the co-operation of these processes in the completed immune response. The paper also emphasises the need for great care in extracting haemocytes for subsequent study as appreciation of their fragile nature is often overlooked with the non-sterile media, smearing techniques and excessive centrifugation sometimes used. The potential vital role of eicosanoids in the instigation of many of the immune reactions described is also discussed. Finally, the priming of the immune system, mainly in mosquitoes, is considered and one possible mechanism is presented.

High climatic ancestral affinity between the lineages of the Leishmania vector Psathyromyia shannoni sensu stricto (Diptera: Phlebotominae).

Psathyromyia (Psathyromyia) shannoni sensu stricto (Dyar) is a vector of Leishmania parasite and the second sandfly of medical importance with a wide geographical but discontinuous distribution in America. Preliminary genetic structure analysis using a mitochondrial marker shows that the species integrated by at least four lineages could be the result of ecological adaptations to different environmental scenarios, but this hypothesis had never been proven. The aim of the present study was to analyse whether the genetic structure that detected Pa. shannoni ss. is associated with divergence or conservatism niche. Using Ecological Niche Models (ENMs) theory, we estimated the potential distribution for each genetic lineage, and then, we evaluated the equivalency niche for assessing whether climatic niche was more different than expected. The ENMs identify different suitable distribution areas but the same climatic or ecological conditions for the genetic lineages of Pa. shannoni (conservatism niche). Our findings allow us to speculate that other potential processes or events could be related to the genetic differentiation of Pa. shannoni. These studies are important because they allow us to identify the factors that could restrict the potential distribution of the different lineages whose vectorial competence is still unknown.

First evidence of experimental genetic hybridization between cutaneous and visceral strains of Leishmania donovani within its natural vector Phlebotomus argentipes.

Leishmaniasis is a neglected tropical disease caused by protozoan parasites of genus Leishmania, and transmitted by different species of Phlebotomine sand flies. More than 20 species of Leishmania are known to cause disease in humans and other animals. Leishmania donovani species complex is known to have a vast diversity of clinical manifestations in humans, but underlying mechanisms for such diversity are yet unknown. Long believed to be strictly asexual, Leishmania have been shown to undergo a cryptic sexual cycle inside its sandfly vector. Natural populations of hybrid parasites have been associated with the rise of atypical clinical outcomes in the Indian subcontinent (ISC). However, formal demonstration of genetic crossing in the major endemic sandfly species in the ISC remain unexplored. Here, we investigated the ability of two distinct variants of L. donovani associated with strikingly different forms of the disease to undergo genetic exchange inside its natural vector, Phlebotomus argentipes. Clinical isolates of L. donovani either from a Sri Lankan cutaneous leishmaniasis (CL) patient or an Indian visceral leishmaniasis (VL) patient were genetically engineered to express different fluorescent proteins and drug-resistance markers and subsequently used as parental strains in experimental sandfly co-infection. After 8 days of infection, sand flies were dissected and midgut promastigotes were transferred into double drug-selective media. Two double drug-resistant, dual fluorescent hybrid cell lines were recovered, which after cloning and whole genome sequencing, were shown to be full genomic hybrids. This study provides the first evidence of L. donovani hybridization within its natural vector Ph. argentipes.

On the systematics of Phlebotomus betisi and two new related species from Laos with proposal of the new subgenus Lewisius.

Phlebotomus betisi was described from Malaysia and classified after its description in the subgenus Larroussius. It was the only species to have a pharyngeal armature composed of dot-like teeth and an annealed spermatheca whose head is carried by a neck in females. Males were characterized by having a style bearing five spines and a simple paramere. The study of sandflies originating from a cave in Laos enabled us to discover and describe two sympatric species close to Ph. betisi Lewis & Wharton, 1963 and new for Science: Ph. breyi Vongphayloth & Depaquit n. sp., and Ph. sinxayarami Vongphayloth & Depaquit n. sp. They were characterized morphologically, morphometrically, geomorphometrically, molecularly, and proteomically (MALDI-TOF). All approaches converged to validate the individualization of these species whose morphological differential characters lay in the two genders by the observation of the interocular suture and by the length of the last two segments of the maxillary palps. In males, the length of the genital filaments discriminates these species. Females are distinguished by the length of the ducts of the spermathecae as well as by the narrow or enlarged shape of the neck bearing their head. Lastly, the particular position of the spines of the gonostyle coupled with molecular phylogeny led us to remove these three species from the subgenus Larroussius Nizulescu, 1931 and to classify them in a new subgenus: Lewisius Depaquit & Vongphayloth n. subg.

Title: Sur la systématique de Phlebotomus betisi et de deux nouvelles espèces apparentées du Laos avec proposition du nouveau sous-genre Lewisius. Abstract: Phlebotomus betisi a été décrit de Malaisie et fut classé après sa description dans le sous-genre Larroussius. C'était la seule espèce à posséder chez la femelle une armature pharyngienne composée de dents en forme de points et à avoir une spermathèque annelée dont la tête est portée par un cou. Les mâles se caractérisaient par un style porteur de cinq épines et par un paramère simple. L'étude de Phlébotomes originaires d'une grotte du Laos nous a permis de découvrir et de décrire deux espèces sympatriques proches de Ph. betisi Lewis & Wharton, 1963 et nouvelles pour la Science : Ph. breyi Vongphayloth & Depaquit n. sp., et Ph. sinxayarami Vongphayloth & Depaquit n. sp. Elles ont été caractérisées morphologiquement, morphométriquement, géomorphométriquement, moléculairement et protéomiquement (MALDI-TOF). Toutes ces approches convergent pour valider l'individualisation de chacune de ces espèces dont les caractères morphologiques différentiels reposent dans les deux sexes par l'observation de la suture interoculaire et par la longueur des deux derniers segments des palpes maxillaires. Chez les mâles, la longueur des filaments génitaux discrimine ces espèces. Les femelles sont distinguées par la longueur des conduits des spermathèques ainsi que par la forme étroite ou élargie du cou portant la tête de ces spermathèques. Enfin, la position particulière des épines sur le gonostyle couplée à une phylogénie moléculaire nous amène à extraire ces trois espèces du sous genre Larroussius Nitzulescu, 1931 pour les classer dans un nouveau sous-genre : Lewisius Depaquit & Vongphayloth n. subg.

Molecular phylogeny of Psychodopygina (Diptera, Psychodidae) supporting morphological systematics of this group of vectors of New World tegumentary leishmaniasis.

New World sandflies are vectors of leishmaniasis, bartonellosis, and some arboviruses. A classification based on 88 morphological characters was proposed 27 years ago when the New World phlebotomines were organized into two tribes Hertigiini and Phlebotomini. The latter was structured into four subtribes (Brumptomyiina, Sergentomyiina, Lutzomyiina, and Psychodopygina) and 20 genera. The subtribe Psychodopygina, including most of the American vectors of tegumentary Leishmania comprises seven genera from which no molecular work has been produced to support this classification. Here, we carried out a molecular phylogeny based on combined sequences (1,334 bp) of two genes: partial 28S rDNA and mtDNA cytochrome b from 47 taxa belonging to the Psychodopygina. The Bayesian phylogenetic reconstruction agreed with the classification based on morphological characters, supporting the monophyly of the genera Psychodopygus and Psathyromyia, whereas Nyssomyia and Trichophoromyia seemed to be paraphyletic. The paraphylies of the two latter groups were exclusively caused by the doubtful position of the species Ny. richardwardi. Our molecular analysis provides additional support to adopt the morphologic classification of Psychodopygina.

Title: La phylogénie moléculaire des Psychodopygina (Diptera, Psychodidae) soutient la systématique morphologique de ce groupe de vecteurs de leishmaniose cutanée du Nouveau Monde. Abstract: Les phlébotomes du nouveau monde sont des vecteurs de leishmaniose, de bartonellose et de certains arbovirus. Une classification basée sur 88 caractères morphologiques a été proposée il y a 27 ans lorsque les phlébotomes du nouveau monde ont été organisés en deux tribus Hertigiini et Phlebotomini. Cette dernière était structurée en quatre sous-tribus (Brumptomyiina, Sergentomyiina, Lutzomyiina et Psychodopygina) et 20 genres. La sous-tribu des Psychodopygina, qui inclut la plupart des vecteurs américains de la leishmaniose cutanée, comprend sept genres mais aucun travail moléculaire n'a été produit pour soutenir cette classification. Dans cet article, nous avons réalisé une phylogénie moléculaire basée sur des séquences combinées (1334 pb) de deux gènes : ADNr 28S partiel et cytochrome b (ADNmt) chez 47 taxons appartenant aux Psychodopygina. La reconstruction phylogénique bayésienne est en accord avec la classification basée sur les caractères morphologiques, soutenant la monophylie des genres Psychodopygus et Psathyromyia, alors que Nyssomyia et Trichophoromyia semblent être paraphylétiques. Les paraphylies des deux derniers groupes sont exclusivement causées par la position douteuse de la seule espèce Ny. richardwardi. Notre analyse moléculaire fournit une raison supplémentaire pour adopter la classification morphologique des Psychodopygina.

Arthropod vectors of disease agents: Their role in public and veterinary health in Turkiye and their control measures.

Mosquitoes, sandflies, and ticks are hematophagous arthropods that pose a huge threat to public and veterinary health. They are capable of serving as vectors of disease agents that can and have caused explosive epidemics affecting millions of people and animals. Several factors like climate change, urbanization, and international travel contribute substantially to the persistence and dispersal of these vectors from their established areas to newly invaded areas. Once established in their new home, they can serve as vectors for disease transmission or increase the risk of disease emergence. Turkiye (formerly Turkey) is vulnerable to climate change and has experienced upward trends in annual temperatures and rising sea levels, and greater fluctuations in precipitation rates. It is a potential hotspot for important vector species because the climate in various regions is conducive for several insect and acari species and serves as a conduit for refugees and immigrants fleeing areas troubled with armed conflicts and natural disasters, which have increased substantially in recent years. These people may serve as carriers of the vectors or be infected by disease agents that require arthropod vectors for transmission. Although it cannot be supposed that every arthropod species is a competent vector, this review aims to (1) illustrate the factors that contribute to the persistence and dispersal of arthropod vectors, (2) determine the status of the established arthropod vector species in Turkiye and their capability of serving as vectors of disease agents, and (3) assess the role of newly-introduced arthropod vectors into Turkiye and how they were introduced into the country. We also provide information on important disease incidence (if there's any) and control measures applied by public health officials from different provinces.

Identification of cuticle and midgut fungal microflora of phlebotomine sandflies collected in Tunisia.

Phlebotomine sand flies (Diptera: Psychodidae) are the proven vectors of Leishmaniases which are widespread parasitosis in many tropical and subtropical countries. The development of infective metacyclic Leishmania (Kinetoplastida: Trypanosomatidae) promastigotes stage is restricted to the vector midgut. Recently, several studies have assessed the influence of the sand fly midgut fungal microflora on the development of invective Leishmania stage. The aim of this study was to identify the fungal microflora from the cuticle and midgut of wild caught sandflies. A total of 50 sandflies were caught in two different leishmaniasis foci of center Tunisia and analyzed using an in vitro isolation of fungi followed by a morphological and molecular identification of fungal isolates. The morphological identification of sandflies specimens revealed five Species: Phlebotomus (P.) papatasi (n = 25), P. perniciosus (n = 15) P. riouxi (n = 6), P. longicuspis (n = 3) and P. sergenti (n = 1). Forty positive fungal cultures were isolated from 34 sand flies (19 males and 15 females) distributed as following: P. papatasi (n = 16), P. perniciosus (n = 11), P. riouxi (n = 4), P. longicuspis (n = 2) and P. sergenti (n = 1). Thirty-five cultures were isolated from the cuticles and five from the guts. A total of 15 fungi genera belonging to 8 families were identified with the predominance of Aspergillus genus followed by Penicillium genus. Among the 15 fungi genera, five were common between males and females specimens. Lecytophora canina and Leishmania major co-infection was detected in the gut of a female P. papatasi. Our preliminary findings highlight the high diversity of fungal microflora from the sand flies midguts.

Mapeando a leishmaniose visceral e tegumentar americana em Birigui-Sp: uma análise epidemiológica retrospectiva e geoespacial / Mapping american visceral and tegumentary leishmaniasis in Birigui-Sp: a retrospective and geospatial epidemiological analysis / Mapeo de la leishmaniasis visceral y tegumentaria americana en Birigui-Sp: un análisis epidemiológico retrospectivo y geoespacial

A leishmaniose é uma doença tropical infecciosa negligenciada com dois espectros clínicos, a Leishmaniose Visceral (LV) e a Leishmaniose Tegumentar Americana (LTA). O objetivo foi determinar a evolução epidemiológica e a distribuição espacial de LV e LTA em Birigui/SP, entre 2010 a 2020. Foi realizado um estudo retrospectivo e observacional, de LV e LTA registrados pelo Serviço de Vigilância Epidemiológica e os endereços foram utilizados para a análise espacial e distribuição dos casos dentro do município. Identificaram-se 233 casos de LV, com maior número em 2012 (15,45%) e 14 óbitos por LV (letalidade de 6,0%), com predomínio de indivíduos do sexo masculino (60,1%), brancos (71,7%), com idades entre 40 e 59 anos (28,8%), ensino médio completo (8,58%) e alta taxa de cura (65,7%) após tratamento. Verificou- se 25 casos de LTA sendo 2015 o ano com maior número de casos (22,2%) e com predomínio em homens (63%), maiores de 60 anos (44,4%), brancos (92,6%) e com ensino fundamental incompleto (18,5%). A manifestação clínica mais observada foi a forma cutânea (59,3%) com mais de 77% de cura após tratamento. Padrões significativos de agregação espacial foram detectados e áreas de cluster acumulam-se na região oriente do município de Birigui, com maior frequência em bairros mais periféricos. Conclui-se que, apesar de ser observada uma leve diminuição nos casos de LV e LTA, reforça-se a necessidade do monitoramento frequente, principalmente nas regiões com maior número de casos.

Leishmaniasis is a neglected tropical infectious disease with two clinical spectrums, Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL). The objective was to determine the epidemiological evolution and spatial distribution of VL and ATL in Birigui/SP, between 2010 and 2020. spatial analysis and distribution of cases within the municipality. A total of 233 cases of VL were identified, with the highest number in 2012 (15.45%) and 14 deaths from VL (6.0% mortality), with a predominance of males (60.1%), whites (71.7%), aged between 40 and 59 years (28.8%), complete high school (8.58%) and high cure rate (65.7%) after treatment. There were 25 cases of ACL, with 2015 being the year with the highest number of cases (22.2%) and with a predominance of men (63%), over 60 years old (44.4%), white (92.6%) and with incomplete primary education (18.5%). The most observed clinical manifestation was the cutaneous form (59.3%) with more than 77% of cure after treatment. Significant patterns of spatial aggregation were detected, and cluster areas accumulated in the eastern region of the municipality of Birigui, with greater frequency in more peripheral neighborhoods. It is concluded that, despite a slight decrease in cases of VL and ATL, the need for frequent monitoring is reinforced, especially in regions with a higher number of cases.

La leishmaniasis es una enfermedad infecciosa tropical desatendida con dos espectros clínicos, la leishmaniasis visceral (LV) y la leishmaniasis cutánea americana (LCA). El objetivo fue determinar la evolución epidemiológica y distribución espacial de la LV y LCA en Birigui/SP, entre 2010 y 2020. análisis espacial y distribución de los casos dentro del municipio. Fueron identificados 233 casos de LV, con el mayor número en 2012 (15,45%) y 14 muertes por LV (6,0% de mortalidad), con predominio del sexo masculino (60,1%), raza blanca (71,7%), edad entre 40 y 59 años (28,8%), enseñanza media completa (8,58%) y alta tasa de cura (65,7%) después del tratamiento. Hubo 25 casos de LCA, siendo 2015 el año con mayor número de casos (22,2%) y con predominio de hombres (63%), mayores de 60 años (44,4%), blancos (92,6%) y con estudios primarios incompletos (18,5%). La manifestación clínica más observada fue la forma cutánea (59,3%) con más del 77% de curación tras el tratamiento. Se detectaron patrones significativos de agregación espacial, acumulándose áreas de conglomerados en la región este del municipio de Birigui, con mayor frecuencia en barrios más periféricos. Se concluye que, a pesar de una leve disminución de los casos de LV y ATL, se refuerza la necesidad de monitoreo frecuente, especialmente en las regiones con mayor número de casos.

Retrospective Analysis of Leishmaniasis in Sicily (Italy) from 2013 to 2021: One-Health Impact and Future Control Strategies.

Leishmaniasis is an important vector-borne disease that represents a serious public health problem, including in Sicily (Italy), which is considered an endemic area. We collected canine, feline and human data from 2013 to 2021 in Sicily, while entomological surveys were conducted only in 2013 and 2021. Overall, 23,794/74,349 (34.4%) of dogs and 274/4774 (11.8%) of cats were positive in one or more diagnostic tests. A total of 467 cases of human Leishmaniasis were reported, with 71% showing cutaneous and 29% visceral involvement. The provinces with the largest number of patients were Agrigento (45.4%) and Palermo (37%). In 2013, Phlebotomus perfiliewi was the dominant sandfly species in Sicily (68.7%), followed by Phlebotomus perniciosus (17.2%) and Sergentomya minuta (14%). In 2021, Phlebotomusperfiliewi was confirmed as the most common species (61.6%), followed by Phlebotomusperniciosus (33.1%) and Sergentomyaminuta (4.7%). Of particular interest was the identification of Phlebotomus papatasi (0.41%) in Agrigento. Our retrospective study can inform health authorities for the development of appropriate screening, treatment and control strategies to reduce Leishmania incidence rate. This study examined the present state of Leishmaniasis control, surveillance, and prevention in Sicily, but also highlighted deficiencies that could be addressed through the application of One-Health principles.

Sandfly-Borne Phleboviruses in Portugal: Four and Still Counting.

According to ICTV, there are currently 66 known phlebovirus species. More than 40 of these viruses were isolated or detected in phlebotomine sandflies and some of them are known pathogens. In Portugal, information about sandfly-borne phleboviruses is scarce and scattered sandfly-borne diseases are neglected and often not considered in differential diagnoses. The main objective of this work was to gather the existing information and to raise awareness about the circulating phleboviruses in this country. To date, Massilia and Alcube phleboviruses have been isolated from sandflies in southern Portugal. Human infections with Toscana and Sicilian phleboviruses have been reported, as well as seroprevalence in cats and dogs. More studies are needed in order to understand if the viruses isolated during the entomological surveys have an impact on human health and to fully understand the real importance of the already recognized pathogens in our country.

From Insect Bites to a Skin Autoimmune Disease: A Conceivable Pathway to Endemic Pemphigus Foliaceus.

In the endemic variants of pemphigus foliaceus (PF), in Brazil and Tunisia, patients generate pathogenic IgG4 anti-desmoglein 1 autoantibodies. Additionally, these patients possess antibodies against salivary proteins from sand flies that react with Dsg1, which may lead to skin disease in susceptible individuals living in endemic areas. This minireview focuses on recent studies highlighting the possible role of salivary proteins from Lutzomyia longipalpis (L. longipalpis) in EPF from Brazil and Phlebotomus papatasi (P. papatasi) in EPF from Tunisia. We will briefly discuss the potential mechanisms of molecular mimicry and epitope spreading in the initiation and development of endemic PF (EPF) in Brazil and Tunisia.

Definition of the main vector of cutaneous leishmaniasis: Ecology and mapping in endemic area of Northeast Brazil.

Cutaneous leishmaniasis is endemic in Pernambuco. Aiming to determine the vector species of cutaneous leishmaniasis in an endemic area of the Northeast region of Brazil, this study aimed to use the spatial mapping of human cases of CL and correlate with ecological studies of the vectors in the municipality of Timbaúba, Pernambuco, Brazil. Individuals infected with CL were recruited through active search in their homes and clinically and serologically diagnosed during the period from 2018 to 2019. Sandflies were captured with CDC-type light traps in peridomiciliary environments and these were identified at the species level. Females were separated for DNA extraction and subsequent analysis by PCR. The points of collection of phlebotomes and the residences of individuals with lesions were marked with GPS. During the study period, 60 cases of CL were diagnosed. A higher concentration of CL cases was observed in proximity to Atlantic forest remnants confirmed by heat map. A total of 3744 sandflies was captured and five distinct species were identified, with the predominance of Nyssomyia whitmani. From the females separated for the identification of Leishmania braziliensis DNA, a rate of 0.68% of infected sandflies was obtained. It was concluded that cutaneous leishmaniasis continues to be a rural feature of the area. And from this study, it is concluded that Ny. whitmani is the carrier species of CL in the municipality of Timbaúba, Pernambuco. This is due to abundance in catching, specialization of species and PCR positivity for Leishmania braziliensis.

Leishmaniasis tegumentaria y flebótomos en la localidad de Colonia Santa Rosa del norte de Argentina / Tegumentary leishmaniasis and sandflies in Colonia Santa Rosa locality in northern Argentina

Resumen La leishmaniasis tegumentaria (LT) es causada por parásitos del género Leishmania y transmitida por especies de flebótomos, insectos pertenecientes al orden Diptera, familia Psychodidae. El objetivo de este trabajo fue analizar la presencia de casos de LT y su variación temporal entre 1985 y 2019, las formas clínicas, la ubicación de las lesiones en los pacientes y la presencia de flebótomos en la localidad de Colonia Santa Rosa, del norte de Argentina. Los pacientes se diagnosticaron mediante frotis e intradermorreacción de Montenegro. Se colocaron trampas CDC en 14 sitios en el horario de 19 a 7. Se diagnosticaron 120 casos de LT y la prevalencia global fue del 0,75% (^16.000 habitantes). Los pacientes presentaron formas cutáneas simples y múltiples (88,79%) y mucocutáneas (10,83%). Las lesiones cutáneas fueron más frecuentes en extremidades inferiores. Del total de flebótomos, Nyssomyia neivai fue la especie predominante (95%), y, en menor proporción, estuvieron presentes Migonemyia migonei (1,9%), complejo cortelezzii (1,3%) y Evandromyia sallesi (0,09%). La persistente ocurrencia de casos y la presencia de flebótomos en la citada localidad nos sugiere la transmisión endémica en la zona. Esto senala la necesidad de planificar medidas preventivas y de control de la LT en el norte de Argentina.

Abstract Tegumentary leishmaniasis (TL) is caused by parasites of the genus Leishmania and transmitted by the sandfly species, insects belonging to the order Diptera, family Psychodidae. Historically, the most endemic area of TL in Argentina has been the northern region. The aim of this work was to analyze the presence and temporal variation of TL cases reported between 1985 and 2019 in Colonia Santa Rosa locality, northern Argentina. Furthermore, its clinical forms were characterized and sandflies were captured. Patients were diagnosed by smear and the Montenegro skin test. For sampling, CDC light traps were placed at 14 sites from 7pm to 7am. The correlation between vegetation cover and sandfly abundance was also studied. One hundred and twenty TL cases were diagnosed and the overall prevalence was 0.75% (M6 000 inhabitants). Patients presented simple and multiple cutaneous leishmaniasis (88.79%) and the mucocutaneous form (10.83%). Skin lesions were more frequent on the lower extremities (46.73%). Of the total number of sandflies, Nyssomyia neivai (95%) was the predominant species followed by Migonemyia migonei (1.9%), cortelezzii complex (1.3%) and Evandromyia sallesi (0.09%). The persistent occurrence of cases and the presence of sandflies in the locality suggest the existence of endemic transmission in the area. This highlights the need to design prevention and control measures for TL in northern Argentina.

Transmission Dynamics of Punique Virus in Tunisia.

A novel phlebovirus, Punique virus (PUNV), was discovered and isolated in 2008 from sandflies from Northern Tunisia. PUNV is now classified as a unique member of the Punique phlebovirus species within the Phlebovirus genus in the Phenuiviridae family (order bunyavirales). In this study, we aimed to investigate the transmission dynamics of PUNV in Tunisia. Sandflies were collected during two consecutive years, 2009 and 2010, by CDC light traps. In 2009, a total of 873 sandflies were collected and identified to the species level. Phlebotomus perniciosus was the most abundant species. One pool of P. perniciosus females collected in autumn contained PUNV RNA, yielding an infection rate of 0.11%. The population densities of circulating sandfly species were assessed during May-November 2010 in Northern Tunisia by using sticky traps. Phlebotomus (Larroussius) perniciosus (71.74%) was the most abundant species, followed by Phlebotumus (Larroussius) longicuspis (17.47%), and Phlebotumus (Larroussius) perfiliewi (8.82%). The densities of dominant sandfly species were found to peak in early spring and again in the autumn. In 2010, species identification was not performed, and sandflies were only discriminated on the basis of sex and collection date. Out of 249 pools, three contained PUNV RNA. Each positive pool allowed virus isolation. The three pools of female sandflies containing PUNV RNA were collected in autumn with an infection rate of 0.05%. These findings provide further evidence that P. perniciosus is the main vector of PUNV in Tunisia, and this phlebovirus is endemic in Tunisia. Our findings provided strong evidence of intensive circulation of PUNV in sandflies and hosts through a viral infection buildup process between sandfly vectors and hosts starting at the beginning of the activity of sandflies in spring to reach a maximum during the second main peak in autumn.

Diversity of cave Phlebotomines (Diptera: Psychodidae) from a Colombian cave.

Sandflies are vector species of Leishmania, among many other pathogens, with a global distribution and a variety of ecological niches. Previous samplings have found that karstic formations (i.e., caves, grottos, and folds formed by the erosion of limestone) serve as a natural habitat to sandfly species. The majority of samplings of cave sandfly diversity have occurred in Brazil and to date none have studied the species composition in a cave in the Northern Andes. We collected sandflies in the Cave "Los Guácharos", in the state of Antioquia, Colombia. The sampling was carried out during two consecutive nights in September 2019. CDC-type light traps were installed inside the cavern and in other surrounding karst systems (caves, rock-breaks, and folds). In total, we identified 17 species of sandfly from the cave and surrounding karst systems, including a new record for Colombia (Bichromomyia olmeca), and provide the first karstic reports for four other species (Lutzomyia gomezi, Lutzomyia hartmanni, Pintomyia ovallesi, and Psychodopygus panamensis). We then used the results of our survey and published literature to test two hypotheses. First, that sandfly diversity in Neotropical caves is richest nearer to the equator, and second that there is a phylogenetic signal of karstic habitat use in sandflies. Counter to our predictions, we found no evidence that diversity follows a latitudinal gradient. Further, we find no evidence of a phylogenetic signal of karstic habitat use, instead finding that the use of caves likely evolved multiple times across several genera. Our results highlight the importance of a wide sampling to understand the natural habitat of sandflies and other disease vectors.

Overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors.

This article presents an overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors. It first briefly summarises some of the disease-causing pathogens vectored by insects and emphasises the need for innovative control methods to counter the threat of resistance by both the vector insect to pesticides and the pathogens to therapeutic drugs. Subsequently, the state of art of paratransgenesis is described, which is a particularly ingenious method currently under development in many important vector insects that could provide an additional powerful tool for use in integrated pest control programmes. The requirements and recent advances of the paratransgenesis technique are detailed and an overview is given of the microorganisms selected for genetic modification, the effector molecules to be expressed and the environmental spread of the transgenic bacteria into wild insect populations. The results of experimental models of paratransgenesis developed with triatomines, mosquitoes, sandflies and tsetse flies are analysed. Finally, the regulatory and safety rules to be satisfied for the successful environmental release of the genetically engineered organisms produced in paratransgenesis are considered.

Robust comparative performance of genomic DNA extraction methods from non-engorged phlebotomine sandflies.

The present study is a comparative analysis of DNeasy Blood & Tissue Qiagen® kit (Qiagen®, Hilden, Alemanha), salting out, HotShot and phenol-chloroform protocols to extract DNA from sandflies. In addition, a comparative test using sandflies with and without eyes evaluated the potential inhibitory effect in the cPCR. An inhibition test was performed using an exogenous DNA added to the qPCR. The genomic DNA quality of each sample was evaluated by cPCR based on the cytochrome c oxidase subunit I (cox1) gene. The DNA extraction protocols showed the following percentage of amplification: HotShot (91.6% [55/60]), salting out (71.6% [43/60]), phenol-chloroform (95% [57/60]) and kit DNeasy Blood & Tissue Qiagen® (73.3% [44/60]). The phenol-chloroform method achieved a significantly higher frequency of cox1 gene amplification. The pigment present in the phlebotomine's eyes seems to inhibit cPCR reactions since the frequency of amplification of the cox1 gene increased in the sandflies without eyes (p < 0.0001). The HotShot method showed the highest inhibitory potential. These manual extraction techniques can be an inexpensive and effective alternative to study vector-pathogen interactions.

Exposure to Phlebotomus perniciosus sandfly vectors is positively associated with Toscana virus and Leishmania infantum infection in human blood donors in Murcia Region, southeast Spain.

Antibodies against Phlebotomus perniciosus sandfly salivary gland homogenate (SGH) and recombinant protein rSP03B, sandfly-borne Toscana virus (TOSV), Sandfly Fever Sicilian virus (SFSV) and Leishmania, as well as DNA of the latter parasite, were investigated in 670 blood samples from 575 human donors in Murcia Region, southeast Spain, in 2017 and 2018. The estimated SGH and rSP03B seroprevalences were 69% and 88%, respectively, although correlation between test results was relatively low (ρ = 0.39). Similarly, TOSV, SFSV and Leishmania seroprevalences were 26%, 0% and 1%, respectively, and Leishmania PCR prevalence was 2%. Prevalences were significantly greater in 2017, overdispersed and not spatially related to each other although both were positively associated with SGH but not to rSP03B antibody optical densities, questioning the value of the latter as a diagnostic marker for these infections in humans.