Leishmaniases in the European Union and Neighboring Countries.

A questionnaire survey of animal and human health authorities in Europe revealed that leishmaniases are not notifiable in all countries with autochthonous cases. Few countries implement surveillance and control targeting both animal and human infections. Leishmaniases are considered emergent diseases in most countries, and lack of resources is a challenge for control.

Determination of sand fly fauna and molecular detection of Leishmania in sand flies in Antalya Province, Southern Turkey.

Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are diseases transmitted by infected female sand flies. Since the eradication of malaria in Turkey, CL is the main vector-borne disease in the country, with more than 2000 cases per year, making it a significant public health problem. The aims of this study were to carry out an entomological survey in Antalya Province, an endemic area for CL in the Mediterranean Region of Turkey, to identify sand fly fauna and to screen female specimens for the presence of Leishmania parasites (Leishmania infantum, L. tropica, L. major, and L. donovani) using molecular analysis. Sand flies were collected in 42 localities of seven districts in Antalya Province using CDC miniature light traps in two different periods, June 2012 and September 2013. The specimens were kept in 96% ethanol until the dissection was done. The head and genitalia of the specimens were cut for preparing individual slides for species identification. The rest of the body of female specimens was kept separately. The specimens were identified at the species level, and 27 pools were generated according to the locations and species for screening the presence of Leishmania. A commercial kit was used for DNA extractions. Real-time and conventional polymerase chain reaction (PCR) targeting the internal transcribed spacer region (ITS1) were then performed. In total, 1306 specimens comprising nine species belonging to the Phlebotomus genus were collected in the study region, with Phlebotomus neglectus/syriacus (38.82%) the most abundant, followed by P. alexandri (21.67%) and P. tobbi (20.44%). In the 27 pools, Leishmania infantum DNA was detected in four pools containing P. neglectus/syriacus and one pool containing P. tobbi. In conclusion, the sand fly fauna in the Antalya Province is diverse. The probable vector sand fly species are P. neglectus/syriacus and P. tobbi with high dominance (59.26%), which indicates a high risk of CL transmission. The data presented here may help to shed more light on the transmission cycles of the Leishmania parasite in this CL endemic area.

First detection of voltage-gated sodium channel mutations in Phlebotomus argentipes collected from Bangladesh.

BACKGROUND & OBJECTIVES: Phlebotomus argentipes is the main vector of visceral leishmaniasis in Bangladesh and is controlled using deltamethrin, a synthetic pyrethroid, through indoor residual spraying (IRS). A mutation at L1014 (leucine at codon 1014) of the voltage-gated sodium channel (VGSC), known as a knockdown resistance (kdr) gene, is thought to be an important pyrethroid resistance mechanism. This study detected mutations at codon 1014, and at codons 1011, 1016, and 1020, which are kdr sites in other insects. The kdr relationship with deltamethrin resistance in P. argentipes from an IRS-targeted site in Bangladesh was also evaluated. METHODS: Sand flies were collected from Magurjora village, Mymensingh district, Bangladesh in November 2012. A WHO cone bioassay test using deltamethrin was conducted and specimens were grouped as 'live' or 'dead'. After morphological identification, genomic DNA was used to genotype a partial VGSC gene from P. argentipes. The kdr/ pyrethroid resistance relationship was evaluated using Fisher's exact test. RESULTS: Targeted codons were genotyped from 8 'live' and 63 'dead' P. argentipes. All 'live' specimens had mutant alleles (L1014F and L1014S) at codon 1014. The mutant allele rate was 94% for 'live' specimens and 55% for 'dead' specimens. The mutant allele survival odds were higher for the wild-type L1014L allele, and L1014F odds were lower for L1014S. There were no mutations at codons 1011, 1016, and 1020. INTERPRETATION & CONCLUSION: The L1014 mutations suggested that pyrethroid resistance had appeared in Bangladesh. Further research on kdr mutations in P. argentipes is important for the appropriate IRS.

First molecular detection and identification of Leishmania species in small wild rodents from Turkey.

Leishmaniasis is a parasitic disease infecting animals and humans. Two clinical forms (Visceral and cutaneous leishmaniasis) and four species are reported to be present in Turkey. Several studies have investigated canine and human leishmaniasis in Turkey but no study was performed to screen the infection among wild rodents, so far. The present study aims to investigate the role of small wild rodents as reservoir animals for Leishmania spp. in different regions of Turkey. Formalin-preserved tissue samples (spleen, liver, lung) of 712 rodents from 30 provinces were screened for the presence of Leishmania spp. DNA. Before DNA extraction, tissues were dried, rehydrated, and homogenated. Leishmania screening in rodent tissues and species determination was performed with a combination of real-time kDNA and ITS1 polymerase chain reaction protocols. Eight (1.12%) out of 712 animals were found to be positive for Leishmania spp. DNA and species typing revealed five L. infantum, two L. tropica and one L. major among positives. Leishmania major and L. infantum DNA were detected in Apodemus spp. from Zonguldak province located in the Western Black Sea Region, while L. tropica DNA was found in Meriones sp. and Gerbillus dasyurus from Adana and Hatay provinces located in Eastern Mediterranean Region of Turkey. The present study is first to report natural infection of L. infantum, L. major and L. tropica in small wild rodents in Turkey, suggesting their possible roles as reservoirs. Further studies are needed for planning epidemiological studies and also for developing rodent control measures in risky endemic areas to break the transmission cycle.

[Cutaneous Leishmaniasis Cases Caused by Leishmania infantum in Sanliurfa Province, Turkey]. / Sanliurfa'da Leishmania infantum'un Etken Oldugu Kutanöz Leysmanyazis (Sark Çibani) Olgulari.

Leishmaniases are a group of vector-borne diseases, and two clinical forms, visceral (VL) and cutaneous leishmaniasis (CL, Oriental sore), are seen in Turkey. While VL cases are recorded as 20-25 per year, CL cases are reported around 2000 per year, and nearly half of CL cases were recorded in Sanliurfa province. Therefore, by knowing the epidemiology of the disease in Sanliurfa province, it is possible to develop control measures and reduce the total number of cases across the country. Although Leishmania tropica is known as the main causative agent in Sanliurfa, other Leishmania species have also been identified as a result of mass human movements in the last 10 years. In this study, we aimed to present the first CL cases caused by Leishmania infantum in Sanliurfa. A total of 14 cases, which were admitted with the suspicion of CL and diagnosed as positive by direct microscopy and/or real-time ITS1-PCR using lesion aspiration samples are included in the study. Two or more smears were prepared from the samples taken from the lesions of the patients by fine needle aspiration. One of the smears was stained with Giemsa stain after fixation with methyl alcohol and examined under the light microscope at x1000 magnification for the presence of Leishmania amastigotes. DNA isolation was made from the other unstained preparations with a commercial kit (Qiagen DNeasy, Germany) according to the recommendations of the manufacturer. The real-time ITS1-PCR method was performed by using the Old World species-specific primers and probes. As a result, by the identification of the species with real-time ITS1-PCR, it was determined that the causative agent was L.infantum in five cases, L.major in one case and L.tropica in eight cases. It was learned that four of the cases in which L.infantum was detected as the causative agent were local, one was Syrian and they lived in the city center. Also two of the eight cases, which were identified as L.tropica, were Syrian and six of them were domestic cases and all of them lived in the city center. While all 14 patients included in the study were positive with real-time ITS1-PCR, amastigotes were detected in 10 cases only. The cases of CL presented in this study are the first cases caused by L.infantum reported from Sanliurfa, and are important in terms of concretely demonstrating the effect of mass human mobility and migration on the epidemiology of the infection.

[Determination of Antimony Resistance Mechanism of Leishmania tropica Causing Cutaneous Leishmaniasis in Turkey]. / Türkiye'de Kutanöz Leysmanyazis Etkeni Leishmania tropica'da Antimon Direnç Mekanizmasinin Belirlenmesi.

World Health Organization reported that approximately one billion people are at risk in endemic areas, one million cases of cutaneous leishmaniasis (CL) and approximately 300,000 cases of visceral leishmaniasis (VL) were reported per year in the last five years. The number of deaths due to VL is reported to be approximately 20,000 per year. Approximately 2500 cases/year have been reported as CL, caused by Leishmania tropica and Leishmania infantum, in Turkey. The significant increase observed in many cities mainly in the provinces of Mediterranean and Aegean regions in cases and foci in recent years, suggests that there may be an increase in this infections in the following years as well. In Turkey, the causative agent of CL is L.tropica and meglumine antimoniate is used in the treatment of CL. We aimed to determine antimony resistance genes specific for L.tropica by comparing the gene and protein expressions of antimony-resistant and non-resistant L.tropica strains. L.tropica isolates obtained from 3 CL patients without antimonate resistance from Aegean, Mediterranean and Southeastern regions of Turkey were provided to transform into 3 resistant isolates against meglumine antimony in the laboratory conditions. Gene expression alterations by microarray method; protein profiles by two-dimensional gel electrophoresis (2D-PAGE) and relevant proteins by MALDI-TOF/TOF MS of these isolates were accomplished and compared. L.tropica isolates from 10 CL patients who did not respond to antimony therapy were analyzed for resistance to antimonial compounds and quantitative real-time polymerase chain reaction was performed to detect the expression of genes responsible for resistance development. Moreover, differences in protein expression levels in isolates with and without antimony resistance were determined by comparing protein profiles and identification of proteins with different expression levels was carried out. Enolase, elongation factor-2, heat shock protein 70, tripanthione reductase, protein kinase C and metallo-peptidase proteins have been shown to play roles in L.tropica isolates developing resistance to antimonial compounds and similar expression changes have also been demonstrated in naturally resistant isolates from patients. In conclusion, it was revealed that L.tropica strains in our country may gain resistance to meglumine antimoniate in a short time. It is foreseen that if the patients living in our country or entering the country are treated inadequately and incompletely, there may be new, resistant leishmaniasis foci that may increase the number of resistant strains and cases rapidly.

[Comparison of polymerase chain reaction using kinetoplast DNA specific primers and other parasitological methods in the diagnosis of clinical samples of suspected patients with cutaneous leishmaniasis in Sanliurfa]. / Sanliurfa'da kutanöz leysmanyazis süpheli hastalardan alinan klinik örneklerin tanisinda kinetoplast DNA'sina özgül primerlerin kullanildigi polimeraz zincir reaksiyonu ile diger parazitolojik yöntemlerin karsilastirilmasi.

Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are seen endemically in Turkey and CL caused by Leishmania tropica is an important public health problem in southeastern as well as other regions of Turkey. The diagnosis has been usually made by clinical view of lesion and/or parasitologically using lesion aspiration smears. Histological examination does not, always reveal the parasite in the skin biopsy, particularly in chronic lesions. Besides this, due to CL infections caused by different species in endemic areas, diagnostic methods enabling species identification are in great need. Species identification, in the time of diagnosis, is an important procedure for helping the clinicians in the planning of treatment as well as control measures. Polymerase chain reaction (PCR) is a specific and sensitive diagnostic tool that can also identify the parasite at species level. Kinetoplast DNA (kDNA) is one of the genetic regions that can be used for the detection of Leishmania parasites in clinical specimens, kDNA PCR is reported as one of the most sensitive methods related to species-specific variable regions in mini-circle long time ago. It has been considered as one of the most ideal targets for the diagnosis of leishmaniasis. The aim of the study was to perform PCR targeting kDNA by using the primers of Uni21/Lmj4 in clinical samples and compare the results with other parasitological methods like smear and culture, for the diagnosis of CL. The kDNA PCR, parasite culture and microscopical evaluation of stained smears of 62 specimens from suspected CL cases who have referred to Cutaneous Leishmaniasis Diagnosis and Treatment Center in Sanliurfa, Turkey were included in the study. The kDNA PCR showed the highest sensitivity 100% of the samples (35/35) among all diagnostic assays, followed by the microscopy (25/35 positive, 71.4% sensitivity) and culture (19/35 positive, 54.3% sensitivity). The sensitivity of combination of culture and microscopy was 88.6% (31/35 positive). These results suggested that performing kDNA PCR in addition to conventional techniques is important for improving the true diagnosis of CL to the species level and also important for establishing treatment regimens and designing appropriate precautions in highly endemic area like the southeastern region of Turkey.

Isolation, genetic characterization, and seroprevalence of Adana virus, a novel phlebovirus belonging to the Salehabad virus complex, in Turkey.

UNLABELLED: A new phlebovirus, Adana virus, was isolated from a pool of Phlebotomus spp. (Diptera; Psychodidae) in the province of Adana, in the Mediterranean region of Turkey. Genetic analysis based on complete coding of genomic sequences indicated that Adana virus belongs to the Salehabad virus species of the genus Phlebovirus in the family Bunyaviridae. Adana virus is the third virus of the Salehabad virus species for which the complete sequence has been determined. To understand the epidemiology of Adana virus, a seroprevalence study using microneutralization assay was performed to detect the presence of specific antibodies in human and domestic animal sera collected in Adana as well as Mersin province, located 147 km west of Adana. The results demonstrate that the virus is present in both provinces. High seroprevalence rates in goats, sheep, and dogs support intensive exposure to Adana virus in the region, which has not been previously reported for any virus included in the Salehabad serocomplex; however, low seroprevalence rates in humans suggest that Adana virus is not likely to constitute an important public health problem in exposed human populations, but this deserves further studies. IMPORTANCE: Until recently, in the genus Phlebovirus, the Salehabad virus species consisted of two viruses: Salehabad virus, isolated from sand flies in Iran, and Arbia virus, isolated from sand flies in Italy. Here we present the isolation and complete genome characterization of the Adana virus, which we propose to be included in the Salehabad virus species. To our knowledge, this is the first report of the isolation and complete genome characterization, from sand flies in Turkey, of a Salehabad virus-related phlebovirus with supporting seropositivity in the Mediterranean, Aegean, and Central Anatolia regions, where phleboviruses have been circulating and causing outbreaks. Salehabad species viruses have generally been considered to be a group of viruses with little medical or veterinary interest. This view deserves to be revisited according to our results, which indicate a high animal infection rate of Adana virus and recent evidence of human infection with Adria virus in Greece.

Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions.

OBJECTIVE: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014. METHODS: Initial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L. major. Developing lesions were observed, and the thickening of footpads was measured weekly. RESULTS: Melting curve analyses of 18 isolates showed a peak concordant with L. major, and two of them were confirmed by isoenzyme analyses as L. major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3 weeks, which even progressed to extremity amputation. CONCLUSION: Cutaneous leishmaniasis-causing L. major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L. major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L. major infections in Turkey, contrary to the infections in neighbouring countries.

Seroprevalence of Leishmania infection and molecular detection of Leishmania tropica and Leishmania infantum in stray cats of Izmir, Turkey.

Leishmaniasis caused by more than 20 species of genus Leishmania is transmitted by the bite of infected phlebotomine sand flies. The studies on Leishmania infection in cats is very few in Turkey and therefore we aimed to screen stray cats living in city of Izmir located in western Turkey using nested PCR targeting kinetoplast DNA and serological techniques (ELISA and IFA). Leishmania DNA positive samples were also studied by ITS1 real time PCR. Whole blood and serum samples were obtained from stray cats (n: 1101) living in different counties of Izmir. In serological assays, a serum sample was considered positive in 1:40 dilution in IFA and for ELISA a serum sample was accepted positive when the absorbance value (AV) exceeded the mean AV + Standard Deviation (SD) of the negative control serum samples. According to the results, the seropositivity rates were 10.8% (119/1101) and 15.2% (167/1101) by in house ELISA and IFA, respectively. Among serology coherent samples, the seropositivity rate was 11.1% (116/1047) as detected by both assays after discordant samples (n: 54) were discarded. Of the 1101 stray cats, six (0.54%) were positive by nested PCR while only one of these six samples was positive by ITS1 real time PCR. During PCR, three controls designated as Leishmania infantum, Leishmania tropica, and Leishmania major were used for species identification. According to nested PCR results, L. tropica was identified in two cats (no.76 and 95). In another cat (no. 269), there were two bands in which one of them was well-matched with L. infantum and the other band had ∼850 bp size which does not match with any controls. Remaining three cats (no. 86, 514, and 622) also had the ∼850 bp atypical band size. ITS1 real time PCR detected L. tropica in only one cat (no. 622) which showed an atypical band size in nested PCR. These results indicated that three cats with only one atypical band (no. 86, 514, and 622) and the cat with mixed infection (no. 269) were infected with L. tropica. Altogether, L. tropica was detected in all six DNA positive cats and L. infantum was detected in one cat with mixed infection. In conclusion, although the reservoir role of cats in nature is still unclear the high seroprevalence rate against Leishmania parasites and detecting parasite DNA in stray cats in Izmir indicates that the stray cats are frequently bitten by infected sand flies. Further research activities are required to reveal the frequency of leishmaniasis in cats in different regions of Turkey where Leishmania species are endemic.

Treatment of head lice (Pediculus humanus capitis) infestation: is regular combing alone with a special detection comb effective at all levels?

Head lice infestation (HLI) caused by Pediculus humanus capitis has been a public health problem worldwide. Specially designed combs are used to identify head lice, while anti-lice products are applied on the scalp for treatment. In the present study, we aimed to test whether combing only by precision detection comb (PDC) or metal pin comb (MPC) could be effective alternatives to the use of anti-lice products in children. A total of 560 children from two rural schools in Turkey were screened. In the PDC trial, children were combed every second day for 14 days, while in the MPC trial, combing was performed once in every four days for 15 days. Children were divided into two groups (dry combing and wet combing) for both trials and results were compared. The results showed no significant differences between dry and wet combing strategies for both combs for the removal of head lice (p > 0.05). The number of adult head lice declined significantly on each subsequent combing day in both approaches, except on day 15 in the MPC trial. In the end, no louse was found in 54.1 and 48.9% of children in the PDC and MPC trials, respectively. Since family members of infested children were not available, they were not checked for HLI. Four times combing within 2 weeks with MPC combs was found effective for both treatment of low HLI and prevention of heavy HLI. In conclusion, regular combing by special combs decreases HLI level in children and is safely applicable as long-term treatment.

Immunoinformatics Approach to Design a Multi-Epitope Vaccine against Cutaneous Leishmaniasis.

Cutaneous Leishmaniasis (CL), a neglected vector-borne disease caused by protozoan parasite Leishmania major (L. major), is a major public health concern, and the development of new strategies to reduce the disease incidence has become a top priority. Advances in immunoinformatics and in-silico epitope prediction could be a promising approach to designing a finest vaccine candidate. In this study, we aimed to design a peptide-based vaccine against CL using computational tools and identified ten B-cell-derived T-cell epitopes from the glycoprotein gp63 of L. major. All of the potential immunodominant epitopes were used to design a vaccine construct along with a linker and an adjuvant at the N-terminal for enhancing its immunogenicity. Additionally, many characteristics of the proposed vaccine were examined, and it was confirmed to be non-allergenic, non-toxic, and thermally stable. To assess the vaccine interaction with the innate immune toll-like receptor-4 (TLR-4), a 3D structure of the vaccine construct was developed. Molecular docking and molecular dynamic simulation were used to confirm the binding and to assess the stability of the vaccine-TLR4 complex and interactions, respectively. In conclusion, our multi-epitope vaccine will provide a gateway to analyze the protein function of a potential vaccine candidate against CL.

Covid-19 vaccination acceptance and hesitancy among the Turkish adult population.

Objective: The aim of this study was to determine the acceptance of Covid-19 vaccine among the Turkish adult population. Methods: A total of 2023 persons participated in this cross-sectional study between October 2020 and January 2021. The questionnaire, which was delivered via social media, was filled out by the participants over "Google Forms". Results: Questionnaire results showed that 68.7% of the participants might agree to vaccinated against COVID-19. According to univariate analysis, the age group of 50-59, urban residents, healthcare workers, non-smokers, and those with chronic diseases, those who were vaccinated against influenza, pneumonia, and tetanus were all willing to be vaccinated against COVID-19. Conclusions: It is very important to determine a community's willingness to be vaccinated against COVID-19 so that interventions can be made to solve related problems. Risk of exposure and importance of Prevention play a critical role in vaccination acceptance.

Comparative analysis of the severity and progression of cutaneous leishmaniasis caused by Leishmania tropica in untreated and glucantime-treated patients.

Millions of people worldwide are affected by cutaneous leishmaniasis (CL), a disease that has a significant impact on morbidity and mortality. Understanding the immune responses responsible for tissue damage or the process of lesion healing plays a pivotal role in shaping optimal treatment strategies. In this study, we investigated immunological phenotypes for three groups: glucantime treated (n = 30) and untreated (n = 30) CL patients infected with Leishmania tropica (L. tropica), and healthy controls (n = 20). T-lymphocytes (CD4+ and CD8+), and B lymphocytes (CD14+ and CD19+) were isolated using antibody-conjugated microbeads and magnetic field isolation to achieve high purity. A higher significant difference was observed between T-lymphocytes (CD4+ and CD8+), and B-lymphocytes (CD14+ and CD19+) cells in CL-infected groups before and after treatment (p < 0.0001). When compared, there was also a significant difference among T-lymphocytes (CD4+ and CD8+), B lymphocytes (CD14+ and CD19+) p < 0.0001, p < 0.0005, and p < 0.0007, respectively between CL-infected individuals (before and after treatment) to controls. Our findings suggest that an increased proportion of these cells seen in treated patients may mediate healing, while it is also possible that they may contribute to tissue injury. Understanding the immune system and lesion size of CL can help develop immunotherapies and comprehend the evolution of this parasitic disease.

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.

Entomological surveillance and spatiotemporal risk assessment of sand fly-borne diseases in Cyprus.

Visceral and cutaneous leishmaniases are important public health concerns in Cyprus. Although the diseases, historically prevalent on the island, were nearly eradicated by 1996, an increase in frequency and geographical spread has recently been recorded. Upward trends in leishmaniasis prevalence have largely been attributed to environmental changes that amplify the abundance and activity of its vector, the phlebotomine sand flies. Here, we performed an extensive field study across the island to map the sand fly fauna and compared the presence and distribution of the species found with historical records. We mapped the habitat preferences of Phlebotomus papatasi and P. tobbi, two medically important species, and predicted the seasonal abundance of P. papatasi at unprecedented spatiotemporal resolution using a climate-sensitive population dynamics model driven by high-resolution meteorological forecasting. Our compendium holds a record of 18 species and the locations of a subset, including those of potential public and veterinary health concern. We confirmed that P. papatasi is widespread, especially in densely urbanized areas, and predicted that its abundance uniformly peaks across the island at the end of summer. We identified potential hotspots of P. papatasi activity even after this peak. Our results form a foundation to inform public health planning and contribute to the development of effective, efficient, and environmentally sensitive strategies to control sand fly populations and prevent sand fly-borne diseases.

The estimated distribution of autochthonous leishmaniasis by Leishmania infantum in Europe in 2005-2020.

BACKGROUND: This study describes the spatial and temporal distribution between 2005 and 2020 of human and animal leishmaniasis by Leishmania infantum in European countries reporting autochthonous cases, and highlights potential activities to improve disease control. METHODOLOGY/PRINCIPAL FINDINGS: It was based on a review of the scientific literature and data reported by the World Health Organization (WHO), the World Organization for Animal Health (WOAH) and the Ministries of Health, including hospital discharges in some countries. Autochthonous infections were reported in the scientific literature from 22 countries, including 13 and 21 countries reporting human and animal infections, respectively. In contrast, only 17 countries reported autochthonous human leishmaniasis cases to the WHO and 8 countries animal infections to the WOAH. The number of WOAH reported cases were 4,203, comprising 4,183 canine cases and 20 cases in wildlife. Of 8,367 WHO reported human cases, 69% were visceral leishmaniasis cases-of which 94% were autochthonous-and 31% cutaneous leishmaniasis cases-of which 53% were imported and mostly in France. The resulting cumulative incidence per 100,000 population of visceral leishmaniasis between 2005-2020, was highest in Albania (2.15 cases), followed by Montenegro, Malta, Greece, Spain and North Macedonia (0.53-0.42), Italy (0.16), Portugal (0.09) and lower in other endemic countries (0.07-0.002). However, according to hospital discharges, the estimated human leishmaniasis incidence was 0.70 in Italy and visceral leishmaniasis incidences were 0.67 in Spain and 0.41 in Portugal. CONCLUSIONS/SIGNIFICANCE: Overall, there was no evidence of widespread increased incidence of autochthonous human leishmaniasis by L. infantum in European countries. Visceral leishmaniasis incidence followed a decreasing trend in Albania, Italy and Portugal, and peaked in Greece in 2013, 2014 and 2017, and in Spain in 2006-2007 and 2011-2013. Animal and human cutaneous leishmaniasis remain highly underreported. In humans, hospital discharge databases provide the most accurate information on visceral leishmaniasis and may be a valuable indirect source of information to identify hotspots of animal leishmaniasis. Integrated leishmaniasis surveillance and reporting following the One Health approach, needs to be enhanced in order to improve disease control.

Fauna, Seasonal Activity, and Altitudinal Distribution of Phlebotomine Sand Flies (Diptera: Psychodidae) in Leishmaniasis Endemic Area, Aydin Mountains, and Surroundings in Western Turkey

Objective: Human and canine leishmaniasis (CanL) are endemic in Turkey, and cutaneous leishmaniasis (CL) is a primary public health problem among vector-borne diseases, with 1500-2000 autochthonous cases per year. In this study, we aimed to perform a comprehensive entomological survey, including the detection of sand fly fauna, seasonal activity, and altitudinal distribution, together with the ecological features of Aydin Mountains and their surroundings that are endemic areas for CL caused by Leishmania tropica and CanL caused by L. infantum MON-1. Methods: The survey was carried out from June 2009 to July 2010. Ten localities with different altitudes were selected. Sand fly collection was done every 15 days in July and August when sand flies are most active and every 30 days between September and June (14 months, 16 samplings) using CDC light traps and sticky traps. During the collection, the temperature, humidity, and ecological and environmental features were recorded. Sand fly specimens were dissected and identified at species level using written keys. Results: A total of 6712 sand fly specimens (3268 females and 3444 males) were collected from 10 localities throughout all study periods. Species identification revealed that nine and three species belonging to Phlebotomus (P. major s.l. 30.38%, P. tobbi 22.93%, P. papatasi 5.88%, P. sergenti s.l. 4.51%, P. alexandri 4.26%, P. simici 3.50%, P. burneyi 0.63%, P. brevis 0.45%, and Transphlebotomus spp. 0.28%) and Sergentomyia (S. dentata 23.17%, S. minuta 2.43%, and S. antennata 1.58%) were found, respectively. The seasonal activities of these 12 sand fly species were determined, and P. major s.l. and P. tobbi, which were probable vectors of VL and CanL, were present in the study area. P. sergenti s.l., a probable vector of CL, was also found in the region and was active for seven months between April and October. P. sergenti s.l. was found in the 400-600 m altitude in July, while P. major and P. tobbi were found in high densities in the 200-400 m altitude in September. Conclusion: Our results showed that leishmaniasis vectors are present in different altitudes in the Aydin mountains and surroundings. The findings revealed the time intervals of parasite transmission in the area and the period of applying protective measures, such as insecticide application.

Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis

Objective: Leishmania RNA virus was detected the first time in the New World Leishmania species. Recent studies were also showed the presence of Leishmania RNA virus 2 (LRV2) in Old Word Leishmania species including Turkish L. major and L. tropica isolates. This study aimed to increase the sensitivity of qPCR with a modification in the denaturation step of cDNA preparation protocol. Methods: In this study, LRV2+ three L. major, two L. tropica strains and L. major control strain (MHOM/SU/73/5-ASKH) were included. Total RNA isolation was done using different numbers of Leishmania promastigotes (108, 105 and 103). Before cDNA synthesis, samples were denatured at 95 °C for 2 min, as a modification of the kit procedure. qPCR was undertaken using 0.5 mM primers (LRV F-HR/LRV R-HR) diluted in SYBR Green Master mix. Results: We observed lower Ct values in amplicons with the modified version than with the classical kit protocol for cDNA synthesis, in all of the strains used in the study. The addition of pre-denaturation step at 95 °C showed lower Ct values meaning the sensitivity increased. Different parasite dilutions showed similar results. Conclusion: It is important to increase the sensitivity especially with the aim for detecting LRV in clinical samples obtained from patients probably have less number of parasites. The presence and burden of the virus can help to understand the relationship between the clinical findings and the pathogenicity of the parasite which may lead to changes in the course of treatment.

Host-Parasite Interactions: Regulation of Leishmania Infection in Sand Fly.

PURPOSE: Sand flies are the only proven vectors of leishmaniases, a tropical neglected disease endemic in at least 92 countries. Vector-parasite interactions play a significant role in vector-borne disease transmission. There are various bottlenecks to Leishmania colonization of the sand fly midgut. Such bottlenecks include the production of innate immune-related molecules, digestive proteases, parasite impermeable peritrophic membrane, and resident gut microbiota. These barriers determine the parasite load transmitted and, consequently, the disease outcome in mammalian host. Therefore, it is important to understand the molecular responses of both sand fly and Leishmania during infection. METHOD: Here, we reviewed the published literature on sand fly-Leishmania interactions bringing together earlier and current findings to highlight new developments and research gaps in the field. CONCLUSION: Recent research studies on sand fly-Leishmania interaction have revealed contrasting observations to past studies. However, how Leishmania parasites evade the sand fly immune response still needs further research. Sand fly response to Leishmania infection can be best understood by analyzing its tissue transcriptome. Better characterization of the role of midgut components could be a game changer in development of transmission-blocking strategies for leishmaniasis.