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Leishmaniasis is endemic in countries of the Mediterranean area. In this study, the information available on leishmaniasis in Navarra in the period 1976-2018 and in the bordering regions was collected, along with an overview of the situation of this disease at the national level, from the vector to humans. Aragón exhibited a lower incidence rate in the last ten years (2008-2018) compared to the previous decade, while Navarra and La Rioja have almost doubled the number of cases per 100,000 inhabitants and, likewise, incidence in the Basque Country has also increased. At the national level, since 2015, such an increase has become significant due to the inclusion of human leishmaniasis as a mandatory notifiable disease. Currently in Spain, although the incidence of human leishmaniasis is not a situation of high concern, it is however a reality. Therefore, it is necessary to monitor its evolution.

Objective: This study aimed to evaluate the polymerase chain reaction (PCR) and immunofluorescence antibody test (IFAT) results of suspected samples with canine leishmaniasis (CanL) that were sent to the Parasitology Department Laboratories of the Veterinary Faculty in Aydın Adnan Menderes University., Methods: The age, gender, and breed of the dogs to be evaluated for CanL were recorded, and IFAT was performed using 80 blood serum samples collected from them. Additionally, after the isolation of genomic DNA of 27 blood samples, PCR of these samples was performed using primers that amplify the 145 bp kDNA region of Leishmania species., Results: Thirty-seven (46.25%) of the serum samples were seropositive in at least one dilution (1/64 or 1/128) according to IFAT. Five (18.5%) of the twenty-seven samples were positive for Leishmania DNA according to PCR. According to IFAT, 38.7% of male dogs and 59% of female dogs were positive. The highest number of seropositive samples were detected in dogs aged 3-5 years (11/27)., Conclusion: Considering the zoonotic potential of leishmaniasis, which is considered endemic in the region, and the high positivity of the IFAT/PCR results, veterinarians should use advanced diagnostic methods, especially serological and molecular tests, in dogs with suspected CanL. The data obtained show that the risk of infection caused by Leishmania spp. is high in the region. Therefore, it is important to routinely ensure the control of CanL to protect both human and animal health.

Leishmaniasis is a zoonotic and neglected disease, which represents an important public health problem worldwide. Different species of Leishmania are associated with different manifestations, and a practical problem that can worsen the condition of hosts infected with Leishmania  is the secondary infection caused by bacteria. This review aims to examine the importance and prevalence of bacteria co-infection during leishmaniasis and the nature of this ecological relationship. In the cases discussed in this review, the facilitation phenomenon, defined as any interaction where the action of one organism has a beneficial effect on an organism of another species, was considered in the Leishmania -bacteria interaction, as well as the effects on one another and their consequences for the host.

Canine leishmaniosis (CanL) is an emerging zoonosis caused by Leishmania infantum and transmitted in southern Europe by phlebotomine sand flies of the subgenus Phlebotomus (Larroussius). Endemic foci of CanL have been recorded in northern continental Italy since early 1990s and attributed to the northward expansion of vector populations due to climatic changes in association with travelling/relocated infected dogs from the southern Mediterranean littoral. In this study, further spread of endemic Leishmania foci was monitored during 2018-2019 in five regions (Aosta Valley, Piedmont, Lombardy, Veneto and Friuli-Venezia Giulia), with focus to territories where investigations were not performed, or they have been inconclusive. Clinical cases of CanL identified by local veterinary practitioners and confirmed by reference diagnosis centers were regarded as autochthonous if their origin from, or travel to, areas endemic for CanL were excluded in the previous ≥2 years. Around these index cases, i) serosurveys for L infantum were carried out where indicated, ii) sampling from potential autochthonous cases in healthy or clinically-suspected resident dogs was intensified by collaborating veterinary practitioners, and iii) suitable sites were investigated for the presence of competent phlebotomine vectors. Fifty-seven municipalities whose enzootic status of CanL was unreported before 2018, were identified as endemic. The stability of 27 foci recorded over the past decade, was also confirmed. Competent phlebotomine vectors, mainly Phlebotomus perniciosus, were collected for the first time in 23 municipalities. The newly recorded endemic municipalities appear to be distributed over a west-to-east decreasing gradient: 30 in Piedmont, 21 in Lombardy, 4 in Veneto and 2 in Friuli-Venezia Giulia. As regards Veneto, it should be noted that a relatively restricted territory was investigated as several municipalities of the region had already been surveyed and detected as endemic for CanL in the past. Cold climate conditions of the easternmost region of Friuli-Venezia Giulia bordering non-endemic territories of Slovenia, are probably less favorable to L infantum transmission., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Being the only established vectors of the protozoan parasites of the genus Leishmania, sand flies have become very important in all countries where leishmaniasis exists. It is caused by a variety of species, each one having specific mammalian reservoir hosts and vectors. Leishmania and sand fly classification has always been a controversial matter, and the increasing number of sand fly species described world complicates the task. Until recently, sand flies distributed in the Old World were known as vectors for few Phleboviruses including two known species (Sandfly fever Naples virus, and alehebad virus), and tentative species such as sand fly fever Sicilian virus and Corfou virus. These infections are emerging in the Mediterranean region and will likely spread in forthcoming decades, posing a complex threat to public health. Here, we reviewed the current literature on phlebotomine sand flies fauna and epidemiology of sand fly‑borne infections in Algeria.

Canine leishmaniasis is a parasitic zoonotic infection of dogs caused by the Leishmania spp. parasites. In Europe, canine leishmaniasis is among the most important vector-borne parasitic diseases of dogs, with majority of data originating from countries of southwestern Europe. With its scarce and outdated records, the Balkan region presents the "blank space" in the European map of canine leishmaniasis knowledge. Numerous records obtained during the past decade and a half indicate the emergence of autochthonous canine leishmaniasis in new settings all across Balkan, a significant increase in the numbers of infected dogs in previously endemic areas, and northward spread of the disease. As the majority of canine leishmaniasis data from Balkan countries are several decades old and generally written in native Balkan languages, hence not available to the broader area of scientists, comprehensive search of scientific literature published from 1934 to January 2021 was conducted in order to provide a better perspective to the past and current canine leishmaniasis situation in Balkan. This review provides exhaustive information on all leishmaniasis records in dogs, wild animals and cats, available from 10 countries of the Balkan region. It identifies dogs as the main disease reservoirs and highlights the importance of wild animals, and potentially cats, as disease reservoirs, which is of great public and veterinary health concern. Additionally, certain shortcomings in canine leishmaniasis research, management, surveillance and control were identified., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: To describe the presence of Leishmania infection within the animal population receiving care from US Army Veterinary Services., Animals: 629 canine, feline, and equine patients of US Army Veterinary Services from 2014 to 2017., Procedures: Personnel at the US Army Public Health Center ran a query within the Remote Online Veterinary Record system using previously validated search terms (eg, liesh, leish, and lesh) and returned data on any patient for which the master problem list included those terms. Next, a query was run to identify all leishmaniasis testing. Records identified by queries were reviewed manually, and data were collected on patient signalment, indication for and type of testing, location of testing, and previous locations or country of the patient., Results: Only dogs (n = 378), not cats or horses, had been tested for leishmaniasis, 54 (14.3%) of which tested positive for Leishmania infection. More specifically, 39 of 104 (37.5%) privately owned dogs tested positive, compared with 15 of 274 (5.6%) government-owned dogs. Overall, 186 dogs had no clinical signs, 12 (6.5%) of which tested positive. Forty-four of the 54 (81%) test-positive dogs were located in or had traveled to an endemic area., Clinical Relevance: The prevalence of leishmaniasis in the various subpopulations of dogs suggested the need for additional prevalence studies. Many animals travel in and out of the US, and repeated introduction of Leishmania spp could lead to this vector-borne disease becoming endemic in the US animal and human populations. Consequently, US veterinarians need to ensure proper testing and follow-up to protect one health.

Introduction: Leishmaniasis is a major public health problem and the second most lethal parasitic disease in the world due to the lack of effective treatments and vaccines. Even when not lethal, leishmaniasis significantly affects individuals and communities through life-long disabilities, psycho-sociological trauma, poverty, and gender disparity in treatment., Areas Covered: This review discusses the most relevant and recent research available on Pubmed and GoogleScholar highlighting leishmaniasis' global impact, pathogenesis, treatment options, and lack of effective control strategies. An effective vaccine is necessary to prevent morbidity and mortality, lower health care costs, and reduce the economic burden of leishmaniasis for endemic low- and middle-income countries. Since there are several forms of leishmaniasis, a pan- Leishmania vaccine without geographical restrictions is needed. This review also focuses on recent advances and common challenges in developing prophylactic strategies against leishmaniasis., Expert Opinion: Despite advances in pre-clinical vaccine research, approval of a human leishmaniasis vaccine still faces major challenges - including manufacturing of candidate vaccines under Good Manufacturing Practices, developing well-designed clinical trials suitable in endemic countries, and defined correlates of protection. In addition, there is a need to explore Challenge Human Infection Model to avoid large trials because of fluctuating incidence and prevalence of leishmanasis.

Background: Canine leishmaniosis (CanL) is a common infectious disease. Age, sex and breed might influence the type of clinical and pathological manifestations that dogs develop. The main objective of this retrospective cross-sectional study was to determine if an association between age, sex, breed and size and the clinical findings of CanL exists., Material and Methods: Dogs with a diagnosis of leishmaniosis were enrolled (n = 123). Clinical information, including signalment, clinical signs and laboratory abnormalities, was retrieved from medical records from different veterinary facilities from Catalonia., Results: Young dogs developed less frequently systemic signs (p = 0.0046), renal (p = 0.0019) and haematologic (p = 0.0275) abnormalities, while dermatologic signs were more common in young and adult dogs compared with old ones (p = 0.0451). Young dogs showed proteinuria less often than adult and old dogs (p = 0.0029). Young dogs did not present renal azotemia, while old dogs showed occasionally renal azotemia (p = 0.0478). Young dogs were mainly classified as mild-moderate LeishVet clinical stages of the disease, and very rarely as severe-very severe LeishVet clinical stages, compared with adult and old dogs (p = 0.0457). Purebred dogs significantly developed ulcerative dermatitis more frequently than crossbred dogs (p = 0.0460)., Conclusion: This study describes that age is associated with differences in clinicopathological findings of CanL., (© 2021 The Authors. Veterinary Record published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

Canine leishmaniasis (CanL) is a zoonosis caused by protozoa of the genus Leishmania and remains an important public health concern in tropical areas. In Brazil, domestic dogs are considered the most relevant reservoir of the parasite and one of the main targets of the disease control actions. Considering this, we aimed herein to evaluate the CanL infection in different canine groups and distribution of cases in the state of Sergipe, an endemic region in Northeastern Brazil. The evaluated 467 animals were classified into four groups: hunting (n = 50), company (n = 64), guard (n = 140), and wandering (n = 213). Samples (blood, bone marrow, conjunctival swab, and lymph node aspirate) were collected from animals in nine municipalities of Sergipe. First, all animals were submitted to general and ophthalmic clinical examination. Next, they were tested serologically by TR-DPP®, and for the presence of Leishmania, amastigotes in samples of bone marrow, conjunctival swab, and lymph node aspirate were diagnosed by PCR and parasitological techniques. It was observed that 34.69% (162) of the evaluated dogs were seropositive. The highest rates of positivity were found in hunting 54% (27/50; OR = 3.52; p-value = 0.001) and guard dogs 42.14% (59/140; OR = 2.18; p-value = 0.01). Otherwise, the highest percentage of symptomatic dogs was observed in wandering animals (85%; OR = 9.63; p-value < 0.0001). The distribution of case analysis showed that the highest positivity rates occurred in inland municipalities situated in arid regions. Taken together, our data demonstrate that hunting and guard dogs are among the animals most exposed and affected by clinical manifestations of CanL, mainly in the inland municipalities of Sergipe State., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Leishmaniasis is endemic and a mandatory reporting disease in Spain since 1982. However, between 1996 and 2014, surveillance on public health was decentralized and only some autonomous regions monitored the disease. The aim of this study is to estimate the incidence of leishmaniasis and to evaluate the extent of underreporting in Spain. A capture-recapture (CRC) study was conducted to calculate the incidence of human leishmaniasis using reports from the National Surveillance Network (RENAVE) and the Hospital Discharge Records of the National Health System (CMBD) for 2016 and 2017. During the study period, 802 cases were reported to RENAVE and there were 1,149 incident hospitalizations related to leishmaniasis. The estimated incidence rates through the CRC study were 0.79 per 100,000 inhabitants for visceral leishmaniasis (VL), 0.88 (cutaneous leishmaniasis (CL)) and 0.12 (mucocutaneous leishmaniasis (MCL)) in 2016 and 0.86 (VL), 1.04 (CL) and 0.12 (MCL) in 2017. An underreporting of 14.7-20.2% for VL and 50.4-55.1% for CL was found. The CRC method has helped us to assess the sensitivity and representativeness of leishmaniasis surveillance in Spain, being a useful tool to assess whether the generalization of leishmaniasis surveillance throughout the Spanish territory achieves a reduction in underreporting., Competing Interests: The authors have declared that no competing interests exist.

Background: The Leishmaniases are on the top of the global list of tropical neglected diseases. The number of infected dogs in South America is estimated in millions and correlated to disease cases in humans, especially in Brazil. Equines may get infected too and can play a role in the epidemiological chain. Thus, the aim of the present study was to evaluate risk and protective factors of leishmaniasis in rural areas of the western border region of Rio Grande do Sul state, Brazil by Leishmania spp. protozoa molecular detection and serological evaluation (ELISA) in equine and canine blood samples. This work included nine farms around the city of Uruguaiana. Epidemiologic information regarding farm characteristics and biologic material collection of canine (22) and equine (91), totalizing 113 samples was collected. The polymerase chain reaction (PCR) technique was used to detect Leishmania spp. in biological samples. Variables related to the farm were collected and evaluated through descriptive analysis followed by chi-square test and a logistic regression analysis., Results: Nineteen positive samples (19/113 - 16,81%) were detected, being 18 equines and 1 canine, in six of the nine farms included in the study. No animal showed clinical signs of the disease. According to the variables analyzed, when compared each characteristic separately, the presence of abundant vegetation and poor hygiene demonstrated to be risk factors to Leishmania infection in rural areas. The logistic regression showed that excellent general hygiene, proximity to the weir and trimmed grass were protective factors (p=0.038, p=0.001 and p=0.014, respectively). Having excellent hygiene represents a 70% lower chance of getting infected, keeping the grass cut protects the animal by more than 90% and the proximity of the weir represents a protective factor of 96%., Conclusions: The presence of Leishmania infection in the western border region of Rio Grande do Sul was 16,81% and it was influenced by farm characteristics. The role of the excellent general hygiene as a protective factor is extremely relevant in the leishmaniases prevention., (© 2021. The Author(s).)

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., Competing Interests: The authors have declared that no competing interests exist.

Objective: To provide an updated overview of the leishmaniasis epidemiology in the Balkans, as this region presents "blank space" on the map of leishmaniasis knowledge in Europe., Methods: Following the PRISMA guideline, literature on the presence and circulation of human leishmaniasis in Balkan countries was screened in PubMed, Google Scholar, Science Direct, Scopus, Web of Science, Matica Srpska Library and the National Library of Serbia., Results: Sixty nine publications from 1919 until September 2020 were reviewed. Historical records indicate that leishmaniasis was previously present mainly in the southern parts of the Balkans, while newer data demonstrate its northward spread., Conclusions: Human leishmaniasis is re-emerging in the Balkans and in some areas is already widespread., (© 2021 John Wiley & Sons Ltd.)

Canine leishmaniosis (CanL) is a severe, potentially life-threatening disease of dogs and is considered one of the most important vector-borne zoonoses in Europe, where there is evidence that it is currently spreading into previously unaffected areas. Veterinary practitioners represent the first-line defence against CanL and in recent years, several guidelines have been developed to assist veterinarians in the correct diagnosis, management and prevention of the disease. The present study was aimed at surveying Italian practitioners on how they manage CanL and compare results to recommendations from guidelines developed by the European Scientific Council for Companion Animal Parasites (ESCCAP®), by the Leishvet® organization and by the Canine Leishmaniasis Working Group (C.L.W.G.). Approximately 14.0 % of veterinarians contacted replied to the survey. Results suggest that Italian practitioners are aware of currently available guidelines for the management of CanL and adherence to recommendations is good. Interestingly, 16.0 % carry out year-long prevention, even though recent data suggest that transmission in Italy continues to be seasonal. In conclusion, the high awareness among practitioners reported here should be considered an excellent opportunity for guideline developers to continue updating their recommendations to include any new information that can support the correct management of CL., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Sandflies are active vectors of several diseases, including leishmaniasis, which Morocco hopes to eliminate by 2030. Despite efforts to limit their spread, they still remain a public health problem in the country, as the behaviour of individuals in relation to sandflies plays an important role in the sustainability of the epidemiological cycle., Aims: To explore and determine the knowledge and behaviours related to sandfly diseases., Methods: A quantitative method was adopted using a questionnaire assisted by a personal interview. Based on the epidemiological situation of leishmaniasis cases reported in recent years in Al-Hajeb province, we conducted a field survey among 281 persons in April and May 2019 residing in the communities where the cases of the disease are registered., Results: 61.6% of Moroccans know sandflies by the name "Chniwla"; 44.1% thought that sandflies do not transmit diseases; 41.3% thought they multiplied in contaminated water; 52.7% thought sandfly bites could not be avoided; and 6.4% recognized the role of individuals in the fight against vectors., Conclusions: The need to raise public awareness of the risks of sandflies, using the popular concepts obtained to simplify scientific terms and formulate targeted health education strategies that make the individual an active player in vector control., (Copyright © World Health Organization (WHO) 2021. Open Access. Some rights reserved. This work is available under the CC BY-NC-SA 3.0 IGO license (https://creativecommons.org/licenses/by-nc-sa/3.0/igo).)

The development of vaccines against one or all forms of human leishmaniasis remains hampered by a paucity of investment, at least in part resulting from the lack of well-evidenced and agreed estimates of vaccine demand. Starting from the definition of 4 main use cases (prevention of visceral leishmaniasis, prevention of cutaneous leishmaniasis, prevention of post-kala-azar dermal leishmaniasis and treatment of post-kala-azar dermal leishmaniasis), we have estimated the size of each target population, focusing on those endemic countries where incidence levels are sufficiently high to justify decisions to adopt a vaccine. We assumed a dual vaccine delivery strategy, including a wide age-range catch-up campaign before the start of routine immunisation. Vaccine characteristics and delivery parameters reflective of a target product profile and the likely duration of the clinical development effort were considered in forecasting the demand for each of the four indications. Over a period of 10 years, this demand is forecasted to range from 300-830 million doses for a vaccine preventing visceral leishmaniasis and 557-1400 million doses for a vaccine preventing cutaneous leishmaniasis under the different scenarios we simulated. In a scenario with an effective prophylactic visceral leishmaniasis vaccine, demand for use to prevent or treat post-kala-azar dermal leishmaniasis would be more limited (over the 10 years ~160,000 doses for prevention and ~7,000 doses for treatment). Demand would rise to exceed 330,000 doses, however, in the absence of an effective vaccine for visceral leishmaniasis. Because of the sizeable demand and potential for public health impact, a single-indication prophylactic vaccine for visceral or cutaneous leishmaniasis, and even more so a cross-protective prophylactic vaccine could attract the interest of commercial developers. Continuous refinement of these first-of-their kind estimates and confirmation of country willingness and ability to pay will be paramount to inform the decisions of policy makers and developers in relation to a leishmaniasis vaccine. Positive decisions can provide a much-needed contribution towards the achievement of global leishmaniasis control., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: PMK is co-author of a patent protecting the gene insert used in Leishmania candidate vaccine ChAd63-KH (Europe 10719953.1; India 315101) and is funded by the UK Medical Research Council / Department for International Development to develop a controlled human infection model for sand fly-transmitted cutaneous leishmaniasis. MMGH Consulting (SM, MM, CFM) were appointed to lead the work under contract from the University of York.

Background: Leishmania tarentolae is a protozoan isolated from geckoes (Tarentola annularis, Tarentola mauritanica), which is considered non-pathogenic and is transmitted by herpetophilic Sergentomyia spp. sand flies. This species occurs in sympatry with Leishmania infantum in areas where canine leishmaniasis is endemic. In the present study, we investigated the circulation of L. tarentolae and L. infantum in sand flies, dogs and lizards in a dog shelter in southern Italy, where canine leishmaniasis by L. infantum is endemic., Methods: Sheltered dogs (n = 100) negative for Leishmania spp. (March 2020) were screened by immunofluorescence antibody test (IFAT) using promastigotes of both species at two time points (June 2020 and March 2021). Whole blood from dogs, tissues of Podarcis siculus lizards (n = 28) and sand flies (n = 2306) were also sampled and tested by a duplex real-time PCR (dqPCR). Host blood meal was assessed in sand flies by PCR., Results: Overall, 16 dogs became positive for L. infantum and/or L. tarentolae by IFAT at one or both sampling periods. One canine blood sample was positive for L. infantum, whilst two for L. tarentolae by dqPCR. At the cytology of lizard blood, Leishmania spp. amastigote-like forms were detected in erythrocytes. Twenty-two tissue samples, mostly lung (21.4%), scored molecularly positive for L. tarentolae, corresponding to 10 lizards (i.e., 35.7%). Of the female Sergentomyia minuta sampled (n = 1252), 158 scored positive for L. tarentolae, four for L. infantum, and one co-infected. Two Phlebotomus perniciosus (out of 29 females) were positive for L. tarentolae. Engorged S. minuta (n = 10) fed on humans, and one P. perniciosus, positive for L. tarentolae, on lagomorphs., Conclusions: Dogs and lacertid lizards (Podarcis siculus) were herein found for the first time infected by L. tarentolae. The detection of both L. tarentolae and L. infantum in S. minuta and P. perniciosus suggests their sympatric circulation, with a potential overlap in vertebrate hosts. The interactions between L. tarentolae and L. infantum should be further investigated in both vectors and vertebrate hosts to understand the potential implications for the diagnosis and control of canine leishmaniasis in endemic areas., (© 2021. The Author(s).)

The study of tropical diseases like leishmaniasis, a parasitic disease, has not received much attention even though it is the second-largest infectious disease after malaria. As per the WHO report, a total of 0.7-1.0 million new leishmaniasis cases, which are spread by 23 Leishmania species in more than 98 countries, are estimated with an alarming 26,000-65,000 death toll every year. Lack of potential vaccines along with the cost and toxicity of amphotericin B (AmB), the most common drug for the treatment of leishmaniasis, has raised the interest significantly for new formulations and drug delivery systems including nanoparticle-based delivery as anti-leishmanial agents. The size, shape, and high surface area to volume ratio of different NPs make them ideal for many biological applications. The delivery of drugs through liposome, polymeric, and solid-lipid NPs provides the advantage of high biocomatibilty of the carrier with reduced toxicity. Importantly, NP-based delivery has shown improved efficacy due to targeted delivery of the payload and synergistic action of NP and payload on the target. This review analyses the advantage of NP-based delivery over standard chemotherapy and natural product-based delivery system. The role of different physicochemical properties of a nanoscale delivery system is discussed. Further, different ways of nanoformulation delivery ranging from liposome, niosomes, polymeric, metallic, solid-lipid NPs were updated along with the possible mechanisms of action against the parasite. The status of current nano-vaccines and the future potential of NP-based vaccine are elaborated here., (Copyright © 2021. Published by Elsevier Masson SAS.)

Green periurban residential areas in Mediterranean countries have flourished in the last decades and become foci for leishmaniasis. To remedy the absence of information on vector ecology in these environments, we examined phlebotomine sand fly distribution in 29 sites in Murcia City over a 3-year period, including the plots of 20 detached houses and nine non-urbanized sites nearby. We collected 5,066 specimens from five species using "sticky" interception and light attraction traps. The relative frequency of the main Leishmania infantum vector Phlebotomus perniciosus in these traps was 32% and 63%, respectively. Sand fly density was widely variable spatially and temporally and greatest in non-urbanized sites, particularly in caves and abandoned buildings close to domestic animal holdings. Phlebotomus perniciosus density in house plots was positively correlated with those in non-urbanized sites, greatest in larger properties with extensive vegetation and non-permanently lived, but not associated to dog presence or a history of canine leishmaniasis. Within house plots, sand fly density was highest in traps closest to walls. Furthermore, the study provides a guideline for insect density assessment and reporting and is envisioned as a building block towards the development of a pan-European database for robust investigation of environmental determinants of sand fly distribution., (© 2021. The Author(s).)

Brazil is one of the countries that concentrates 90% of all tegumentary and visceral leishmaniases cases and Bahia is one of the highly affected states. In the present report, we consolidated secondary data from several complementary databases that allowed us to record the sand fly species identified including areas of Leishmania spp. transmission in the state of Bahia. We then overlayed the geographical distribution data onto maps of vegetational aspects found across the state. Overall, 21 602 records of phlebotomine sand flies occurrence between 1949 and 2016 were analysed, encompassing 85% of Bahia's municipalities. Seventy-six sand fly species under 17 genera were enlisted. Among described species, 27 were proven or putative Leishmania spp. vectors and three were considered exclusively endemic in the state. Lutzomyia longipalpis, Nyssomyia intermedia and Nyssomyia whitmani were found in 74, 29 and 27% of municipalities, respectively. Salvador, the state capital and major city presented records for 21 different sand fly species, including known vectors for leishmaniasis. In particular, a wide distribution of Evandromyia sallesi was detected for this city. This consolidated account on phebotomine fauna and distribution may be explored for improving the planning and deployment of vector-focused leishmaniasis control measures in affected areas of Bahia., (© 2021 The Royal Entomological Society.)

Background: In urban settings, the presence of a high density of the human population and contact with domestic and/or stray animals such as dogs and cats can be risk factors for the transmission of zoonotic protozoa parasites. Toxoplasma gondii (T. gondii) and Leishmania spp. are zoonotic protozoon parasites with significant health burdens worldwide., Methods: A cross-sectional study was used to investigate the antibody prevalence and risk factors of T. gondii and Leishmania spp. infections in 385 randomly selected dogs of Ambo, Bako, and Gojo towns of West Shewa Zone, Oromia regional state, Ethiopia. A questionnaire survey was administered to households to collect data on potential risk factors. Dog sera samples were assayed for T. gondii IgG antibodies using the direct agglutination test while Leishmania spp. specific antibodies tested using an indirect enzyme-linked immunosorbent assay (ELISA). Logistic regression was used for data analysis., Results: Overall, T. gondii and Leishmania spp. infection seroprevalence was found to be 82.86% (95% confidence interval (CI): 78.71-86.49%) and 92.47% (95% CI: 89.36-94.90%), respectively. Seropositivity for both T. gondii and Leishmania spp. was found in 82.58% of the dogs. None of the investigated factors were associated with Leishmania spp. seropositivity (p > 0.05). The seroprevalence of T. gondii was significantly different among the study towns (p = 0.003). The risk of T. gondii infection was 2.71 times higher in adult dogs than juvenile dogs (p = 0.043). Dogs kept simultaneously with other domestic animals had increased odds of T. gondii seropositivity compared to those with no other domestic animals (Adjusted Odds ratio: 1.96, p = 0.021). However, altitude, sex, breed, housing, feeding, educational level of head of the household, and dog's living area were not significantly associated with T. gondii seropositivity (p > 0.05)., Conclusions: The high seropositivity and the simultaneous presence of antibodies of T. gondii and Leishmania spp. in dogs suggest the widespread nature of these parasites in the environment and the high potential of transmission to other animals and humans. Further epidemiological studies, isolation and molecular characterization of the parasites, and educational campaigns are suggested., (© 2021. The Author(s).)

The scarce information about breeding sites of phlebotomines limits our understanding of the epidemiology of tegumentary leishmaniasis. Identifying the breeding sites and seasons of immature stages of these vectors is essential to propose prevention and control strategies different from those targeting the adult stage. Here we identified the rural breeding environments of immature stages of Ny. whitmani, vector species of Leishmania braziliensis in the north of Misiones province, Argentina; then we determined and compared the environmental and structural characteristics of those sites. We also identified the season of greatest emergence and its relationship with adult abundance. During a first collection period, between 28 and 48 emergence traps were set continuously for 16 months in six environments of the farm peridomicile and domicile: below house, chicken shed, experimental chicken shed, forest edge, pigsty and under fruit tree. Traps were checked and rotated every 40 nights. A total of 146 newly emerged individuals were collected (93.8% of them were Ny. whitmani), totaling an effort of 23,040 emergence trap-nights. The most productive environments were chicken shed and below house, and the greatest emergence was recorded in spring and summer. During a second collection period, emergence traps and light traps for adult capture were placed in the chicken shed and below house environments of eight farms. Emergence traps were active continuously during spring, summer, and early autumn. Environmental and structural characteristics of each environment were recorded. A total of 84 newly emerged phlebotomines (92.9% Ny. whitmani; 72,144 emergence trap-nights) and 13,993 adult phlebotomines (147 light trap-nights) were recorded in the chicken shed and below house environments. A positive correlation was also observed between trap success of newly emerged phlebotomines and of adults after 120 days. A high spatial variability was observed in the emergence of Ny. whitmani, with the number of newly emerged individuals being highest in soils of chicken sheds with the highest number of chickens and closest to forest edge. Moreover, below house was found to be as important as chicken sheds as breeding sites of Ny. whitmani. Management of the number of chickens in sheds, soil moisture and pH, and the decision of where to localize the chicken sheds in relation to the houses and the forest edge, might contribute to reduce the risk of human vector exposure and transmission of Leishmania., Competing Interests: The authors have declared that no competing interests exist.

Background: Phlebotomine sand flies are prominent vectors of Leishmania parasites that cause leishmaniasis, which comes second to malaria in terms of parasitic causative fatalities globally. In the absence of human vaccines, sand fly chemical-based vector control is a key component of leishmaniasis control efforts., Methods and Findings: We performed a literature review on the current interventions, primarily, insecticide-based used for sand fly control, as well as the global insecticide resistance (IR) status of the main sand fly vector species. Indoor insecticidal interventions, such as residual spraying and treated bed nets are the most widely deployed, while several alternative control strategies are also used in certain settings and/or are under evaluation. IR has been sporadically detected in sand flies in India and other regions, using non-standardized diagnostic bioassays. Molecular studies are limited to monitoring of known pyrethroid resistance mutations (kdr), which are present at high frequencies in certain regions., Conclusions: As the leishmaniasis burden remains a major problem at a global scale, evidence-based rational use of insecticidal interventions is required to meet public health demands. Standardized bioassays and molecular markers are a prerequisite for this task, albeit are lagging behind. Experiences from other disease vectors underscore the need for the implementation of appropriate IR management (IRM) programs, in the framework of integrated vector management (IVM). The implementation of alternative strategies seems context- and case-specific, with key eco-epidemiological parameters yet to be investigated. New biotechnology-based control approaches might also come into play in the near future to further reinforce sand fly/leishmaniasis control efforts., Competing Interests: The authors have declared that no competing interests exist.

Visceral leishmaniasis (VL) in patients receiving immunosuppressant drugs for autoimmune disease has been on the rise. It is important-but difficult-to know when cure has been achieved in these patients since the withdrawal of immunosuppressants during antileishmania treatment is commonly required, and there is a risk of relapse when immunosuppression is restored. The prevalence of asymptomatic infection among those immunosuppressed for autoimmune disease is also uncertain. The present work describes how cytokine release assays can be used to confirm the cure of VL, and to determine the prevalence of asymptomatic infection, in such patients. After collection of blood from volunteers (n = 108), SLA-stimulation of peripheral blood mononuclear cell cultures and of whole blood was found to induce the production of different combinations of cytokines that served to confirm recovery from VL, and asymptomatic Leishmania infection. Indeed, cure was confirmed in 14 patients, all of whom showed a specific Th1 immune response against Leishmania, and the prevalence of asymptomatic infection was determined as 21.27%. Cytokine profiles could be used to manage VL in patients with autoimmune disease, and to identify and better protect those with asymptomatic infection who are at risk of developing this disease., Competing Interests: The authors have declared that no competing interests exist.

Phlebotomine sand flies are the main natural vectors of Leishmania, which cause visceral and tegumentary tropical diseases worldwide. However, their taxonomy and evolutionary history remain poorly studied. Indeed, as for many human disease vectors, their small size is a challenge for morphological and molecular works. Here, we successfully amplified unbiased copies of whole genome to sequence thousands of restriction-site associated DNA (RAD) markers from single specimens of phlebotomines. RAD markers were used to infer a fully resolved phylogeny of the subgenus Paraphlebotomus (11 species + 5 outgroups, 32 specimens). The subgenus was not recovered as monophyletic and we describe a new subgenus Artemievus subg. nov. Depaquit for Phlebotomus alexandri. We also confirm the validity of Ph. riouxi which is reinstated as valid species. Our analyses suggest that Paraphlebotomus sensu nov. originated ca 12.9-8.5 Ma and was possibly largely distributed from peri-Mediterranean to Irano-Turanian regions. Its biogeographical history can be summarized into three phases: i) a first split between Ph. riouxi + Ph. chabaudi and other species that may have resulted from the rise of the Saharan belt ca 8.5 Ma; ii) a Messinian vicariant event (7.3-5.3 Ma) during which the prolonged drought could have resulted in the divergence of main lineages; iii) a recent radiation event (3-2 Ma) that correspond to cycles of wet and dry periods in the Middle East and the East African subregions during the Pleistocene. Interestingly these cycles are also hypothetical drivers of the diversification of rodents, in the burrows of which Paraphlebotomus larvae develop. By meeting the challenge of sequencing pangenomics markers from single, minute phlebotomines, this work opens new avenues for improving our understanding of the epidemiology of leishmaniases and possibly other human diseases transmitted by arthropod vectors., Competing Interests: The authors have declared that no competing interests exist

Background: Leishmaniasis is an emerging infectious disease reported in the north and south of Thailand of which patients with HIV/AIDS are a high risk group for acquiring the infection. A lack of information regarding prevalence, and the risk association of Leishmania infection among asymptomatic immunocompetent hosts needs further investigation. Information on potential vectors and animal reservoirs in the affected areas is also important to control disease transmission., Methods: An outbreak investigation and a cross-sectional study were conducted following one index case of cutaneous leishmaniasis (CL) caused by L. martiniquensis in an immunocompetent male patient reported in August 2015, Chiang Rai Province, Thailand. From September to November 2015, a total of 392 participants at two study areas who were related to the index case, 130 students at a semi-boarding vocational school and 262 hill tribe villagers in the patient's hometown, were recruited in this study. The nested internal transcribed spacer 1-PCR (ITS1-PCR) was performed to detect Leishmania DNA in buffy coat, and nucleotide sequencing was used to identify species. Antibody screening in plasma was performed using the Direct Agglutination Test (DAT), and associated risk factors were analyzed using a standardized questionnaire. Captured sandflies within the study areas were identified and detected for Leishmania DNA using nested ITS1-PCR. Moreover, the animal reservoirs in the study areas were also explored for Leishmania infection., Results: Of 392 participants, 28 (7.1%) were positive for Leishmania infection of which 1 (4.8%) was L. martiniquensis, 12 (57.1%) were L. orientalis and 8 (38.1%) were Leishmania spp. Of 28, 15 (53.6%) were DAT positive. None showed any symptoms of CL or visceral leishmaniasis. Risk factors were associated with being female (adjusted odds ratio, AOR 2.52, 95%CI 1.01-6.26), increasing age (AOR 1.05, 95%CI 1.02-1.08), having an animal enclosure in a housing area (AOR 3.04, 95%CI 1.13-8.22), being exposed to termite mounds (AOR 3.74, 95%CI 1.11-12.58) and having domestic animals in a housing area (AOR 7.11, 95%CI 2.08-24.37). At the semi-boarding vocational school, six Sergentomyia gemmea samples were PCR positive for DNA of L. orientalis and one S. gemmea was PCR positive for DNA of L. donovani/L. infantum. Additionally, one Phlebotomus stantoni was PCR positive for DNA of L. martiniquensis, and one black rat (Rattus rattus) was PCR positive for DNA of L. martiniquensis., Conclusion: This information could be useful for monitoring Leishmania infection among immunocompetent hosts in affected areas and also setting up strategies for prevention and control. A follow-up study of asymptomatic individuals with seropositive results as well as those with positive PCR results is recommended., Competing Interests: The authors have declared that no competing interests exist.

Both cutaneous (zoonotic cutaneous leishmaniasis [ZCL] and anthroponotic cutaneous leishmaniasis [ACL]) and visceral leishmaniasis (VL) are prevalent in Pakistan, although the cutaneous form is comparatively more widespread. The present study aimed to spatially map different forms of leishmaniasis in Pakistan. In addition, the risk for acquiring leishmaniasis was assessed in different locations of the country. Previously published literature on leishmaniasis distribution and associated risk factors in Pakistan was obtained from PubMed/NCBI and GoogleScholar using specific keywords. Further, 50 articles that focused on geographical distribution and risk factors of the disease in Pakistan were obtained and systematically reviewed. Data regarding location (used in ArcGIS for spatial analysis), number of cases, and risk factors were identified. The present study revealed a high burden of cutaneous leishmaniasis in the northern and western foci of Pakistan. Leishmania major, chiefly a causative agent of ZCL, was observed to be prevalent in the arid and semi-arid climatic zones of the country with elevations between 45 and 2,837 m (maximum number at low elevation). Leishmania tropica cases, associated with ACL, were recorded at elevations ranging from 551 to 2,837 m (majority cases at high elevation). Visceral leishmaniasis (VL) (reportedly caused by Leishmania infantum) was sporadically distributed in the northern areas at elevations 1,432-2,873 m. For VL, particularly, mountainous communities, farming communities, and the presence of dogs were the most commonly reported risk factors. On the other hand, poor preventive measures, migration, presence of domesticated animals, presence of cattle dung, living in mud houses, and outdoor sleeping habits were the principal risk factors associated with cutaneous leishmaniasis (CL). This investigation can guide the health experts and medical entomologists toward targeted and cost-effective surveillance and control of leishmaniasis in Pakistan., (© American Society of Parasitologists 2021.)

The presence of DNA and anti- Leishmania spp. antibodies in the serum of 112 healthy horses was investigated by evaluating the physical examination, from a rural society located in the north central region of Paraná. The antigens of Leishmania amazonensis , Leishmania braziliensis , and Leishmania infantum were used to perform the indirect enzyme-linked immunosorbent assay, where it was possible to detect the reaction in 27.67% of the samples. These were also subjected to the real-time quantitative PCR, which confirmed the presence of Leishmania spp. DNA in 67.34% of the tested samples. The results show that the tested animals were previously exposed to the protozoan. Thus, these animals can be considered environmental bioindicators of the presence of Leishmania spp. at the study site. The material used in this study (serum), although not ideal, proved to be effective and less invasive. Taking into account the importance of the disease, the absence of more in-depth information on the species, the high zootechnical value of these animals, and their strictly close contact with the urban area and the human species, it is essential that further studies are carried out to elucidate the epidemiological profile of them in the face of the disease, as well as the possibility of them acting not only as hosts but also as reservoirs.

Background: Currently, various zoonotic diseases are classified as emerging or reemerging. Because equids have a direct relationship with various vectors, they are possibly more frequently exposed to zoonotic agents than are humans. The undeniable importance of diseases such as human granulocytic anaplasmosis, spotted fever, and leishmaniasis for both public and animal health, as well as the possibility of equids acting as sources, reservoirs, or even sentinels for these pathogens, justifies the detection of their frequency and factors associated with infection in equids from northeastern Brazil., Methods: Blood samples were collected from 569 equids (528 horses, 33 donkeys, and 8 mules), 516 from a rural area and 53 from an urban area. Pathogen detection was carried out as follows: Borrelia spp. and Rickettsia spp., serological analysis; Leishmania spp., serological analysis and polymerase chain reaction (PCR); Anaplasma phagocytophilum, PCR. Determination of associated factors was carried out through generalized linear models., Results: The frequencies of positivity for the pathogens observed in equids were as follows: Borrelia spp., 13.9% (79/569); Leishmania spp., 3.5% (20/569); Rickettsia spp. 33.4% (190/569). Regarding factors associated with infection, male sex was associated with protection against Borrelia spp.; donkeys and mules were associated with protection against Rickettsia spp., while a younger age was a risk factor. The infection of A. phagocytophilum was not detected in the sampled population. Co-infection was detected in 5.1% (29/569) of the animals., Conclusions: Most of the studied pathogenic agents are present in the prospected area, indicating a possible risk for both human and animal health. This demonstrates that equids can be considered important sentinels in the assessment of pathogens with zoonotic potential in the region.

The present study aimed to evaluate a methodology for active surveillance of visceral leishmaniasis by detecting Leishmania DNA in organs of wild road-killed animals from November 2016 to October 2018 in the North of Paraná, Brazil. The collection points of road-killed wild animals were georeferenced. The animals were autopsied and samples of bone marrow, lymph node, liver, spleen, and ear skin were collected. Genomic DNA was extracted and subjected to PCR for amplification of Leishmania spp. 18S, kinetoplastic DNA (kDNA), HSP70, and ITS1 genes, and DNA sequencing was performed. The primers used for the amplification of kDNA, ITS1, and HSP70 genes presented non-specific results. Of the 66 mammals collected from 24 different municipalities, one Southern Tamandua (Tamandua tetradactyla) presented DNA of Leishmania spp. in lymph nodes by 18S PCR. DNA sequencing confirmed the results of the subgenus, Viannia, identification. We suggest using the methodology showed in the present study in the active and early surveillance of visceral leishmaniasis in a non-endemic area.

Introduction: The first visceral and cutaneous leishmaniasis cases were reported in Cameroon since more than six decades. However, interest in the disease has decreased over time and data on its epidemiology across the country are scanty. This systematic review aims to update data on what is known and done so far on leishmaniasis in Cameroon., Methods and Analysis: PubMed/MEDLINE, EMBASE and Web of Science will be searched from inception onwards. Grey literature will be identified through Google Scholar searches, dissertation databases and other relevant documents such as report of the National Control Program. Searches will be conducted between January and February 2021. All studies reporting endemicity, distribution, infecting species, vectors and reservoirs will be eligible. The main outcomes will be epidemiological data (infection rate, distribution, infecting species, vectors and animal reservoir), while the secondary outcomes will be the cases management (diagnostic, treatment, reporting, intervention…). Two reviewers will independently screen eligible papers, and potential conflicts will be resolved by involving a third reviewer as an adjudicator. Methodological quality including bias will be appraised using a methodological quality critical appraisal checklist proposed in the Joanna Briggs Institute systematic review methods manual. A narrative synthesis will describe quality and content of the epidemiological evidence. Data on prevalence and vectors will be used to draw thematic maps of the distribution of leishmaniasis in Cameroon., Ethics and Dissemination: This study will not require ethical approval as it will be based on already published or unpublished data. The final report of this review will be published in a peer-reviewed journal, and the outcomes will be used (1) as baseline information to design further studies that will help to better refine the epidemiological situation of leishmaniasis in Cameroon, and (2) to inform both programme managers and policy-makers of the situation of leishmaniasis in the country., Systematic Review Registration: This protocol was registered with the International Prospective Register of Systematic reviews (PROSPERO; registration number: CRD42020211864) database., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Due to its geographical location, Muğla province is one of the most frequently used places by refugees. Although leishmaniasis have been previously reported in this region, there is a lack of information on the etiological agent and possible vectors. The main objectives of this study were; i) to investigate the sand fly fauna, ii) to reveal the natural Leishmania spp. infection in wild caught sand flies using molecular tools, and iii) to determine the annual seasonal dynamics of the sand flies in Muğla region. Totally, 2093 specimens belonging to 15 species [12 Phlebotomus, three Sergentomyia; 51 unidentified] were collected during the one-year (June 2016- June 2017) period. Of the collected sand flies, 1928 (92.12%) were caught by the Centers for Disease Control (CDC) light traps, while 165 (7.88%) of them were caught by sticky traps. Phlebotomus major sensu lato (s.l.), the potential vector of visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean and Aegean region, was detected in all sampling locations and found as the dominant taxon (n=1035; 49.45%) of the study area and followed by Phlebotomus tobbi (n=371; 17.72%). During the sampling period, sand fly activity was started in March and peaked in August. Sand fly population size reduced dramatically between mid-September and early October. The number of collected specimens was peaked in August, while there is only one sample collected both in November and March. The majority of the sand flies (78.66%) were collected at an altitude range of 200-400 m. Seventy-two monospecific pools were screened for the presence of Leishmania DNA by real time ITS1 PCR and 24 (nine P. major s.l., eight P. tobbi, two P. papatasi, two S. minuta, one P. alexandri, one P. similis, and one Phlebotomus (Transphlebotomus spp.) of them (33.8%) were found positive (L. infantum, L. tropica, and L. major). To the best of our knowledge, the presence of fifteen sand fly species and their distribution, seasonal dynamics, molecular detection of Leishmania parasites in Muğla province was reported for the first time. The presence of vector species in the study area, appropriate temperature and humidity conditions, long sand fly activity season, and presence of Leishmania parasite suggests that there is a serious risk in the transmission of leishmaniasis in Muğla., (Copyright © 2021. Published by Elsevier B.V.)

Asymptomatic leishmaniasis is believed to play important role in maintaining the transmission of Leishmania spp. within endemic communities. Therefore, the efforts to eliminate leishmaniasis are daunting if we cannot manage asymptomatic leishmaniasis well. To clarify the global prevalence and factors associated with the asymptomatic Leishmania infection, we assessed the prevalence of asymptomatic leishmaniasis by a systematic review followed by meta-analyses. In addition, factors associated with the asymptomatic leishmaniasis versus symptomatic were also analyzed. We included all of the original articles alluding to the human asymptomatic leishmaniasis that was confirmed by at least one laboratory diagnosis method regardless of age, sex, race, and ethnicity of the patients, study design, publication date or languages. In total, 111 original articles were chosen for the data extraction. Based on our meta-analyses of the original articles reporting asymptomatic leishmaniasis mostly in endemic areas, the prevalence of asymptomatic leishmaniasis was 11.2% [95% confidence interval (CI) 8.6%-14.4%] in general population, 36.7% [95% CI 27.6%-46.8%] in inhabitants living in the same or neighboring household to the symptomatic patients, and 11.8% [95% CI 7.1%-19%] in HIV infected patients. Among individuals with leishmaniasis, 64.9% [95% CI 54.7%-73.9%] were asymptomatic and males were more susceptible to develop symptoms, with OR=1.88, 95% CI 1.19-2.99, P=0.007. Meta-regression analysis showed no significant change in the prevalence of asymptomatic leishmaniasis during the last 40 years., (Copyright © 2020. Published by Elsevier B.V.)

Background: Recent climate and environmental changes have resulted in the geographical expansion of Mediterranean Leishmania infantum vectors towards northern latitudes and higher altitudes in different European countries, including Italy, where new foci of canine leishmaniasis have been observed in the northern part of the country. Northern Italy is also an endemic area for mosquito-borne diseases. During entomological surveillance for West Nile virus, mosquitoes and other hematophagous insects were collected, including Phlebotomine sand flies. In this study, we report the results of Phlebotomine sand fly identification during the entomological surveillance conducted from 2017 to 2019., Methods: The northeastern plain of Italy was divided by a grid with a length of 15 km, and a CO 2 -CDC trap was placed in each geographical unit. The traps were placed ~ 15 km apart. For each sampling site, geographical coordinates were recorded. The traps were operated every two weeks, from May to November. Sand flies collected by CO 2 -CDC traps were identified by morphological and molecular analysis., Results: From 2017 to 2019, a total of 303 sand flies belonging to the species Phlebotomus perniciosus (n = 273), Sergentomyia minuta (n = 5), P. mascittii (n = 2) and P. perfiliewi (n = 2) were collected, along with 21 unidentified specimens. The trend for P. perniciosus collected during the entomological surveillance showed two peaks, one in July and a smaller one in September. Sand flies were collected at different altitudes, from -2 m above sea level (a.s.l.) to 145 m a.s.l. No correlation was observed between altitude and sand fly abundance., Conclusions: Four Phlebotomine sand fly species are reported for the first time from the northeastern plain of Italy. Except for S. minuta, the sand fly species are competent vectors of Leishmania parasites and other arboviruses in the Mediterranean Basin. These findings demonstrate the ability of sand flies to colonize new environments previously considered unsuitable for these insects. Even though the density of the Phlebotomine sand fly population in the plain areas is consistently lower than that observed in hilly and low mountainous areas, the presence of these vectors could herald the onset of epidemic outbreaks of leishmaniasis and other arthropod-borne diseases in areas previously considered non-endemic.

Competing Interests: The authors have declared that no competing interests exist.

Leishmaniasis is a public health problem worldwide. We aimed to predict ecological niche models (ENMs) for visceral (VL) and cutaneous (CL) leishmaniasis and the sand flies involved in the transmission of leishmaniasis in São Paulo, Brazil. Phlebotomine sand flies were collected between 1985 and 2015. ENMs were created for each sand fly species using Maximum Entropy Species Distribution Modeling software, and 20 climatic variables were determined. Nyssomyia intermedia (Lutz & Neiva, 1912) and Lutzomyia longipalpis (Lutz & Neiva, 1912), the primary vectors involved in CL and VL, displayed the highest suitability across the various regions, climates, and topographies. L. longipalpis was found in the border of Paraná an area currently free of VL. The variables with the greatest impact were temperature seasonality, precipitation, and altitude. Co-presence of multiple sand fly species was observed in the cuestas and coastal areas along the border of Paraná and in the western basalt areas along the border of Mato Grosso do Sul. Human CL and VL were found in 475 of 546 (86.7%) and 106 of 645 (16.4%) of municipalities, respectively. Niche overlap between N. intermedia and L. longipalpis was found with 9208 human cases of CL and 2952 cases of VL. ENMs demonstrated that each phlebotomine sand fly species has a unique geographic distribution pattern, and the occurrence of the primary vectors of CL and VL overlapped. These data can be used by public authorities to monitor the dispersion and expansion of CL and VL vectors in São Paulo state.

Ensuring access to essential medicines for leishmaniasis control is challenging, as leishmaniasis is a very small and unattractive market for pharmaceutical industry. Furthermore, control programmes are severely underfunded. We conducted an analysis of global procurement of leishmaniasis medicines for the past 5 years in order to shed light on the current leishmaniasis market landscape and supply and demand dynamics. We estimated global demand of each leishmaniasis medicines, the amount of each medicine required to treat all reported cases, based on the number of cases reported to WHO and the first-line treatment regimen used in each country. Procurement data were obtained from procurement agencies, international organizations, WHO, national leishmaniasis programmes and manufacturers. Expert interviews were conducted to have a better understanding of how medicines were procured and used. The comparison of estimated need and procurement data indicated discrepancies in supply and demand at global level as well as in the most endemic countries. The extent of the gap in supply was up to 80% of the needs for one of the leishmaniasis medicines. Mismatch between supply and demand was much wider for cutaneous leishmaniasis than visceral leishmaniasis. This study presents a current picture of procurement patterns and imbalance in global supply and demand. Addressing improved access and supply barriers requires concerted and coordinated efforts at the global and national levels. Priority actions include setting up a procurement coordination mechanism among major procurers, partners and national programmes where forecasting and supply planning is jointly developed and communicated with manufacturers. In addition, continuous engagement of manufacturers and advocacy is critical to diversify the supplier base and ensure quality-assured and affordable generic medicines for leishmaniasis., Competing Interests: The authors have declared that no competing interests exist.

Leishmaniasis remains one of the world's leading infectious diseases and a public health problem for Bahia and Brazil. We made a retrospective cohort study of leishmaniasis cases reported between 2007 and 2016 in the Barreiras city, an important agribusiness city whose urbanization process was recent. Leishmaniasis presented epidemiological data with similar characteristics of the disease in other regions of the Brazil, that is, with the highest prevalence of visceral leishmaniasis found in children, while cutaneous leishmaniais was found in adults. The disease presented an urban configuration with wide distribution in the Barreiras city. We found the vectors of the different clinical forms of the disease occurring in the urban area of Barreiras city, particularly the sand fly of the Lutzmyia longipalpis, Evandromyia sallesi and Nyssomyia intermedia species. More studies will be necessary to indentify whether these species are really incriminated in the urban transmission of the parasites in the urban areas. This study indicates the places of the spatio-temporal greatest occurrence and transmission of the disease in the urban zones of Barreiras city., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Visceral leishmaniasis (VL) in HIV-positive individuals is a global health problem. HIV-Leishmania coinfection worsens prognosis and mortality risk, and HIV-Leishmania coinfected individuals are more susceptible to VL relapses. Early initiation of antiretroviral therapy can protect against Leishmania infection in individuals living in VL-endemic areas, and regular use of antiretrovirals might prevent VL relapses in these individuals. We conducted a cross-sectional study in Petrolina, Brazil, an VL-endemic area, to estimate the prevalence of asymptomatic Leishmania cases among HIV-positive outpatients., Methods: We invited any HIV-positive patients, aged ≥ 18-years-old, under antiretroviral therapy, and who were asymptomatic for VL. Patients were tested for Leishmania with enzyme-linked immunosorbent assays (ELISA)-rK39, immunochromatographic test (ICT)-rK39, direct agglutination test (DAT), latex agglutination test (KAtex), and conventional polymerase chain reaction (PCR). HIV-Leishmania coinfection was diagnosed when at least one VL test was positive., Results: A total of 483 patients were included. The sample was predominantly composed of single, < 48-years-old, black/pardo, heterosexual males, with fewer than 8 years of schooling. The prevalence of asymptomatic HIV-Leishmania coinfection was 9.11% (44/483). HIV mono-infected and HIV-Leishmania coinfected groups differed statistically significantly in terms of race (p = 0.045), marital status (p = 0.030), and HIV viral load (p = 0.046). Black/pardo patients, married patients, and those with an HIV viral load up to 100,000 copies/ml presented higher odds for HIV-Leishmania coinfection., Conclusions: A considerable number of asymptomatic Leishmania cases were observed among HIV-positive individuals in a VL-endemic area. Given the potential impact on transmission and health costs, as well as the impact on these coinfected individuals, studies of asymptomatic Leishmania carriers can be useful for guiding public health policies in VL-endemic areas aiming to control and eliminate the disease., Competing Interests: The authors have declared that no competing interests exist.

Background: Zoonotic visceral leishmaniasis by Leishmania infantum is a first-order pathology in canine veterinary clinics in endemic areas. Moreover, canine infections are considered the main reservoir for human disease; despite their importance in the control of the disease within a One Health approach, no scientometric study has been published. Aims of the study included analyzing the impact of canine leishmaniasis (CanL) on the scientific literature, drugs or combinations used, trends in the period from 2000 to 2020 and efficacy criteria employed., Methods: A Web of Science (WOS)-based analysis of publications on CanL and chemotherapy of the disease in the period 2000-2020 was carried out using a stepwise methodology. Data were analyzed by year, geographical origin, chemical groups, drugs and combinations, and efficacy criteria., Results: Reports on CanL (n = 3324) represented < 16% of all publications on leishmaniasis (n = 20,968), and of these around 18% (n = 596) were related to chemotherapy. Publication records on CanL followed the distribution of the infection by L. infantum in endemic areas although Mediterranean countries were overrepresented in the reports on chemotherapy of CanL. Publications on the main antileishmanial drugs used in clinical practice showed a sustained tendency in the period analyzed. Pentavalent antimonials (Sb V ), alone or in combination with allopurinol, represented > 50% of all publications on chemotherapy of CanL despite the availability of more recently marketed drugs., Conclusions: Chemotherapy of CanL still relies on Sb V and combinations and to a lesser extent on miltefosine (MIL). Reports on chemotherapy are scarce and mostly publicly funded, and the variability of experimental conditions hampers the direct comparison of the efficacy of drugs, combinations and schedules. The vast majority of reports on efficacy do not include any information on supportive therapy; this reduces the actual value of the studies if intended for the practical management of the disease. Complete reports on the chemotherapy (etiological + symptomatic) would add value to the trials performed.

Background: Cutaneous leishmaniasis caused by Leishmania species ( L. spp ) is one of the most important parasitic diseases in humans. To gain information on the metabolite variations and biochemical pathways between L. spp , we used the comparative metabolome of metacyclic promastigotes in the Iranian isolates of L. major and L. tropica by proton nuclear magnetic resonance ( 1 H-NMR)., Methods: L. tropica and L. major were collected from three areas of Iran, namely Gonbad, Mashhad, and Bam, between 2017 and 2018, and were cultured. The metacyclic promastigote of each species was separated, and cell metabolites were extracted. 1 H-NMR spectroscopy was applied, and the data were processed using ProMatab in MATLAB (version 7.8.0.347). Multivariate statistical analyses, including the principal component analysis and the orthogonal projections to latent structures discriminant analysis, were performed to identify the discriminative metabolites between the two L. spp . Metabolites with variable influences in projection values of more than one and a P value of less than 0.05 were marked as significant differences., Results: A set of metabolites were detected, and 24 significantly differentially expressed metabolites were found between the metacyclic forms of L. major and L. tropica isolates. The top differential metabolites were methionine, aspartate, betaine, and acetylcarnitine, which were increased more in L. tropica than L. major (P<0.005), whereas asparagine, 3-hydroxybutyrate, L-proline, and kynurenine were increased significantly in L. major (P<0.01). The significantly altered metabolites were involved in eight metabolic pathways., Conclusion: Metabolomics, as an invaluable technique, yielded significant metabolites, and their biochemical pathways related to the metacyclic promastigotes of L. major and L. tropica . The findings offer greater insights into parasite biology and how pathogens adapt to their hosts., (Copyright: © Iranian Journal of Medical Sciences.)

Objectives: The protracted and violent conflict in Syria has resulted in large-scale displacement of people and destruction of health and sanitation infrastructure. The aim of this study was to examine epidemiological trends in vector-borne disease (VBD) outbreaks before and following the onset of the Syrian conflict (2011)., Methods: ProMED, a digital disease surveillance tool, was queried for VBD outbreak reports affecting humans and animals in Syria and select bordering countries between 2003 and 2018. Data were normalized by dividing the number of unique VBD events by the total number of unique outbreak events reported by ProMED for each year. Suspected and confirmed case counts and deaths were manually extracted., Results: Reports on VBDs increased from a mean of 2.9/year pre-2011 to 12.8/year post-2011, a 343.5% (p < 0.05) increase. After normalization, reports increased by 485.5% (p < 0.05) over the time periods. Post-2011, the most commonly reported VBDs were leishmaniasis, Crimean-Congo hemorrhagic fever, and lumpy skin disease. Reported numbers of suspected and confirmed cases and deaths increased during the conflict period., Conclusions: VBD outbreak events in ProMED increased in Syria and select bordering countries after the onset of the Syrian conflict in 2011. Enhanced disease surveillance is critical to detect and manage outbreaks in conflict settings., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Leishmaniasis is a widespread, vector-borne parasitosis causing clinical manifestations in animals and in humans. In dogs, Canine Leishmaniasis has been reported in as much as 50 countries and the Mediterranean basin is known to be one of the most affected zones. Within these areas, the Island of Sardinia (Italy) has long been considered endemic for leishmaniasis and the presence of two arthropod vectors has recently been reported there. Nevertheless, to date, no epidemiological surveys regarding CanL have been carried out on the island. Hence, for the first time, the seroprevalence and the risk factors were investigated. Blood samples, as well as clinical and general information from 1.147 dogs, were collected and analyzed. Dogs consisted of two distinct populations, namely "owned dogs" and "kennel dogs." Anti-Leishmania IgG antibodies were detected using IFAT and samples were scored as positive at a cut-off dilution of 1:80. Data was analyzed using a Chi-squared test and bivariate and multivariate analyses were performed. Overall, 15.4% of dogs were found to be infected with CanL while only 44.1% of these animals exhibited clinical signs. Owned dogs (27.2%) were found to be infected more often than kennel dogs (10.6%); male dogs were found to be more frequently infected than female dogs and the number of infected animals increases with age. The present survey confirmed the endemic nature of leishmaniasis in Sardinia with a similar seroprevalence as mainland Italy. The results obtained serve as validation for the hypothesis that, in endemic areas, clinical CanL representations constitute only a fraction of the leishmaniasis cases.

Human leishmaniosis caused by Leishmania infantum is a zoonotic disease, with dogs as the main reservoir in Mediterranean Basin countries. The largest European outbreak of human leishmaniosis declared in the southwestern Madrid region (Spain) is characterized by unusual epidemiological and clinical features, such as the emergence of new wild reservoirs (hares and rabbits), whereas the seroprevalence, infection, and severity of canine leishmaniosis have not substantially changed since the first studies conducted in Madrid before the outbreak. Previous studies reported that L. infantum isolates from the Madrid leishmaniosis focus displayed elevated virulence in in vivo models of infection and increased infectivity in murine target cells. With the aim of studying whether changes in the host-parasite interaction and virulence profile have developed, we first assessed the behaviour of one circulating isolate of the outbreak, IPER/ES/2012/BOS1FL1 (BOS1FL1), compared to that of a well-characterized strain from canine leishmaniosis, MCAN/ES/1996/BCN150 (BCN150), in terms of infection capacity (percentage of infected cells, representing infectivity, and number of amastigotes per infected cell, representing the intensity of infection) in canine monocytes and macrophages. BCN150 displayed significantly higher infectivity (76.82 ± 4.40 vs 38.58 ± 2.19; P <  0.0001) and intensity of infection (3.64 ± 0.13 vs 1.83 ± 0.12; P <  0.0001) than BOS1FL1 when interacting with canine cells. Our ROS induction results did not differ significantly between the two isolates or with the responses previously described for other L. infantum isolates. Paradoxically, increased resilience to hydrogen peroxide exposure was observed for BOS1FL1 (% viability 40.62 ± 5.54 vs 26.37 ± 2.93; P = 0.039). Finally, we demonstrated that a decreased intracellular load of BOS1FL1 was associated with increased IFN-γ (261.21 ± 26.29 vs 69.80 ± 9.02; P = 0.0151) and decreased IL-10 production (165.06 ± 23.87 vs 264.41 ± 30.58; P = 0.0002). In this study, we provide the first detailed insight into the differences between the isolate BOS1FL1 from the outbreak in Madrid and the well-characterized strain BCN150 MON-1 obtained from a dog in their response to interacting with canine cells. However, further studies are necessary to shed light on the immune mechanisms resulting in BOS1FL1 exhibiting less virulent behaviour in canine cells than in cells derived from other host species., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The American cutaneous (CL) and visceral leishmaniasis (VL) are zooanthroponoses transmitted by sand flies. Brazil records thousands of human leishmaniasis cases annually. Dogs are reservoirs of Leishmania infantum , which causes VL, but their role in the transmission cycle of CL is debatable. Wild mammals are considered reservoirs of the aetiological agents of CL ( Leishmania spp.)., Objective: To describe the aetiology of leishmaniasis in dogs in an endemic area for CL and VL in the Amazon, Brazil., Methods: Clinical evaluation and blood collection of 40 dogs from the villages Ubim (20) and Socorro (20), city of Tomé-Açu, state of Pará, were carried out. The DNA extracted from the blood was used for PCR with Leishmania -specific primers targeting the hsp70-234 gene sequence. Products were sequenced (ABI3500XL), and the sequences were aligned, edited (BioEdit), and analyzed (Blastn)., Results: Of the 34 amplified samples, 21 were sequenced, namely Leishmania infantum (12), L. guyanensis (5), L. braziliensis (3), and Leishmania sp. (01)., Conclusion: Given the diversity of circulating pathogens, elucidation of the role of the dog in the Leishmania spp. cycle in Amazonian villages is imperative to the surveillance of CL in the region. We present the first report in Brazil, confirmed by sequencing, of canine infection by L. guyanensis , a species highly resistant to treatment in humans, with the drug of first choice (Glucantime ® ).

The aim of this study was to explore Leishmania infantum epidemiology through a One Health approach that promotes a better estimation of leishmaniasis burden and a deeper understanding of the spatial distribution of the key actors of the parasite life cycle (vectors, reservoirs and humans). We conducted a 14-year mixed retrospective and prospective study of leishmaniasis cases in an endemic area in southern Spain (Granada province), to estimate the human incidence and its association with the vector presence, cryptic leishmaniasis rates and canine leishmaniasis prevalence. We found an annual linear increase in the incidence that cannot be fully explained by active case surveillance and the improvement of PCR diagnostic techniques. 49.4% of cases were not reported to the surveillance system. Approximately half of the human cases correspond to the visceral form that occurred more frequently in men; cutaneous, mucosal and cryptic forms were also detected. Leishmaniasis is no longer a disease of young children, accounting for a quarter of immunocompetent patients and most infected people remained asymptomatic. Human and canine leishmaniasis, cryptic or symptomatic, are present in the whole province, where there is a medium/high risk of the presence of Phlebotomus perniciosus, the main vector. We found association between the incidence of human leishmaniasis and the presence of the vector, but not with the prevalence of canine leishmaniasis and cryptic human leishmaniasis. A potential hot spot was also found, where high leishmaniasis incidence may be associated to the involvement of host species other than dogs., (© 2020 Blackwell Verlag GmbH.)

Objectives: The aims of this study were to investigate the prevalence of Leishmania species infection in cats in Northern Italy and to evaluate the associations between infection and signalment and clinicopathological data., Methods: The study was carried out in a veterinary university hospital from June to November 2017. Blood, urine, conjunctival swabs and hair were collected from all randomly selected cats. Leishmania species infection was evaluated using the indirect fluorescent antibody test (IFAT), setting a cut-off value of 1:80, and using real-time PCR on blood, conjunctival and hair samples. A complete blood count, serum chemistry profile, serum electrophoresis and urinalysis were also carried out. The cats were grouped on the basis of the results of the diagnostic criteria adopted in positive, negative and unconfirmed Leishmania cases. Non-parametric variables and continuous data were compared among the study groups using the χ 2 test and the Mann-Whitney U-test, respectively., Results: One hundred and fifty-two cats were included. Nineteen of the 152 (12.5%) cats were positive (18/152 [11.8%] showed an IFAT titre of ⩾1:80 and 1/152 [0.7%] was real-time PCR-positive from a hair sample); 106/152 (69.7%) cats were negative; and 27/152 (17.8%) cats were unconfirmed for Leishmania species. Total proteins, beta 2 -globulin and gamma-globulin were significantly increased in the positive Leishmania group compared with the negative group., Conclusions and Relevance: The results of the present study demonstrated the spread of Leishmania infantum infection in cats in Northern Italy. Hyperproteinaemia and hypergammaglobulinaemia appeared to be significant clinicopathological abnormalities in this population of cats with L infantum infection.