Revisiting epidemiology of leishmaniasis in central Asia: lessons learnt.

In this work we reviewed historical and recent data on Leishmania spp. infection combining data collected in Turkmenistan, Uzbekistan, Kazakhstan, Kyrgyzstan, Iran, China and Mongolia. We specifically focused on a complex of co-existing species (Leishmania major, Leishmania turanica and Leishmania gerbilli) sharing the same animal reservoirs and vectors. In addition, we analysed the presence of dsRNA viruses in these species and discussed future research directions to identify species-specific traits, which may determine susceptibility of different Leishmania spp. to viral infection.

Microsatellite based molecular epidemiology of Leishmania infantum from re-emerging foci of visceral leishmaniasis in Armenia and pilot risk assessment by ecological niche modeling.

BACKGROUND: Visceral leishmaniasis (VL) is re-emerging in Armenia since 1999 with 167 cases recorded until 2019. The objectives of this study were (i) to determine for the first time the genetic diversity and population structure of the causative agent of VL in Armenia; (ii) to compare these genotypes with those from most endemic regions worldwide; (iii) to monitor the diversity of vectors in Armenia; (iv) to predict the distribution of the vectors and VL in time and space by ecological niche modeling. METHODOLOGY/PRINCIPAL FINDINGS: Human samples from different parts of Armenia previously identified by ITS-1-RFLP as L. infantum were studied by Multilocus Microsatellite Typing (MLMT). These data were combined with previously typed L. infantum strains from the main global endemic regions for population structure analysis. Within the 23 Armenian L. infantum strains 22 different genotypes were identified. The combined analysis revealed that all strains belong to the worldwide predominating MON1-population, however most closely related to a subpopulation from Southeastern Europe, Maghreb, Middle East and Central Asia. The three observed Armenian clusters grouped within this subpopulation with strains from Greece/Turkey, and from Central Asia, respectively. Ecological niche modeling based on VL cases and collected proven vectors (P. balcanicus, P. kandelakii) identified Yerevan and districts Lori, Tavush, Syunik, Armavir, Ararat bordering Georgia, Turkey, Iran and Azerbaijan as most suitable for the vectors and with the highest risk for VL transmission. Due to climate change the suitable habitat for VL transmission will expand in future all over Armenia. CONCLUSIONS: Genetic diversity and population structure of the causative agent of VL in Armenia were addressed for the first time. Further genotyping studies should be performed with samples from infected humans, animals and sand flies from all active foci including the neighboring countries to understand transmission cycles, re-emergence, spread, and epidemiology of VL in Armenia and the entire Transcaucasus enabling epidemiological monitoring.

History of the E.I. Martsinovsky Institute of Medical Parasitology and Tropical Medicine: research on malaria and leishmaniasis.

This review presents the 100-year history of the Martsinovsky Institute of Medical Parasitology and Tropical Medicine in Moscow, Russia, starting with its foundation and early activities, and also describes the impact of its leading scientists, some of whom became internationally known. The institute headed a network of nine tropical institutes in the various Soviet republics from the 1920s to 1990. The extensive body of literature on the history and research accomplishments of this institute has mainly been published in Russian; our goal here is to introduce these achievements and this expertise to the international scientific and medical community, focusing on malaria and leishmaniasis and the development of measures to control and monitor these diseases in the USSR.

History of the E.I. Martsinovsky Institute of Medical Parasitology and Tropical Medicine: research on malaria and leishmaniasis / História do Instituto E.I. Martsinovsky de Parasitologia Médica e Medicina Tropical: pesquisa sobre malária e leishmaniose

Abstract This review presents the 100-year history of the Martsinovsky Institute of Medical Parasitology and Tropical Medicine in Moscow, Russia, starting with its foundation and early activities, and also describes the impact of its leading scientists, some of whom became internationally known. The institute headed a network of nine tropical institutes in the various Soviet republics from the 1920s to 1990. The extensive body of literature on the history and research accomplishments of this institute has mainly been published in Russian; our goal here is to introduce these achievements and this expertise to the international scientific and medical community, focusing on malaria and leishmaniasis and the development of measures to control and monitor these diseases in the USSR.

Resumo O artigo analisa a história centenária do Instituto Martsinovsky de Parasitologia Médica e Medicina Tropical em Moscou, Rússia, desde sua fundação e primeiras atividades, e descreve a influência de seus principais cientistas, alguns dos quais viriam a conquistar renome internacional. O instituto liderou uma rede de nove institutos tropicais em diversas repúblicas soviéticas entre as décadas de 1920 e 1990. A vasta literatura sobre o trabalho de história e pesquisa desse instituto foi publicada sobretudo em russo; nosso objetivo aqui é apresentar esse trabalho e conhecimento à comunidade médica e científica internacional, concentrando-se na malária e na leishmaniose e no avanço de medidas de controle e monitoramento dessas doenças na URSS.

History of the E.I. Martsinovsky Institute of Medical Parasitology and Tropical Medicine: research on malaria and leishmaniasis

This review presents the 100-year history of the Martsinovsky Institute of Medical Parasitology and Tropical Medicine in Moscow, Russia, starting with its foundation and early activities, and also describes the impact of its leading scientists, some of whom became internationally known. The institute headed a network of nine tropical institutes in the various Soviet republics from the 1920s to 1990. The extensive body of literature on the history and research accomplishments of this institute has mainly been published in Russian; our goal here is to introduce these achievements and this expertise to the international scientific and medical community, focusing on malaria and leishmaniasis and the development of measures to control and monitor these diseases in the USSR

Re-Emerging foci of visceral leishmaniasis in Armenia - first molecular diagnosis of clinical samples.

Visceral leishmaniasis (VL) was firstly reported in Armenia in 1913. Following a considerable increase of the number of cases until the mid 1950s, the disease disappeared after 1969 and re-emerged in 1999. Scientific literature about VL in Armenia is available only in Russian or Armenian. This paper presents a historical overview about leishmaniasis in Armenia based on this literature as well as an epidemiological update since the re-emergence of the disease. In 1999-2016, 116 indigenous VL cases were recorded mainly in children in 8 of the 11 districts, however, VL is underreported because of lack of trained medical personal and diagnostic facilities. The aim of this work was to apply for the first time molecular diagnosis of VL in Armenia. Out of 25 VL suspected patients, 22 were positive by microscopy and polymerase chain reaction (PCR). Genotyping using internal transcribed spacer 1-PCR-restriction fragment length polymorphism and sequencing identified the causative agent of VL in Armenia as Leishmania infantum. The present work is an important step towards the inclusion of molecular techniques in the current diagnosis of VL in Armenia and the establishment of local molecular diagnostic facilities.

A narrative review of visceral leishmaniasis in Armenia, Azerbaijan, Georgia, Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, Uzbekistan, the Crimean Peninsula and Southern Russia.

There is an extensive body of medical and scientific research literature on visceral leishmaniasis (VL) in the Caucasus, Central Asia, the Crimean Peninsula and the southern part of The Russian Federation that is written in Russian, making it inaccessible to the majority of people who are interested in the leishmaniases in general and VL in particular. This review and summary in English of VL in what was Imperial Russia, which then became the Soviet Union and later a number of different independent states intends to give access to that majority. There are numerous publications in Russian on VL and, mostly, those published in books and the main scientific journals have been included here. The vast geographical area encompassed has been subdivided into four main parts: the southern Caucasus, covering Armenia, Azerbaijan and Georgia; Central Asia, covering Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan and Uzbekistan; the Crimean Peninsula and the northern Caucasus, which is part of The Russian Federation. Only rare cases of VL have been recorded in the northern Caucasus and Crimean Peninsula. In the other countries mentioned, human VL has been more intense but epidemics like those associated with L. donovani in India and East Africa have not occurred. For most of the countries, there are sections on the distribution, clinical aspects, the causative agent, the reservoirs and the vectors. Serological surveys and research into therapy are also covered. Recent studies on VL in Uzbekistan covered the application of serological, biochemical and molecular biological methods to diagnose human and canine VL, to identify the leishmanial parasites causing them in Uzbekistan and neighbouring Tajikistan and the epidemiology of VL in the Namangan Region of the Pap District, Eastern Uzbekistan. More recently, two studies were carried out in Georgia investigating the prevalence of human and canine VL, and the species composition of phlebotomine sand flies and their rates of infection with what was probably L. infantum in Tbilisi, eastern Georgia and Kutaisi, a new focus, in western Georgia. Though published in English, summaries of this information have been included where relevant to update the parts on VL in Uzbekistan and Georgia.

Selective amplification of maxicircle classes during the life cycle of Leishmania major.

The kinetoplast genome contains several thousands of minicircles of various sequence classes and several scores of maxicircles. We demonstrated that maxicircles are heterogeneous in clonal cultures of Leishmania major, and, therefore, probably heterogeneous (heteroplasmic) within the kinetoplast. Sequence heterogeneity was observed in a non-coding fragment upstream of the 12S rRNA gene. We identified about 20 stable variants of this fragment, which were composed of one to five non-identical repeats 200-300bp in length. Promastigote-to-amastigote and amastigote-to-promastigote differentiation was often accompanied by shifts in abundance of some maxicircle classes. Reversion to promastigote-specific maxicircle patterns was usually observed in the life cycle (promastigote-amastigote-promastigote), however there were many exceptions.

Identification of geographically distributed sub-populations of Leishmania (Leishmania) major by microsatellite analysis.

BACKGROUND: Leishmania (Leishmania) major, one of the agents causing cutaneous leishmaniasis (CL) in humans, is widely distributed in the Old World where different species of wild rodent and phlebotomine sand fly serve as animal reservoir hosts and vectors, respectively. Despite this, strains of L. (L.) major isolated from many different sources over many years have proved to be relatively uniform. To investigate the population structure of the species highly polymorphic microsatellite markers were employed for greater discrimination among it's otherwise closely related strains, an approach applied successfully to other species of Leishmania. RESULTS: Multilocus Microsatellite Typing (MLMT) based on 10 different microsatellite markers was applied to 106 strains of L. (L.) major from different regions where it is endemic. On applying a Bayesian model-based approach, three main populations were identified, corresponding to three separate geographical regions: Central Asia (CA); the Middle East (ME); and Africa (AF). This was congruent with phylogenetic reconstructions based on genetic distances. Re-analysis separated each of the populations into two sub-populations. The two African sub-populations did not correlate well with strains' geographical origin. Strains falling into the sub-populations CA and ME did mostly group according to their place of isolation although some anomalies were seen, probably, owing to human migration. CONCLUSION: The model- and distance-based analyses of the microsatellite data exposed three main populations of L. (L.) major, Central Asia, the Middle East and Africa, each of which separated into two sub-populations. This probably correlates with the different species of rodent host.

Conserved repeats in the kinetoplast maxicircle divergent region of Leishmania sp. and Leptomonas seymouri.

The maxicircle control region [also termed divergent region (DR)] composed of various repeat elements remains the most poorly studied part of the kinetoplast genome. Only three extensive DR sequences demonstrating no significant similarity were available for trypanosomatids (Leishmania tarentolae, Crithidia oncopelti, Trypanosoma brucei). Recently, extensive DR sequences have been obtained for Leishmania major and Trypanosoma cruzi. In this work we have sequenced DR fragments of Leishmania turanica, Leishmania mexicana, Leishmania chagasi and two monogenetic trypanosomatids Leptomonas seymouri and Leptomonas collosoma. With the emergence of the additional extensive sequences some conserved features of DR structure become evident. A conserved palindromic sequence has been revealed in the DRs of the studied Leishmania species, L. seymouri, and T. cruzi. The overall DR structure appears to be similar in all the Leishmania species, their relative L. seymouri, and T. brucei: long relatively GC-rich repeats are interspersed with clusters of short AT-rich repeats. C. oncopelti, L. collosoma, and T. cruzi have a completely different DR structure. Identification of conserved sequences and invariable structural features of the DR may further our understanding of the functioning of this important genome fragment.

The Leishmania major maxicircle divergent region is variable in different isolates and cell types.

The maxicircle divergent region (DR) was partially sequenced in several isolates of Leishmania major. The sequence contains various repeated elements: two types of long GC-rich repeats alternating with clusters of short AT-rich repeats. The arrangement of repeats appears to be similar in the studied Leishmania species and their relative Leptomonas seymouri. Furthermore, a conserved sequence containing putative promoters within a palindrome was revealed in the DRs of these species. Unexpectedly, the DR sequence proved to be dissimilar in promastigotes and amastigotes of the same isolate perhaps through selection of parasites with particular maxicircle variants in the course of the promastigote-amastigote differentiation. Different number of repeats and numerous single nucleotide polymorphisms are observed in the compared sequences. We have also investigated the DR structure in 21 L. major isolates by PCR and demonstrated its great variability. We suppose, however, that different variants of the DR structure are generated by combination of several highly conserved domains.

Genetic and biological diversity among populations of Leishmania major from Central Asia, the Middle East and Africa.

Evidence is provided for genetic and biological variation among Leishmania major strains that correlates with their geographical origin. The host-parasite relationship also appears to be specific. Great gerbils, Rhombomys opimus, and fat sand rats, Psammomys obesus, are the main reservoir hosts in Central Asia and the Middle East, respectively. However, the Central Asian parasite failed to infect the Middle Eastern rodent host in the laboratory, and vice versa. A permissively primed intergenic polymorphic (PPIP)-PCR and a single-stranded conformation polymorphism (SSCP)-PCR exposed genetic polymorphism among 30 strains of L. major from different geographical regions. This was verified by subsequent sequencing of DNA from the same strains using four genomic targets: (a) the NADH-dehydrogenase (NADH-DH) gene, (b) the 6-phosphogluconate dehydrogenase (6PGD) gene, (c) the ribosomal internal transcribed spacers, and (d) an anonymous DNA sequence originally amplified with random primers. All the genetic markers indicated that the nine Central Asian strains were a separate homogenous genetic group. The Middle Eastern strains formed another geographical group that displayed heterogeneity corresponding with their different Middle Eastern locations. Molecular markers and host-parasite relationships confirmed that Central Asian and Middle Eastern strains are genetically and biologically distinct sub-populations of L. major. Three African strains of L. major were genetically closer to the Middle Eastern strains, and a representative one did infect fat sand rats, but they had distinct permissively primed inter-genic polymorphic PCR patterns and internal transcribed spacer 2 types.

Comparison of some molecular-genetic techniques for identification of Leishmania circulating in natural foci of zoonotic cutaneous leismaniasis in the central Asia region

Different molecular-genetic methods were used to identify a cohort of Leishmania strains from natural foci of zoonotic cutaneous leishmaniasis located in Central Asia on the former USSR territory. The results obtained using isoenzymes, PCR, restriction fragment length polymorphisms of kDNA and molecular hybridization techniques are discussed in terms of their applicability, discrimination power and feasibility for answering questions related to molecular epidemiological research and for detecting mixed Leishmania infections.