[THE EPIDEMIOLOGICAL SITUATION OF LEISHMANIASIS IN THE RUSSIAN FEDERATION: THE FIRST VALID CASES OF LOCAL TRANSMISSION].

The authors have analyzed the data available in the literature and statistical offices and the results of their observations of the epidemiological situation of leishmaniasis in the Russian Federation (RF). In October 1991 to December 2014, the RF registered 89, leishmaniasis cases, including 83 imported cases from 24 countries and 6 local cases. Out of the 83 imported cases, there were those diagnosed with anthroponotic cutaneous leishmaniasis (n=31), zoonotic cutaneous leishmaniasis (n=20), visceral leishmaniasis (n=31), and mucocutaneous leishmaniasis (n=1). The Republic of Dagestan and Eastern Crimea are visceral Ieishmaniasis-enderpic, areas. The obtained data on the local cases of visceral leisimaniasis in Dagestan and on its sporadic cases in Crimea suggest that it is necessary to conduct epidemiological, epizootological, and entomological surveys in the above areas and to develop a system for visceral leishmaniasis epidemiological surveillance encompassing the whole package of antiepidemic measures. The effective treatment of leishmaniases requires that the RF Ministry of Health should register some current antileishmanial drugs (such as Glucantime) as soon as possible.

[The species composition and epidemiological significance of mosquitoes (Diptera, Psychodidae, Phlebotominae) in the foci of visceral leishmaniasis in the Papsky District, Namangan Region, Uzbekistan].

In 2008, mosquito observations were made in 4 populated areas of the Papsky District, Namangan Region, Uzbekistan (Fergana Valley), where visceral leishmaniasis cases had been registered. The mosquitoes were caught in Oltinkan, Gulistan, Chodak, and Chorkesar in July and in Oltinkan in September. A total of 7245 mosquitoes were caught in the living and utility premises during the observation period. The mosquito fauna of this focus was found to represent 10 species: P. papatasi, P. sergenti, P. alexandri, P. caucasicus, P. nuri, P. keshishiani, P. angustus, P. longiductus, S. grecovi, and S. sumbarica. It also contained P. papatasi, a vehicle for transmission of zoonotic cutaneous leishmaniasis, P. sergenti, an anthroponotic cutaneous leishmaniasis vehicle, and P. longiductus, a visceral leishmaniasis one. The major site of hatching and habitat for mosquitoes were utility premises for large and small cattle. A polymerase chain reaction was used to determine mosquito infestation with L. infantum. A total of 38 female pools of 5 species: P. papatasi, P. sergenti, P. keshishiani, P. angustus, and P. longiductus were tested. Testing of female mosquitoes for L. infantum yielded a negative result.

[The epidemiological features of visceral leishmaniasis, revealed on examination of children by polymerase chain reaction, in the Papsky District, Namangan Region, Uzbekistan].

Patients with visceral leishmaniasis (VL) have been registered in the Papsky District, Namangan Region, Uzbekistan, over the past 23 years. A total of 95 patients were notified in 1987 to 2009. In 2007-2008, a mass survey using the polymerase chain reaction (PCR) within the international INTAS project 05-100006-8043 was conducted in 5 population aggregates of the Papsky District, Namangan Region, Uzbekistan, where VL cases had been regularly registered in the last years. Bone marrow and venous and peripheral blood smears were used as a test material. A total of 234 samples, including 3 bone marrow biopsy specimens, 9 venous blood samples and 222 peripheral blood ones, were tested. All the samples were on the glass slides. Three groups were identified among the examinees. Group 1 consisted of 13 subjects who had been ill at different times. Group 2 comprised 27 children treated at hospital for various diagnoses. Group 3 (the largest one, n=190) included apparently healthy children. All the children of this group felt well and had no symptoms of any illnesses at the examination. In this group, 85 (44.7%) subjects were PCR-positive. Twenty-four (55.8%) of 43 children in the age group of 0-3 years were PCR-positive; the 4-7-year age group comprised 66 subjects and 33 (50%) of them were PCR-positive. Group over 7 years of age included 81 subjects; 45 (55.5%) were PCR-positive. The results of the mass survey with PCR, which covered the 5 population aggregates in the Papsky District, Namangan Region, Uzbekistan, suggest the epidemic activity of a synathropic focus of VL and make us look at many fixed notions of its epidemiology in new contexts.

[Human and canine visceral leishmaniasis in the Papsky District, Namangan Region, Uzbekistan: seroepidemiological and seroepizootological surveys].

In 2007 - 2008, four (Chodak, Oltinkan, Gulistan, and Chorkesar) of 9 population aggregates in the Papsky District, Namangan Region, Uzbekistan, where visceral leishmaniasis (VL) cases had been registered in the last years were selected to make seroepidemiological and seroepizootological surveys within the international project funded by INTAS grant 05-100006-8043. The surveys of the populations were conducted visiting their homesteads. These additionally included children's and health care facilities where all children aged less than 14 years were examined. On examining the children, their peripheral blood (approximately 0.1 ml) was taken on filter paper for serological assays. Canine blood was sampled from the vein. Enzyme-linked immunosorbent assay (ELISA) was carried out to detect antibodies to VL pathogens. A total of 521 children were examined for two years, by applying ELISA. Five hundred and fourteen blood samples from children younger than 14 years, 162 dogs, 4 foxes, and 1 cat were tested. Testing 514 children's blood samples for VL pathogen antigen ascertained that in the 4 population aggregates there was an average of 10% VL-seropositive children, including those who were ill with VL at the moment of the examination and had been ill. The highest number of VL-seropositive samples (14.9%) was found in the settlement of Chodak. VL pathogen antibodies were detected in 26 (61.9%) of 42 dogs with the clinical signs of VL. VL-positive tests were found in 26 (21.6%) of 120 apparently healthy dogs. The samples from 4 foxes and 1 cat were negative. Immunological findings indicated that 0-3-year-old children were a group that is most susceptible to VL in the study focus of this disease. The high proportion of dogs with VL may account for the rise in infant morbidity and suggests the epizootic strain in the focus of VL in the Papsky District.

[Investigations on leishmaniases at the E.I. Martsinovsky Institute of Medical Parasitology and Tropical Medicine].

The paper assesses the investigations on leishmaniases at the E.I. Martsinovsky Institute of Medical Parasitology and Tropical Medicine in 1920 to 2009. The analysis includes papers on biology, ecology, taxonomy, and experimental transmission of the agents of leishmaniases via the bites of sand flies, the principle in the control of zoonotic cutaneous leishmaniasis (ZCL) during the agricultural development of extensive territories in the Karshin steppe, on quantitative approaches in the epidemiology of ZCL, a search for Russian effective medicaments to treat patients with this disease and the development of criteria for selecting L. major strains for individual and mass vaccinations against ZCL, the revision of Leishmania circulating in great gerbil populations, and the description of the new species L. turanica, an important parasite for L. major persistence from one transmission season to the next. The first investigations on leishmaniasis were made by Prof. E.I. Martsinovsky, the founder and the first director of the Institute in the early 20th century.

[Visceral leishmaniasis in the Commonwealth of Independent States (CIS): results and basic lines of further study].

The results of studies of visceral leishmaniasis (VL) made in the 20th century in the Commonwealth of Independent States that are VL-endemic, such as Azerbaijan, Armenia, Georgia, Kazakhstan, Kyrghyzstan, Tadjikistan, Turkmenistan, and Uzbekistan are summed up. The magnitude of studies of VL in different regions is different. The authors analyze the epidemiological and epizootological situation and define the basic lines of further studies of VL, which include the present view of the prevalence of VL in Central Asia and Transcaucasia; identification of strains of the pathogen by molecular genetic methods; study of its vectors; detection of natural reservoirs of the pathogen, improvement of methods for VL diagnosis; and their introduction into laboratories' work.

[Ecology and the genetic structure of sympatric leishmania species circulating in the intra-continental deserts of the south Palaearctic region]. / Ekologia i geneticheskaia struktura populiatsii simpatrichnykh vidov leishmanii, tsirkuliruiushchikh vo vnutrikontinental'nykh pustyniakh iuzhnoi Palearktiki.

A PCR fingerprinting approach with single non-specific primers--(GTG)5 and T3B--was apply to investigate variations in the genotyping of three species of Leishmania species within the Rhombomys opimus area. Forty-three strains of Leishmania major, L. turanica, and L. gerbilli circulating among great gerbils in Turkmenistan, Uzbekistan, Kazakhstan, and Mongolia were examined. PCR fingerprint revealed a high genetic intraspecific heterogeneity among L. turanica strains. Three groups of strains were clearly identified. The strains from Mongolia greatly differed from other L. turanica ones. The second group was formed by strains from Kazakhstan, they also demonstrated rather different patterns. L. turanica strains from Turkmenistan and Uzbekistan showed only minor differences, but greatly different from those from Kazakhstan and Mongolia. The groups identified by the PCR fingerprint correlate with the conditions of circulation: the duration of a transmission season and as the result of different periods of retention of Leishmania in the skin of great gerbils, as well as the presence or absence of L. major as coexisting species. There were no differences between L. turanica strains isolated from different hosts in the same geographical region, as well as between L. turanica strains isolated in the hyper- or meso and hypoendemic foci. There was no correlation between the genotypic and phenotypic characteristics of L. turanica. No intraspecific polymorphism was found among L. major and L. gerbilli strains from different geographical regions within the great gerbil area.

[Glass slide smears and imprints from infected organs for identification of Leishmania major by polymerase chain reaction methods]. / Vozmozhnost' ispol'zovaniia mazkov i otpechatkov na stekle ot porazhennykh organov dlia identifikatsii Leishmaniia major metodami PTSR analiza.

The paper presents data showing that the DNA isolated from the smears and imprints of L. major-infected hamsters is suitable for use in the polymerase chain reaction (PCR) to detect the causative agent of leishmaniasis. The most solid data have been obtained with the smears unexposed to staining and examination by using immersion oil and to benzene treatment. The DNA isolated from these smears infected may preserve for at least 1.5 months in a domestic refrigerator. The immersion oil-treated smears may be also used to identify leishmanias, but DNA should be isolated from the infected specimens of these smears just before PCR. The original primer pair L-unit/L-mail that has shown itself well in the experiments on cultured promastigotes may be, if required, used to differentiate L. major and L. turanica in the infected material collected from infected rodents.

Mixed leishmanial infections in Rhombomys opimus: a key to the persistence of Leishmania major from one transmission season to the next.

An important feature of the foci of zoonotic cutaneous leishmaniasis (ZCL) in Turkmenistan and Uzbekistan is a 6-10-month break in transmission when Leishmania parasites persist in great gerbils (Rhombomys opimus)--the main host for three species (L. major, L. turanica and L. gerbilli). Almost all (95%) of the laboratory-maintained R. opimus experimentally infected with L. major cured their infections within 6 months, a situation which, if mirrored in field conditions, cannot provide reliable persistence of the infection to the next transmission season. However, infections with L. turanica alone persisted for a mean of 15 months, and mixed infections of L. major and L. turanica persisted even longer (mean = 25 months), parasites of both species remaining detectable in the skin for at least 18 months. Isoenzyme identification of 664 isolates obtained from wild-caught R. opimus, and of 58 cloned strains developed from them, showed that L. turanica, which is non-pathogenic for humans, tends to predominate in the gerbils from all types of natural ZCL foci, including those which are hyper-endemic; in June, L. turanica may be present in 80%-100% of the R. opimus in the foci. In contrast, infections with L. major alone occur far less commonly, and are especially hard to find at the beginning of the transmission season. However, 5%-25% of great gerbils in these foci are each infected with a mixture of L. major and L. turanica. In hyper- and meso-endemic foci, the proportion of L. major within mixed infections of Leishmania increases significantly towards the end of transmission season (August-September). It would appear, therefore, that mixed L. major/L. turanica infections in R. opimus promote the persistence of L. major between transmission seasons.

Comparison of some molecular-genetic techniques for identification of Leishmania circulating in natural foci of zoonotic cutaneous leishmaniasis 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.

[A dry nutrient medium for the cultivation of Leishmania promastigotes]. / Sukhaia pitatel'naia sreda dlia kul'tivirovaniia promastigot leishmanii.

A dry medium formulation which provides the growth and reproduction of Leishmania promastigotes was designed. The medium has been prepared by using nondietary protein raw material, it contains domestic-production ingredients of that in high supply. The nutrient basis of the medium is enzymatic peptone (a source of amine nitrogen). The growth factors of Leishmania (a yeast extract, vitamins, amino acids, a dry blood derivative), carbohydrates (an energy components of the medium), inorganic salts (a source of trace elements), which are selected in strictly defined quantitative ratios, meet the physiological requirements of Leishmania. The agent rules out the necessity of adding ex tempore blood, making the process for preparing the medium from the dry powder simple and easy-to-use.

[The use of radioisotope labelling for studying the feeding of sandflies (Phlebotominae) and their move into colonies of the greater gerbil (Rhombomys opimus Licht.)]. / Ispol'zovanie radioizotopnogo mecheniia dlia izucheniia prokormleniia moskitov (Phlebotominae) i ikh peremeshchenii v poseleniiakh bol'shoi peschanki (Rhombomys opimus Licht.).

The fly off and distribution of sandflies, vectors of zoonotic cutaneous leishmaniasis after meal on great gerbils were studied by means of radioisotope techniques. The experiments were carried out near the town of Mubarek (Uzbekistan). Three Rhombomys opimus were marked by intraperitoneal injection of 14C-glycine solution. Then they were allowed to come back to their burrows. The sandflies were radioactively labelled during bloodsucking on the rodents. Fifty-seven labeled female sandflies were caught on the surface of burrows with the marked R. opimus and on 5 colony burrows at a distance of 70-280 m from it for 6 nights. Seventeen females were Phlebotomus papatasi (0.9% of all the caught females of this species) and 40 females belonged to the Paraphlebotomus subgenus (7.0%). The fly off occurred both before and against the wind. The labeled females were found on all burrows where sticky papers were placed, but their density was 16 times lower at a distance of 280 m than in the burrow colony of their release.

Progress in studies on Central Asian foci of zoonotic cutaneous leishmaniasis: a review.

In this paper, studies on zoonotic cutaneous leishmaniasis (ZCL) are reviewed that were performed during the last ten years largely by scientists of the Martsinovsky Institute of Medical Parasitology and Tropical Medicine. New data on the taxonomy of Leishmania circulating in populations of Rhombomys opimus, their main host, and the results of field and laboratory studies allowed revision of certain concepts generally accepted in epidemiology and epizootology of ZCL in Central Asia.

[The host-opportunistic protozoa system. The dissemination of Pneumocystis infection in white rats under the influence of drug-induced and biological immunosuppression]. / Sistema "khoziain--uslovno-patogennye prosteishie". Disseminatsiia pnevmotsistnoi infektsii u belykh krys pod vliianiem medikamentoznoi i biologicheskoi immunosupressii.

The main purpose of the present study is the investigation of relationships of Pneumocystis carinii with the organism of white rat Wistar, which is natural carrier of this parasite. The series of experiments has shown that the immunosuppressor Tricort-40 (corticosteroid of prolonged activity) in a short time reactivates the Pneumocystis infection. The parasites have been observed in a great number in the lungs and rarely in the liver. The reactivation and dissemination of the Pneumocystis infection have been achieved constantly and with great expression by the combined immunosuppression of rats, by the medicamentous immunosuppression (injection of T-40) and biological one (infection with amastigotes Leishmania infantum). The developing mixed-infection (with pneumocysts and leishmania) could be the basis for the parasitocenological relationships.

[The development of a new experimental model of pneumocystosis]. / Razrabotka novoi éksperimental'noi modeli pnevmotsistoza.

The corticosteroid tricort-40 (triamcinolone acetonide) was found to induce the reactivation of endogenous pneumocystic injection caused by P. carinii in Wistar rats given 5-9 infections subcutaneously in median doses of 0.5-4.0 mg/kg at 6-22 days intervals each. Manifest P. carinii infection in the animals was characterized by early detection of its agent (starting from week 2 of immunosuppression), a progression of the infection and its transition to the generalized stage. The combined use of tricort-40 and infection of the animals with Leishmania infantum as an immunosuppressive cofactor ensured an increase in reproducibility of a manifest pneumocystosis model and its more rapid transition to the generalized stage. The experimental model of mixed infection of two AIDS-associated parasite infections--generalized pneumocystosis and visceral leishmaniasis--was first reproduced in inbred rats, which may be suitable for simultaneous screening of new antipneumocystic and antileishmanial drugs.

[The isoenzyme identification of Leishmania isolates taken from greater gerbils, sandflies and human patients in foci of zoonotic cutaneous leishmaniasis in Turkmenistan]. / Izoénzimnaia identifikatsiia izoliatov leishmanii, vydelennykh ot bol'shikh peschanok, moskitov i bol'nykh liudei v ochagakh zoonoznogo kozhnogo leishmanioza v Turkmenistane.

In 1991-1992, 230 isolates were obtained in the Tedzhen oasis and its adjacent desert areas: 172 isolates from great gerbils, 39 from P. papatasi, and 19 from human cutaneous leishmaniasis patients. All the isolates were identified by the isoenzyme polyacrylamide gel electrophoresis by 8 enzymes. The characteristics of Leishmania circulation in the hyperendemic foci of Turkmenistan were similar to those previously studied in the mesoendemic areas of Uzbekistan and Kazakhstan. L. turanica which is non-pathogenic for man prevailed among infected great gerbils in winter, spring, and early summer, making the natural foci epidemiologically safe in that period of time. It was only in August-September that the great gerbil infection rate by L. major appeared to increase, occasionally reaching 100%. Epizootics due to L. major are developing in the presence of L. turanica, therefore most isolates are clone mixtures of L. major and L. turanica. P. papatasi is the only vector in the Tedzhen oasis; there has been strong evidence for its transmission of both L. major and L. turanica, which makes the concept inconsistent that P. papatasi is associated only with L. major. The overall analysis of recent findings of the distribution of L. major in the populations of great gerbils makes it possible to limit the former endemic zoonotic cutaneous leishmaniasis areas to 40 degrees N latitude and the southern borders of Turkmenistan and Uzbekistan. Within this area, the distribution of L. major is uneven and associated basically with rivers, valleys, oases, and foothill desert plains.

[The identification of marker strains of Leishmania major, L. turanica and L. gerbilli by the polymer chain reaction with a universal primer]. / Identifikatsiia markernykh shtammov Leishmania major, L. turanica, L. gerbilli metodom polimeraznoi tsepnoi reaktsii s universal'nym praimerom.

The Leningrad Nuclear Physics Institute, Academy of Science of the USSR, and Martsinovskii Institute of Medical Parasitology and Tropical Medicine, USSR Ministry of Health, developed polymerase chain reaction (PCR) technique with universal primer 3-2 for Leishmania identification. The primers were patented in the USSR (Patent No 4757254, 1989). Reference strains of three Leishmania species were identified: L. major--MRHO/SU/59/Neal P; L. gerbilli--MRHO/CN/60/gerbilli; L. turanica--MRHO/SU/80/Cl 3720 and MRHO/SU/83/KD 051. Each Leishmania species is specific and different in its PCR pattern whereas the two L. turanica strains have identical PCR patterns of the given primer.

[The characteristics of the epidemic activation of a natural focus of zoonotic cutaneous leishmaniasis in places with a sympatric dissemination of Leishmania major, L. turanica and L. gerbilli]. / Osobennosti épidemicheskoi aktivizatsii prirodnogo ochaga zoonoznogo kozhnogo leishmanioza v mestakh simpatrichnogo rasprostraneniia Leishmania major, L. turanica i L. gerbilli.

In 1975-1978 and in 1985-1988 studies of species composition and number of Phlebotomus, R. opimus infestation with L. major, L. turanica and L. gerbilli as well as epidemic activity of the natural foci were performed in the Karshi steppe in Uzbekistan. Typical areas have been compared in the desert, oasis and desert land irrigated for cotton growing. A correlation has been established between the epidemic activity of the natural focus and the nature of epizootic development in R. opimus and species composition of vectors. On desert territories epizootic transmission throughout the whole season develops with the domination of one leishmania species (L. turanica) which is not pathogenic for human beings. Paraphlebotomus are the main vectors transmitting leishmania. In oases epizootic process assumes a two-phase course, with accumulation by the middle of the season L. major species pathogenic for human beings. It is transmitted by P. papatasi. With desert irrigation, the former Phlebotomus species is transformed into the latter one, with the predominance of P. papatasi. Epizootics acquire a stable two-phase nature.

[Susceptibility to and the characteristics of the course of experimental leishmaniasis in different species of mammals infected with Leishmania major, L. turanica and L. gerbilli]. / Vospriimchivost' i osobennosti techeniia éksperimental'nogo leishmanioza u raznykh vidov mlekopitaiushchikh pri zarazhenii Leishmania major, L. turanica i L. gerbilli.

40 R. opimus and 69 M. libycus were infected and 59 human subjects were vaccinated in laboratory conditions. 13 cultures of 3 leishmania species and their mixtures were used. R. opimus turned to be sensitive to 3 leishmania species. Leishmaniasis developed in the form of infiltrates irrespective of the leishmania species and pathogenic activity. Ulceration and visceralization were always absent. Differences in the duration of leishmania preservation in R. opimus have been noted. In L. major infection the mean disease duration was 7 months, in L. turanica infection it was 15 months, in L. gerbilli infection it was 18 months and in combined L. major and L. turanica infection the disease duration was 39 months. M. libycus proved to be sensitive to L. major and L. major/L. turanica mixture and resistant to L. turanica and L. gerbilli. In L. major infection ulceration (mean duration, 4 months) was observed. In combined L. major and L. turanica infection the signs varied from infiltrates to ulceration, with the disease duration increasing to 8.5 months. In human subjects, the typical course of leishmaniasis was observed only in L. major infection, when L. turanica with artificially increased pathogenic activity was administered, the course of skin lesions was abortive, which, however, in 1 volunteer led to the development of immunity to subsequent administration of L. major.