Genetic features of the Puumala virus (Hantaviridae: Orthohantavirus) identified in the Moscow region.

INTRODUCTION: Puumala virus (family Hantaviridae, genus Orthohantavirus) is distributed in most regions of the European part of Russia. However, information about its genetic variants circulating on the territory of the Central Federal District is extremely scarce. MATERIALS AND METHODS: Rodents' tissue samples were tested after reverse transcription by PCR for the presence of hantaviral RNA. The amplified fragments of the L segment were sequenced by the Sanger method. For two samples, sequences of all three segments were obtained using the NGS method. Phylogenetic trees were built in the MEGA-X software. RESULTS: Puumala virus was found in six samples. Based on the phylogenetic analysis of sequences of three segments, the obtained genetic variants belong to the sublineage previously designated as W-RUS. CONCLUSION: A genetic variant of the Puumala virus, belonging to the subline W-RUS, circulates on the territory of the Volokolamsk district of Moscow region.

[(ON THE NECESSITY AND WAYS TO IMPROVE ENTOMOLOGICAL MONITORING IN THE EPIDEMIOLOGICAL SURVEILLANCE FOR WEST NILE FEVER)].

West Nile fever (WNF) is a natural focal vector-borne disease; the main vectors are mosquitoes. Specific means for its prevention are not available; therefore, one of the main areas of WNF epidemiological surveillance is entomological monitoring that encompasses the control of the size ofvector populations arid the determination of infection rate of mos- quitoes with the virus in order to predict the epidemic situation. An analysis of normative documents on the collection and control of mosquito populations showed it necessary to improve entomological monitoring. Based on the literary and their own data, the authors propose to catch only Culix mosquitoes in the foci of WNF and to analyze their infection rates. The mosquitoes should be caught in mid-June to mid-September in the stationary points of urban and rural biotopes once every 7-10 days, by using automatic traps with CO2 as an attractant. At least 500 specimens should be examined for virus RNA by PCR every 7-10 days to predict the epidemic situation.

[Detection of West Nile virus RNA and specific antibodies in blood donors of Volgograd region].

AIM: Study infection level by West Nile virus (WNV) of donors living on the territory of West Nile fever (WNF) foci with long-term activity, and evaluate the possibility of non-transmissible transfer of the causative agent during use of donor blood. MATERIALS AND METHODS: Blood sera (432) collected in August and September 2012 in Volgograd and Volzhsky cities were studied by PCR and EIA. RESULTS: Antibodies to WNV were detected in 16.2% (70/432); 8.8% (38/432) donors had IgG; 0.93% (4/432)--IgM and 6.5% (28/432)--IgG and IgM antibodies simultaneously. WNV RNA was detected in one of the samples in the last group. Thus, 5 donors (1.1%, 5/432) were at the early stages after infection. CONCLUSION: The data obtained give evidence on the necessity to execute control of donor blood by PCR and EIA in WNF foci with long-term activity.

[The spatial distribution and infection of Culex pipiens L. mosquitoes with Western Nile virus in the Volgograd region].

The infection of Culex pipiens f. pipiens and Cx. pipiens f. molestus mosquitoes with Western Nile virus was studied. Their taxonomic status was determined from the type of mitochondrial DNA. Among 424 females individually examined, there were 4 infected specimens: 3 pipens females gathered in the urban and suburban open biotopes and one molestus female caught in the open urban biotope. Molestus mitotype females were first detected in the suburban open biotopes. The possible reasons for the wide colonization of molestus mosquitoes and its epidemic consequences are discussed.

[Persistence of antibodies specific to West Nile virus in blood of reconvalescents of Volgograd region].

AIM: Determine the duration of persistence of IgM and IgG in reconvalescents of West Nile fever (WNF) 1 year after the disease in southern regions of Russia and evaluate effectiveness of PCR method for acute infection diagnostics. MATERIALS AND METHODS: Blood sera of 87 patients with WNF diagnosis was studied for the presence of West Nile virus (WNV) RNA and IgM and IgG by PCR and EIA. Samples of the first sera were collected in 2010 at days 2 - 30 after the onset of the disease, samples of the second sera--at days 5 - 23 and third--264 - 385 days later. RESULTS: During the first 2 weeks of the disease WNV RNA was detected in more than 50% of patients. In all the first sera IgM at titers of ≥ 1:800 were detected. Seroconversion of IgG titers of 4 and more times was observedin 83% (30/36) of patients. In 2011 IgG were detected in 91% of reconvalescents (79/87), IgM--in 57% (50/87), and in 25% (22/87) IgM titers were ≥ 1:800. CONCLUSION: The results obtained give evidence on the necessity of using several diagnostic criteria simultaneously for the confirmation of WNF clinical diagnosis.

[Role of the mosquitoes Culex pipienis f. pipiens and Cx. pipiens f. molestus (Diptera, Culicidae) in the spread of West Nile virus in the south of Russia].

Cx. pipiens is one of the major vectors of West Nile virus in the south of Russia and it is represented by the autogenic form, molestus, and the non-autogenic form, pipiens. The spatial distribution of its larvae and food preferences of females was investigated to assess the potential role of each form in the spread of the virus. The taxonomic status of the mosquitoes was determined from their capacity for autogenicity (543 specimens) and the type of mitochondrial DNA (348 specimens). The mosquitoes of the pipiens form were shown to develop in the open urban and rural reservoirs and the females were non-autogenic. Cx. pipiens form molestus was found only in the urban biotopes, mainly in the flooded basements and, in some years, also in the open reservoirs where it formed a mixed population with Cx. pipiens form pipiens. Both autogenic (85-95.2%) and non-autogenic specimens were identified among the females with the mitotype in form molestus. Genetic analysis of Cx. pipiens females collected under the Berezantsev bell and in the trap with a bird showed that both forms were attracted to man and the bird. The findings suggest that Cx. pipiens form pipiens can transmit West Nile virus to humans both in the town and in its suburb and Cx. pipiens form molestus can only in urban areas.

[Correlations between placental morphological and functional parameters and neonatal status after normal full-term, prolonged pregnancy, and truly protracted pregnancy].

To solve the problem of differentiating protracted pregnancy, a special morphometric study was undertaken to examine three placental groups: 1) after normal full-term pregnancy (n = 35); 2) after prolonged pregnancy (n = 40); 3) after truly protracted pregnancy with partial or complete Clifford's syndrome (n = 30). A semiquantitative score (14 most important placental indicators), placenta and birth weights, Apgar scores, the infant's weight and height at the end of the first year of life were used so as to estimate the mild, moderate, and severe degree at 0.5, 1.5, and 3 points, respectively. One hundred and twenty-six possible pairs of structural and functional parameters were made up in all the groups. Pearson's correlation coefficient (r > 0.4) was applied to graphically display the pairs. Normal full-term pregnancy was characterized by few positive correlations mainly between the terminal villi and weight-height indices without associations with the Apgar scale. Prolonged pregnancy with the multiple placental tissue structural parameters along with organometric indices being involved was intermediate. The total scores permitted grades 1 and 2 chronic placental insufficiency (CPI) to be diagnosed in 27.5 and 2.5%, respectively. The severest form was truly protracted pregnancy that was distinguished by the maximum positive and negative correlations between all the parameters and the highest rate of diagnosis of CPI of grades 1 (45%) and 2 (15%). The findings strongly suggest that it is essential to identify prolonged and truly protracted pregnancy as important risk factors, by following up the infants for a year.

[Genotyping of West Nile fever virus strains circulating in southern Russia as an epidemiological investigation method: principles and results].

AIM: Characteristic of West Nile fever (WNF) virus strains circulating in southern Russia. MATERIALS AND METHODS: WNF RNA was amplified directly from clinical samples, mosquitoes and bird tissues by PCR, nucleotides were sequenced directly and analyzed comparatively. RESULTS: Related but different genovariants of WNF lineage 1a--"Volgograd" and "Astrakhan"--circulated during WNF outbreaks of 1999 and 2000-2003 in Volgograd and Astrakhan regions. In 2005 "Volgograd" WNF variant emerged in Astrakhan region and along with "Astrakhan" variant caused a new morbidity increase. In 2004 in sera of 2 WNF patients from Rostov region WNF lineage 2 RNA was detected, this was the first WNF clinical case caused by WNF lineage 2 outside of Africa. WNF outbreak in Volgograd region in 2007 was caused by this unique WNF lineage that may preliminary be called Russian. Finally, during a major WNF outbreak in 2010 in Volgograd and Rostov regions in clinical samples only russian genovariant WNF lineage 2 RNA was detected again. CONCLUSION: After emergence of a certain WNF genovariant the virus is capable of persisting in natural foci in southern Russia. A near disappearance of one of the WNF clones by substitution or displacement with another maybe possible. Determination of genetic characteristics of WNF strains circulating in Russia is an important element of WNF epidemiological surveillance and control of this disease.

[Mosquito complex (Diptera, Culicidae) in a West Nile fever focus in the Volgograd Region. II. Host-feeding patterns of mosquitoes in different habitats].

Host preference of the mosquitoes collected in the urban and rural habitats of Volgograd and its suburbs was studied by the precipitation reaction test. Human and avian blood was detected in Cx. pipiens, Cx. modestus, Ae. vexans, Ae. behningi, Ae. caspius, Ae. sticticus, and females of the Anopheles maculipennis. The proportion of the mosquitoes fed on birds was similar in the urban and rural biotopes whereas that of the mosquitoes feeding on humans was significantly higher in Volgograd than in its environs. The increase in the number of human blood-fed mosquitoes in the city resulted mainly from the females collected in its multi-storied buildings.

[Mosquito complex (Diptera, Culicidae) in a West Nile fever focus in the Volgograd region. I. Species diversity and relative abundance in different habitats].

Potential West Nile virus (WNV) vectors were assessed during 2003 and 2004 at the indoor and outdoor collection sites of Volgograd and surrounding rural areas. A total of 26 387 female mosquitoes comprising 16 species in 5 genera of the subfamily Culicinae and 4 species of the subfamily Anophelinae were collected. The seasonal abundance and the dominance structure greatly differed in the multi-storied buildings, farms, and open habitats whereas the similarity of species diversity remained high throughout the season. In the porches of buildings, Aedes vexans (58%) and Cx. pipiens (37%) prevailed in early July (the beginning of a transmission period), Cx. pipiens (92%) was an absolute dominant species in August. The autogenous form of Cx. pipiens was most common in the flooded-basement houses of the city's outskirts. The number of anautogenous females in the indoor collections increased by August. Only the anautogenous form of Cx. pipiens was sampled in the open rural and urban areas. The findings suggest that the attraction of potential vectors, including anautogenous Cx. pipiens, from the outdoor biotopes to the porches of apartment houses is one of the main reasons for the spread of the disease in the urban population.

[A complex of blood-sucking mosquitoes (Diptera, Culicidae) in the focus of West Nile fever in the Volgograd Region. III. Species feeding on birds and man and the rhythms of their nocturnal activity].

The rate and nocturnal rhythm of mosquito attacks of birds and human beings were studied in the open biotopes of Volgograd and its vicinity in 2004. Thirteen and 11 species of the subfamily Culicinae were collected under the Berezantsev bell and from the traps containing a chicken (a hen), respectively; of them 9 species were common. The mosquitoes of an Anopheles maculipennis complex were caught in a small portion to the traps of both types. Most species of Aedes were highly anthropophilic, showed the minimum activity at night and their abundance considerably decreased by the early transmission period. Among the species that were active during the transmission period, Ae. vexans, Coq. richiardii, and Cx. modestus more intensively attacked a human being than birds and Cx. pipiens was frequently attracted into the hen traps. The attraction of each species of the caught varied during the transmission period. The maximum attacks of Cx. modestus and Cx. pipiens on man and birds coincide and those of Coq. Richiardii and Cx. pipiens on man was observed earlier than on birds. A possible role of mosquitoes of different species in the epizootic and epidemiological processes is discussed.

[Analysis of competitive capacity of male mosquitoes (Diptera, Culicidae)].

The reproductive behavior of insects is characterized by two types of sexual encounter systems: 1) systems in which males have a resource required for a female and 2) the so-called resource-free systems in while a male has nothing other than sperm to give a female. Selection of the most competitive males in the first-type systems is usually considered from the sexual selection concept, i.e. male resource competition and female selection of a partner that has a better a resource, and appreciated from the individual reproductive success of individuals. The application of this approach to the second-type systems offers apparent problems. The data on possible mechanisms of male competition in the resource-free sexual encounter systems are analyzed using the swarming types of mosquitoes. It is shown that selection of competitive males may occur at several stages of reproductive activity and is determined by both the physiological and behavioral signs of individuals.

[Uterine pathomorphology in abruptio placentae].

15 uteri removed surgically because of abruptio placentae (AP) were studied morphologically. AP develops as a process due to a combination of maternal (endometritis, gestosis, adenomyosis), placental (pathology of microvilli, hypercoagulation of venous blood) and hemorheological (thrombosis of placental bed veins, partial blockade of venous circulation) factors. These factors result in the formation of retroplacental hematoma and progressing intrauterine fetal hypoxia.

[The study of mosquito fauna (Diptera, Culicidae) in Volgograd city in light of the outbreak of West Nile fever in Volgograd region, 1999]. / Izuchenie fauny krovososushchikh komarov (Diptera,Culicidae) g.Volgograda v sviazi so vspyshkoi likhoradki Zapadnogo Nila v Volgogradskoi oblasti v 1999 g.

Mosquito sampling was carried out in Volgograd city and its vicinity in August 2001 and 2002. In total 16,000 individuals belonging to 6 genera and 12 species were collected. Nine species were anthropophilic. Culex modestus and Aedes vexans dominated in all outdoor samples collected in Volgograd city. In addition to these species, Coquillettidia richiardii and Ae. caspius were abundant in the vicinity of Volgograd. Autogenous Cx. popiens dominated among six species sampled indoors in Volgograd city. The possible role of different mosquito species in West Nile virus circulation in Volgograd city is discussed.

[Nocturnal biting activity of mosquitoes (Diptera, Culicidae) in Volgograd and its environs]. / Nochnaia aktivnost' komarov (Diptera, Cilicidae) v Volgograde i ego okrestnostiakh.

The nocturnal biting activity of mosquitoes was investigated by a Berezansev's bell and bird traps as baits in August 2003. Among 7 man-attacking species, C. modestus, C. pipiens, and Coq. richiardii were most abundant. C. pipens showed two peaks of biting activity at night while both C. modestus and Coq. richiardii displayed one peak of this activity. Bird traps attracted the mosquitoes C. pipiens and C. modestus. The results are discussed in terms of the possible involvement of the above species in the transmission of WNF virus in Volgograd and its environs.

[Specific features of performing anesthesia and artificial circulation in cardiosurgical interventions during pregnancy]. / Osobennosti provedeniia anestezii i iskusstvennogo krovoobrashcheniia pri kardiokhirurgicheskikh vmeshatel'stvakh na fone beremennosti.

The authors summed up the main principles of performing anesthesia and artificial blood circulation in cardiosurgical interventions in 5 women with 20-27 week pregnancy. In pregnant women with progressing cardiac failure due to a heart disease, thrombosis of the cardiac valve prosthesis, acute infectious endocarditis with the altered valve, large size tumor of the heart with disturbed valve function an emergency operation must be performed under conditions of artificial circulation. Operative treatment is the only effective method for saving life of the mother and fetus. The proposed complex preoperative preparation, intra- and postoperative management of the patients are thought to be an effective method of supporting operations on the heart under conditions of artificial blood circulation in pregnant women. In order to prevent possible complications antioxidants should be included in the complex of anesthesiologic means. During operations with artificial circulation in pregnant women it is necessary to strive for performing normothermal perfusions with high volume velocities.