Трансмиссивные паразитарные зоонозы Калужской области

the purpose of the research is monitoring population and species composition of common blood-sucking ectoparasites of the Kaluga Region, mosquitoes and ixodid ticks, and parasitic zoonoses, in the circulation of which they are involved.Materials and methods. The records were made in all districts of the Kaluga Region and in the city of Kaluga. We studied open meadow-field and forest-shrub stations, closed meadow-field and near-water stations, and settlement stations. The species was identified using the atlases of ixodid ticks by I. M. Ganiev, A. A. Aliverdiev (1968) and V. N. Shevkoplyas (2008), and the guidance of R. M. Gornostaeva (1999). The situation with transmissible parasitic zoonoses in the Kaluga Region was assessed based on the analysis of information from open sources of the Hygienic and Epidemiological Center of the Kaluga Region and the Veterinary Committee under the Government of the Kaluga Region.Results and discussion. There are two species of ixodid ticks in the Kaluga region: Ixodes ricinus and Dermacentor reticulatus. The abundance index of ixodid ticks of the species I. ricinus is 16.8±1.32 individuals per 1 flag-hour in forest biotopes, and 11.6±1.12 individuals per 1 flag-hour in meadow biotopes, and the abundance index of D. reticulatus is 10,8±1.14 and 15.9±1.30 individuals per 1 flag-hour, respectively. Three mosquito species prevail, namely, Stegomya communis, S. vexans, and Culex pipiens. The maximum number was 8 thousand per 1 m3. During the study period, the causative agent of infectious tick-borne borreliosis was found on average in 10.6% of ixodid ticks collected from humans and in 13.2% of ticks collected in natural biotopes. The causative agent of human granulocytic anaplasmosis was isolated on average in 3.1% of ixodid ticks collected from humans and 6.4% of those collected in natural biotopes. Infection rate for I. ricinus ticks infected with pathogen of infectious tick-borne borreliosis was 16.9%, and D. reticulatus 12.3%.Цель исследований: провести мониторинг численности, видового состава распространенных кровососущих эктопаразитов Калужской области – комаров и иксодовых клещей и паразитарных зоонозов, в циркуляции которых они участвуют.Материалы и методы. Учеты проводили на территории всех районов Калужской области и в г. Калуге. Были исследованы открытые луго-полевые, лесокустарниковые стации, закрытые луго-полевые, околоводные стации и стации населенных пунктов. Видовую принадлежность определяли, используя атласы иксодоидных клещей И. М. Ганиева, А. А. Аливердиева (1968), В. Н. Шевкопляса (2008) и руководство Р. М. Горностаевой (1999). Ситуацию по паразитарным трансмиссивным зоонозам Калужской области оценивали по результатам анализа информации открытых источников Центра гигиены и эпидемиологии Калужской области и Комитета ветеринарии при Правительстве Калужской области.Результаты и обсуждение. В Калужской области обитают два вида иксодовых клещей: Ixodes ricinus и Dermacentor reticulatus. Индекс обилия иксодовых клещей вида I. ricinus в лесных биотопах составляет 16,8±1,32 особей на 1 флаго-час, в луговых биотопах 11,6±1,12 особей на 1 флаго-час, индекс обилия D. reticulatus соответственно 10,8±1,14 и 15,9±1,30 особей на 1 флаго-час. Преобладают три вида комаров: Stegomya communis, S. vexans, Culex pipiens. Максимальная численность составила 8 тыс. экз. на 1 м3. В период исследований возбудитель инфекционного клещевого боррелиоза обнаружен в среднем у 10,6% иксодовых клещей, собранных с людей и у 13,2% клещей, собранных в природных биотопах. Возбудитель гранулоцитарного анаплазмоза человека выделен в среднем у 3,1% иксодовых клещей, собранных с людей и 6,4%, собранных в природных биотопах. Зараженность клещей I .ricinus возбудителем инфекционного клещевого боррелиоза составила 16,9%, D. reticulatus – 12,3%

Особенности паразитирования комаров вида Culex pipiens Culex Linnaeus, 1758 (Diptera, Culicidae) на территории Калужской области

The purpose of the research is to study and systematize data on the biological, fauna-ecological characteristics of mosquitoes Culex pipiens Culex Linnaeus species in the Kaluga Region.Materials and methods. To clarify the fauna and ecological characteristics of mosquitoes Culex pipiens Culex Linnaeus species in the Kaluga Region, we caught adults, larvae and pupae of mosquitoes in the basements of the city of Kaluga, on the control day of mosquitoes, in the natural land biotopes of the Kaluga region. The larvae of blood-sucking mosquitoes were observed on natural and artificial reservoirs of the city and region, in puddles, trenches. Mosquitoes were caught during an attack on a person or animal with an entomological net and placed in a specially prepared cage. Larvae and pupae were collected at breeding sites (from aquatic plants, in the water column). The study of the biotopic distribution and daily activity was carried out using the method of accounting for the number of insects attacking the human forearm for 20 minutes in the period from 20:00–24:00 h.Results and discussion. Two forms of mosquitoes Culex pipiens Culex Linnaeus, 1758 (Diptera, Culicidae) are found in the Kaluga Region: Cx. pipiens f. pipiens L. (non-autogenous form) and Cx. p. f. molestus fors. (autogenous form). Form Cx. pipiens f. pipiens L. is very aggressive, lives in urban and agricultural areas. Cx p. f. molestus fors. is an urbanized form; prefers to live and develop in urban environments. A feature is also the possibility of the first laying of eggs without bloodsucking, and the larvae can develop without sunlight. In late August, female C. pipiens mosquitoes can enter diapause before spring. A temperature below 4°C and above 35°C is critical for the development of larvae. In a year, 3–4 mosquito generations develop in the Kaluga Region. Цель исследований: изучить и систематизировать данные о биологических, фауно-экологических особенностях комаров вида Culex pipiens Culex Linnaeus в Калужской области.Материалы и методы. Для выяснения фауно-экологических особенностей комаров вида Culex pipiens Culex Linnaeus на территории Калужской области проводили отлов имаго, личинок и куколок комаров в подвальных помещениях г. Калуги, на контрольной дневке комаров, в природных наземных биотопах Калужской области. За личинками кровососущих комаров вели наблюдения на естественных и искусственных водоемах города и области, в лужах, траншеях. Комаров отлавливали во время нападения на человека или животное энтомологическим сачком и помещали в специально приготовленный садок. Личинок и куколок собирали в местах выплода (с водных растений, в толще воды). Изучение биотопического распределения и суточной активности проводили, используя метод учета числа насекомых, нападающих на предплечье человека в течение 20 мин. в промежуток времени 20:00–24:00 ч.Результаты и обсуждение. На территории Калужской области встречаются две формы комаров вида Culex pipiens Culex Linnaeus, 1758 (Diptera, Culicidae): Cx. pipiens f. pipiens L. (неавтогенная форма) и Cx. p. f. molestus Fors. (автогенная форма). Форма Cx. pipiens f. pipiens L. очень агрессивна, обитает на городских и сельскохозяйственных территориях. Cx. p. f. molestus Fors. является урбанизированной формой; предпочитает жить и развиваться в городских условиях. Особенностью является также возможность первой откладки яиц без кровососания, а личинки способны развиваться без солнечного света. В конце августа самки комаров Culex pipiens могут вступать в диапаузу до весны. Температура ниже 4 оС и выше 35 оС является критической для развития личинок. За год на территории Калужской области развивается 3–4 генерации комаров.

Regression mathematical modeling of the population of mouse-like rodents, hosts of blood-sucking arthropods in the Non-Black Earth Zone by the example of the Kaluga Region

The purpose of the research is a regression mathematical modeling of the population of small mammals, hosts of ixodid ticks in the Kaluga Region, which allows assessing the likelihood of zoonotic and vector-borne diseases.Materials and methods. For 10 years (from 2009 to 2019), a 2k multifactorial experiment was performed in the field by the method of V. V. Kalmykov (2016). The study object was mouse-like rodents that are the most common in the Kaluga Region. Rodents were counted by standard methods using techniques of V. N. Shnitnikov (1929), P. B. Yurgenson (1934) and A. N. Formozov (1937). The generally accepted steel spring traps were used. Animals were counted throughout all habitats.Results and discussion. Regression mathematical models of the mouse-like rodent population were obtained depending on the average monthly temperature, precipitation and atmospheric pressure for the year in the Non-Black Earth Zone by the example of the Kaluga Region. The specific nature of the obtained analytical model is that the greatest influence on the population of small mammals is made by the interaction effect of two factors, temperature and atmospheric pressure. Their impact is more significant than the influence of each of the three study factors separately, in particular, it is stronger than the only influence of temperature by 1.02, the only impact of precipitation by 2.58, and the only impact of atmospheric pressure by 2.72. The analytic mathematical model allows us to calculate, without significant material or time costs, populations of mouse-like rodents not only in the Kaluga Region, but also in regions with similar climate conditions

Фауно-экологические особенности паразитирования иксодовых клещей Центральной части Восточно-Европейской равнины

The purpose of the research is finding out features of fauna and ecology of ixodid ticks parasitizing in the Non-Black Earth Region of the central part of the East European Plain, which inhabit the Kaluga Region. Materials and methods. Ixodid ticks were collected and recorded according to generally accepted methods in all districts of the Kaluga Region and the city of Kaluga in 2009–2019 during their activity (in the spring from the melting of snow and until late autumn before the snow cover formation). A total of 11,282 ticks were collected in 412 flag-hours, of which 7,421 (65.7%) were collected from vegetation and 3,861 (34.3%) from animals. We studied open meadow and field areas, forest and shrub areas, closed meadow and field areas, and wetland stations and settlements. The species was identified using the Atlas of ixodid ticks by Ganiev and Aliverdiev (1968) and the Atlas by Shevkoplyas (2008). Results and discussion. There are two species of ixodid ticks in the Kaluga Region, Ixodes ricinus and Dermacentоr reticulatus. The number of D. reticulatus slightly exceeds (by 6%) I. ricinus, 53 and 46% respectively, which is explained by the even distribution of forest and meadow (pasture) biotopes in the Region. The abundance index of I. ricinus was 16.8±1.32 individuals per 1 flag-hour in forest biotopes, and 11.6±1.12 individuals per 1 flag-hour in meadow biotopes. The abundance index of D. reticulatus was 10.8±1.14 individuals per 1 flag-hour in forest biotopes, and 15.9±1.30 individuals per 1 flag-hour in meadow biotopes.Цель исследований – выяснить фауно-экологические особенности паразитирования иксодовых клещей, обитающих в Нечерноземной зоне Центральной части Восточно-Европейской равнины. Материалы и методы. Сбор и учет иксодовых клещей проводили по общепринятым методикам на территории всех районов Калужской области и г. Калуги в 2009–2019 гг. в период их активности (весной со схода снега и до поздней осени до установления снежного покрова). Всего собрано 11 282 клеща за 412 флаго-часов, из них 7 421 экз. (65,7%) собрано с растительности и 3861 экз. (34,3%) – с животных. Были исследованы открытые луго-полевые, лесокустарниковые стации, закрытые луго-полевые, околоводные стации и стации населенных пунктов. Видовую принадлежность определяли, используя атлас иксодовых клещей Ганиева, Аливердиева (1968) и атлас Шевкопляса (2008). Результаты и обсуждение. В Калужской области обитает два вида иксодовых клещей: Ixodes ricinus и Dermacentоr reticulatus. Численность D. reticulatus незначительно превышает (на 6%) таковую I. ricinus: 53 и 46% соответственно, что объясняется равномерным распределением лесных и луговых (пастбищных) биотопов на территории области. Индекс обилия вида I. ricinus в лесных биотопах составил 16,8±1,32 экз. на 1 флаго-час, в луговых биотопах – 11,6±1,12 экз. на 1 флаго-час. Индекс обилия вида D. reticulatus составил в лесных биотопах 10,8±1,14 экз. на 1 флаго-час, в луговых биотопах – 15,9±1,30 особей на 1 флаго-час

Fenfluramine hydrochloride for the treatment of seizures in Dravet syndrome: a randomised, double-blind, placebo-controlled trial

BACKGROUND: Dravet syndrome is a rare, treatment-resistant developmental epileptic encephalopathy characterised by multiple types of frequent, disabling seizures. Fenfluramine has been reported to have antiseizure activity in observational studies of photosensitive epilepsy and Dravet syndrome. The aim of the present study was to assess the efficacy and safety of fenfluramine in patients with Dravet syndrome. METHODS: In this randomised, double-blind, placebo-controlled clinical trial, we enrolled children and young adults with Dravet syndrome. After a 6-week observation period to establish baseline monthly convulsive seizure frequency (MCSF; convulsive seizures were defined as hemiclonic, tonic, clonic, tonic-atonic, generalised tonic-clonic, and focal with clearly observable motor signs), patients were randomly assigned through an interactive web response system in a 1:1:1 ratio to placebo, fenfluramine 0·2 mg/kg per day, or fenfluramine 0·7 mg/kg per day, added to existing antiepileptic agents for 14 weeks. The primary outcome was the change in mean monthly frequency of convulsive seizures during the treatment period compared with baseline in the 0·7 mg/kg per day group versus placebo; 0·2 mg/kg per day versus placebo was assessed as a key secondary outcome. Analysis was by modified intention to treat. Safety analyses included all participants who received at least one dose of study medication. This trial is registered with ClinicalTrials.gov with two identical protocols NCT02682927 and NCT02826863. FINDINGS: Between Jan 15, 2016, and Aug 14, 2017, we assessed 173 patients, of whom 119 patients (mean age 9·0 years, 64 [54%] male) were randomly assigned to receive either fenfluramine 0·2 mg/kg per day (39), fenfluramine 0·7 mg/kg per day (40) or placebo (40). During treatment, the median reduction in seizure frequency was 74·9% in the fenfluramine 0·7 mg/kg group (from median 20·7 seizures per 28 days to 4·7 seizures per 28 days), 42·3% in the fenfluramine 0·2 mg/kg group (from median 17·5 seizures per 28 days to 12·6 per 28 days), and 19·2% in the placebo group (from median 27·3 per 28 days to 22·0 per 28 days). The study met its primary efficacy endpoint, with fenfluramine 0·7 mg/kg per day showing a 62·3% greater reduction in mean MCSF compared with placebo (95% CI 47·7-72·8, p<0·0001); fenfluramine 0·2 mg/kg per day showed a 32·4% reduction in mean MCSF compared with placebo (95% CI 6·2-52·3, p=0·0209). The most common adverse events (occurring in at least 10% of patients and more frequently in the fenfluramine groups) were decreased appetite, diarrhoea, fatigue, lethargy, somnolence, and decreased weight. Echocardiographic examinations revealed valve function within the normal physiological range in all patients during the trial and no signs of pulmonary arterial hypertension. INTERPRETATION: In Dravet syndrome, fenfluramine provided significantly greater reduction in convulsive seizure frequency compared with placebo and was generally well tolerated, with no observed valvular heart disease or pulmonary arterial hypertension. Fenfluramine could be an important new treatment option for patients with Dravet syndrome. FUNDING: Zogenix

Регрессионное математическое моделирование популяции мышевидных грызунов – хозяев кровососущих членистоногих зоны Нечерноземья на примере Калужской области

The purpose of the research is a regression mathematical modeling of the population of small mammals, hosts of ixodid ticks in the Kaluga Region, which allows assessing the likelihood of zoonotic and vector-borne diseases.Materials and methods. For 10 years (from 2009 to 2019), a 2k multifactorial experiment was performed in the field by the method of V. V. Kalmykov (2016). The study object was mouse-like rodents that are the most common in the Kaluga Region. Rodents were counted by standard methods using techniques of V. N. Shnitnikov (1929), P. B. Yurgenson (1934) and A. N. Formozov (1937). The generally accepted steel spring traps were used. Animals were counted throughout all habitats.Results and discussion. Regression mathematical models of the mouse-like rodent population were obtained depending on the average monthly temperature, precipitation and atmospheric pressure for the year in the Non-Black Earth Zone by the example of the Kaluga Region. The specific nature of the obtained analytical model is that the greatest influence on the population of small mammals is made by the interaction effect of two factors, temperature and atmospheric pressure. Their impact is more significant than the influence of each of the three study factors separately, in particular, it is stronger than the only influence of temperature by 1.02, the only impact of precipitation by 2.58, and the only impact of atmospheric pressure by 2.72. The analytic mathematical model allows us to calculate, without significant material or time costs, populations of mouse-like rodents not only in the Kaluga Region, but also in regions with similar climate conditions.Цель исследований – провести регрессионное математическое моделирование популяции мелких млекопитающих – прокормителей иксодовых клещей Калужской области, позволяющее оценить вероятность вспышек зоонозных, трансмиссивных болезней.Материалы и методы. На протяжении 10 лет (с 2009 по 2019 гг.) проводили многофакторный эксперимент типа 2к в полевых условиях по методике В. В. Калмыкова (2016). Объектом исследования были мышевидные грызуны, наиболее распространенные на территории Калужского региона. Учет численности грызунов проводили стандартными методами с использованием техник В. Н. Шнитникова (1929), П. Б. Юргенсона (1934) и А. Н. Формозова (1937). Использовали общепринятые ловушки Геро. Животных учитывали на всей территории области всех стаций обитаний.Результаты и обсуждение. Получены регрессионные математические модели количества популяции мышевидных грызунов в зависимости от среднемесячных показателей температуры, уровня осадков и атмосферного давления за год в зоне Нечерноземья на примере Калужской области. Особенность полученной аналитической модели заключается в том, что наибольшее влияние на популяцию мелких млекопитающих оказывает эффект взаимодействия двух факторов: температуры и атмосферного давления. Сила его влияния значительнее, чем влияние каждого из трех исследуемых факторов в отдельности, в частности, сильнее, чем степень влияния одной температуры в 1,02 раза, чем сила влияния одного фактора количества осадков в 2,58 раз, чем сила влияния одного атмосферного давления в 2,72 раз. Расчетная математическая модель дает возможность без существенных материальных, временных затрат рассчитывать численность популяций мышевидных грызунов не только на территории Калужской области, но и в регионах с похожими условиями климата

Testing association of rare genetic variants with resistance to three common antiseizure medications

OBJECTIVE: Drug resistance is a major concern in the treatment of individuals with epilepsy. No genetic markers for resistance to individual antiseizure medication (ASM) have yet been identified. We aimed to identify the role of rare genetic variants in drug resistance for three common ASMs: levetiracetam (LEV), lamotrigine (LTG), and valproic acid (VPA). METHODS: A cohort of 1622 individuals of European descent with epilepsy was deeply phenotyped and underwent whole exome sequencing (WES), comprising 575 taking LEV, 826 LTG, and 782 VPA. We performed gene- and gene set-based collapsing analyses comparing responders and nonresponders to the three drugs to determine the burden of different categories of rare genetic variants. RESULTS: We observed a marginally significant enrichment of rare missense, truncating, and splice region variants in individuals who were resistant to VPA compared to VPA responders for genes involved in VPA pharmacokinetics. We also found a borderline significant enrichment of truncating and splice region variants in the synaptic vesicle glycoprotein (SV2) gene family in nonresponders compared to responders to LEV. We did not see any significant enrichment using a gene-based approach. SIGNIFICANCE: In our pharmacogenetic study, we identified a slightly increased burden of damaging variants in gene groups related to drug kinetics or targeting in individuals presenting with drug resistance to VPA or LEV. Such variants could thus determine a genetic contribution to drug resistance

Mutations in GABRB3

Objective: To examine the role of mutations in GABRB3 encoding the b3 subunit of the GABAA receptor in individual patients with epilepsy with regard to causality, the spectrum of genetic variants, their pathophysiology, and associated phenotypes. Methods: We performed massive parallel sequencing of GABRB3 in 416 patients with a range of epileptic encephalopathies and childhood-onset epilepsies and recruited additional patients with epilepsy with GABRB3 mutations from other research and diagnostic programs. Results: We identified 22 patients with heterozygous mutations in GABRB3, including 3 probands frommultiplex families. The phenotypic spectrum of the mutation carriers ranged from simple febrile seizures, genetic epilepsies with febrile seizures plus, and epilepsy withmyoclonic-atonic seizures to West syndrome and other types of severe, early-onset epileptic encephalopathies. Electrophysiologic analysis of 7 mutations in Xenopus laevis oocytes, using coexpression of wild-type or mutant beta(3), together with alpha(5) and gamma(2s) subunits and an automated 2-microelectrode voltage-clamp system, revealed reduced GABA-induced current amplitudes or GABA sensitivity for 5 of 7 mutations. Conclusions: Our results indicate that GABRB3 mutations are associated with a broad phenotypic spectrum of epilepsies and that reduced receptor function causing GABAergic disinhibition represents the relevant disease mechanism

Rare coding variants in genes encoding GABA_A receptors in genetic generalised epilepsies: an exome-based case-control study

BACKGROUND: Genetic generalised epilepsy is the most common type of inherited epilepsy. Despite a high concordance rate of 80% in monozygotic twins, the genetic background is still poorly understood. We aimed to investigate the burden of rare genetic variants in genetic generalised epilepsy. METHODS: For this exome-based case-control study, we used three different genetic generalised epilepsy case cohorts and three independent control cohorts, all of European descent. Cases included in the study were clinically evaluated for genetic generalised epilepsy. Whole-exome sequencing was done for the discovery case cohort, a validation case cohort, and two independent control cohorts. The replication case cohort underwent targeted next-generation sequencing of the 19 known genes encoding subunits of GABAA receptors and was compared to the respective GABAA receptor variants of a third independent control cohort. Functional investigations were done with automated two-microelectrode voltage clamping in Xenopus laevis oocytes. FINDINGS: Statistical comparison of 152 familial index cases with genetic generalised epilepsy in the discovery cohort to 549 ethnically matched controls suggested an enrichment of rare missense (Nonsyn) variants in the ensemble of 19 genes encoding GABAA receptors in cases (odds ratio [OR] 2·40 [95% CI 1·41-4·10]; pNonsyn=0·0014, adjusted pNonsyn=0·019). Enrichment for these genes was validated in a whole-exome sequencing cohort of 357 sporadic and familial genetic generalised epilepsy cases and 1485 independent controls (OR 1·46 [95% CI 1·05-2·03]; pNonsyn=0·0081, adjusted pNonsyn=0·016). Comparison of genes encoding GABAA receptors in the independent replication cohort of 583 familial and sporadic genetic generalised epilepsy index cases, based on candidate-gene panel sequencing, with a third independent control cohort of 635 controls confirmed the overall enrichment of rare missense variants for 15 GABAA receptor genes in cases compared with controls (OR 1·46 [95% CI 1·02-2·08]; pNonsyn=0·013, adjusted pNonsyn=0·027). Functional studies for two selected genes (GABRB2 and GABRA5) showed significant loss-of-function effects with reduced current amplitudes in four of seven tested variants compared with wild-type receptors. INTERPRETATION: Functionally relevant variants in genes encoding GABAA receptor subunits constitute a significant risk factor for genetic generalised epilepsy. Examination of the role of specific gene groups and pathways can disentangle the complex genetic architecture of genetic generalised epilepsy. FUNDING: EuroEPINOMICS (European Science Foundation through national funding organisations), Epicure and EpiPGX (Sixth Framework Programme and Seventh Framework Programme of the European Commission), Research Unit FOR2715 (German Research Foundation and Luxembourg National Research Fund)