vniigisРоссийский паразитологический журналRussian Journal of Parasitology1998-84352541-7843ВНИИП – филиал ФГБНУ ФНЦ ВИЭВ РАН10.31016/1998-8435-2024-18-2-170-178vniigis-1169Research ArticleБИОХИМИЯ, БИОТЕХНОЛОГИЯ И ДИАГНОСТИКАBIOCHEMISTRY, BIOTECHNOLOGY AND DIAGNOSTICSАнализ наличия резистентности у паразитических нематод Haemonchus contortus к антигельминтным препаратам бензимидазольного ряда методом гнездовой изотермической амплификации (ПЦР)Analysis of benzimidazole anthelmintic resistance in parasitic nematodes Haemonchus contortus using nested isothermal amplification (PCR)ПименовИ. А.PimenovI. A.

Пименов Илья Александрович, аспирант

117218 Москва, ул. Б. Черемушкинская, 28

Pimenov Ilya A., Postgraduate Student

28, Bolshaya Cheremushkinskaya st., Moscow, 117218

mr.pimenov123@yandex.ruhttps://orcid.org/0000-0001-8364-5055ВарламоваА. И.VarlamovaA. I.

Варламова Анастасия Ивановна, доктор биологических наук

117218 Москва, ул. Б. Черемушкинская, 28

Varlamova Anastasia I., Doctor of Biological Sciences

28, Bolshaya Cheremushkinskaya st., Moscow, 117218

arsphoeb@mail.ruАфанасьевА. Д.AfanasyevA. D.

Афанасьев Алексей Дмитриевич, аспирант

117218 Москва, ул. Б. Черемушкинская, 28

Afanasyev Alexey D., Postgraduate Student

28, Bolshaya Cheremushkinskaya st., Moscow, 117218

a.afanasyev@avgust.comhttps://orcid.org/0000-0002-3644-5592ОдоевскаяИ. М.OdoevskayaI. M.

Одоевская Ирина Михайловна, кандидат биологических наук

117218 Москва, ул. Б. Черемушкинская, 28

Odoevskaya Irina M., Candidate of Biological Sciences

28, Bolshaya Cheremushkinskaya st., Moscow, 117218

odoevskayaim@rambler.ruВсероссийский научно-исследовательский институт фундаментальной и прикладной паразитологии животных и растений – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр – Всероссийский научно-исследовательский институт экспериментальной ветеринарии имени К. И. Скрябина и Я. Р. Коваленко Российской академии наук» (ВНИИП – фил. ФГБНУ ФНЦ ВИЭВ РАН)РоссияAll-Russian Scientific Research Institute for Fundamental and Applied Parasitology of Animals and Plant – a branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV” (VNIIP – FSC VIEV)Russian Federation202428052024182170178Copyright © Пименов И.А., Варламова А.И., Афанасьев А.Д., Одоевская И.М., 20242024Пименов И.А., Варламова А.И., Афанасьев А.Д., Одоевская И.М.Pimenov I.A., Varlamova A.I., Afanasyev A.D., Odoevskaya I.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vniigis.elpub.ru/jour/article/view/1169

Цель исследования – провести мониторинг фермерских хозяйств, расположенных на территории Европейской части РФ, на предмет выявления резистентности к воздействию антигельминтных препаратов из группы бензимидазолов в популяциях нематод Haemonchus contortus, паразитирующих в желудочно-кишечном тракте у мелкого рогатого скота.

Материалы и методы

Материалы и методы. Исследования проводили в 2023–2024 гг. на убойных пунктах, расположенных в Московской области. На первом этапе была проведена таксономическая идентификация паразитических нематод и личинок (L3), определена видовая принадлежность стронгилят от овец. Материалом для исследования служили сычуги с фрагментами 12-перстной кишки и дистальный фрагмент прямой кишки с содержащимися в ней фекалиями. Для молекулярных исследований были использованы половозрелые нематоды и личинки L3 H. contortus, выделенные из сычугов и фекалий мелкого рогатого скота, привезённого на убойные пункты в Московской области из 8 регионов Европейской части РФ – Московской, Астраханской, Орловской, Липецкой, Тульской, Брянской областей, Ставрополья и Дагестана. Исследования проводили на базе лаборатории молекулярной биологии Всероссийского НИИ фундаментальной и прикладной паразитологии животных и растений – фил. ФГБНУ ФНЦ ВИЭВ РАН. Проведена статистическая обработка полученных данных, определены средние показатели заражённости паразитическими нематодами (ИИ и ЭИ). Было исследовано 56 проб ДНК нематод H. contortus методом гнездовой изотермической амплификации (ПЦР) на предмет выявления наличия аллелей генов, определяющих наличие резистентности к препаратам из группы бензимидазолов.

Результаты и обсуждение

Результаты и обсуждение. При проведении молекулярно-генетических исследований ДНК H. contortus, отобранных от овец, прибывших из разных регионов, гомозиготные особи (100%), резистентные к бензимидазолу, были обнаружены только в популяции паразитических нематод из Орловской области. В остальных областях были выявлены только гомозиготные и гетерозиготные особи, восприимчивые к бензимидазолу.

The purpose of the research is to monitor farms located in the European part of the Russian Federation to identify resistance to effects of benzimidazole anthelmintics in populations of nematodes Haemonchus contortus dwelling in the gastrointestinal tract of small cattle.

Materials and methods

Materials and methods. The studies were conducted in slaughterhouses located in the Moscow Region in 2023–2024. At the first stage, taxonomic identification of parasitic nematodes and larvae (L3) was made, and Strongylata species was determined from sheep. The study material was the abomasum with duodenum fragments and a distal rectum fragment with feces. For molecular studies, we used mature nematodes and H. contortus L3 larvae isolated from the abomasum and feces of small cattle brought to slaughterhouses in the Moscow Region from 8 regions of the European part of the Russian Federation: Moscow, Astrakhan, Oryol, Lipetsk, Tula, Bryansk regions, Stavropol and Dagestan. The studies were conducted at the premises of the Laboratory of Molecular Biology, the VNIIP – FSC VIEV. Statistical processing of the obtained data was made, and mean infection rates of parasitic nematodes (infection intensity and prevalence) were determined. Fifty-six DNA samples of nematodes H. contortus were examined using nested isothermal amplification (PCR) to identify gene alleles that determine resistance to benzimidazole drugs.

Results and discussion

Results and discussion. Molecular genetic studies of H. contortus DNA sampled from sheep brought from different Regions only detected homozygous individuals (100%) resistant to benzimidazole in the parasitic nematode population from the Oryol Region. Other regions identified only homozygous and heterozygous individuals susceptible to benzimidazole.

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The authors declare that there are no conflicts of interest present.