Prevalence, Infection Intensity and Molecular Diagnosis of Mixed Infections with Metastrongylus spp. (Metastrongylidae) in Wild Boars in Uzbekistan

The aim of the present study was to characterize the diversity of Metastrongylus spp. in wild boars and the earthworm intermediate host species contributing to the maintenance of the life cycle. Here, wild boars were subjected to parasitological necropsies, and lungworm species were identified morphologically, followed by confirmation using ITS-2 sequencing and a phylogenetic analysis. Earthworms were collected from wild boar habitats and investigated for the presence of larvae. The prevalence of Metastrongylus spp. in wild boars was 78.8%, and many individuals were positive for all three detected species, Metastrongylus pudendotectus, Metastrongylus salmi and Metastrongylus elongatus. The phylogenetic analysis did not clearly resolve all species, except for M. pudendotectus. Age group and season had no influence on prevalence, while intensity was significantly higher in autumn than in spring and summer (Kruskal–Wallis followed by Dunn’s test). Three out of six investigated earthworm species were positive for metastrongyloid larvae (prevalence of 10.4–16.7%), but neither their phylogenetic relationship nor ecological microhabitats were able to explain these differences. Further sequence data should be used to improve the resolution in phylogenetic trees to determine potential cryptic species in the genus, while the application of deep sequencing approaches might provide insights into species-specific epidemiology and pathology

Molecular analysis of polymorphic species of the genus Marshallagia (Nematoda: Ostertagiinae)

Background: The genus Marshallagia (Family Haemonchidae, subfamily Ostertagiinae) contains multiple species of nematodes parasitising the abomasum (or duodenum) of ruminants, in particular of Caprinae. Male specimens have been described to be polymorphic with the frequent/major morphotype initially described in the genus Marshallagia while the minor/rare morphotype was initially often placed in the genus Grossospicularia. Due to common morphological features, certain pairs of morphotypes were suggested to belong to the same species such as Marshallagia marshalli/M. occidentalis. However, molecular evidence to confirm these pairs of morphotypes belonging to the same species is missing. Methods: In the present study, Marshallagia sp. were collected from domestic sheep in Uzbekistan. Male specimens were morphologically described with particular emphasis on the structure of the bursa copulatrix. After DNA isolation from morphologically identified specimens, PCRs targeting the ribosomal internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (cox1) regions were conducted. After Sanger sequencing, maximum likelihood phylogenetic analyses and pairwise identities between sequences were calculated. Results: The major morphotypes of M. marshalli, M. schumakovitschi and M. uzbekistanica and the minor morphotypes M. occidentalis, M. trifida and M. sogdiana were identified and their morphology was documented in detail. ITS2 sequences showed little variation and did not allow diagnosing species. In contrast, phylogenetic analysis of cox1 sequences identified highly supported clusters and verified that M. marshalli, M. occidentalis and M. uzbekistanica are different morphotypes of the species M. marshalli while M. schumakovitschi and M. trifida represent distinct morphotypes of M. trifida. For M. sogdiana no corresponding major morphotype could be identified in the present study. Due to a large barcoding gap, comparison of cox1 sequences in terms of percent identity was sufficient to reliably assign the sequences to a particular species without phylogenetic analysis. Conclusions: The data presented here create a framework that will allow the classification of other members of the genus in the future and underline that parallel morphological and molecular analysis of specimens is crucial to improve the taxonomy of polymorphic species

НАЗЕМНЫЕ МОЛЛЮСКИ УЗБЕКИСТАНА - ПРОМЕЖУТОЧНЫЕ ХОЗЯЕВА ПРОТОСТРОНГИЛИД (NEMATODA: PROTOSTRONGYLIDAE)

Objective of research: To determine the intermediate hosts of nematodes and their role in the distribution of causative agents of animal protostrongylidosis in Uzbekistan. Materials and methods: Research was conducted in 2010-2015 in biogeocenoses of Fergana valley (Namangan, Fergana, Andijan regions), in the north-west of Uzbekistan (Tashkent, Syrdarya and Zhizzakh regions). Natural invasion of land mollusks - intermediate hosts of helminths was investigated under field conditions. To identify the species belonging of land mollusks, we carried out a standard anatomization of reproductive system of mollusks using Magnifier MBS-9. 10554 individuals of land mollusks were investigated by the method of Azimov et al. and by compressor method of Boev. To conduct a morphological study of third-stage protostrongylid larvae, feet of infected mullusks were separated and placed into artificial gastric juice where the cap was destroyed and infected larvae released. Results and discussion: Terrestrial mollusks of 11 genera Vallonia, Gibbulinopsis, Pupilla, Pseudonapaeus, Bradybaena, Angiomphalia, Xeropicta, Deroceras, Candaharia, Maсrochlamys and Succinea were identified as intermediate hosts of Protostrongylidae in Uzbekistan. Their infestation with protostrongylid larvae was on average 21,0 %. Among the mollusks examined, X. candaсharica (33,3 %) proved to be the most infected with nematode larvae. In plain and foothill-mountain zones, protostrongylid larvae are detected in these mollusks in May. In plain, the peak of invasion was registered in July (18.3%), in foothills and mountain range characterized by two rises in July (38.2%) and October (43.5).Цель исследования - установление промежуточных хозяев нематод и их роли в распространении возбудителей протостронгилидозов животных Узбекистана. Материалы и методы. Исследования проводили в 2010-2015 гг. в биогеоценозах Ферганской долины (Наманганской, Ферганской и Андижанской областях), северо-востока Узбекистана (Ташкентской, Сырдарьинской и Жиззахской областях). В полевых условиях изучена естественная зараженность наземных моллюсков - промежуточных хозяев гельминтов. Для установления видовой принадлежности наземных моллюсков проводили стандартное анатомирование репродуктивной системы моллюсков с использованием бинокулярной лупы МБС-9. Исследовано 10554 особей наземных моллюсков по методу Азимова и др. и компрессорным методом Боева. Для морфологического изучения личинок третьей стадии протостронгилид отделяли ножки зараженных моллюсков и помещали их в искусственный желудочный сок, в котором разрушался чехлик и освобождались инвазионные личинки. Результаты и обсуждение. В качестве промежуточных хозяев протостронгилид Узбекистана зарегистрированы наземные моллюски 11 родов: Vallonia, Gibbulinopsis, Pupilla, Pseudonapaeus, Bradybaena, Leucozonella, Xeropicta, Deroceras, Candaharia, Macrochlamys и Succinea. Зараженность их личинками протостронгилид составила, в среднем, 21,0 %. Среди исследованных моллюсков широко распространенными и наиболее зараженными личинками нематод оказались X. candaсharica (33,3 %). В равнинном и предгорно-горном поясе личинки протостронгилид в этих моллюсках обнаруживают в мае. Пик инвазии в равнинном поясе приходится на июль (18,3 %), в предгорно-горном поясе характеризуется двумя подъемами: в июле (38,2 %) и октябре (43,5 %)

НЕМАТОДЫ ПИЩЕВАРИТЕЛЬНОЙ СИСТЕМЫ ДОМАШНИХ ЖВАЧНЫХ УЗБЕКИСТАНА

Objective of research. A study of species composition of nematodes parasitizing the digestive system of domestic ruminants in Uzbekistan, analysis of seasonal dynamics of infestation and features of the nematode fauna in plains and foothill-mountain areas.Material and methods. In the period of 2014–2015, the digestive tracts of 206 head of cattle and 878 head of small cattle from the farms of Tashkent, Syr-Darya, Dzhizak, Namangan, Kashkadarya, Surkhandarya, Navoiy and Bukhara regions as well as Karakalpakstan autonomous republic were investigated by the method of full helminthological dissection.Results and discussion. 30 nematode species were found in digestive tracts of domestic ruminants of Uzbekistan; 28 nematode species in sheep, 26 species in goats, and 22 species in cattle. The highest rates of extensity of infection (EI) and intensity of infection (II) were detected in Ostertagia ostertagi, Marshallagia marshalli, Teladorsagia circumcincta and Parabronema skrjabini. An increase of EI and II in summer and autumn period was typical for the most of the discovered nematode species.However, EI in O. ostertagi, T. circumcincta, Marshallagia spp. and P. skrjabini was relatively stable in all seasons, but II increased to autumn. 26 nematode species were registered in plains; only 3 of them with high EI and II.28 nematode species were registered in foothill-mountain areas, 8 of them showed high rates of EI and II. 24 species of nematodes are common in both areas.The obtained results allow us to conclude that it is necessary to improve helminth control of ruminants in Uzbekistan. Alternation of pastures and regulation of grazing, as well as preventive dehelmintization should not be ignored; observe the optimal requirements for feeding and management of animals.Цель исследования — изучение видового состава нематод пищеварительного тракта домашних жвачных Узбекистана, анализ сезонной динамики зараженности, выявление особенностей нематодофауны в равнинной и предгорно-горной зонах.Материалы и методы. В 2014–2015 гг. в хозяйствах Ташкентской, Сырдарьинской, Джизакской, Наманганской, Кашкадарьинской, Сурхандарьинской, Навоийской, Бухарской областей и Автономной Республики Каракалпакстан по методу полного гельминтологического вскрытия отдельных органов исследовано 878 голов мелкого и 206 голов крупного рогатого скота.Результаты и обсуждение. У домашних жвачных Узбекистана зарегистрировано 30 видов нематод пищеварительного тракта, в том числе: у овец 28, у коз 26, у крупного рогатого скота 22 вида. Наиболее высокие показатели, как по экстенсивности инвазии (ЭИ), так и по интенсивности инвазии (ИИ), отмечены у Ostertagia ostertagi, Marshallagia marshalli, Teladorsagia circumcincta, Parabronema skrjabini. Для большинства обнаруженных видов нематод было характерно нарастание ЭИ и ИИ в летние и осенние месяцы. Однако у O. ostertagi, T. circumcincta, Marshallagia spp., P. skrjabini ЭИ была относительно стабильна во все сезоны, а ИИ нарастала к осени. В равнинной зоне зарегистрировано 26 видов нематод, из них высокие показатели ЭИ и ИИ имеют лишь 3 вида. В предгорно-горной зоне зарегистрировано 28 видов нематод, высокие показатели ЭИ и ИИ имеют 8 видов. Для равнинной и предгорно-горной зон общими являются 24 вида нематод. Полученные результаты позволяют сделать вывод о необходимости более активной борьбы с гельминтозами жвачных в Узбекистане. Нельзя игнорировать мероприятия по смене пастбищ, регулированию выпаса, необходимо применять профилактические дегельминтизации, соблюдать оптимальные условия содержания и кормления животных

FAUNA AND ECOLOGY OF NEMATODE OF THE GENUS HAEMONCHUS (NEMATODA: HAEMONCHIDAE) - ENDOPARASITES OF ANIMALS

Currently 13 species of the considered genus parasitizing the abomasums of ungulate animals have been recorded. The ungulates of the family of Cervidae (1) Antilocapridae (1), Giraffidae (1), Bovidae (12) and Camelidae (2) were recorded as definitive hosts of this parasite. Individual populations of the Haemonchus species were recorded in Asia, Europe, America, Africa and Australia. Sex ratio in Haemonchus contortus between females and males is 1:5. One female lays from 150 to 10.000 eggs per da

Prevalence, Infection Intensity and Molecular Diagnosis of Mixed Infections with Metastrongylus spp. (Metastrongylidae) in Wild Boars in Uzbekistan

The aim of the present study was to characterize the diversity of Metastrongylus spp. in wild boars and the earthworm intermediate host species contributing to the maintenance of the life cycle. Here, wild boars were subjected to parasitological necropsies, and lungworm species were identified morphologically, followed by confirmation using ITS-2 sequencing and a phylogenetic analysis. Earthworms were collected from wild boar habitats and investigated for the presence of larvae. The prevalence of Metastrongylus spp. in wild boars was 78.8%, and many individuals were positive for all three detected species, Metastrongylus pudendotectus, Metastrongylus salmi and Metastrongylus elongatus. The phylogenetic analysis did not clearly resolve all species, except for M. pudendotectus. Age group and season had no influence on prevalence, while intensity was significantly higher in autumn than in spring and summer (Kruskal–Wallis followed by Dunn’s test). Three out of six investigated earthworm species were positive for metastrongyloid larvae (prevalence of 10.4–16.7%), but neither their phylogenetic relationship nor ecological microhabitats were able to explain these differences. Further sequence data should be used to improve the resolution in phylogenetic trees to determine potential cryptic species in the genus, while the application of deep sequencing approaches might provide insights into species-specific epidemiology and pathology

LARVAE OF HELMINTHES OF TERRESTRIAL MOLLUSKS OF THE FERGANA VALLEY: FAUNA AND TAXONOMY

In this article \u27rovides information on the systematic com\u27osition, distribution and intermediate host of helminths larvae found on terrestrial mollusks of Fergana Valley. During 2016-2018 in Namangan, Fergana and Andijan regions, investigations were conducted on the infection of terrestrial mollusks by helminths larvae. Result of this study, 21 s\u27ecies of helminth larvae are identified from 23 terrestrial mollusks. Of these, 12 s\u27ecies are trematodes, 2 cestodes, and 7 nematode

MORPHOLOGICAL FEATURES OF HAEMONCHUS CONTORTUS AND H. PLACEI SPECIES (NEMATODA: TRICHOSTRONGYLIDAE)

The morphological features of the nematodes of the genus Haemonchus Cobbold, 1898, which is represented by two species of H. contortus Rudolphi, 1803 and H. placei Place, 1893, were studied. Based on morphological studies, the independence of two species of gemonchus, Haemonchus contortus and Haemonchus placei parasitizing in sheep and cattle, was determined

Identification of nematodes of the genus

The present study delves into a methodological framework aimed at establishing species-specific markers via the utilization of sequences derived from the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA. This method, in conjunction with polymerase chain reaction (PCR) testing, serves as a diagnostic tool for discerning species belonging to the genus Teladorsagia Andreeva et Satubaldin, 1954. These species, constituents of the subfamily Ostertagiinae (Nematoda: Trichostrongylidae), exhibit wide distribution within the gastrointestinal tracts of ruminants across the geographic expanse of Uzbekistan. The heart of this endeavor is the development of species-specific primers, a pioneering creation in its own right. These primers are crafted using sequences emanating from the ITS2 region of the ribosomal DNA, an innovative approach that facilitates the precise identification of morphospecies within the Teladorsagia genus. Notably, the primers exhibit a nucleotide length of 153 base pairs, an attribute instrumental in their capacity to accurately distinguish and diagnose eggs and larvae of three distinct morphspecies: T. circumcincta, T. trifurcata, and T. davtiani. The potential implications of this method are significant, with ramifications reverberating across the field of veterinary diagnostics. Through the application of these primers, practitioners and researchers alike can effectively ascertain the presence of specific Teladorsagia morphospecies in ruminant animals. This holds the promise of not only enhancing diagnostic precision but also contributing to the broader comprehension of the prevalence and distribution of these nematode species within the local ruminant populations