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

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

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

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

Occurrence of Larval Dicrocoelium dendriticum and Brachylaima sp. in Gastropod Intermediate Hosts from Fergana Valley, Uzbekistan

The occurrence of larval Dicrocoelium dendriticum and Brachylaima sp. is described with molecular evidences in gastropod intermediate hosts from Fergana Valley, Uzbekistan. Larvae of D. dendriticum were detected in 28 (10.7%) out of 262 Xceropicta candacharica, and 8 (9.7%) of 82 Angiomphalia gereliana. Brachylaima sp. larvae were found in 3 (1.6%) of 95 Pseudonapaeus sogdiana. The total number of larvae per snail varied from 8 to 110 individuals. Alignment of the first four sequences of 28S rDNA was revealed a 99-100% similarity to D. dendriticum. Larvae from P. sogdiana snails were 98% similar to Brachylaima sp. In this study, it was confirmed that 2 species of terrestrial snail, X. candacharica and A. gereliana, act as the first intermediate hosts of D. dendriticum, and P. sogdiana snail play a role of intermediate host of Brachylaima sp. in the Fergana Valley, Uzbekistan

ウズベク共和国における Protostrongylidae 科線虫の中間宿主 : 腹足類

Protostrongylid species are causative agents of pulmonary protostrongyliasis in caprine species in Uzbekistan. These nematodes typically require one intermediate host, terrestrial mollusks, to complete their life cycle. In this study, eight species of gastropods were found to be positive for protostrongylid larvae. Haplotypes of larvae corresponding to sequences of Protostrongylus rufescens and Muellerius capillaris were detected. Morphological identification of gastropods, based on shell characteristics, revealed six different morphotypes. Anatomic-morphological and molecular results confirmed the membership of these gastropods to the Buliminidae, Hygromiidae, Agriolimacidae and Parmacellidae and revealed eight different species: Pseudonapaeus maydanika, P. sogdiana, P. albiplicatus, Pseudonapaeus sp., Angiomphallia regeliana, Xeropicta candacharica, Candaharia levanderi and Deroceras reticulatum. This study displays the first report of P. maydanika, C. levanderi and D. reticulatum as natural intermediate hosts of M. capillaris. The infection rate of snails with protostrongylids was 27.9%(616/2,207) and the infection rate of slugs was 6.5%(18/279).ウズベク共和国における羊の肺虫症の原因寄生虫プロトストロンギルス科線虫は陸棲腹足類を中間宿主としているので、この線虫症の疫学では腹足類の調査が必須であった。そこで、同国において貝殻の形態で 6種に分類された軟体類を対象にプロトストロンギルス科の感染幼虫の保有状況を調査した。その結果、分子生物学的にProtostrongylus rufescens および Muellerius capillaris のハプロタイプに一致した感染幼虫を得た。また、今回、幼虫が得られた腹足類について解剖学および分子生物学的な検討によりこれらは 4つの科 Buliminidae, Hygromiidae, Agriolimacidae あるいは Parmacellidae の次のような 8種に同定された; Pseudonapaeus maydanika, P. sogdiana, P. albiplicatus, Pseudonapaeus sp. Angiomphallia regeliana, Xeropicta candacharica, Candaharia levanderi, Deroceras reticulatum. 本研究により P. maydanika, Candaharia levanderi および D. reticulatum が M. capillaris の自然中間宿主であることが証明された。全体的なプロトストロンギルス科線虫の感染幼虫の保有率はカタツムリ類で 27.9%(616/2,207)、一方、ナメクジ類では 6.5%(18/279)と低かった