US-wide equine strongylid egg count data demonstrate seasonal and regional trends.

Equine strongylid parasites are ubiquitous around the world and are main targets of parasite control programmes. In recent years, automated fecal egg counting systems based on image analysis have become available allowing for collection and analysis of large-scale egg count data. This study aimed to evaluate equine strongylid fecal egg count (FEC) data generated with an automated system over three years in the US with specific attention to seasonal and regional trends in egg count magnitude and sampling activity. Five US regions were defined; North East, South East, North Central, South Central and West. The data set included state, region and zip code for each FEC. The number of FECs falling in each of the following categories were recorded: (1) 0 eggs per gram (EPG), (2) 1 ⩽ 200 EPG, (3) 201 ⩽ 500 EPG and (4) >500 EPG. The data included 58 329 FECs. A fixed effects model was constructed fitting the number of samples analysed per month, year and region, and a mixed effects model was constructed to fit the number of FECs falling in each of the 4 egg count categories defined above. The overall proportion of horses responsible for 80% of the total FEC output was 18.1%, and this was consistent across years, months and all regions except West, where the proportion was closer to 12%. Statistical analyses showed significant seasonal trends and regional differences of sampling frequency and FEC category. The data demonstrated that veterinarians tended to follow a biphasic pattern when monitoring strongylid FECs in horses, regardless of location.

Characterization of the complete mitochondrial genomes of the zoonotic parasites Bolbosoma nipponicum and Corynosoma villosum (Acanthocephala: Polymorphida) and the molecular phylogeny of the order Polymorphida.

Acanthocephalans of the order Polymorphida mainly parasitic in birds and mammals, are of veterinary, medical and economic importance. However, the evolutionary relationships of its 3 families (Centrorhynchidae, Polymorphidae and Plagiorhynchidae) remain under debate. Additionally, some species of Polymorphida (i.e. Bolbosoma spp. and Corynosoma spp.) are recognized as zoonotic parasites, associated with human acanthocephaliasis, but the mitochondrial genomes for representatives of Bolbosoma and Corynosoma have not been reported so far. In the present study, the complete mitochondrial genomes B. nipponicum and C. villosum (Acanthocephala: Polymorphidae) are reported for the first time, which are 14 296 and 14 241 bp in length, respectively, and both contain 36 genes [including 12 PCGs, 22 tRNA genes and 2 rRNA genes] and 2 non-coding regions (NCR1 and NCR2). The gene arrangement of some tRNAs in the mitogenomes of B. nipponicum and C. villosum differs from that found in all other acanthocephalans, except Polymorphus minutus. Phylogenetic results based on concatenated amino acid (AA) sequences of the 12 protein-coding genes (PCGs) strongly supported that the family Polymorphidae is a sister to the Centrorhynchidae rather than the Plagiorhynchidae, and also confirmed the sister relationship of the genera Bolbosoma and Corynosoma in the Polymorphidae based on the mitogenomic data for the first time. Our present findings further clarified the phylogenetic relationships of the 3 families Plagiorhynchidae, Centrorhynchidae and Polymorphidae, enriched the mitogenome data of the phylum Acanthocephala (especially the order Polymorphida), and provided the resource of genetic data for diagnosing these 2 pathogenic parasites of human acanthocephaliasis.

Efficacy of a single-dose albendazole against lancet liver fluke Dicrocoelium dendriticum and liver enzymes activity in naturally infected sheep.

Infections with D. dendriticum are distributed worldwide and mostly associated with ruminant livestock. Depending on the length and strength of the infection it can be manifested with losses in milk production, reductions in milk and wool quality, decreased weight gains, reproductive performance and poor carcass quality. The objective of this study was to determine the efficacy of albendazole (ABZ) against the lancet liver fluke Dicrocoelium dendriticum in naturally infected sheep using parasitological methods. Twenty-four sheep were divided into four groups: two untreated control groups (C1, C2) and two treated groups (T1, T2), with six animals in each group. The sheep in the treated groups were administered a single oral dose (15 mg/kg bwt) of ABZ suspension. After ABZ treatment the animals were slaughtered on Day 14 (groups C1, T1) and Day 30 (groups C2, T2) and were necropsied. Coprological therapeutic ABZ efficacy reached 92.4% on Day 14 (P < 0.001) and 88.5% on Day 30 (P < 0.001). On Day 30, the serum activities of hepatic and cholestatic enzymes including serological analysis of total protein concentration (TP) and protein fractions were evaluated. Significant decrease of aspartate aminotransferase (AST) (P < 0.01) and gamma-glutamyltransferase (GGT) (P < 0.05) activity by 36.9% and 34.6%, respectively, were detected for sheep in T2 group. These enzymes showed a strong positive correlation to fluke burden: AST (r = 0.654) and GGT (r = 0.768), respectively (P < 0.05). Additionally, the electrophoretic analysis of serum total protein and protein fraction concentrations revealed minimal hypoproteinemia and hyperalbuminemia after ABZ treatment. The decrease of liver enzyme activities and their correlation with fluke burden may indicate recovery of hepatocellular and biliary damage following the reduction of fluke burdens after ABZ therapy. A decline in AST and GGT activity could serve as a valuable adjunct bioindicator of liver damage and fluke reduction after treatment of dicrocoeliosis in naturally infected sheep.

Equine tapeworm (Anoplocephala spp.) infection: evaluation of saliva- and serum-based antibody detection methods and risk factor analysis in Slovak horse populations.

A lack of accurate information on the prevalence and distribution of Anoplocephala spp. infections on horse farms has led to insufficient attention to tapeworm control and increasing horse anoplocephaloses in Europe. Our study aimed to examine the occurrence of Anoplocephala spp. infection using coprological, serum- and saliva-based antibody detection methods and to analyze the risk factors associated with tapeworm infection in domestic horses in Slovakia. Fecal, serum, and saliva samples were collected from 427 horses from 31 farms in Slovakia. Additionally, a questionnaire study was conducted to collect information on tapeworm distribution on horse farms and analyze risk factors associated with infection. Fecal samples were examined by the mini-FLOTAC and the double centrifugation/combined sedimentation-flotation techniques. Serum and saliva samples were analyzed by ELISA to determine antibody levels against Anoplocephala spp. The effects of variables associated with an individual horse were tested for the positive result of the saliva ELISA test on Anoplocephala spp. Cestode eggs were detected in 1.99% of fecal samples (farm prevalence 12.90%), with no differences between the two coprological methods. Serum-based tapeworm ELISA results revealed that 39.39% of horses tested positive (farm prevalence 83.87%); while saliva-based tapeworm ELISA results revealed 56.95% positive horses (farm prevalence 96.77%). Binary logistic regression analysis revealed four meaningful predictors that significantly impacted the likelihood of detecting tapeworm infection in horses: horse age, pasture size, anthelmintic treatment scheme, and access to pasture. The influences of other variables associated with an individual horse were not significantly associated with detecting tapeworm infection.

Infection patterns of helminth community in black rockcod Notothenia coriiceps in West Antarctica over a 6-year term.

Patterns of the rockcod Notothenia coriiceps infection with helminths were analysed to understand the dynamics of parasite communities in this Antarctic fish and to test their stability over time. The study was performed using helminth samples collected from 183 N. coriiceps in 2014-2015 and 2020-2021 in the vicinity of the Ukrainian Antarctic station (UAS) "Akademik Vernadsky", Galindez Island, Argentine Islands, West Antarctica. Overall, 25 helminth taxonomical categories (nine trematodes, four cestodes, five nematodes, and seven acanthocephalans) were subjected to analysis. A direct comparison of the helminth population characteristics showed that nine species significantly changed their infection parameters during the 6 years between the samples. Seven of them (Pseudoterranova sp., Contracaecum sp., Ascarophis nototheniae, monolocular metacestodes, bilocular metacestodes, Metacanthocephalus rennicki, and Diphyllobothrium sp.) were found to have a significant impact on the differences between helminth infracommunities in 2014-2015 and 2020-2021. Most studied patterns of helminth component community appeared to show a stable tendency, and observed fluctuations were close to the steady trend. Slight but significant changes in the infection patterns observed in this study might have been caused by changes in the populations of intermediate, paratenic, and definitive hosts of helminths (marine invertebrates, mammals, and birds), which participate in helminth transmission in Antarctic ecosystems.

Effect of sainfoin (Onobrychis viciifolia) on cyathostomin eggs excretion, larval development, larval community structure and efficacy of ivermectin treatment in horses.

Alternative strategies to chemical anthelmintics are needed for the sustainable control of equine strongylids. Bioactive forages like sainfoin (Onobrychis viciifolia) could contribute to reducing drug use, with the first hints of in vitro activity against cyathostomin free-living stages observed in the past. We analysed the effect of a sainfoin-rich diet on cyathostomin population and the efficacy of oral ivermectin treatment. Two groups of 10 naturally infected horses were enrolled in a 78-day experimental trial. Following a 1-week adaptation period, they were either fed with dehydrated sainfoin pellets (70% of their diet dry matter) or with alfalfa pellets (control group) for 21-days. No difference was found between the average fecal egg counts (FECs) of the two groups, but a significantly lower increase in larval development rate was observed for the sainfoin group, at the end of the trial. Quantification of cyathostomin species abundances with an ITS-2-based metabarcoding approach revealed that the sainfoin diet did not affect the nemabiome structure compared to the control diet. Following oral ivermectin treatment of all horses on day 21, the drug concentration was lower in horses fed with sainfoin, and cyathostomin eggs reappeared earlier in that group. Our results demonstrated that short-term consumption of a sainfoin-rich diet does not decrease cyathostomin FEC but seems to slightly reduce larval development. Consumption of dehydrated sainfoin pellets also negatively affected ivermectin pharmacokinetics, underscoring the need to monitor horse feeding regimes when assessing ivermectin efficacy in the field.

Trematodes from Antarctic teleost fishes off Argentine Islands, West Antarctica: molecular and morphological data.

In 2014-2015 and 2019-2021, teleost fishes off Galindez Island (Antarctic Peninsula) were examined for trematodes. Combined morphological and molecular analyses revealed the presence of eight trematode species of four families (Hemiuridae, Lecithasteridae, Opecoelidae, Lepidapedidae) from five fish species. Only adult trematodes were found and all of them are Antarctic endemics with their congeners occurring on other continents. The hemiuroids, Elytrophalloides oatesi (Leiper & Atkinson, 1914), Genolinea bowersi (Leiper & Atkinson, 1914), and Lecithaster macrocotyle Szidat & Graefe, 1967 belong to the most common Antarctic species and together with Lepidapedon garrardi (Leiper & Atkinson, 1914) and Neolebouria georgiensis Gibson, 1976 they were recorded as the least host-specific parasites. The originally sub-Antarctic Neolepidapedon macquariensis Zdzitowiecki, 1993 is a new record for the Antarctic Peninsula and Parachaenichthys charcoti (Vaillant), is a new host record. Neolebouria terranovaensis Zdzitowiecki, Pisano & Vacchi, 1993 is considered a synonym of N. georgiensis because of identical morphology and dimensions. The currently known phylogenetic relationships within the studied families are supported, including the polyphyly of Macvicaria Gibson & Bray, 1982 with the future need to accommodate its Antarctic species in a new genus. The validity of M. georgiana (Kovaleva & Gaevskaja, 1974) and M. magellanica Laskowski, Jezewski & Zdzitowiecki, 2013 needs to be confirmed by further analyses. Genetic sequence data are still scarce from Antarctica, and more studies applying integrative taxonomic approaches and large-scale parasitological examinations of benthic invertebrates are needed to match sequences of larval stages to those of well-characterised adults and to elucidate trematode life-cycles.

Review of metazoan parasites of the northern fur seal (Callorhinus ursinus) and the analysis of the gastrointestinal helminth community of the population on St. Paul Island, Alaska.

The northern fur seal (NFS), Callorhinus ursinus (Mammalia: Otariidae), is a marine mammal species included into the IUCN Red List as the vulnerable species which population is dramatically declining. A significant amount of parasitological data collected previously and our recent data allowed us to clarify the list of NFS metazoan parasites and to perform a comprehensive analysis of the gastrointestinal helminth community. Gastrointestinal tracts from 756 NFSs (3- to 4-year-old males) were collected during the annual Aleut subsistence harvests in July-August of 2011-2014 from five separate rookeries on St. Paul Island, Alaska. Totally, 27,625 specimens of helminths and approximately 1000 nasal mites were collected and identified. Detailed analysis of the previously published and newly obtained data revealed 32 species of metazoan parasites, including trematodes (6 species), cestodes (4), nematodes (9), acanthocephalans (9) and arthropods (4). The gastrointestinal helminth community of newly studied NFSs comprised 19 species including trematodes (4), cestodes (3), nematodes (5) and acanthocephalans (7). Temporal changes in the helminth community structure were small but statistically significant. Gastrointestinal helminth infracommunities comprised from 1 to 10 species (average of 4). Small but significant correlation was found between the abundances of acanthocephalans (Corynosoma similis and C. strumosum), nematodes (Contracaecum osculatum, Pseudoterranova spp.) and cestode Diphyllobothrium tetrapterum.

New morphological and molecular perspectives about Macracanthorhynchus hirudinaceus (Acanthocephala: Oligacanthorhynchidae) from wild boar, Sus scrofa Linn., in Ukraine.

We make new morphological observations not previously reported for the old acanthocephalan Macracanthorhynchus hirudinaceus (Pallas, 1781) Travassos, 1917, described for the first time about 240 years ago. Our specimens were collected from the wild boar, Sus scrofa Linn., in Ukraine in 2005. We provide comparative morphometrics with other populations from Ukraine, Union of Soviet Socialist Republics, Europe, Brazil and the United States of America. Our specimens from the Ukraine had the smallest trunk (110-120 mm long), longest and thickest hooks, with the third hook being the largest, largest eggs, and a proboscis wider than long. We document the morphology of the proboscis, apical organ, hooks, hook roots, sensory pores, micropores, and eggs with scanning electron microscopy for the first time. We also provide chemical analysis of hooks and eggs using energy-dispersive X-ray analysis, discuss its micropores, and provide a new molecular profile based on 18S rDNA from a European population for the first time. Edge of hook tips feature high levels of calcium and phosphorous but the high level of sulphur is mostly found in the cortical layer of eggs. One new partial 18S rDNA sequence (482 nt length) was generated from an adult specimen of M. hirudinaceus. We present the first 18S rDNA published sequence for this cosmopolitan acanthocephalan obtained from Europe. The amplified region corresponded to the approximate middle region of the small subunit ribosomal ribonucleic acid gene, which is ~1800 nt in length. This molecular contribution is especially valuable in light of the extreme scarcity of genetic information about species of Macracanthorhynchus and of the family Oligacanthorhynchidae as a whole.

Effects of life history stage and climatic conditions on fecal egg counts in plains zebras (Equus quagga) in the Serengeti National Park.

In wildlife, endoparasite burden can be affected by host life history stage, environmental conditions, host abundance, and parasite co-infections. We tested the effects of these factors on gastrointestinal parasite infection in plains zebras (Equus quagga) in the Serengeti ecosystem, Tanzania, using fecal egg counts of two nematode families (Strongylidae and Ascarididae) and the presence/absence of cestode (Anoplocephalidae) eggs. We predicted higher egg counts of Strongylidae and Ascarididae, and increased likelihood of Anoplocephalidae infection in individuals (1) during energetically costly life history stages when resource allocation to immune processes may decrease and in young zebras after weaning because of increased uptake of infective stages with forage, (2) when climatic conditions facilitate survival of infective stages, (3) when large zebra aggregations increase forage contamination with infective stages, and (4) in individuals co-infected with more than one parasite group as this may indicate reduced immune competence. Strongylidae egg counts were higher, and the occurrence of Anoplocephalidae eggs was more likely in bachelors than in band stallions, whereas Ascarididae egg counts were higher in band stallions. Strongylidae and Ascarididae egg counts were not increased in lactating females. Strongylidae egg counts were higher in subadults than in foals. Regardless of sex and age, Ascarididae infections were more likely under wet conditions. Co-infections did not affect Strongylidae egg counts. Ascarididae egg counts in adult females were higher when individuals were co-infected with Anoplocephalidae. We present evidence that parasite burdens in plains zebras are affected by life history stage, environmental conditions, and co-infection.

Concurrent Proteomic Fingerprinting and Molecular Analysis of Cyathostomins.

Rapid, cost-effective, efficient, and reliable helminth species identification is of considerable importance to understand host-parasite interactions, clinical disease, and drug resistance. Cyathostomins (Nematoda: Strongylidae) are considered to be the most important equine parasites, yet research on this group is hampered by the large number of 50 morphologically differentiated species, their occurrence in mixed infections with often more than 10 species and the difficulties associated with conventional identification methods. Here, MALDI-TOF MS, previously successfully applied to identify numerous organisms, is evaluated and compared with conventional and molecular genetic approaches. A simple and robust protocol for protein extraction and subsequent DNA isolation allowing molecular confirmation of proteomic findings is developed, showing that MALDI-TOF MS can discriminate adult stages of the two closely related cyathostomin species Cylicostephanus longibursatus and Cylicostephanus minutus. Intraspecific variability of proteomic profiles within morphospecies demonstrated an identification of morphospecies with an accuracy of close to 100%. In contrast, three genospecies within C. minutus and sex-specific profiles within both morphospecies could not be reliably discriminated using MALDI-TOF MS. In conclusion, MALDI-TOF MS complemented by the molecular protocol is a reliable and efficient approach for cyathostomin species identification.

Multifocal Necrotizing Myopathy in Northern Elephant Seal ( Mirounga angustirostris) Pups, San Miguel Island, California.

A field study addressing causes of mortality in freshly dead northern elephant seals ( Mirounga angustirostris, Gill, 1866) was conducted on San Miguel Island, California, in February 2015. Necropsies were performed on 18 pups ranging in age from stillbirths to approximately 7 to 8 weeks. The primary gross diagnoses in these pups included trauma, myopathy, starvation/emaciation, infections, congenital anomalies, and perinatal mortality. However, 6 (33%) had a previously unrecognized myopathy characterized by multiple white streaks that were most obvious within the inner layer of the abdominal wall and the small innermost ventral intercostal muscles. Following histological examination, 2 more pups from San Miguel Island and 6 pups from The Marine Mammal Center (Sausalito, California) were found to have similar lesions. Histologically, the lesions within the skeletal muscles were characterized by a multifocal polyphasic, mild to severe, acute to subacute necrotizing myopathy with mineralization. Acute necrosis and degeneration characterized by pyknotic nuclei, eosinophilic cytoplasm and cytoplasmic vacuolization were found in smooth muscle myocytes within the urinary bladder and digestive system. Degeneration of myocytes was present in the tunica media of a few small- to medium-sized vessels and was characterized by a vacuolar degeneration and occasionally necrosis. This condition has been termed multifocal necrotizing myopathy. A cause of this myopathy was not identified.

Morphological and molecular evidence for synonymy of Corynosoma obtuscens Lincicome, 1943 with Corynosoma australe Johnston, 1937 (Acanthocephala: Polymorphidae).

Corynosoma obtuscens Lincicome, 1943 (Acanthocephala: Polymorphidae) is synonymised with Corynosoma australe Johnston, 1937 based on combined morphological and molecular evidence. Morphological comparison of C. obtuscens (24 males and 27 females) collected from a California sea lion Zalophus californianus (Lesson) in California, USA, with the type-specimens of C. obtuscens and C. australe, and with published data on C. australe collected from different hosts and regions showed no significant differences. The levels of genetic divergence in the cox1 sequences obtained from C. obtuscens from a California sea lion in the present study and C. australe from otariid seals from Argentina and penguins from Brazil ranged between 1.4-1.6% and was considered to represent intraspecific variability. Additionally, cox1 sequences were generated for Andracantha phalacrocoracis (Yamaguti, 1939), Corynosoma semerme (Forssell, 1904), C. strumosum (Rudolphi, 1802), C. validum Van Cleave, 1953 and C. villosum Van Cleave, 1953. Our results revealed inconsistency in the identification of material used as a source of the previously published sequence data for C. obtuscens and C. magdaleni Montreuil, 1958.

Macrocyclic lactones differ in interaction with recombinant P-glycoprotein 9 of the parasitic nematode Cylicocylus elongatus and ketoconazole in a yeast growth assay.

Macrocyclic lactones (MLs) are widely used parasiticides against nematodes and arthropods, but resistance is frequently observed in parasitic nematodes of horses and livestock. Reports claiming resistance or decreased susceptibility in human nematodes are increasing. Since no target site directed ML resistance mechanisms have been identified, non-specific mechanisms were frequently implicated in ML resistance, including P-glycoproteins (Pgps, designated ABCB1 in vertebrates). Nematode genomes encode many different Pgps (e.g. 10 in the sheep parasite Haemonchus contortus). ML transport was shown for mammalian Pgps, Pgps on nematode egg shells, and very recently for Pgp-2 of H. contortus. Here, Pgp-9 from the equine parasite Cylicocyclus elongatus (Cyathostominae) was expressed in a Saccharomyces cerevisiae strain lacking seven endogenous efflux transporters. Pgp was detected on these yeasts by flow cytometry and chemiluminescence using the monoclonal antibody UIC2, which is specific for the active Pgp conformation. In a growth assay, Pgp-9 increased resistance to the fungicides ketoconazole, actinomycin D, valinomycin and daunorubicin, but not to the anthelmintic fungicide thiabendazole. Since no fungicidal activity has been described for MLs, their interaction with Pgp-9 was investigated in an assay involving two drugs: Yeasts were incubated with the highest ketoconazole concentration not affecting growth plus increasing concentrations of MLs to determine competition between or modulation of transport of both drugs. Already equimolar concentrations of ivermectin and eprinomectin inhibited growth, and at fourfold higher ML concentrations growth was virtually abolished. Selamectin and doramectin did not increase susceptibility to ketoconazole at all, although doramectin has been shown previously to strongly interact with human and canine Pgp. An intermediate interaction was observed for moxidectin. This was substantiated by increased binding of UIC2 antibodies in the presence of ivermectin, moxidectin, daunorubicin and ketoconazole but not selamectin. These results demonstrate direct effects of MLs on a recombinant nematode Pgp in an ML-specific manner.

Description of Uncinaria lyonsi n. sp. (Nematoda: Ancylostomatidae) from the California sea lion Zalophus californianus Lesson (Carnivora: Otariidae).

A new species of hookworm, Uncinaria lyonsi n. sp., is described based on morphological studies of the nematodes collected by Dr. E. T. Lyons from the California sea lion Zalophus californianus (Lesson) on San Miguel Island, California, USA. The new species is morphologically similar to three other species of the genus Uncinaria Frölich, 1789 parasitising pinnipeds, U. lucasi Stiles, 1901, U. hamiltoni Baylis, 1933 and U. sanguinis Marcus, Higgins, Slapeta & Gray, 2014, in the body dimensions, the structure of the buccal capsule, the shape and structure of the male caudal bursa and female genital system. Uncinaria lyonsi n. sp. is differentiated from U. lucasi by having longer spicules and gubernaculum, larger buccal capsule and more slender oesophagus. The new species differs from U. hamiltoni and U. sanguinis in having shorter spicules and narrower buccal capsule. The latter two species also occur in the Southern Hemisphere and are geographically separated from U. lyonsi n. sp. The present study confirms the existence of a host-specific species of Uncinaria in the California sea lion, previously revealed by molecular and biological investigations.

Strongylids of Domestic Horses in Eastern Slovakia: Species Diversity and Evaluation of Particular Factors Affecting Strongylid Communities.

PURPOSE: This study aimed to examine the species diversity and structure of the strongylid community in domestic horses in Eastern Slovakia. Also, an analysis of the impact of age, sex, and collection location factors on the strongyid communities was performed. METHODS: Twenty-seven horses 1.5-21 years old from two farms in eastern Slovakia with different horse-management conditions were studied. Strongylids were collected after horse treatments with Noromectin (0.2 mg ivermectin); 66,170 specimens were collected and identified. Faecal egg count reduction test (FECRT) was performed following fenbendazole (FBZ) and ivermectin (IVM) treatments. RESULTS: Twenty-four strongylid species were found; horses were infected with 6 to 16 (average = 11.7) species. Six cyathostomin species (Cylicocyclus nassatus, Cyathostomum catinatum, C. pateratum, Cylicostephanus longibursatus, C. goldi, C. calicatus) were the most prevalent; C. catinatum was the dominant species in both farms (Berger-Parker index 0.34 and 0.42). The structure of the strongylid community was multimodal with dominant, subdominant, background, and rare species. The Mantel test showed that horse age and sex did not significantly affect the nematode infracommunity composition (p > 0.05), while the impact of the collection location (farm) was significant (p = 0.03). Additionally, C. longibursatus was identified as the species contributing significantly to the observed farm differences. Strong resistance to FBZ was documented on both farms (FECRT was 36.4% and 22.7%); IVM resistance was not observed (FECRT = 100%). CONCLUSION: This study presents the first report on the strongylids parasitizing domestic horses in Eastern Slovakia and gives basic information for further studies of horse parasites and their control in the region.

Equine Strongylidae and other parasitic nematodes described by Arthur Looss during 1895-1911 in the collections of the Swedish Museum of Natural History.

Prof. Arthur Looss (1861-1923) was a prolific German parasitologist, who, among other things, authored descriptions of 22 new species of nematodes and 115 new species of trematodes. After his death, his collection (including type material) was split between several institutions: Smithsonian National Museum of Natural History in Washington (USA), Natural History Museum in Berlin and the Natural History Museum in Leipzig (Germany), Gothenburg Museum of Natural History and Swedish Museum of Natural History (Sweden). Here we revise all type specimens of nematodes from the A. Looss collection that are currently preserved in the Swedish Museum of Natural History (Strongylus subtilis, Sclerostomum edentatum, S. vulgare, Cyathostomum labratum, C. coronatum, C. bicoronatum, C. calicatum, C. alveatum, C. catinatum, C. nassatum, C. radiatum, C. elongatum, C. auriculatum, Triodontus minor, T. serratus, C. labiatum and Uncinaria polaris), designate and describe lectotypes wherever deemed necessary and provide catalogue access numbers to all type materials. We also revise all notes and drawings associated with new species that A. Looss described and provide previously unpublished pencilled sketches and ink print-ready drawings of some of these species (Strongylus subtilis, Cyathostomum poculatum, C. radiatum, C. elongatum, C. calicatum, C. auriculatum, Triodontus serratus, Trichostrongylus vitrinus and possibly Necator africanus).

Patterns of variation in equine strongyle community structure across age groups and gut compartments.

BACKGROUND: Equine strongyles encompass more than 64 species of nematode worms that are responsible for growth retardation and the death of animals. The factors underpinning variation in the structure of the equine strongyle community remain unknown. METHODS: Using horse-based strongyle community data collected after horse deworming (48 horses in Poland, 197 horses in Ukraine), we regressed species richness and the Gini-Simpson index upon the horse's age, faecal egg count, sex and operation of origin. Using the Ukrainian observations, we applied a hierarchical diversity partitioning framework to estimate how communities were remodelled across operations, age groups and horses. Lastly, strongyle species counts collected after necropsy (46 horses in France, 150 in Australia) were considered for analysis of their co-occurrences across intestinal compartments using a joint species distribution modelling approach. RESULTS: First, inter-operation variation accounted for > 45% of the variance in species richness or the Gini-Simpson index (which relates to species dominance in communities). Species richness decreased with horse's age (P = 0.01) and showed a mild increase with parasite egg excretion (P < 0.1), but the Gini-Simpson index was neither associated with parasite egg excretion (P = 0.8) nor with horse age (P = 0.37). Second, within-host diversity represented half of the overall diversity across Ukrainian operations. While this is expected to erase species diversity across communities, community dissimilarity between horse age classes was the second most important contributor to overall diversity (25.8%). Third, analysis of species abundance data quantified at necropsy defined a network of positive co-occurrences between the four most prevalent strongyle genera. This pattern was common to necropsies performed in France and Australia. CONCLUSIONS: Taken together, these results show a pattern of ß-diversity maintenance across age classes combined with positive co-occurrences that might be grounded by priority effects between the major species.

Multidrug resistance in Haemonchus contortus in sheep - can it be overcome?

Introduction: Gastrointestinal nematodes pose a threat to animal health and affect farmers by negatively impacting farm management. Material and Methods: The study was conducted on a sheep farm with suspected reductions in the efficacies of anthelmintics. Efficacy was determined using in vivo faecal egg count reduction, in vitro egg hatch (EHT) and larval development (LDT) tests. In the first phase, 60 sheep were equally split into six groups. Group 1 received the recommended dose of albendazole (ALB), group 2 received the same after fasting for 24 h, group 3 received the dose divided into two halves at 6 h intervals, group 4 received a double dose of ALB, and group 5 received the recommended dose of ivermectin (IVM). Group 6 served as a control. The second phase of the experiment had two groups: one treated with levamisole (LEV) and a control group. Faecal samples were collected from all sheep. Results: No reduction of egg output was observed in the groups treated with single, double, or divided doses of ALB, but one of 13.7-16.9% was noted in the fasting group. Efficacy in the IVM group ranged from 31.50 to 39.97%. The mean concentrations sufficient to prevent 50% of the eggs from hatching in the in vitro EHT and the mean concentrations in which the development of larvae to the L3 stage was inhibited by 50% in the LDT exceeded established thresholds for benzimidazoles and IVM. Haemonchus contortus was the only species identified after treatment. The LDT did not indicate the presence of resistance to LEV. All animals treated with LEV were negative for eggs 10 d after treatment. Conclusion: Resistance to ALB and IVM in Haemonchus contortus was confirmed. Alternative approaches to improve the efficacies of benzimidazole did not sufficiently increase the efficacy, but LEV was an efficient anthelmintic treatment.

Comparison of two molecular barcodes for the study of equine strongylid communities with amplicon sequencing.

Basic knowledge on the biology and epidemiology of equine strongylid species still needs to be improved to contribute to the design of better parasite control strategies. Nemabiome metabarcoding is a convenient tool to quantify and identify species in bulk samples that could overcome the hurdle that cyathostomin morphological identification represents. To date, this approach has relied on the internal transcribed spacer 2 (ITS-2) of the ribosomal RNA gene, with a limited investigation of its predictive performance for cyathostomin communities. Using DNA pools of single cyathostomin worms, this study aimed to provide the first elements to compare performances of the ITS-2 and a cytochrome c oxidase subunit I (COI) barcode newly developed in this study. Barcode predictive abilities were compared across various mock community compositions of two, five and 11 individuals from distinct species. The amplification bias of each barcode was estimated. Results were also compared between various types of biological samples, i.e., eggs, infective larvae or adults. Bioinformatic parameters were chosen to yield the closest representation of the cyathostomin community for each barcode, underscoring the need for communities of known composition for metabarcoding purposes. Overall, the proposed COI barcode was suboptimal relative to the ITS-2 rDNA region, because of PCR amplification biases, reduced sensitivity and higher divergence from the expected community composition. Metabarcoding yielded consistent community composition across the three sample types. However, imperfect correlations were found between relative abundances from infective larvae and other life-stages for Cylicostephanus species using the ITS-2 barcode. While the results remain limited by the considered biological material, they suggest that additional improvements are needed for both the ITS-2 and COI barcodes.