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Purpose: The present paper reports the nematodes of the suborder Trichostrongylina collected from the common spiny bandicoot, Echymipera kalubu, in Arso, Papua Indonesia. The description of Kalubustrongylus arsoensis gen. et sp. n. (Trichostrongylidae: Filarinematinae) is given herein., Materials and Methods: The specimens were collected from three common spiny bandicoots at Arso, Papua Indonesia, captured using traditional snap traps in August 1993. The new taxon is described and illustrated by light and scanning electron microscopy. Type and voucher specimens were deposited in the Museum Zoologicum Bogoriense (MZB), Bogor, Indonesia., Results: The new genus Kalubustrongylus is classified as a member of the subfamily Filarinematinae in the family Trichostrongylidae by having a bilaterally symmetrical synlophe and an uncoiled body, and by lacking a cephalic vesicle. It resembles Peramelistrongylus but is distinguished by having additional ridges in the synlophe, an elliptical bursa, an apically divided dorsal ray and minute extra-dorsal rays. Besides Ka. arsoensis, Mackerrastrongylus biakensis, Peramelistrongylus skedastos, Beveridgiella spp., Dessetostrongylus sp. and Herpetostrongylinae sp. were collected. Dessetostrongylus is recorded for the first time outside of Australia and from non-dasyurid marsupials., Conclusions: The present findings add a genus to the three genera previously known in the subfamily Filarinematinae in the family Trichostrongylidae., Competing Interests: Declarations. Ethical Approval: The specimen is not under the listed category of experimental animals, which needs ethics approval. Consent for Publication: All the authors consent to the publication of this manuscript. Competing Interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Faecal samples were obtained from 77 first season grazers from 20 Norwegian dairy herds in autumn 2020 for analysis of Cooperia oncophora and Ostertagia ostertagi infection. Strongylid eggs per gram of faeces (EPG) were determined for each sample and the samples underwent larval culture. DNA was extracted from the faeces at different stages of the culture preparation: from faecal slurry (FS), direct extraction before culture (DBC), and direct extraction after culture (DAC). Extracted DNA was subject to molecular assay for both C. oncophora and O. ostertagi using both a published ddPCR assay and a modified qPCR duplex assay targeting the ITS-2 gene. For C. oncophora, DBC samples contained significantly less ITS-2 copies than DAC and FS samples (p ≤ 0.0001 for both) for qPCR, but not between DAC and FS samples (p = 0.339). Droplet digital PCR with DBC samples yielded significantly fewer ITS-2 copies than DAC and FS samples (p ≤ 0.0001). For O. ostertagi, the ITS-2 copies in DBC samples differed significantly from levels in DAC and FS samples on the qPCR platform (p=0.002 and 0.044, respectively) as was the case with ddPCR (p ≤ 0.0001 and 0.0137). Overall, across the various processing techniques, C. oncophora results obtained on both ddPCR and qPCR platforms correlated with EPG better than was found for equivalent results for O. ostertagi and results are strongly indicative of poor correlation when EPG counts are low. Results from this study suggest that DAC faecal processing was of limited practical use., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

We describe a new species of the genus Alippistrongylus (Nematoda: Trichostrongylina) in the small intestine of Delomys dorsalis (Rodentia: Sigmodontinae) found in Itatiaia National Park (Parque Nacional do Itatiaia, PNI), state of Rio de Janeiro, Brazil. The new species can be distinguished between the other two species previously described by the following morphological characteristics: pattern of the rays from the caudal bursa rays in males and the spherical-shaped appendage in the female. Additionally, we provide molecular genetic data from the new species. The discovery of Alippistrongylus itatiaiaensis sp. n. expands our understanding of nematode diversity and underscores the importance of parasite studies in biodiversity conservation. Its occurrence in a preserved area like the PNI emphasises the role of such habitats in maintaining unique ecological interactions.

Background: This study was carried out to investigate the prevalence and analyze the molecular characteristics based on the internal transcribed spacer (ITS) 2 region of the ribosomal RNA (RNA) gene of trichostrongylid nematodes in different ruminants from Guilan province, northern of Iran., Methods: The gastrointestinal tracts of 144 ruminants including 72 cattle, 59 sheep, and 13 goats were collected from an abattoir in Guilan province during July to September 2018. After isolation the helminths, male specimens were identified based on morphological parameters. PCR and partial sequencing of the ITS2 fragment were conducted. After phylogenetic analysis, the intraspecific and interspecific differences were calculated., Results: The prevalence of total infections with the nematodes was 38.9, 74.6 and 84.6% among cattle, sheep and goats, respectively. Eleven species of trichostrongylid nematodes including Haemonchus contortus, Marshallagia marshalli, Trichostrongylus axei, T. colubriformis, T. vitrinus, Ostertagia trifurcata, Teladorsagia circumcincta, Marshallagia occidentalis, O. lyrata, O. ostertagi, and Cooperia punctate were recovered from the ruminants. The most prevalent trichostrongyloid nematodes in cattle, sheep and goats were O. ostertagi (26.4%), M. marshalli (64.4%) and T. circumcincta (69.2%), respectively. Phylogenetic tree was discriminative for Trichostrongylidae family, while phylogenetic analysis of the ITS2 gene represented low variations and no species identification of Haemonchidae and Cooperiidae families., Conclusions: This study suggests the high prevalence and species diversity of trichostrongyloid nematodes in different ruminants, indicating the importance of implement antiparasitic strategies in north regions of Iran. As well, this study showed that the ITS2 fragment is not a discriminative marker for Haemonchidae and Cooperiidae families, and investigation of other genetic markers such as mitochondrial genes would be more valuable for better understanding of their phylogenetic relationships., (© 2021. The Author(s).)

European beaver (Castor fiber L. 1758) is the biggest rodent living in Europe. It is a semi-aquatic animal known for building dams and burrows. European beaver is a potential host for a wide range of parasites and other infectious diseases. In Slovakia, there is an increasing number of beavers but the data about their parasitic fauna are missing. Our work is the first documentation about the beaver's parasitofauna in Slovakia. In a 1-year study, we collected and examined 19 beaver fecal samples from the vicinity of beaver burrows inhabiting three particular localities at the Danube, Topľa, and Laborec rivers in Slovakia. In these fecal samples, 4 different species of intestinal endoparasites were detected as follows: oocysts of Cryptosporidium, cysts of Giardia, eggs of Stichorchis subtriquetrus, and eggs and larvae of Travassosius rufus. Parasites were confirmed only in samples collected at river Topľa. Based on our results, we can conclude that European beaver can be an important source of parasitic contamination of surface waters especially in the localities shared by people.

Gastro-intestinal tracts were examined from thirteen Gudali zebu cattle, ten goats and ten sheep from the Adamawa highland in Northern Cameroon. A total of 28,325 adult helminths were recovered from the abomasa, small and large intestines. Five trichostrongylid genera were identified by their morphology: Haemonchus, Trichostrongylus and Oesophagostomum were predominant in both cattle and small ruminants, whilst Cooperia was only found in cattle both in the abomasum and small intestines. The molecular species identification and the inference of their phylogenetic relationships was based on the analysis of the hypervariable region I of the small subunit 18S rDNA (SSU) and the Second Internal Transcribed Spacer (ITS-2) of 408 adult trichostrongylid worms, which were PCR-amplified, sequenced, and compared with available database entries. Consistent with earlier findings, the SSU was invariable within the Haemonchus and Trichostrongylus genera, confirming the prior classification based on the morphology of the worms, but the ITS-2 was highly inter- and intraspecifically variable and thus allowed to distinguish individual species and to study the haplotype diversity within the different species. In cattle, we report for the first time in Cameroon co-infection with two species of Haemonchus (H. placei and H. similis), together with two species of Cooperia (C. punctata and C. pectinata) and one species of Trichostrongylus (T. axei). In goats and sheep, we found one highly polymorphic clade of Haemonchus contortus and two Trichostrongylus species (T. axei and T. colubriformis). When compared with other Trichostrongylidae from different regions of the world and wildlife, the analysis of haplotypes did not indicate any host and geographical isolation, but a very high haplotype diversity among H. contortus. These findings illustrate the complexity of trichostrongylid populations in domestic ruminants and suggest grazing overlap between domestic and wildlife hosts., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Gastrointestinal nematode (GIN) infections in cattle cause appetite suppression which leads to poor feed conversion, reduced weight gain and reduced milk production. Overuse and exclusive reliance on anthelmintic drugs has resulted in widespread resistance in many parasitic nematode species infecting livestock making control increasingly difficult. Wild ruminants are competent hosts of a number of nematode species that typically infect and are best adapted for cattle, sheep, and goats. Thus, the potential exists for wild ruminants to act as reservoirs in the translocation of domestic GIN, including those carrying anthelmintic resistance mutations as well as susceptible genotypes. The potential for parasite exchange is heightened by interfaces or ecotones between managed and wild rangelands, and by perturbations linked to climate warming that can increasingly alter the distributions of wild ungulates and their interactions with domestic and free-ranging ruminants. To investigate the extent to which wild ruminants harbour parasites capable of infecting domestic ruminants we first performed an epidemiological study of feces from wildlife hosts that spanned 16 states and included white-tailed deer (85 % of the samples), pronghorn, elk, mule deer, bighorn sheep, moose, cattle, and caribou across the United States. All samples were cultured to third stage larvae and nematode DNA was isolated and PCR amplified. Among the 548 wild ruminant samples received, 33 % (181 samples) were positive for nematode DNA, among which half (84 samples) contained DNA from GIN species commonly found in cattle. DNA from cattle GIN species was detected in 46 % of samples from the Northeast, 42 % from the Southeast, 10 % from the Midwest, 0 % from the Southwest and 11 % from the West. Deep amplicon sequencing of the ITS-2 rDNA indicated that Ostertagia and Trichostrongylus were present in 90 % and 69 % of the nematode DNA positive samples, respectively, whereas Haemonchus, Cooperia and Oesophagostomum were present in 26 %, 2 % and 10 % of the samples, respectively. These data clearly show that wild ruminants commonly harbour multiple parasite species whose primary hosts are domestic cattle, and suggest that further work is warranted to investigate their specific roles in the management of anthelmintic resistance., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Standard diagnostic methods currently in use for the identification of helminth infections in ruminants are based on the morphological analysis of immature and adult stages of parasites. This paper describes a method for the semiquantitative identification of nematodes, mainly Trichostrongyloidea, at species-level resolution. The method is based on amplification and fragment analysis followed by minisequencing of the ITS-2 region (internal transcribed spacer 2) of the ribosomal DNA of parasite eggs or larvae. This method allows for the identification of seven genera (Chabertia, Cooperia, Haemonchus, Oesophagostomum, Ostertagia, Teladorsagia, and Trichostrongylus) and 12 species (Chabertia ovina, Cooperia curticei, Cooperia punctata, Cooperia oncophora/Cooperia surnabada, Haemonchus contortus, Haemonchus placei, Haemonchus longistipes, Oesophagostomum asperum, Oesophagostomum radiatum, Ostertagia ostertagi, Trichostrongylus axei, and Trichostrongylus colubriformis) of infectious nematodes of domestic ruminants. The concordance between the morphological and molecular analyses in the detection of genera ranged from 0.84 to 0.99, suggesting the proposed detection method is specific, semiquantitative, less laborious, and highly cost-efficient.

Gastrointestinal nematodes are a significant threat to the economic and environmental sustainability of keeping livestock, as adequate control becomes increasingly difficult due to the development of anthelmintic resistance in some systems and climate-driven changes to infection dynamics. To mitigate any negative impacts of climate on gastrointestinal nematode epidemiology and slow anthelmintic resistance development, there is a need to develop effective, targeted control strategies that minimise the unnecessary use of anthelmintic drugs and incorporate alternative strategies such as vaccination and evasive grazing. However, the impacts climate and gastrointestinal nematode epidemiology may have on the optimal control strategy are generally not considered, due to lack of available evidence to drive recommendations. Parasite transmission models can support control strategy evaluation to target field trials, thus reducing the resources and lead-time required to develop evidence-based control recommendations incorporating climate stochasticity. Gastrointestinal nematode population dynamics arising from natural infections have been difficult to replicate and model applications have often focussed on the free-living stages. A flexible framework is presented for the parasitic phase of gastrointestinal nematodes, GLOWORM-PARA, which complements an existing model of the free-living stages, GLOWORM-FL. Longitudinal parasitological data for two species that are of major economic importance in cattle, Ostertagia ostertagi and Cooperia oncophora, were obtained from seven cattle farms in Belgium for model validation. The framework replicated the observed seasonal dynamics of infection in cattle on these farms and overall, there was no evidence of systematic under- or over-prediction of faecal egg counts. However, the model under-predicted the faecal egg counts observed on one farm with very young calves, highlighting potential areas of uncertainty that may need further investigation if the model is to be applied to young livestock. The model could be used to drive further research into alternative parasite control strategies such as vaccine development and novel treatment approaches, and to understand gastrointestinal nematode epidemiology under changing climate and host management., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Background: Parasitic trichostrongyloid nematodes have a worldwide distribution in ruminants and frequently have been reported from humans in Middle and Far East, particularly in rural communities with poor personal hygiene and close cohabitation with herbivorous animals. Different species of the genus Trichostrongylus are the most common trichostrongyloids in humans in endemic areas. Also, Ostertagia species are gastrointestinal nematodes that mainly infect cattle, sheep and goats and in rare occasion humans. The aim of the present study was to identify the trichostrongyloid nematodes obtained from a familial infection in Guilan province, northern Iran, using morphological and molecular criteria., Methods: After anthelmintic treatment, all fecal materials of the patients were collected up to 48 h and male adult worms were isolated. Morphological identification of the adult worms was performed using valid nematode keys. Genomic DNA was extracted from one male worm of each species. PCR amplification of ITS2-rDNA region was carried out, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 6.0 software., Results: Adult worms expelled from the patients were identified as T. colubriformis, T. vitrinus and Teladorsagia circumcincta based on morphological characteristics of the males. Phylogenetic analysis illustrated that each species obtained in current study was placed together with reference sequences submitted to GenBank database., Conclusions: The finding of current study confirms the zoonotic aspect of Trichostrongylus species and T. circumcincta in inhabitants of Guilan province. The occurrence of natural human infection by T. circumcincta is reported for the first time in Iran and the second time in the world.

Objective: The aim of the current study was to determine the presence and prevalence of Eimeria and helminth species in sheep raised in Erzurum province by using fecal examination., Methods: Faecal samples were collected from a total of 784 sheep raised in Aziziye, Yakutiye and Palandöken districts between February-March 2019. The samples were examined by Fulleborn’s flotation, Benedect sedimentation, and Baermann-Wetzel methods., Results: Eimeria spp. and helminths were found in 49.36% (387/784) and 74.11% (581/784) of the samples, respectively. Identified Eimeria species were as follows: E. parva (59.68%), E. ovina (51.67%), E. faurei (47.80%), E. ahsata (39.27%), E. granulosa (36.62%), E. punctata (28.42%), E. pallida (26.09%), E. ovinoidalis (18.34%), E. crandallis (16.79%), E. intricata (15.76%), E. weybridgensis (11.36%) and E. marsica (6.20%). Helminth species identified at genus/species level were Dicrocoelium spp. (33.91%), Fasciola spp. (5.68%), Paramphistomum spp. (2.58%), Moniezia spp. (5.85%), Trichostrongylid type egg (49.05%), Marshallagia spp. (38.73%), Nematodirus spp. (20.98%), Trichuris spp. (14.46%), Protostrongylus spp. (18.42%), Dictyocaulus filaria (2.41%) and Muellerius capillaris (1.38%)., Conclusion: Parasitic diseases cause important economic losses in livestock industry. In following years, it is aimed to plan prevention and control strategies for the parasites detected in this area in line with the data of this study and to share this data with the animal breeders.

The nematode genus Bidigiticauda has 2 species ( Bidigiticauda vivipara and Bidigiticauda embryophilum ), which are parasites of bats from the Neotropical region. The present paper describes a new species of Bidigiticauda from a male Artibeus planirostris specimen collected in the Pratigi Environmental Protection Area in Bahia state, Brazil. The new species, Bidigiticauda serrafreirei n. sp., differs from B. embryophilum by having longer spicules, rays 5 and 6 arising from a common trunk and bifurcating in its first third, rays 3 and 4 emerging slightly separated from each other, and dorsal rays reaching the margin of the caudal bursa. The new species also differs from B. vivipara by the dorsal ray bifurcating at the extremity of the trunk. A molecular phylogenetic analysis was conducted to determine the evolutionary affinities of Bidigiticauda serrafreirei n. sp. within the Strongylida, which identified a clade that grouped Bidigiticauda with the other members of the Anoplostrongylinae. However, the molineid subfamilies did not group together, indicating that the family Molineidae is polyphyletic. Further analyses, which include additional taxa and genetic markers, should elucidate the complex relationships within the Molineidae, in particular its subfamilies and the evolution of the traits that define these groups.

Little is known about the diversity of many parasites infecting camels, with most relying on morphological parameters. DNA extracted from different tissues (n = 90) and fecal samples (n = 101) from dromedary camels (Camelus dromedarius) in Egypt were screened for multiple parasites using different molecular markers. Screening of tissue samples (heart) for Toxoplasma gondii and Sarcocystis spp. was performed using B1 and 18S rRNA gene markers, respectively. T. gondii was further genotyped using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP). Sarcocystis was analyzed using PCR-RFLP characterization (XbaI and MboI restriction enzymes). A taxonomically challenging but important group of nematodes (Trichostrongylidae family) were screened using the ITS-2 ribosomal DNA (rDNA) species-specific markers. Furthermore, nested PCR was used for the detection of Cryptosporidium spp. (SSU rRNA gene) and positive samples were genotyped after RFLP (SspI and VspI) and sequencing. Cryptosporidium parvum isolates were subtyped by sequence analysis of the 60-kDa glycoprotein gene. This study revealed that many parasites infect the investigated camels, including T. gondii (1.1%), Sarcocystis spp. (64.4%), Cryptosporidium spp. (5.9%) and Trichostrongylidae nematodes (22.7%). The species contribution for nematodes was as follows: Haemonchus spp. (95.6%), Trichostrongylus axei (26%), Trichostrongylus colubriformis (65.2%) and Cooperia oncophora (60.8%). Mn-PCR-RFLP typing for Toxoplasma was only successful for three markers: 5'-SAG2 (type II), 3'-SAG2 (type II) and alt. SAG2 (type II). PCR-RFLP using XbaI showed possible mixed Sarcocystis infection. Moreover, the Cryptosporidium genotypes detected were C. parvum (IIdA19G1 and IIaA15G1R1), Cryptosporidium rat genotype IV and a novel genotype (camel genotype). This approach revealed the unique Cryptosporidium genotypes infecting the investigated camels, and the high genetic diversity of the investigated parasites., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Mycoplasma haemolamae (Mhl) and gastrointestinal nematodes can cause anemia in camelids. Control programs aim to suppress parasitism without promoting anthelminthic resistance, but few evidence-based guidelines define acceptable parasite loads in camelids., Hypothesis/objectives: In clinically healthy nonanemic camelids, compare erythrocyte variables to Mhl real-time PCR status and to fecal egg count (FEC). Determine the FEC threshold above which erythrocyte variables are consistently below reference interval medians., Animals: One hundred fourteen client-owned adult alpacas and llamas., Methods: In a cross-sectional study, whole blood in ethylenediaminetetraacetic acid (EDTA) was assessed for packed cell volume (PCV) by centrifugation, erythrocyte count (RBC), and hemoglobin concentration (HGB) using an ADVIA120 analyzer, and Mhl using real-time PCR. Trichostrongyle eggs per gram (epg) were counted by modified McMaster test on freshly collected feces. Significant differences in erythrocyte variables based on Mhl status and FEC thresholds were assessed by independent t test and one-way ANOVA, respectively., Results: Packed cell volume, RBC, and HGB were not significantly different between Mhl-positive and Mhl-negative animals, but were significantly lower in animals with FEC >1000 epg compared to those with <500 epg. All animals with FEC >600 epg had RBC and HGB below the reference interval median. All animals with FEC >750 epg had PCV below the reference interval median., Conclusions and Clinical Importance: In healthy nonanemic camelids, positive Mhl PCR is not associated with lower erythrocyte variables and such animals may not warrant treatment. Fecal egg count >600-750 epg has a negative effect on erythrocyte variables, and may be a guide for deworming protocols., (© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Trichostrongylid nematodes are a common cause of gastroenteritis in sheep. Despite its worldwide distribution, Teladorsagia circumcincta has not been included in reports listing the various trichostrongyles infecting sheep from Egypt. Herein, we describe the presence of 2 T. circumcincta haplotypes infecting small ruminants from Egypt. For this study, fresh fecal samples were collected from 340 sheep and 115 goats reared at 5 districts in Dakahlia governorate and its surroundings, Egypt. Trichostrongyle eggs were harvested from the samples, and then subjected to DNA isolation and analysis. Polymerase chain reaction (PCR) amplification was carried out for the second internal transcribed spacer of ribosomal DNA ( ITS2 rDNA ). Purified PCR products of T. circumcincta were sequenced, and the revealed sequences were subjected to the nucleotide and phylogenetic analysis. A relatively high prevalence of trichostrongyles eggs was identified in sheep (33.2%) and a lower prevalence was found in goats (14.7%). Molecular analysis revealed, for the first time, 2 sheep herds from Egypt that were infected with T. circumcincta . Both infected herds were raised by the Bedouins in rural areas of El Mahalla El Kubra city. No T. circumcincta infections were found in any of the goats. Nucleotide sequence analysis revealed 2 haplotypes (Te1 and Te2) from 7 successfully sequenced samples (5 from the first and 2 from the second herd). Te1 was the major haplotype in both herds, and Te2 was retrieved from a single sample. Phylogenetic analysis displayed that the Te1 haplotype clustered with one from Cyprus, which might have been introduced to Egypt via goats imported from Cyprus due to a program to improve meat and milk production in Egypt. The present results could be beneficial in understanding the epidemiology of T. circumcincta and other trichostrongyles in Egypt, and have implementations in the effective control strategies used in this region.

A new species of Nippostrongylinae (Nematoda: Heligmonellidae), Stilestrongylus rolandoi n. sp., is described from specimens collected from the small intestine of the rodent Euryoryzomys russatus in the Atlantic Forest (Santo Amaro da Imperatriz, Santa Catarina state, southern Brazil). The genus Stilestrongylus includes 23 species, which parasitize rodents occurring in the Neotropical region. Stilestrongylus aureus (Durette-Desset & Sutton, 1985) from Argentina, S. azarai (Durette-Desset & Sutton, 1985) from Argentina, S. flavescens (Sutton & Durette-Desset, 1991) from Uruguay, S. franciscanus (Digiani & Durette-Desset, 2002) from Argentina, S. gracielae (Digiani & Durette-Desset, 2006) from Argentina, and S. oryzomysi (Sutton & Durette-Desset, 1991) from Argentina are closely related to Stilestrongylus rolandoi n. sp., all having caudal bursa patterns of types 1-4 in one of the lobes. Stilestrongylus rolandoi n. sp. is distinguished from the aforementioned species by its ray 6 being short in relation to rays 4 and 5, which are long and robust, and by having caudal bursa patterns of types 1-4 in both lobes. The new species has 27 ridges in the mid-body in males, and 24 in females, and has one of the highest ratios of spicule length to body length (21-33%) in this genus.

Spent mushroom compost (SMC) is a residue generated in edible mushrooms production, such as Hypsizygus marmoreus. Its genome was recently sequenced, demonstrating cuticle-degrading protease genes. The present work aims to investigate the proteases from H. marmoreus spent mushroom compost (SMC) by verifying its action on nematode larvae. The extraction of the crude extract directly with water from H. marmoreus SMC proved to be efficient for proteases obtainment, with proteolytic activity of 195.36 ± 18.38 U g -1 of compound. Moreover, the zymogram and SDS-PAGE indicated the presence of two proteases with estimated molecular weights of 30.2 and 33.7 kDa. Due to the protease activity present in H. marmoreus SMC extract, there was a significant reduction in the number of Panagrellus redivivus and L 3 in treated group compared to control group (p < 0.01), with 52% and 26% of reduction, respectively. A0A151VWY3 mature protein is composed of 296 amino acid residues, exhibiting molecular weight and pI of 29.5 kDa and 6.72. A0A151WD28 mature protein is composed of 343 amino acid residues, exhibiting molecular weight and pI of 34.4 kDa and 8.04. In the present work it was demonstrated that SMC from H. marmoreus has easily extracted protease content, presenting two proteases, possibly with cuticle-degrading activity, which had significant nematicidal effect on P. redivivus and bovine infective larvae., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Ashworthius sidemi is a blood-sucking nematode, which has spread out among wild ruminants in several European countries during last decades. The nematode has recently been detected in cattle as well. The distribution of A. sidemi in Russia has not been sufficiently clarified yet. In European part of Russia A. sidemi was formerly registered in sika deer (Cervus nippon) and maral (Cervus elaphus sibiricus) introduced from Asia, and also in aboriginal elks (Alces alces). Taking into consideration the presence of other species of ruminants susceptible to A. sidemi in European Russia, it is necessary to control the spread of this parasite. The specimens of males and females of A. sidemi were found during the autopsies of three naturally infected roe deer (Capreolus capreolus) from Voronezh and Tver regions (European Russia). The species affiliation of the discovered nematodes was determined according to morphological features and confirmed using molecular techniques. The intensity of infection with A. sidemi in two roe deer from Voronezh was 11 and 63 nematodes, and it was 17 nematodes in roe deer from Tver. All of the discovered specimens of A. sidemi were referred to juvenile forms based on features of male bursa morphology and weak development of female reproductive system. In Russia, A. sidemi has not previously been detected in C. capreolus and the present report constitutes the first record of the parasite occurrence in this species of ruminant., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The genus Cooperia includes important parasites of ruminants and currently contains 34 accepted species. However, even for those species infecting livestock, there is a considerable lack of molecular information and many species are only identifiable using subtle morphological traits. The present study aimed to provide molecular data to allow diagnosis of Cooperia species infecting cattle. Partial sequences of two mitochondrial (cytochrome oxidase 2, 12S rRNA gene) and two nuclear genes (isotype 1 β tubulin gene including two introns, internal transcribed spacers (ITS) were obtained from morphologically identified specimens of Cooperia pectinata, Cooperia punctata and Cooperia spatulata as well as from larvae of pure Cooperia oncophora and C. punctata laboratory isolates. Pairwise identity of ITS-2 sequences was very high and it was the only region able to identify a specimen as Cooperia sp. However, the ITS-2 was unreliable for diagnosis at the species level. All other marker sequences could not unequivocally be allocated to the genus Cooperia but allowed clear species identification with the exception of the pair C. punctata/C. spatulata for which no significant differences were found for any marker sequence. Maximum-likelihood phylogenetic analyses of individual genes as well as a multi-locus analysis covering all four sequences confirmed that specimen identified as C. spatulata were randomly distributed throughout the C. punctata cluster and formed no group of their own. In contrast, the other Cooperia species formed clearly separated and statistically supported clusters. These data indicate that C. spatulata is most likely only a morphotype of C. punctata and the name should be considered a synonym. Combinations of nuclear and mitochondrial markers should be used to identify morphotypes or cryptic species to benefit from excellent barcoding properties of the latter but allowing proper phylogenetic analyses and controlling for lineage sorting that might occur for mitochondrial genotypes within a species., Competing Interests: The authors have declared that no competing interests exist.

Gizzard helminths were examined in 100 (50 adult, 50 juvenile) female northern pintails ( Anas acuta). Sixty-three individual helminths, representing 5 species ( Amidostomum acutum, Echinuria uncinata, Epomidiostomum uncinatum, Streptocara crassicauda, and Gastrotaenia cygni) were found. Twenty-seven northern pintails were infected with 1-3 helminth species and averaged 1.4 species. Overall, A. acutum and G. cygni were the most prevalent and abundant species (20%, n = 31 and 10%, n = 25, respectively), followed by S. crassicauda (5%, n = 5), E. uncinata (1%, n = 1), and E. uncinatum (1%, n = 1). Intensity of infection for A. acutum, E. uncinata, E. uncinatum, S. crassicauda, and G. cygni was 1.6 ± 0.3 [SE], 1.0 ± 0, 1.0 ± 0, 1.0 ± 0, and 2.5 ± 0.6, respectively. Our findings represent new information about gizzard helminth infections in northern pintails wintering along the Texas coast.

Background: Many trichostrongylid nematode species are reported to infect bison, some of which are major causes of disase and production loss in North American bison herds. However, there is little information on the species distribution and relative abundance of these parasites in either commercial or conservation herds. This is largely because trichostrongylid nematode species cannot be distinguished by visual microscopic examination of eggs present in feces. Consequently, we have applied ITS2 rDNA nemabiome metabarcoding to describe the trichostrongyle parasite species diversity in 58 bison production groups derived from 38 commercial North American plains bison (Bison bison bison) herds from across western Canada, and two bison conservation herds located in Elk Island National Park (EINP) [plains bison and wood bison (Bison bison athabascae)] and one in Grasslands National Park (GNP) (plains bison)., Results: We report much higher infection intensities and parasite species diversity in commercial bison herds than previously reported in beef cattle herds grazing similar latitudes. Predominant trichostrongyle parasite species in western Canadian commercial bison herds are those commonly associated with Canadian cattle, with Ostertagia ostertagi being the most abundant followed by Cooperia oncophora. Combined with high fecal egg counts in many herds, this is consistent with significant clinical and production-limiting gastrointestinal parasitism in western Canadian bison herds. However, Haemonchus placei was the most abundant species in five of the production groups. This is both surprising and important, as this highly pathogenic blood-feeding parasite has not been reported at such abundance, in any livestock species, at such northerly latitudes. The presence of Trichostrongylus axei as the most abundant parasite in four herds is also unusual, relative to cattle. There were striking differences in parasite communities between the EINP and commercial bison herds. Most notably, Orloffia bisonis was the predominant species in the wood bison herd despite being found at only low levels in all other herds surveyed., Conclusions: This study represents the most comprehensive description of parasite communities in North American bison to date and illustrates the power of deep amplicon sequencing as a tool to study species diversity in gastrointestinal nematode communities.

Nematodes of the Libyostrongylus genus are hematophagous parasites found under the ostrich's proventriculus membrane: they are frequent and can cause 50% of mortality in chicks and occasionally death in adults. With the aim of determining the presence of the Libyostrongylus genus in two private collections, one located in Ayapango, State of Mexico and the other in Amacuzac, Morelos, a total of 27 fecal samples were collected between August and December of 2016. Samples were analyzed using the flotation technique. The eggs were quantified using the McMaster technique. Positive samples were cultured to obtain infective larvae and to be identified by their morphometric characteristics. Only 18 samples collected in Ayapango were positive to Libyostrongylus douglassii. This is the first time that L. douglassii is reported in Mexico., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Libyostrongylus douglassii, Libyostrongylus dentatus and Libyostrongylus magnus are nematodes that infect ostriches. The first species has been identified in ostriches from Africa, Europe, Americas and Oceania. Although the natural range of ostriches is Africa, L. dentatus was first described in birds from the USA and later identified in Brazil, where co-infections with L. douglassii have been commonly reported. Libyostrongylus magnus is known from the original description only. There are a few reports on infections with L. douglassii in ostriches from Africa and all farmed birds examined are from the southern region of the continent. The aim of this report was to verify Libyostrongylus spp. infections in wild ostriches from Ethiopia. Fecal samples from ostriches, Struthio molybdophanes, were collected and submitted to coproculture. Infective larvae were identified to the species level based on general morphology and morphometry. In addition, phylogenetic analysis of the first and second internal transcribed spacer (ITS1 and ITS2) of the nuclear ribosomal DNA was performed., Results: Infective larvae from Ethiopian ostriches had the morphological characteristics of L. dentatus. Confidence interval estimate for sheath tail length from Ethiopian Libyostrongylus sp. isolates overlapped one for Brazilian L. dentatus. Neighbor-joining and Maximum Likelihood phylogenetic trees based on sequences of the ITS1 and ITS2 regions revealed that the Ethiopian samples belong to the L. dentatus species clade. Monospecific infections with L. dentatus were confirmed in Ethiopian wild ostriches, opposed to the co-infections typically found in the Americas., Conclusions: To our knowledge, this is the first record of L. dentatus from African ostriches, the region from which this parasite originated.

A new species of Heligmonellidae (Trichostrongylina, Heligmosomoidea), Stilestrongylus kaaguyporai n. sp. is described from the small intestine of Euryoryzomys russatus (Rodentia, Cricetidae, Sigmodontinae) from the Argentine Atlantic Forest, in the Misiones province. The new species was found at Campo Anexo Manuel Belgrano, Reserva de Vida Silvestre Urugua-í and Parque Provincial Urugua-í, with a prevalence of 73% in 15 hosts examined. Stilestrongylus includes 24 Neotropical species, all parasitic in rodents, mostly Sigmodontinae. Stilestrongylus kaaguyporai n. sp. can be differentiated from its congeners by the following characters: caudal bursa dissymmetrical with right lobe larger and pattern of type1-4 in both lobes, rays 6 not forming a lateral trident with rays 4 and 5, rays 8 with dissymmetrical pathway, genital cone hypertrophied with a conspicuous hood-like projection and females with a marked dorso-ventral torsion of the posterior end. This report is the second record of a Stilestrongylus species in E. russatus, increasing to nine the number of parasitic species known from this host.

Marshallagia marshalli (Nematoda: Trichostrongylidae) infection can lead to serious parasitic gastroenteritis in sheep, goat, and wild ruminant, causing significant socioeconomic losses worldwide. Up to now, the study concerning the molecular biology of M. marshalli is limited. Herein, we sequenced the complete mitochondrial (mt) genome of M. marshalli and examined its phylogenetic relationship with selected members of the superfamily Trichostrongyloidea using Bayesian inference (BI) based on concatenated mt amino acid sequence datasets. The complete mt genome sequence of M. marshalli is 13,891 bp, including 12 protein-coding genes, 22 transfer RNA genes, and 2 ribosomal RNA genes. All protein-coding genes are transcribed in the same direction. Phylogenetic analyses based on concatenated amino acid sequences of the 12 protein-coding genes supported the monophylies of the families Haemonchidae, Molineidae, and Dictyocaulidae with strong statistical support, but rejected the monophyly of the family Trichostrongylidae. The determination of the complete mt genome sequence of M. marshalli provides novel genetic markers for studying the systematics, population genetics, and molecular epidemiology of M. marshalli and its congeners.

Maternal dietary protein deficiency and gastrointestinal nematode infection during early pregnancy have negative impacts on both maternal placental gene expression and fetal growth in the mouse. Here we used next-generation RNA sequencing to test our hypothesis that maternal protein deficiency and/or nematode infection also alter the expression of genes in the developing fetal brain. Outbred pregnant CD1 mice were used in a 2×2 design with two levels of dietary protein (24% versus 6%) and two levels of infection (repeated sham versus Heligmosomoides bakeri beginning at gestation day 5). Pregnant dams were euthanized on gestation day 18 to harvest the whole fetal brain. Four fetal brains from each treatment group were analyzed using RNA Hi-Seq sequencing and the differential expression of genes was determined by the edgeR package using NetworkAnalyst. In response to maternal H. bakeri infection, 96 genes (88 up-regulated and eight down-regulated) were differentially expressed in the fetal brain. Differentially expressed genes were involved in metabolic processes, developmental processes and the immune system according to the PANTHER classification system. Among the important biological functions identified, several up-regulated genes have known neurological functions including neuro-development (Gdf15, Ing4), neural differentiation (miRNA let-7), synaptic plasticity (via suppression of NF-κβ), neuro-inflammation (S100A8, S100A9) and glucose metabolism (Tnnt1, Atf3). However, in response to maternal protein deficiency, brain-specific serine protease (Prss22) was the only up-regulated gene and only one gene (Dynlt1a) responded to the interaction of maternal nematode infection and protein deficiency. In conclusion, maternal exposure to GI nematode infection from day 5 to 18 of pregnancy may influence developmental programming of the fetal brain., (Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Nematodes Ashworthius sidemi are typical parasites of Asiatic cervids, mostly sika deer (Cervus nippon). Migrating red deer have probably introduced this parasite to the area of Poland from neighbouring countries. The aim of the study was to determine prevalence and intensity of infection of red deer and roe deer from the Lower Silesian Wilderness with A. sidemi after one year from previous studies and to examine the influence of these parasites on other species of gastrointestinal nematodes. 31 abomasa of red deer and 3 abomasa of roe deer from Forest Dictrict of Ruszów have been examined on the presence of parasites. In comparison with previous studies, red deer showed significant, over triple increase of prevalence of A. sidemi and over fivefold increase of mean intensity of infection. Maximum intensity of infection has increased over 24 times. These results indicate on dynamic spread of this new aswortiosis foci. Apart from previously registered parasite species, four new nematodes have been found in examined red deer and roe deer during this year studies. Red deer have been infected with Spiculopteragia assymmetrica, typical for fallow deer and Haemonchus placei - typical for bovine, whereas roe deer have been infected with Trichostrongylus axei and Trichostrongylus capricola, parasites of sheep, goats and bovine. Prevalence of red deer infection with nematodes S. boehmi and Ostertagia leptospicularis was lower than two years ago while prevalence of infection with S. mathevossiani and O. kolchida was higher. Intensity of infection with these nematodes has increased.

The spread of parasites through a host population is based on the variation in behavior and immune function between individuals and is rarely uniform. We studied the gastrointestinal parasites of common mole-rats ( Cryptomys hottentotus hottentotus, Lesson 1826) from 2 sites and assessed the levels of infection based on host sex, breeding status, and season. Only nematode species were found: Neoheligmonella sp. and Mammalakis macrospiculum (Ortlepp, 1939) and a single specimen of Trichuris sp., all of which have direct life cycles. Parasite burden and species richness was greater in the mesic habitat. The abundance of Neoheligmonella sp. differed significantly between seasons, and the season of peak abundance differed between sites, perhaps due to differences in host densities between sites. In addition, parasite burden did not differ between the sexes, but breeding animals had higher infections of Neoheligmonella sp. and M. macrospiculum than non-breeding animals. This and previous studies thus suggest that the subterranean environment is beneficial in reducing parasite diversity, although the restrictions on movement may lead to certain individuals suffering higher parasite burdens.

We report the finding of 2 species of Pudica (Nematoda: Heligmonellidae: Pudicinae) in 2 rodents endemic to Chile, the common degu Octodon degus (Octodontidae) and the Bennett's chinchilla rat Abrocoma bennettii (Abrocomidae). Pudica degusi ( Babero and Cattan, 1975 ) n. comb., originally described as a species of Longistriata (Heligmosomidae), was found in the common degu; through the study of its synlophe, the species is reassigned to the Heligmonellidae: Pudicinae and the genus Pudica, and it is revalidated through comparison with the remaining species of the genus. Pudica cattani n. sp. is described from both O. degus and A. bennettii. It is characterized by its large body size, bursal pattern of type 1-3-1 on right lobe, 1-3-1 tending to 1-4 on left lobe, synlophe with 11 ridges including a careen, dorsal ray of the bursa dividing proximally and bursal rays 9 and 10 relatively short. Pudica degusi n. comb. and Pudica cattani n. sp. were found in the same host species but not as coparasitic in the same individuals. The common degu is confirmed as the sole and primary host of Pudica degusi n. comb. It is unlikely that it is the primary host for Pudica cattani n. sp., whose host affinities are less clear mainly due to the scarcity of data. Pudica cattani n. sp. is the first helminth reported from the Bennett's chinchilla rat. Both findings enlarge the host range of the Pudicinae to the families Octodontidae and Abrocomidae, i.e., 9 out of the 11 extant families of caviomorphs, thereby establishing the presence of this nematode subfamily as typical parasites of the Neotropical Hystricognathi.

Haemonchus contortus is a major parasite of small ruminants and its blood-feeding behaviour causes effects ranging from mild anaemia to death. Knowledge of the genetic variation within and among H. contortus populations can provide the foundation for understanding transmission patterns and aid in the control of haemonchosis. Adult male H. contortus were collected from three geographical regions in Egypt. The second internal transcribed spacer (ITS2) of nuclear ribosomal DNA was amplified using the polymerase chain reaction (PCR) and sequenced directly. The population genetic diversity and sequence variations were determined. Nucleotide sequence analyses revealed one genotype (ITS2) in all worms, without genetic differentiation. The similarity in population genetic diversity and genetic patterns observed among the three geographical regions could be attributed to possible movement between the sites. This is the first study of genetic variation in H. contortus in Egypt. The present results could have implications for the rapid characterization of H. contortus and other trichostrongyloid nematodes, and evaluation of the epidemiology of H. contortus in Egypt.

Background: Traditional methods of detecting and identifying gastrointestinal nematode infections in small ruminants, including sheep and goats, are time-consuming and lack in sensitivity and specificity. Recently, we developed an automated multiplexed-tandem (MT)-PCR platform for the diagnosis and identification patent infections with key genera/species of gastrointestinal nematodes of sheep and validated this approach in detailed experiments carried out in Australia. In the present study, we deployed this diagnostic platform in Scotland and Belgium to test samples from naturally infected sheep in these environments and to validate the MT-PCR platform relative to traditional diagnostic methods routinely used by local laboratories., Results: MT-PCR detected all microscopy positive samples and there was a significant agreement between the results of the different test methods in terms of the species detected and their relative proportion in a test sample, however, for some samples, there were discrepancies between the results of the different test methods. Selective sequencing of purified MT-PCR products demonstrated the results to be 100% specific., Conclusions: The MT-PCR platform is an advanced method for the species/genus-specific diagnosis of gastrointestinal nematode infections in small ruminants and has demonstrated utility when deployed in different countries and climatic zones. The platform is user-friendly due to the largely automated procedure and has high versatility in that it can achieve a specific diagnosis from different types of sample templates, including larval culture and faecal samples. With appropriate modifications of the primers used, the MT-PCR platform also provides potential for the diagnosis of a variety of other pathogens of veterinary or medical importance.

Ostrich farming is an important livestock industry in different world regions with a diverse offer of products and services. In Colombia, as in other countries, this market led the importation of animals from countries like Canada, United States of America and South Africa for breeding objectives. With the animals, specific pathogens for these ratites could be introduced. Libyostrongylus spp. is a strongylid nematode with worldwide distribution, which can induce a severe disease and mortality in infected animals. Limited studies in Colombia have identified parasites in ostrich farming systems. The aim of this study was to identify parasites of the genus Libyostrongylus to a species level in faecal samples from ostrich farms in three departments of Colombia. Five ostrich farms from Boyacá, Meta and Tolima were sampled in 2011 and in 2013 to obtain fresh faecal samples which were further processed by flotation tests for egg visualization and faecal culture for infective larvae identification by morphological and morphometric parameters. One from the five farms, located in Meta department, was positive for strongylid eggs in both sampling periods. After faecal culture, infective larvae were identified as Libyostrongylus douglassii. These results corroborate previous records of Libyostrongylus in ostrich farms from Meta and confirms, for the first time, infection by L. douglassii in ratites from this region. Further studies must identify associated determinants for infection and its effects on the flock health and production., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Resistance to benzimidazoles (BZs) in trichostrongyloid nematodes is a worldwide problem for livestock production, particularly regarding small ruminants. Sensitive and reliable methods are required to assess anthelmintic resistance status. Currently available methods for BZ resistance detection can be divided into three main groups, in vivo (e.g. faecal egg count reduction test), in vitro (e.g. egg hatch assay) and molecular tests. Three single nucleotide polymorphisms (SNPs) in the isotype-1 β-tubulin gene of various nematode species correlate with BZ resistance. While PCR-based methods have been reported for the three most economically important nematodes of sheep, namely, Trichostrongylus, Haemonchus and Teladorsagia, pyrosequencing assays are so far only available for the latter two. Here, the design and evaluation of pyrosequencing assays for isotype-1 and isotype-2 β-tubulin genes of Trichostrongylus colubriformis are described. PCR fragments carrying the susceptible and corresponding resistant genotype were combined in defined ratios to evaluate assay sensitivity and linearity. The correlation between the given and the measured allele frequencies of the respective SNPs (codons F167Y, E198A and F200Y) was very high. Pyrosequencing assays for Haemonchus, Teladorsagia and Trichostrongylus were subsequently used for a BZ resistance survey, carried out in the three European countries, namely Ireland, Italy and Switzerland. Larval cultures obtained from field survey samples in 2012 and 2013 were used for pyrosequencing. The test was applied when the target species represented at least 10% of the sample. Trichostrongylus and Teladorsagia were detected in all countries' samples whereas Haemonchus was not detected in samples from Ireland. SNPs in isotype-1 associated with resistance were detected for all three species, with frequencies at codon F200Y far exceeding those at codons F167Y and E198A. Elevated SNP frequencies in isotype-2 of Trichostrongylus were only rarely detected. Farms with BZ resistance-associated SNP frequencies above 10% were most often found in Switzerland followed by Ireland and Italy., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The efficacy of ivermectin (IVM) against gastrointestinal nematodes in Danish cattle was assessed by faecal egg count reduction test (FECRT). Six cattle farms with history of clinical parasitism and avermectin use were included. On the day of treatment (Day 0), 20 naturally infected calves per farm (total n = 120) were stratified by initial faecal egg counts (FEC) and randomly allocated to a treatment group dosed with 0.2 mg IVM kg -1 body weight s.c. (IVM; n = 10) or an untreated control group (CTL; n = 10). Individual FEC were obtained at Day 0 and Day 14 post-treatment and pooled faeces by group were cultured to isolate L3 for detection of Ostertagia ostertagi and Cooperia oncophora by qPCR. Treatment efficacies were analysed using the recommended WAAVP method and two open-source statistical procedures based on Bayesian modelling: 'eggCounts' and 'Bayescount'. A simulation study evaluated the performance of the different procedures to correctly identify FEC reduction percentages of simulated bovine FEC data representing the observed real data. In the FECRT, reduced IVM efficacy was detected in three farms by all procedures using data from treated animals only, and in one farm according to the procedures including data from treated and untreated cattle. Post-treatment, O. ostertagi and C. oncophora L3 were detected by qPCR in faeces of treated animals from one and three herds with declared reduced IVM efficacy, respectively. Based on the simulation study, all methods showed a reduced performance when FEC aggregation increased post-treatment and suggested that a treatment group of 10 animals is insufficient for the FECRT in cattle. This is the first report of reduced anthelmintic efficacy in Danish cattle and warrants the implementation of larger surveys. Advantages and caveats regarding the use of Bayesian modelling and the relevance of including untreated cattle in the FECRT are discussed., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: This study aimed to comparatively determine the prevalence of trichostrongylid nematodes and egg excretions in cows that are in the periparturient period (PPP) and pregnant cows in the non-periparturient period (NPPP)., Methods: Animal materials are constituted by cows in PPP and NPPP in 10 ranches in Kars and surrounding areas. Stool samples were collected from the rectum of 125 pregnant cows, which were in NPPP during November 2010 - January 2011 and from the same cows in PPP during April-May 2011. Stool samples were analyzed for the presence of nematode eggs by the flotation method. EPG was determined by the McMaster method., Results: Trichostrongylid nematode eggs were detected in seven of 10 dairy cattle ranches (70%). The prevalence rates of trichostrongylid nematodes were 16.8% (21/125) during NPPP and 27.2% (34/125) during PPP. Nineteen of 21 (90.5%) positive samples during NPPP were positive during PPP. Fifteen negative samples during NPPP were positive during PPP with respect to trichostrongylid nematode eggs. The egg count of trichostrongylid during PPP (total, 4350; mean, 34.8; n, 125) was greater than that during NPPP (total, 1250; mean, 10; n, 125)., Conclusion: Cows had nematode eggs during PPP and played an important role in transmission.

This paper focuses on the species diversity among the Trichostrongylidae Leiper, 1912 (Nematoda: Strongylida), and complexity of the family systematics. Polymorphism (subfamilies: Ostertagiinae, Cooperiinae and Haemonchinae), the presence of cryptic species (genus: Teladorsagia) and hybridization (genera: Cooperia, Haemonchus and Ostertagia) are presented and discussed, considering both morphological and molecular evidence. Some of these phenomena are common, nevertheless not sufficiently understood, which indicates the need for expanding the current state of knowledge thereof. Within the Trichostrongylidae, species distinction supported merely by morphological features is difficult, and requires confirmation by means of molecular methods. The parasitic nematode taxonomy is complicated mainly by the genus Teladorsagia, but complexity may also be expected among other Ostertagiinae (e.g. in the genera Ostertagia and Marshallagia). The data presented here show that the members of the Trichostrongylidae can significantly complicate unambiguous species identification. Hence, it is essential to consider the phenomena mentioned, to gather valid and comparable data on the biodiversity of this family.

In reproducing ewes, a periparturient breakdown of immunity is often observed to result in increased fecal egg excretion, making them the main source of infection for their immunologically naive lambs. In this study, we expanded a simulation model previously developed for growing lambs to explore the impact of the genotype (performance and resistance traits) and host nutrition on the performance and parasitism of both growing lambs and reproducing ewes naturally infected with Teladorsagia circumcincta. Our model accounted for nutrient-demanding phases, such as gestation and lactation, and included a supplementary module to manage the age structure of the ewe flock. The model was validated by comparison with published data. Because model parameters were unknown or poorly estimated, detailed sensitivity analysis of the model was performed for the sheep mortality and the level of infection, following a preliminary screening step. The parameters with the greatest effect on parasite-related outputs were those driving animal growth and milk yield. Our model enables different parasite-control strategies (host nutrition, breeding for resistance and anthelmintic treatments) to be assessed on the long term in a sheep flock. To optimize in silico exploration, the parameters highlighted by the sensitivity analysis should be refined with real data.

North American bison (Bison bison) producers face many challenges, including the potential clinical and economics problems caused by trichostrongyle nematodes within their herds. Little is known about the prevalence, intensity, geographical distribution and clinical significance of these parasites in commercial bison herds, even from regions where bison production has become popular. This study involved a large herd of bison from eastern South Dakota that was experiencing clinical parasitism due to a temporary over-stocking problem. After documenting fecal egg counts (FECs) and trichostrongyle genera present among the 3 main age-categories (i.e. adults, yearlings, calves) of bison during this heavily infected grazing season, the effects of doramectin treatment on the different age groups was also evaluated. This is the first bison study using PCR to identify genera of trichostrongyles in fecal samples. Virtually all 103 bison fecal samples from all 3 age classes were shedding trichostrongyle eggs by the end of the season, and the mean FECs were 34 eggs/g (EPG) among the cows, 125 EPG in the yearlings, and 186 EGP among calves. Based upon this heavily-infected herd, there is evidence that the susceptibility of bison to trichostrongyles is more similar to beef cattle than to sheep. Other parasites such as Moniezia, Nematodirus, Trichuris, and coccidians were also identified in these samples. All but 3 of the 51 samples analyzed with PCR shown at least 1 trichostrongyle genera. Ostertagia was detected in 68.6% of the samples, Cooperia in 80.39%, Haemonchus in at least 73% and Trichostrongylus in 16% of the herd. Most commonly, bison were infected with combinations of Haemonchus/Ostertagia/Cooperia. After treatment with doramectin, the mean FECs dropped by 99.9% for all of the bison age classes., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Resistance to ivermectin and moxidectin was explored by a faecal egg count reduction test in two sheep flocks with suspected anthelmintic resistance. The FECRT confirmed one suspicion, with a mean percentage of reduction in egg excretion within the treated groups of 0% for ivermectin (CI 95%: -228 to 58) and 13% for moxidectin (CI 95%: -152 to 70). This was further explored by a controlled efficacy test. An experimental infection of 18 naïve lambs was set up using infective larvae isolated from this flock (5000 L3/lamb). Compared to the control group, abomasal worm burdens (Teladorsagia circumcincta) were reduced by 90% [CI 95%: 81.5-94.8] and 85% [CI 95%: 72.4-92.2] after ivermectin (p<0.05) and moxidectin (p<0.05) treatment respectively. Again, compared to the control group, there was a reduction for intestinal strongyles (Trichostrongylus colubriformis) of 100% and 99% [CI 95%: 97.5-99.7] for ivermectin and moxidectin respectively. No difference was found between the efficacy of moxidectin and ivermectin. Pharmacokinetic values indicated that the strongyles were submitted to anthelmintic concentrations usually lethal to them. This trial demonstrated the first multiple resistance of ovine strongyles in France., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Refugia based anthelmintic protocols aim to reduce the rate of development of anthelmintic resistance in gastrointestinal nematodes (GIN). Previous studies have illustrated the impact of different drenching regimes on drug efficacy and animal growth; however, the impact on nematode populations has yet to be characterised within natural infections. This study investigated the changes in species composition of GIN throughout the grazing season, following implementation of four different ivermectin drenching regimes over six years: neo-suppressive monthly treatment (NST), targeted selective treatment (TST), strategic prophylactic treatment (SPT) and treatment upon observation of clinical signs (MT)., Methods: Lambs were grazed on one of eight replicate paddocks each grazing season following treatment regimes assigned in year 1. Faecal samples were collected fortnightly from all animals and hatched to first stage larvae (L1). DNA was extracted from individual L1 and a multiplex PCR assay targeting the internal transcribed spacer 2 (ITS2) region of Teladorsagia circumcincta, Trichostrongylus spp. and Haemonchus contortus conducted. Other species were identified using species-specific PCR. Worm-naïve tracer lambs were grazed on the paddocks at the start and end of each grazing season and adult worms recovered at post mortem to investigate the parasite population on pasture., Results: Results showed an overall decrease in species diversity in egg output from the NST group which occurred within a single grazing season and was consistent throughout the experiment. Species diversity was protected over six years in groups implementing TST, SPT and MT treatment regimes, designed to offer refugia. The expected shift in species prevalence throughout the season from Teladorsagia to Trichostrongylus was observed in all but the NST group where only Teladorsagia spp. were recovered from trial lambs by the end of the experiment. Worm burdens indicated the presence of several species at relatively low abundance on pasture in the NST group in 2011. However, these species were not represented in egg output from trial lambs, probably due to the frequent anthelmintic treatment administered throughout the grazing season., Conclusion: The molecular methods utilised here worked well. The comparable results of the three refugia-based treatment regimes suggest that nematode diversity can be maintained using part or whole group treatments if a rich supra-population of parasites are available to re-infect animals post treatment.

Gastrointestinal nematodes within the subfamily Ostertagiinae (Teladorsagia, Ostertagia, and Marshallagia et al.) are among the most common infections of domesticated livestock. These parasites are of particular interest, as many of the species within this group are of economic importance worldwide. Traditionally, nematode species designations have been based on morphological criteria. However, this group possesses poorly defined species. There is an urgent need to develop a reliable technique that can distinguish species of Ostertagiinae. DNA barcoding has been proved to be a powerful tool to identify species of birds, mammals, and arthropods, but this technique has not yet been examined for identifying species of Ostertagiinae. In this study, a total of 138 mitochondrial DNA (mtDNA) cytochrome c oxidase subunit I (COI) sequences from individuals representing 11 species of Ostertagiinae were acquired by PCR for the first time. The specimens were collected from pastoral area of northern China. Genetic divergence analyses showed that mean interspecific Kimura two-parameter distances of COI (13.61 %) were about four times higher than the mean value of the intraspecific divergence (3.69 %). Then, the performance of the COI to identify species of Ostertagiinae was evaluated by identification success rates using nearest neighbor (NN) and BLASTn. The results indicated that the rates of correct sequence identification for COI were high (>80 %) when using the NN and BLASTn methods. Besides, the deep lineage divergences are detected in Teladorsagia circumcincta. Meanwhile, the analyses also detected no genetic differentiation between some species such as Ostertagia hahurica and Ostertagia buriatica. These results indicate that the traditional status of species within Ostertagiinae should be closely examined based on the molecular data.

Based on the number and arrangement of cuticular ridges and configuration of the dorsal ray, nematode specimens collected from the small intestine of eight Guianan arboreal mice, Oecomys auyantepui (Rodentia: Sigmodontinae), in French Guiana are herein described and characterized. Guerrerostrongylus marginalis n. sp. (Heligmosomoidea: Heligmonellidae) shows a synlophe consisting of more than 40 ridges and a unique bursal arrangement with ray 8 (externo-dorsal) extending to the edge of the bursal margin, and appearing more prominent than the dorsal ray. This bursal arrangement is common in members of Hassalstrongylus Durette-Desset, 1971, but uncommon in the other four species in Guerrerostrongylus Sutton & Durette-Desset, 1991. The placement of the new species in Guerrerostrongylus is based on the number and nature of cuticular ridges and the ray arrangement and symmetry of the caudal bursa. Diagnostic characteristics of Guerrerostrongylus marginalis n. sp. include the length of ray 8 relative to bursal margin, the relative size of the spicules and vestibule, and the number of eggs in the uterus. We propose an amendment to the generic diagnosis of Guerrerostrongylus to modify the characters of the long rays 6 (postero-lateral), rays 8 (externo-dorsal), and dorsal ray as diagnostic, since at least ray 6 appears to be short in two different species in the genus, namely G. ulysi Digiani, Notarnicola & Navone, 2012 and G. marginalis n. sp., (© J.M. Weirich et al., published by EDP Sciences, 2016.)

A simple polymerase chain reaction (PCR) test was used to identify Ashworthius sidemi, a blood-sucking gastrointestinal nematode that commonly infects bison, red and roe deer, and moose in Poland. The present study uses this technique to confirm the possibility of transmission of A. sidemi infection from wildlife to domestic animals, such as cattle and sheep, grazing on the same natural pastures. A 406 bp fragment of genomic A. sidemi DNA was actually detected in DNA isolated from larval cultures derived from feces from cattle. A. sidemi DNA has been detected in cattle which represent a new host for this parasite. This is the first evidence of A. sidemi in cattle. The results reveal that a PCR test based on DNA from L3 larvae can be used for in vivo detection of A. sidemi invasions in breeding animals. In conclusion, the transfer of A. sidemi infection from wildlife to the farm animals sharing the same pastures appears possible., (Copyright © 2015. Published by Elsevier B.V.)

A new species of nematode, Torrestrongylus tetradorsalis n. sp., is described herein, based on specimens recovered from the small intestine of the leaf-nosed bat, Macrotus waterhousii, from the Biosphere Reserve "Sierra de Huautla" in the state of Morelos, Mexico. The new species is included in Torrestrongylus because it features a bursa of the type 3 - 2, a divided cephalic vesicle with an anterior half in the shape of an umbrella, and a posterior widened half. The new species can be distinguished from the only other congener T. torrei Pérez-Vigueras, 1935 by four key features: first, by the absence of cervical alae in both males and females; second, by the relatively longer second half of the cephalic cap; third, by the configuration of the dorsal ray, that does not have a medial terminal ray, and finally, by the structure of the spicules. This is the second species in the genus, previously known from bats of the families Phyllostomidae and Molossidae in Cuba, and now in Mexico., (© J.M. Caspeta-Mandujano et al., published by EDP Sciences, 2015.)

The difference between castrated male lambs and entire female lambs in susceptibility to gastrointestinal nematode infection was investigated in a flock of Scottish Blackface sheep. Castrated male lambs had higher faecal egg counts, higher body weights, lower parasite-specific plasma IgA activity and more adult nematodes. There were no detectable differences in the length of adult Teladorsagia circumcincta. Heavier egg shedding by males was associated with greater worm burdens. The difference in plasma IgA activity may have been a consequence of larger numbers of adult nematodes inhibiting the transfer of mucosal IgA to the plasma. At least part of the sex difference in egg production could be a consequence of males ingesting larger amounts of grass and hence numbers of infective larvae because of their larger size. The differences between castrated males and females contribute to the observed variation in nematode egg production in naturally infected lambs., (Copyright © 2014. Published by Elsevier B.V.)

Teladorsagia circumcincta is among the most important gastrointestinal parasites in small ruminants and the predominant species in Southern European goats. Parasite control is largely based on metaphylactic/preventative treatments, which is often seen as non-sustainable anymore. The reasons are increased consumer demand to reduce chemicals in livestock production and anthelmintic resistance against the common drugs. This study aimed at the development of a T. circumcincta-enzyme-linked immunosorbent assay (ELISA) specifically for goats. Samples were obtained from goats raised parasite-free or infected experimentally. Sampling continued during the following pasture season and housing period. The sensitivity for the use in bulk milk samples as an indicator of T. circumcincta infection levels in grazing goats was examined. The ELISA enables clear differentiation of negative and positive animals. With a specificity of 100% negative cut-off values for serum and milk were 0.294 and 0.228 (sensitivity, 95%). Positive cut-off values (sensitivity, 90%) were 0.606 (serum) and 0.419 (milk), while a sensitivity of 95% resulted in 0.509 and 0.363, respectively. The grey-zone between negative/positive cut-offs was introduced to deal with animals in pre-patency and decreasing antibody levels after infection. There was no cross reactivity for Trichostrongylus colubriformis and Cooperia oncophora while for Haemonchus contortus and Fasciola hepatica it cannot be fully excluded currently. In bulk milk samples, 5% of the milk had to be contributed from animals infected with T. circumcincta to be detected as positive. The results derived from experimentally and naturally infected as well as parasite naïve animals indicate the potential of the ELISA to be used in targeted anthelmintic treatment regimes in goats.

Twelve novel polymorphic microsatellite markers were produced and characterized for Spiculopteragia spiculoptera (Nematoda, Trichostrongyloidae) a common parasite of abomasum of Roe and Red deer, using next generation sequencing approach, and two multiplexes PCR were developed with these markers. Polymorphism of each locus was tested in 40 individuals of this species from diverse wild populations of cervids, and was tested for crossed-amplification on four other species of nematodes, close to S. spiculoptera among the Trichostrongyloidea: 20 Spiculopteragia houdemeri, 34 Ostertagia leptospicularis, 16 Ashworthius sidemi, and 25 Trichostrongylus spp. Our new microsatellite markers seem to be specific to Spiculopteragia spiculoptera since no amplifications were obtained for the four other species. The number of alleles per locus ranged from 2 to 12, the average observed and expected heterozygosity per locus ranged from 0.025 to 0.641 and from 0.049 to 0.664, respectively. Four of the 12 microsatellite loci showed significant deviations from Hardy-Weinberg equilibrium (which two slightly significant). One locus pair showed significant linkage disequilibrium (Sspi4 vs. Sspi8). Neither evidence of scoring error due to stuttering nor evidence of large allele dropout was found at all of the 12 loci, but evidence of null alleles was indicated at three loci because of general excess of homozygotes for most allele size classes. These polymorphic loci will be useful markers to study population genetics structure of Spiculopteragia spiculoptera in order to understand transfer and to explain the relationships between deer populations., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Results from worm egg counts (WECs) of cattle from across Australia over an 11 year period have been analysed to provide contemporary data on WEC and worm genus distribution in Australian cattle. A sampling kit consisted of ten sample containers. Faecal samples were collected into each sampling kit from groups of cattle and WEC and larval cultures were conducted using standard procedures. A submission form was completed for each kit with data requested including date of sampling, property address, age, average estimated weight and production type of cattle, last drench used and date of drenching. Regional analysis was done by postcode region and postcode regions were grouped into bioclimatic regions. A total of 5069 submissions were received from 2002 to the end of 2012 representing over 50,000 individual faecal samples. Seventy-seven percent of cattle sampled were no more than 2 years of age with the remainder representing a range of age groups. Samples were collected from all of the significant cattle producing regions of Australia. There was a tendency for higher geometric mean WEC in cattle in northern Australia and in high rainfall areas along the east coast of Australia. Geometric mean WEC for bioclimatic regions varied seasonally with a peak in autumn in regions with summer dominant rainfall, but little seasonal variation in regions with winter dominant rainfall patterns. Worm genera cultured varied throughout Australia with Cooperia spp. being most prevalent across the country, followed by Haemonchus spp. and Oesophagostomum spp. in summer rainfall dominant regions and Ostertagia spp. and Trichostrongylus spp. in winter rainfall regions. In the coastal, tablelands, and temperate rangelands regions of NSW, Haemonchus spp. were as prevalent as Cooperia spp. during autumn months and levels of Haemonchus spp. found in south-western Western Australia were higher than previously documented. These data provide an up-to-date summary of internal parasites in Australian cattle. Regional differences in WEC between northern and southern Australia are evident and should be further explored. These data suggest that there has been a move to Cooperia spp. being the predominant worm genus in Australian cattle, with increasing levels of Haemonchus spp. being evident in southern temperate regions of Australia., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The objectives of the present study were: (1) to report the percentage of cattle farms with gastrointestinal nematodes (GINs) resistant to levamisole in Veracruz, Mexico, (2) to identify the genera of GINs involved in resistance, and (3) to identify factors associated with these resistances. The faecal egg count reduction test (McMaster technique) was used to detect the presence of resistant GINs. A questionnaire was given to owners to understand the history of anthelmintic use. The percentage of cattle farms with GINs resistant to levamisole was 36.4% (4/11). The percentage of faecal egg count reduction on resistant farms was 91%, 82%, 42% and 88%. A similar number of cattle farms (4/11) were identified as potentially having levamisole resistance. Only three farms had GIN populations susceptible to levamisole. Cooperia spp. was the genus most commonly found to be resistant, followed by Haemonchus spp., Ostertagia spp. and Oesophagostomum spp. No factors were identified that influenced the presence of GIN resistance. However, there were identified inappropriate anthelmintic practices in cattle farms that should be improved. None of the farmers weighed their animals in order to dose them correctly with anthelmintics. Six cattle farms (54.5%) applied anthelmintics to new arriving animals. This is the first report of levamisole resistant GINs in Mexico. Improving the use of anthelmintics and measures of quarantine for infected cattle will help control the spread of resistance., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Livestock with access to pasture is generally exposed to infections with parasitic nematode species by uptake of infective third stage larvae (L3) with the grass. L3 can survive on pasture and particularly also in the soil up to several months and sometimes even longer, depending on temperature and humidity. As indicators for health and productivity of grazing animals it is important to determine the intensity and species spectrum of parasitic nematode larvae by analysing grass as well as soil samples. A rapid method for the recovery of L3 using a centrifugal-flotation technique from soil samples of 50-500 g was developed. The method takes advantage of the low specific weight of larvae to separate them from equal sized soil and debris particles by centrifuging them in a saturated sugar solution. A stack of differently sized sieves is used to achieve elimination of larger particles, dust and sugar from the sample to enable easy counting of larvae. Independent of the number of larvae used for inoculation of the samples a mean recovery of 75.3% was obtained. The recovery rates obtained ranged between 60.8% and 88.0% which demonstrates a considerably lower variability compared to earlier approaches and therefore a more precise estimation of the actual numbers of parasite larvae in soil is achieved. Further advantages over already developed methods are the use of easy, affordable and eco-friendly materials, the simplicity of the procedure and a faster processing time with the possibility to examine up to 20 samples per day., (Copyright © 2014 Elsevier Inc. All rights reserved.)