Occurrence of gastrointestinal nematodes in lambs in Norway, as assessed by copromicroscopy and droplet digital polymerase chain reaction.

BACKGROUND: Gastrointestinal nematodes (GINs) have a major impact on sheep production, health, and welfare worldwide. Norway is no exception, but there are only a few studies on the prevalence of GINs in Norwegian sheep. The aim of this study was to investigate the current occurrence of the most important nematodes in sheep flocks in Norway. Faecal samples were collected from flocks in 2021/2022, mainly from three geographical regions in Norway, i.e., northern, eastern, and western. In each of 134 flocks included, individual samples from 10 lambs (autumn) were pooled. Third stage larvae (L3) were cultivated and harvested (Baermann method) from the pooled samples. The DNA was then extracted and further analysed using droplet digital PCR (ddPCR). This enables assessment of the proportions of the three most important nematode species/genera, i.e., H. contortus, T. circumcincta, and Trichostrongylus. The fractional abundance/relative proportion of each species/genus was assessed by performing duplex assays with universal strongyle and species/genus-specific primers and probe sets. In addition, the occurrence of Nematodirus eggs was assessed by standard faecal egg counts (i.e., McMaster method). RESULTS: Of the 134 flocks sampled, 24 were from the northern region, 31 from eastern, and 71 from western Norway. In addition, some flocks from central (n = 7), and southern (n = 1) Norway were included. Among the sampled flocks, T. circumcincta occurred most commonly (94%), followed by H. contortus (60%) and Trichostrongylus (55%), and Nematodirus (51%). In general, mixed infections were observed, with 38% and 18% of flocks infected with three or all four genera, respectively. CONCLUSIONS: The results of this study indicate that GINs are widespread in Norway. Teladorsagia circumcincta seems to be present in most flocks based on this screening. Moreover, the results show that Nematodirus spp. infect lambs throughout the country, predominantly N. battus, and indicate that this nematode has become more abundant, which could lead to an increase in nematodirosis.

A single subcutaneous dose of eprinomectin (Eprecis®) is effective against common gastrointestinal nematodes and lungworms in experimentally infected lactating goats.

BACKGROUND: The health and productivity of dairy goats continue to be impacted by gastrointestinal nematodes (GIN) and lungworms (LW). Eprinomectin (EPN) is frequently selected for treatment because it is generally effective and does not require a milk withdrawal period. However, some factors, such as lactation, can have an impact on EPN pharmacokinetics and potentially its efficacy. To evaluate whether this can alter the efficacy of Eprecis® 2%, an eprinomectin injectable solution, a study was performed in lactating goats using the dose currently registered in cattle, sheep and goats (0.2 mg/kg). METHODS: This study was a blinded, randomized, controlled trial performed according to the VICH guidelines. Eighteen (18) worm-free lactating goats were included and experimentally challenged on day 28 with a mixed culture of infective gastrointestinal and lung nematode larvae (Haemonchus contortus, Trichostrongylus colubriformis, Teladorsagia circumcincta, Dictyocaulus filaria). At D-1, fecal samples were collected to confirm patent infection in all animals. On D0, the goats were randomly allocated into two groups of nine goats; group 1 was treated with Eprecis® 2% at 0.2 mg/kg BW by subcutaneous injection, while group 2 remained untreated. Fecal samples for egg counts were collected from all animals on days 3, 5, 7, 9, 11 and 14. On D14, all goats were killed, and the abomasum, small intestine and lungs were removed, processed and subsampled to record the number and species of worms. RESULTS: The treatment was well tolerated. After treatment, the arithmetic mean FEC decreased in the treated group and remained < 5 EPG until the end of the study, while the arithmetic mean FEC in the control group remained > 849.0 EPG. At D14, goats in the treated group had very limited or zero total worm counts, whereas all animals from the control group had a high worm burden. The measured efficacy was 100.0% against H. contortus and T. colubriformis, 99.9% against T. circumcincta and 98.0% against D. filaria. CONCLUSIONS: Eprinomectin (Eprecis®, 20 mg/ml), administered at the label dose (0.2 mg/kg), is highly effective against gastrointestinal nematodes and lungworms in lactating goats.

The identification of small molecule inhibitors with anthelmintic activities that target conserved proteins among ruminant gastrointestinal nematodes.

Gastrointestinal nematode (GIN) infections are a major concern for the ruminant industry worldwide and result in significant production losses. Naturally occurring polyparasitism and increasing drug resistance that potentiate disease outcomes are observed among the most prevalent GINs of veterinary importance. Within the five major taxonomic clades, clade Va represents a group of GINs that predominantly affect the abomasum or small intestine of ruminants. However, the development of effective broad-spectrum anthelmintics against ruminant clade Va GINs has been challenged by a lack of comprehensive druggable genome resources. Here, we first assembled draft genomes for three clade Va species (Cooperia oncophora, Trichostrongylus colubriformis, and Ostertagia ostertagi) and compared them with closely related ruminant GINs. Genome-wide phylogenetic reconstruction showed a relationship among ruminant GINs structured by taxonomic classification. Orthogroup (OG) inference and functional enrichment analyses identified 220 clade Va-specific and Va-conserved OGs, enriched for functions related to cell cycle and cellular senescence. Further transcriptomic analysis identified 61 taxonomically and functionally conserved clade Va OGs that may function as drug targets for new broad-spectrum anthelmintics. Chemogenomic screening identified 11 compounds targeting homologs of these OGs, thus having potential anthelmintic activity. In in vitro phenotypic assays, three kinase inhibitors (digitoxigenin, K-252a, and staurosporine) exhibited broad-spectrum anthelmintic activities against clade Va GINs by obstructing the motility of exsheathed L3 (xL3) or molting of xL3 to L4. These results demonstrate valuable applications of the new ruminant GIN genomes in gaining better insights into their life cycles and offer a contemporary approach to discovering the next generation of anthelmintics.IMPORTANCEGastrointestinal nematode (GIN) infections in ruminants are caused by parasites that inhibit normal function in the digestive tract of cattle, sheep, and goats, thereby causing morbidity and mortality. Coinfection and increasing drug resistance to current therapeutic agents will continue to worsen disease outcomes and impose significant production losses on domestic livestock producers worldwide. In combination with ongoing therapeutic efforts, advancing the discovery of new drugs with novel modes of action is critical for better controlling GIN infections. The significance of this study is in assembling and characterizing new GIN genomes of Cooperia oncophora, Ostertagia ostertagi, and Trichostrongylus colubriformis for facilitating a multi-omics approach to identify novel, biologically conserved drug targets for five major GINs of veterinary importance. With this information, we were then able to demonstrate the potential of commercially available compounds as new anthelmintics.

Unveiling a novel parasitosis: Trichostrongylus colubriformis infection in captive ring-tailed lemurs (Lemur catta).

Ring-tailed Lemur (Lemur catta) is the only surviving semi-terrestrial diurnal lemur in Madagascar. Despite being the most intensively studied of lemur taxa, only a few helminths have been described in this species. In this study we describe a persistent infection due to Trichostrongylus colubriformis in a captive population of L. catta hosted in a zoological park of northern Italy. In the context of a parasitological survey on zoo animals, we investigated parasites in a captive colony of ring-tailed lemurs within a zoological park. Parasitological analysis included necropsy of a deceased lemur in 2019, subsequent fecal sample collections in 2021-2022, followed by coprological examination and coprocolture. Morphological and molecular analyses were conducted on adult helminths, larvae and eggs, involving microscopy, scanning electron microscopy (SEM), and sequencing of the ITS rDNA region. Trichostrongylidae parasites were primarily found after necropsy in the intestine of the lemur. Morphological and molecular investigations on adults and eggs/larvae recovered from feces collected at different times from lemurs of the same captive population, allowed to properly identify the parasite as T. colubriformis. To the best of our knowledge this is the first description of T. colubriformis in L. catta. Although its presence in wild populations is not necessarily implied by our finding, this parasitosis represent a cause of concern in captive lemurs, considering the possibility of interspecies transmission and the zoonotic implications.

Pathomorphological Findings and Infectious Diseases in Selected European Brown Hare (Lepus europaeus Pallas, 1778) Populations from Schleswig-Holstein, Germany.

In the northernmost German federal state Schleswig-Holstein, populations of European brown hares (Lepus europaeus Pallas, 1778) show diverse densities and varying courses over the years. To examine differences in pathomorphological findings and infectious diseases as possible reasons for varying population dynamics, we assessed 155 hunted hares from three locations in Schleswig-Holstein from 2016 to 2020. We investigated the association of location, year, age, and sex of animals to certain pathomorphological findings and infectious diseases. Frequent pathomorphological findings were intestinal parasites (63.9%), hepatitis (55.5%), nephritis (31.0%), steatitis (23.2%), enteritis (13.5%), and pneumonia (5.2%). Body condition differed significantly between locations, and the prevalence of pneumonia was significantly higher in females. Enteritis was not detected in 2019, when much more juveniles were sampled. Hepatitis and nephritis occurred significantly more often in 2016 and among adults. Additionally, more adults showed hepatitis with concurrent serotitre for European brown hare syndrome virus (EBHSV), while intestinal parasitosis as well as high excretion rates of coccidia were more common in juveniles. Sampled animals showed high infection rates with Eimeria spp. (96.1%), Trichostrongylus spp. (52.0%), Graphidium strigosum (41.2%), and a high seroprevalence (90.9%) for EBHSV, without severe symptoms. This study revealed a low prevalence of infectious pathogens, but a high prevalence of chronic inflammations of unknown origin in the tested brown hare populations. Overall, our results indicate a rather minor importance of infectious diseases for observed population dynamics of analysed hare populations in Schleswig-Holstein.

Efficacy of a fixed-dose combination injectable (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) in Australian cattle against naturally acquired gastrointestinal nematode infections.

Australian producers have long used macrocyclic lactones (MLs) to successfully control cattle gastrointestinal nematodes (GINs) and consequently improve production parameters. However, the trajectory of ML resistance development in cattle GINs is following that of small ruminant nematode populations, highlighting a need for novel treatment options to provide efficacy in the current environment and interrupt the long-term establishment of ML-resistant GIN populations in Australian cattle. Here, we describe three field studies conducted in Australia to evaluate the efficacy of a single administration of a novel fixed-dose combination injectable (FDCI) endectocide against naturally acquired infections of cattle GINs. The FDCI is administered subcutaneously to deliver 0.2 mg/kg doramectin and 6 mg/kg levamisole hydrochloride (HCl). Study sites consisted of three farms in New South Wales (n = 2) and Victoria (n = 1). At each site, cattle were randomly allocated into one of three treatment groups: (1) untreated control (saline), (2) FDCI (0.2 mg/kg doramectin, 6 mg/kg levamisole HCl) or (3) positive control (0.2 mg/kg ivermectin). All treatments were administered on Day 0. Fecal samples were collected prior to treatment on Days -1 (Study 3) or 0 (Studies 1 and 2) and again on Day 14 (post-treatment) to evaluate efficacy via fecal egg count (FEC) and for coproculture. Adequacy of infection was confirmed at all three study sites, with Day 14 geometric mean (GM) FECs for saline-treated cattle ranging from 32.5 eggs per gram (EPG) to 623.7 EPG. FECs for FDCI-treated cattle were significantly reduced compared to saline-treated cattle (p ≤ 0.0001) on Day 14, with GM-based efficacy ≥ 99.7% at all three study sites. In contrast, ivermectin was 97.4% effective against cattle GINs in Study 1 but was only 47.2% and 39.8% effective at study site 2 and 3, respectively. Genus-specific efficacies suggest the presence of ivermectin-resistant Cooperia spp. (Study 1), Haemonchus spp. (Study 2) and Ostertagia spp. (Study 3) populations in the naturally infected cattle used in these studies. The post-treatment FEC and genus-specific efficacy estimations indicate the doramectin + levamisole HCl FDCI was highly efficacious against cattle GINs even in the face of ivermectin LOE at study sites 2 and 3. The efficacy of the new FDCI against both ML-susceptible and ML-resistant economically important cattle GINs in Australia affirms it is a valuable treatment option for producers operating in an environment of ML loss of efficacy.

Investigating intestinal parasitic infections with emphasis on molecular identification of Strongyloides stercoralis and Trichostrongylus colubriformis in north of Iran.

Currently, parasitic infections are one of the important health problems in the world, especially in developing countries. This study aims to investigate intestinal parasites with an emphasis on molecular identification through the analysis of mitochondrial COX1 and ITS2 gene sequences of Strongyloides stercoralis (S. stercoralis) and Trichostrongylus spp. in north of Iran. Five hundred forty stool samples were collected from medical diagnostic laboratories affiliated with Mazandaran University of Medical Sciences in Sari city, north of Iran. First, all the samples were examined using direct smear, formalin-ether sedimentation, and trichrome staining technique. Suspected samples of Strongyloides larvae were cultured in agar plate. Then, DNA was extracted from samples containing Trichostrongylus spp. eggs and Strongyloides larvae. To amplify DNA, PCR was performed and the samples with a sharp band in electrophoresis were sequenced by Sanger method. Overall, the prevalence of parasitic infections in the study population was 5.4%. The highest and the lowest level of infection was observed with Trichostrongylus spp. and S. stercoralis at 3% and 0.2%, respectively. No traces of live Strongyloides larvae were seen in the culture medium of the agar plate. The six isolates obtained from the amplification of the ITS2 gene of Trichostrongylus spp. were sequenced, all of which were Trichostrongylus colubriformis. The sequencing results of COX1 gene indicated S. stercoralis. In the present study, the prevalence of intestinal parasitic infections in north of Iran has relatively decreased that its main reason can be due to the coronavirus epidemic and compliance with health principles. However, the prevalence of Trichostrongylus parasite was relatively high that it requires special attention to apply appropriate control and treatment strategies in this field.

Molecular and phylogenetic characterization of zoonotic Trichostrongylus species from goats for the first time in Bangladesh.

BACKGROUND: Trichostrongylus is one of the most important nematodes infecting animals and humans. The current study was designed to identify the Trichostrongylus species infecting goats by multiplex PCR and phylogenetic analysis. METHODS: A total of 124 goats' viscera were collected from different abattoirs of Mymensingh division. Trichostrongylus species were isolated and characterized based on morphometry, multiplex PCR and phylogenetic analysis. RESULTS: Among 124 viscera of goats, 39 were positive with two species, Trichostrongylus colubriformis and Trichostrongylus vitrinus, revealing an overall 31.45% prevalence. Morphological identification of Trichostrongylus species was confirmed by multiplex PCR amplification of the ITS2 gene and sequencing. Partial sequencing of the ITS2 gene of two species revealed seven single nucleotide polymorphisms (three transitions and four transversions) in this study. The neighbor-joining phylogenetic tree demonstrated that T. colubriformis and T. vitrinus isolates were clustered together with the reference sequences that belong to the clade A and B without any geographical boundaries. CONCLUSIONS: This is the first report on molecular and phylogenetic analysis of Trichostrongylus species from ruminants in Bangladesh. These results provide the baseline data for understanding the zoonosis and epidemiology of this parasite in Bangladesh and global perspectives.

Two in vitro anthelmintic assays of four Paraguayan medicinal plants for proof of concept of the role of polyphenols in their biological activities and LC-HRMS analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Gastrointestinal nematodes (GIN) control in small ruminants has relied on the systematic use of synthetic anthelmintics (AH), their effectiveness has been progressively decreasing due to the rise and diffusion of anthelmintic resistances. The most prevalent genera affecting small ruminants were Haemonchus spp., and Trichostrongylus spp. The investigation of new anthelmintics in plants is a highly studied option, especially when it is linked to ethnobotanical knowledge and phenolic compounds. THE AIMS OF OUR STUDY: Four medicinal plants mentioned in traditional medicine were selected to evaluate their anthelmintic proprieties at different stages of the life cycle of GIN, namely: Kyllinga odorata Valh., Cassia occidentalis L., Artemisia absinthium L, and Verbena litoralis Kunth and to explore the role of polyphenols in the AH activity. MATERIALS AND METHODS: To evaluate the anthelmintic activity in this study, two models of GIN species, namely Haemonchus contortus (Hc) and Trichostrongylus colubriformis (Tc) were selected and tested on two in vitro assays: 1) Larval Exsheathment Inhibition Assay (LEIA) and, 2) Egg Hatch Assay (EHA). To explore the role of tannins and polyphenols in AH activity by comparing the effects of LEIA and EHA with or without polyvinylpolypyrrolidone (PVPP) and to characterize the phytochemical composition of the most active plants using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). RESULTS: C. occidentalis exhibited the highest activity on LEIA (EC50 = 250.42-41.80 µg/mL) and A. absinthium on egg hatching processes (EC50 = 121.70-137.34 µg/mL) in both species of GIN. The inhibition in the development of eggs was from 67.70% to 96.36% on H. contortus, and from 78.87% to 99.65% on T. colubriformis. At the maximal dose, Additionally, it was observed that the AH on eggs varies according to the GIN species: on H. contortus the extracts tested blocked the formation of larvae Ovicidal Effect (% higher OE) and on T. colubriformis they blocked the appearance of L1 larvae, Larvae Failing Eclosion (% higher LFE). After PVPP, a reduction in AH activity on LEIA and EHA was noted, especially with C. occidentalis (87.20-67.00% of larvae exsheathment, (p < 0.05) and 40.51-24.96% of egg hatching, (p > 0.05) of both parasite species. Nine putative features were identified using HRMS and MS/MS after addition of PVPP. CONCLUSIONS: The present study demonstrated that C. occidentalis, A. absinthium, and K. odorata, which parts have been traditionally used as medicinal plants are a valuable source of active compounds with anthelmintic activity. The medicinal use of these plants against GIN parasites was proven by in vitro analysis. Therefore exploration of the secondary metabolites of these plant extracts and testing of isolated fractions of active compounds under in vivo experiments are planned and represent a specific challenge for alternative drug research. Regarding the PVPP, in this study we hypotheses about the standard doses it was not able to completely absorb the polyphenols of extracts of K. odorata, C. occidentalis, and A. absinthium, which would lead to more studies to evaluate the role of this product in the absorption of phenolic compounds.

Molecular Characterization of Mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene from Trichostrongylus Species (Nematoda: Trichostrongylidae) in Northern Iran

Objective: The objective of the present study was to identify Trichostrongylus species by molecular analysis and also phylogenetic relationships of Trichostrongylus species by mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene in Guilan province, northern Iran. Methods: Abomasum and duodenum contents of 144 livestock were collected from sheep, goats, and cattle in Guilan province. Morphological survey was performed for initial screening. Total DNA was extracted, and the partial region of Cox1 gene was amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the data on nucleotide sequence was conducted by MEGA7 software. Results: Three species of Trichostrongylus including T. colubriformis, T. vitrinus, and T. axei were identified by morphological characteristics. The genetic divergence within the species in the present study was observed for T. axei (0-2.5%), T. colubriformis (0.77%), and T. vitrinus (0%). The mean inter-species difference between the three species of Trichostrongylus obtained in this study was 14.4-15.4%. Conclusion: The Cox1 sequences of the members of Trichostrongylus spp. were highly variable and this could be used as a valuable measure to achieve a proper assessment on biodiversity. Sequence data generation from other species of Trichostrongylus will be needed to reconstruct the phylogenetic relationships of this genus of nematodes.

Status of Infectious Diseases in Free-Ranging European Brown Hares (Lepus europaeus) Found Dead between 2017 and 2020 in Schleswig-Holstein, Germany.

The European brown hare (Lepus europaeus) is a quite adaptable species, but populations have been decreasing for several decades in different countries, including Germany. To investigate infectious diseases as possible influences on observed population decline in the German federal state Schleswig-Holstein, 118 deceased free-ranging European brown hares were collected between 2017 and 2020 and underwent detailed postmortem examination with extended sampling. Infectious diseases were a major cause of death (34.7%). The number of juveniles found exceeded the adult ones. The main pathomorphological findings were hepatitis (32.8%), pneumonia (22.2%), nephritis (19.1%), liver necrosis (12.9%), and enteritis (40.7%). An unusual main finding was steatitis (20.9%) of unknown origin. Animals were mainly emaciated and showed high infection rates with Eimeria spp. (91.3%) and Trichostrongylus spp. (36.2%). European Brown Hare Syndrome Virus reached an epidemic status with few fatal infections (4.2%) and high seroprevalence (64.9%), whereas the prevalence of Rabbit Haemorrhagic Disease Virus 2 was very low (0.8%) in hares in Schleswig-Holstein. Pathogens such as Yersinia pseudotuberculosis (5.9%), Pasteurella multocida (0.8%), and Staphylococcus aureus (3.4%) only caused sporadic deaths. This study illustrates the wide distribution of various infectious pathogens with high mortality and even zoonotic potential. Infectious diseases need to be considered as an important influence on population dynamics in Schleswig-Holstein.

Trichostrongylosis: a zoonotic disease of small ruminants.

In the present world a significant threat to human health is posed by zoonotic diseases. Helminth parasites of ruminants are one of the most common zoonotic organisms on the planet. Among them, trichostrongylid nematodes of ruminants, found worldwide, parasitize humans in different parts of the world with varying rates of incidence, particularly among rural and tribal communities with poor hygiene, pastoral livelihood and poor access to health services. In the Trichostrongyloidea superfamily, Haemonchus contortus, Teladorsagia circumcincta, Marshallagia marshalli, Nematodirus abnormalis and Trichostrongylus spp. are zoonotic in nature. Species of the genus Trichostrongylus are the most prevalent gastrointestinal nematode parasites of ruminants that transmit to humans. This parasite is prevalent in pastoral communities around the world and causes gastrointestinal complications with hypereosinophilia which is typically treated with anthelmintic therapy. The scientific literature from 1938 to 2022 revealed the occasional incidence of trichostrongylosis throughout the world with abdominal complications and hypereosinophilia as the predominant manifestation in humans. The primary means of transmission of Trichostrongylus to humans was found to be close contact with small ruminants and food contaminated by their faeces. Studies revealed that conventional stool examination methods such as formalin-ethyl acetate concentration or Willi's technique combined with polymerase chain reaction-based approaches are important for the accurate diagnosis of human trichostrongylosis. This review further found that interleukin 33, immunoglobulin E, immunoglobulin G1, immunoglobulin G2, immunoglobulin M, histamine, leukotriene C4, 6-keto prostaglandin F1α, and thromboxane B2 are vital in the fight against Trichostrongylus infection with mast cells playing a key role. This review focuses on the prevalence, pathogenicity and immunological aspects of Trichostrongylus spp. in humans.

Influence of targeted selective anthelmintic treatment on the productive performance of wool and hair lambs naturally infected with gastrointestinal nematodes in Brazil.

Targeted selective treatment (TST) is an alternative method to reduce the use of anthelmintics and delay the development of resistant nematode populations. However, there is limited information on the actual effects of this type of treatment on livestock productivity. The objective of this study was to evaluate the production performance of Santa Ines (hair) and Ile de France (wool) lambs naturally infected with gastrointestinal nematodes (GIN) under TST based on packed cell volume (PCV) versus suppressive anthelmintic treatments. Thirty-eight lambs were divided into two treatment groups: Suppressive treatment, animals were drenched with monepantel every two weeks and TST, animals were treated with the same anthelmintic when they presented PCV ≤ 20%. Feces, blood, and weight were measured weekly to determine eggs per gram of feces, PCV, total plasma protein, and weight gain. After animals were slaughtered, carcasses were weighed to determine carcass yield. In the TST group, substantial productive losses of approximately 21.3% in the wool and 25.9% in the hair lambs were observed in body weight compared to their counterparts. Significant differences in hematological variables occurred over the experimental period, especially in the wool lambs under TST. Favorable environmental conditions enabled infective larvae to survive and thrive on pasture. Haemonchus contortus and intestinal nematodes were the most common parasites found in the Ile de France lambs and the Santa Ines lambs, respectively. Although TST prevented mortality, it did not prevent production losses. Both breeds showed a significant drop in production due to GIN parasitism.

Comparative analysis of the anthelmintic efficacy of European heather extracts on Teladorsagia circumcincta and Trichostrongylus colubriformis egg hatching and larval motility.

BACKGROUND: Gastrointestinal nematode (GIN) control is traditionally achieved with the use of anthelmintic drugs, however due to regulations in organic farming and the rise in anthelmintic resistance, alternatives are sought after. A promising alternative is the use of bioactive plant feeding due to the presence of plant secondary metabolites (PSMs) such as proanthocyanidins (PAs). This study focussed on the perennial shrub heather (Ericaceae family), a plant rich in PAs, highly abundant across Europe and with previously demonstrated anthelmintic potential. METHODS: In vitro assays were used to investigate heather's anthelmintic efficacy against egg hatching and larval motility. Heather samples were collected from five European countries across two seasons, and extracts were tested against two GIN species: Teladorsagia circumcincta and Trichostrongylus colubriformis. Polyphenol group-specific ultraperformance liquid chromatography-tandem mass spectrometry analysis was performed to identify relevant polyphenol subgroups present, including the PA concentration and size and ratio of the subunits. Partial least squares analysis was performed to associate efficacy with variation in PSM composition. RESULTS: Heather extracts reduced egg hatching of both GIN species in a dose-dependent manner by up to 100%, while three extracts at the highest concentration (10 mg/ml) reduced larval motility to levels that were not significantly different from dead larvae controls. PAs, particularly the procyanidin type, and flavonol derivatives were associated with anthelmintic activity, and the particular subgroup of polyphenols associated with the efficacy was dependent on the GIN species and life stage. CONCLUSIONS: Our results provide in vitro evidence that heather, a widely available plant often managed as a weed in grazing systems, has anthelmintic properties attributed to various groups of PSMs and could contribute to sustainable GIN control in ruminant production systems across Europe.

Gastrointestinal Parasites in Sheep from the Brazilian Pampa Biome: Prevalence and Associated Factors.

Parasitic infections are common in sheep farming worldwide. A cross-sectional study was carried out to determine the prevalence and associated risk factors among gastrointestinal parasites and sheep herds from the Brazilian Pampa biome, Rio Grande do Sul state, Brazil. Twenty-one farms were visited, where rectal faecal samples were individually collected from 309 animals. The sheep industry on the studied farms was characterised by small farms with a low level of technification, an extensive grazing system and meat production. Of all samples analysed, strongyle nematodes had the highest prevalence (77.02%), followed by Eimeria spp. (70.55%), Moniezia expansa (20.39%) and Strongyloides papillosus (17.48%). Mixed infection, between helminths and protozoans, was detected in 68.61% of samples. All herds were positive for strongyle and Eimeria spp. A predominance of Haemonchus spp. and Trichostrongylus spp. nematodes was observed in the herds. Younger animals were significantly more affected by Eimeria and M. expansa. In the semi-intensive and intensive systems, a higher frequency of Eimeria and strongyle infections was observed. Parasite infection was significantly reduced at low animal densities. The Brazilian Pampa region presents a high prevalence of gastrointestinal parasites among sheep; age, breeding system and stocking density were factors associated with parasite infection.

As infecções parasitárias são comuns na criação de ovinos em todo o mundo. Um estudo transversal foi realizado para determinar a prevalência e fatores de risco associados entre parasitoses gastrintestinais e rebanhos ovinos do bioma Pampa brasileiro, estado do Rio Grande do Sul, Brasil. Vinte e uma fazendas foram visitadas, onde foram coletadas amostras de fezes individuais da ampola retal de 309 animais. A produção ovina era caracterizada por pequenas propriedades com baixo nível de tecnificação, os animais eram criados em sistema extensivo e para produção de carne. De todas as amostras analisadas, os estrongilídeos (Nematoda: Strongylida) foram mais prevalentes (77,02%), seguido por Eimeria spp. (70,55%), Moniezia expansa (20,39%) e Strongyloides papillosus (17,48%). Infecção mista, entre helmintos e protozoários, foi detectada em 68,61% das amostras. Todos os rebanhos foram positivos para estrongilídeos e Eimeria spp. Foi observada predominância dos nematódeos Haemonchus spp. e Trichostrongylus nos rebanhos. Animais mais jovens foram significativamente mais afetados por Eimeria e M. expansa. Nos sistemas semi-intensivo e intensivo, observou-se maior frequência de infecções por Eimeria e estrongilídeos. A infecção parasitária foi significativamente menor em fazendas com baixa densidade animal. A região do Pampa brasileiro apresenta alta prevalência de parasitos gastrintestinais em ovinos; idade, sistema reprodutivo e densidade animal foram fatores associados à infecção parasitária.

Eprinomectin and Moxidectin Resistance of Trichostrongyloids on a Goat Farm in Austria.

Anthelmintic resistance (AR) of trichostrongyloids is widespread in Europe, but there is no up-to-date information on the spread of AR in caprine parasites in Austria. Eprinomectin (EPR) is currently the only anthelmintic drug of the macrocyclic lactones registered for goats in Europe. The aim of the present study was to gather information regarding the efficacy of anthelmintics against trichostrongyloids on a dairy goat farm in Austria with reported treatment failure of macrocyclic lactones and to determine the presence of different trichostrongyloid genera. Faecal egg count reduction tests (FECRT) using Mini-FLOTAC were performed with eprinomectin (EPR) and moxidectin (MOX). Egg count reduction, calculated with the R package egg-Counts, was 44% for EPR and 86% for MOX, confirming AR of trichostrongyloids for both compounds. The most frequently detected genus in larval cultures was Haemonchus, followed by Trichostrongylus. This is the first report of MOX resistance in caprine trichostrongyloids in Europe. Failure of EPR and MOX to control trichostrongyloid infections is a severe threat to dairy goat farming, since other compounds must not be applied in goats used for milk production. Haemonchus contortus is one of the most pathogenic parasites of small ruminants and can quickly develop AR. Thus, immediate action should be taken to slow the further spread of AR in this and other roundworm species of ruminants in Austria.

First Report of Multiple Drug-resistant Gastrointestinal Nematodes of Sheep in Arbat District, Sulaymaniyah, Iraq Detected By in Vivo and in Vitro Methods.

INTRODUCTION: The control of gastrointestinal nematodes (GINs) in small ruminants is principally dependent on anthelmintic therapy, which encounters the rising problem of anthelmintic resistance (AR) development. Veterinarians reported anthelmintic failure in several sheep farms in Arbat District, Sulaymaniyah, northern Iraq, which called for a systematic study about the efficacy of three commonly used drugs: albendazole, ivermectin, and levamisole. MATERIAL AND METHODS: A faecal egg count reduction test (FECRT) was conducted to elucidate the anthelmintics' efficacies, with coprocultures to determine the GINs parasitising sheep in the study area. Larval development assays (LDAs) were conducted to determine the drugs' median inhibitory concentrations (IC50). RESULTS: The FECRT revealed that AR was widespread to all three drugs, and ivermectin was the least effective, reducing the faecal egg counts (FECs) by 50.5% to 57.1%. The coprocultures revealed that the GIN genera of sheep in Arbat District were Trichostrongylus, Nematodirus, and Trichuris, and the resistance was mainly due to Trichostrongylus species. The mean IC50 of albendazole, ivermectin, and levamisole were 0.073 ng/mL, 7.97 ng/mL, and 1.43 ng/mL, respectively. CONCLUSION: This study is the first Iraqi report of AR confirmed by both in vivo FECRT and in vitro LDA methods.

Case report: Control of intestinal nematodes in captive Chlorocebus sabaeus.

There are limited data on the efficacy of antiparasitic treatments and husbandry methods to control nematode infections in captive populations of African green monkeys (AGMs), Chlorocebus sabaeus. In faecal egg count (FEC) tests, 10 of the 11 (91%) adult male AGMs captured from the large feral population on the island of St Kitts had evidence of nematode infections, mostly Capillaria (8/11, 73%), Trichuris trichiura (7/11, 64%) and strongylid species (7/11, 64%) specifically (hookworm and Trichostrongylus, 50/50), but also Strongyloides fuelleborni (1/11, 9%). When kept in individual cages with cleaning and feeding regimens to prevent reinfections and treated concurrently with ivermectin (300 µg/kg, given subcutaneously) and albendazole (10 mg/kg, given orally) daily for 3 days, 60% (6/10) of the AGMs were negative at a follow-up FEC at 3 months and by FEC and necropsy at the end of the study 5-8 months later. One monkey appeared to have been reinfected with T. trichiura after being negative by FEC at 3 months post-treatment. Four AGMs were positive for T. trichiura at the 3 month FEC follow-up but were negative at the end of the study after one further treatment regimen. Although initially being cleared of Capillaria following treatment, three AGMs were found to be infected at the end of the study. The ivermectin and albendazole treatment regimen coupled with good husbandry practices to prevent reinfections effectively controlled nematode infections in captive AGMs.

Brote familiar por el nematodo Trichostrongylus colubriformis en una zona rural de la provincia de Valdivia: una zoonosis de rara ocurrencia / Family outbreak caused by the nematode Trichostrongylus colubriformis in a rural area of the province of Valdivia: a rare occurrence zoonoses

Resumen La trichostrongylosis es una zoonosis de rara ocurrencia causada por nematodos intestinales, favorecida por el consumo de vegetales crudos o agua contaminada con larvas infectantes de Trichostrongylus spp. En 2015, se registró un brote familiar transmitido por alimentos en una zona rural de Valdivia, afectando a una mujer de 51 años de edad y su pareja e hija de 56 y 12 años; respectivamente. Solo la mujer adulta, el caso índice, presentó síntomas como diarrea, dolor abdominal, astenia y náuseas. Los pacientes eliminaron huevos de Trichostrongylidae gen. sp. en sus deposiciones, identificándose en dos de ellos, adultos de Trichostrongylus colubriformis. Tuvieron una buena respuesta a albendazol. En Chile, se han registrado prevalencias de infección de 0,1 a 3,5%, incluyendo un total de 93 casos diagnosticados entre las provincias de Concepción y Llanquihue.

Abstract Trichostrongylosis is a rare occurrence zoonosis caused by intestinal nematodes, favored by the consumption of raw vegetables or water contaminated with infective larvae of Trichostrongylus spp. In 2015, a family outbreak of foodborne disease was registered in a rural zone of Valdivia, affecting to 51-year-old woman and her 56-year-old partner and her 12-year-old daughter. Only the adult woman, the index case, presented symptoms such as diarrhea, abdominal pain, asthenia, and nausea. The patients revealed Trichostrongylidae gen. sp. eggs in their stools, identifying Trichostrongylus colubriformis adults in two of them. They had a good response to treatment with albendazol. In Chile, 0.1 to 3.5% prevalence of infection has been reported, including a total of 93 cases recorded between the provinces of Concepción and Llanquihue.

Triple lectin staining of trichostrongyle eggs from naturally infected small ruminants.

Trichostrongyle nematodes can be a major threat to the profitability of small ruminant producers depending of the species and intensity of trichostrongyles parasitizing their herd. Haemonchus contortus, Teladorsagia circumcincta, and Trichostrongylus colubriformis are typically the most common and clinically important species. Three lectins (PNA, LCA and AAL) have been reported to bind specifically to eggs from these three genera and therefore could be used to quantify the intensity of each species in individual animals. Peanut agglutinin (PNA) has been the most commonly tested lectin because it selectively binds intensely to eggs of the most pathogenic species, H. contortus. Lens culinaris agglutinin (LCA) and Aleuria aurantia agglutinin (AAL) have shown specificity to T. circumcincta and Trichostrongylus spp. respectively, however, these lectins have only been evaluated using eggs harvested directly from adult females, and not from fecal samples. The purpose of the present study is to describe a method to sequentially stain H. contortus, T. circumcincta and Trichostrongylus spp. fecal eggs with PNA, LCA and AAL, and then evaluate the resultant staining patterns seen with eggs collected from a naturally infected goat shown with PCR to contain H. contortus, T. circumcincta, Ostertagia leptospicularis, Trichostrongylus colubriformis and Trichostrongylus axei eggs. These results were also compared with patterns observed with eggs stained with single lectins and double combinations of lectins. The various patterns were then compared to those seen with egg samples collected from an ewe shown to only contain H. contortus. PNA bound intensely and uniformly to all eggs from samples containing only H. contortus eggs; however, some eggs additionally bound LCA and AAL in localized patches of varying size, and a few eggs exhibited intense and uniform binding of all three lectins. Single PNA-staining of goat samples containing the five trichostrongyles species identified most eggs as H. contortus, and triple-staining showed patterns consistent with those seen for H. contortus. Binding of AAL to non-Haemonchus eggs was uniform but showed significant variations in intensity. Lesser staining eggs tended to also stain intensely with LCA, which is consistent with published binding pattern for T. circumcincta. Most eggs that AAL bound intensely to did not bind with LCA, which is consistent with published binding pattern for Trichostrongylus spp. Autofluorescence was observed with the DAPI filter-cube among most non-Haemonchus eggs. This study demonstrates the need for additional field studies to further validate the specificity of these three lectins for use in identifying eggs from the three species of trichostrongyles.