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Haemonchus contortus is known for its high pathogenicity in sheep, and the uncontrolled use of anthelmintics resulted in the emergence of multiple drug-resistant populations. Breeding sheep for gastrointestinal nematode resistance is a sustainable alternative to reduce dependence of anthelmintic drugs, and differences in the degree of resistance between breeds have been reported. Here we compare two sheep breeds (Santa Ines and Ile de France), concerning the differences in innate and adaptive immune response involved in the resistance against H. contortus infection. Immunohistochemical analyses of the abomasum were conducted in naïve Santa Ines (n = 14) and Ile de France (n = 12) lambs randomized into four groups: infected Santa Ines (n = 8), non-infected control Santa Ines (n = 6), infected Ile de France (n = 8), and non-infected control Ile de France (n = 4). The infected lambs were initially infected with H. contortus infective larvae at 14 days of age, and multiple infections were conducted every second day until they reached 66 days of age. There was a significant effect (P < 0.001) of the infection with increase in numbers of CD3 + T; CD79α+ B; GATA3+ Th2/ILC2; POU2F3+ tuft cells; FOXP3+ T reg; and IgE + cells in the fundus of the abomasal mucosa in both Santa Ines and Ile de France lambs. Nevertheless, the infected Santa Ines lambs presented the highest averages for CD79α+ B; GATA3+ Th2/ILC; IgE + cells; and POU2F3+ tuft cells and there was a significant association of the breed and infection status with regards to POU2F3+ tuft cells, with the highest mean in the infected Santa Ines group. The infected Santa Ines group had three lambs with high degree of resistance and five lambs that showed a moderate infection. Our results suggest a mechanism of synergistic coordination between different immune-cell types in promoting resistance of suckling lambs under H. contortus infection., Competing Interests: Declaration of competing interest There are no conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome., (Copyright © 2024. Published by Elsevier Ltd.)

Due to the negative impact of Haemonchus contortus in the tropics and subtropics, the detection of serum protein profiles that occur in infected sheep is of high relevance for targeted selective treatment strategies (TST). Herein, we integrated proteomics with phenotypic traits to elucidate physiological mechanisms associated to H. contortus infection in susceptible (Dorper - D) and resistant (Santa Inês - S) sheep breeds. Naïve female lambs were infected with H. contortus third-stage larvae on day zero (D0), and samples were collected weekly, for 28 days. Feces were used for individual fecal egg counts (FEC) blood for packed cell volume (PCV) and serum for specific antibody quantification through ELISA. Sera was collected on D0 (-) and D21 (+), and analyzed using a LC-MS/MS based proteomics approach. FEC, PCV, and anti-H. contortus antibody levels confirmed the absence of infection on D0. On D28 there was a significant difference between the two breeds for logFEC means (D = 3774 and S = 3141, p=0.033) and PCV means (D = 16.3 % and S = 24.3 %, p=0.038). From a total of 754 proteins identified, 68 differentially abundant proteins (DAPs) were noted. Phosphopyruvate hydratase (ENO3) was a DAP in all comparisons, while S+ vs D+ and S- vs D- shared the highest number of DAPs (8). Each of the four experimental groups clustered separately in a principal component analysis (PCA) of protein profile. Among the DAPs, proteins associated with the innate and adaptive immune system were detected when comparing S- vs D- and S+ vs D+. In D-, some proteins were linked to stress response to handling, sampling and heat. Focusing on the consequences of infection in each breed, in the D+ vs D- comparison, upregulated proteins were associated with inflammation control and immune response, where downregulated proteins pointed to a negative impact of infection on tissue anabolism, compromising muscle growth and fat deposition. In the S+ vs S- comparison, upregulated proteins were related to immune response, while the downregulated proteins were possibly linked to muscular development and growth, impaired by infection. Collectively, it can be concluded that ENO3 regulation emerges as a potential factor underlying the differential immune response observed between Santa Inês and Dorper sheep infected with H. contortus. In turn, detected acute phase proteins (APPs) reinforce their relation with infection, inflammation and stress conditions, whereas THEMIS-like may contribute to the immune system in Dorper. GSDMD, Guanylate-binding protein and ACAN warrant further investigation as possible biomarkers for TST strategy development., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ana Carolina S. Chagas reports financial support was provided by State of Sao Paulo Research Foundation. Andre M. Almeida reports financial support was provided by Foundation for Science and Technology. If there are other authors, they 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 Elsevier B.V. All rights reserved.)

Haemonchus contortus and Trichostrongylus colubriformis are the most important gastrointestinal nematodes causing serious losses in sheep production of tropical and subtropical regions. Prophylaxis of gastrointestinal nematode infections is based on anthelmintics use, but their frequent administration selects multiple-resistant parasites. To evaluate how the situation has changed over the last decades, the anthelmintic resistance status of gastrointestinal nematodes in sheep flocks was assessed in the current study and compared to previous surveys. In each one of the 15 flocks evaluated, animals (n ≥ 7) were allocated into at least five groups and treated as follows: 1) untreated control; 2) albendazole; 3) levamisole; 4) ivermectin; and 5) monepantel. If more animals were available, two additional groups were included: 6) closantel, and 7) moxidectin. The faecal egg count reduction test (FECRT) was carried out to evaluate the pre- and post-treatment using the SHINY tool. Haemonchus spp. was the most prevalent nematode from faecal cultures. The mean efficacy of albendazole was 40%. Only in two farms, levamisole presented a relatively high percentage of reduction in the FECRT about 90%, while ivermectin and moxidectin presented the worst mean efficacy of 34% and 21% among all farms, respectively. Like other anthelmintics, closantel demonstrated low efficacy (63%) across all farms evaluated. Monepantel presented an overall mean efficacy of 79%, but it was the only anthelmintic that presented efficacy ≥95%, in five farms. The results revealed that gastrointestinal nematodes with multiple anthelmintic resistance were prevalent in all 15 sheep herds. The research suggests that nematodes are becoming more and more resistant to various anthelmintic compounds, which has made the problem worse. This circumstance highlights the necessity to put into practice sustainable and long-lasting methods to prevent gastrointestinal nematode infections in sheep husbandry., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Although ivermectin (IVM) has a wide spectrum and long half-life, its frequent use as an anthelmintic for the last 42 years led to its worldwide tolerance by Haemonchus contortus. We evaluated the combination of limonene (LIM), a P-glycoprotein (Pgp) modulator, with IVM in lambs infected with a multidrug-resistant H. contortus. Twenty-four male Dorper lambs were artificially infected with two doses (seven days apart) of 8000 infective larvae of a multidrug-resistant isolate of H. contortus. The infection was patent 25 days later. Fifteen days before treatment with IVM (DAY -15), animals were divided into 4 groups: Infected-untreated control (CTL), IVM, LIM, and LIM+IVM. From DAY -15 to DAY + 14, groups LIM and LIM+IVM received 200 mg/kg of body weight/day of LIM via oral. On DAY 0, a single dose of IVM at 200 µg/kg of body weight was administered orally to groups IVM and LIM+IVM. On DAY + 7 and DAY + 14, fecal egg counts (FEC) were performed and on DAY + 14 animals were euthanized for total worm count (TWC), worm length, fecundity of females, and Pgp-9 gene expression. On DAY + 7, group LIM+IVM had 96.29% efficacy based on Fecal Egg Count Reduction TEST (FECRT) and a highly significant reduction in FEC (P = 0.0005) when compared to CTL. On DAY + 14, the efficacy of LIM+IVM was 82.87% on FECRT, although no differences were found among groups for FEC, TWC, worm length, or Pgp-9 gene expression. Female worms from the CTL group had higher egg counts in their uterus when compared to LIM. No differences were found for hematological or biochemical parameters, body weight, or weight gain among groups. Thus, LIM given daily at 200 mg/kg was safe for animals and, when combined with IVM, decreased egg shedding and could reduce pasture contamination, although it was unable to kill multidrug-resistant H. contortus., Competing Interests: Declaration of Competing Interest The authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest or non-fi nancial interest in the subject matter or materials discussed in this manuscript., (Copyright © 2023. Published by Elsevier B.V.)

Santa Ines (SI) and Ile de France (IF) sheep are known to be resistant and susceptible to Haemonchus contortus infection, respectively. Several studies have shown some genes as potential biological markers for sheep resistance against gastrointestinal nematodes using molecular tools, including transcriptomic analysis. In this study, we sequenced the polyadenylated RNA of the abomasal tissue of SI and IF suckling lambs to identify mucosa-specific transcript alterations between breeds artificially infected with H. contortus. Naïve SI (n = 4) and IF (n = 4) lambs were artificially infected every other day, over a period of 52 days, from 14 to 66 days old, with a total of 5,400 H. contortus infective larvae. Fundic abomasal tissue samples were collected at 68 days old, and submitted to high-throughput RNA sequencing (RNA-seq). Differential expression analysis (P value < 0.001 and False Discovery Rate (FDR) < 0.05) between SI and IF samples identified 292 genes, most of which showed greater expression in SI lambs. To help annotate and assign possible function to differentially expressed genes (DEGs), we used previously available single-cell RNA-seq (scRNA-seq) data from ovine abomasal mucosa to putatively identify cell types and possible mechanisms involved in resistance to H. contortus. In particular, genes associated with endothelial and tuft cells showed the greatest increases in expression in SI relative to IF lambs. SI lambs had higher percentages of tuft cells than IF lambs in the fundic abomasal mucosa. Although we found innate immunity (cell-mediated in mucosa) acting as a protagonist in impairing H. contortus infection, a stronger acquired immune response was being modulated at an earlier stage by SI lambs. We suggest that the complex connection between innate and adaptive immunity is via cellular antigen processing and presentation (APP). Based on comparison with scRNA-seq data, SI lambs showed a robust APP mechanism characterized mainly by greater T cell APP, macrophage differentiation, and cytokine signalling. We identified potential mechanisms and markers to advance knowledge for selection of H. contortus resistance at a very early age, in SI as well as in other commercial sheep breeds., (Copyright © 2023 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

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., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

The indiscriminate use of drugs to control gastrointestinal helminths in horses can lead to serious consequences for the animals, which represents a growing problem for health, animal welfare and productivity. Therefore, the present study aimed to evaluate the anthelmintic efficacy of ivermectin in naturally infected horses in the western region of the state of São Paulo. From May 2021 to April 2022, 123 naturally infected adult horses in 12 equine breeding farms (7 to 14 animals per farm) were evaluated with the fecal egg count reduction test. The horses had not been treated with anthelmintic drugs for at least 60 days before the start of the study. The animals were treated with ivermectin (0.2 mg/kg; Eqvalan®, Merial) administered orally according to recommended dosage by the manufacturer. Individual fecal samples were collected directly from the rectal ampulla and used to determine the eggs per gram of feces (EPG) and perform coproculture for identification of larvae on the day of anthelmintic treatment (D0) and 14 days later (D14). The fecal egg count reduction (FECR) on each property was calculated using the program Shiny-egg Counts R version 3.6.1, and the presence of anthelmintic resistance was confirmed when the FECR percentage was less than 95%, and when the lower confidence limit [LCI] was less than 90%. The pre-treatment average EPG count in the 12 properties was 991. In five properties the FECR was lower than 90%; in three properties between 90% and 95%; and in four properties equal to or greater than 95% after treatment with ivermectin. The occurrence of cyathostomins with resistance to ivermectin was recorded in the majority of the farms., 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 © 2023 Elsevier B.V. All rights reserved.)

The aim of this study was to assess the suitability of PCR and ELISA as diagnostic method in young sheep naturally infested by Oestrus ovis larvae. The experiment was carried out from December 2020 to April 2021 with 39 lambs divided into two groups: infested (n = 26) and control treated group (n = 13). The infested group did not receive treatment against oestrosis, and the control group was treated with closantel (10 mg/kg orally) every 28 days in order to keep the animals as free as possible of O. ovis infestation. The clinical signs varied among animals regardless of the number of recovered larvae of each lamb, however, the thick mucus and mucopurulent nasal discharge scores were less frequent in lambs from treated group. There was no correlation between the nasal discharge score and the number of O. ovis recovered larvae (R² = 0.012, P = 0.165). Three control treated animals only presented first instar larvae (L1) (1 - 4 larvae/animal) which were smaller than L1 found in the lambs of the infested group. Ninety-two percent of the lambs from infested group (24/26) were parasitized by O. ovis with number ranging from 1 to 54 larvae per animal. A gradual increase in plasma IgG (anti-antigen of O. ovis larvae) levels of animals from infested group after the third week of the trial was observed, whereas the control lambs had low levels of IgG until the end of the experiment. The PCR had low sensitivity (26 %) and high specificity (100 %), and it presented poor agreement (k = 0.177) with the larvae detection after the lamb slaughter. The oestrosis clinical signs were not related to larvae infestation intensity and ELISA showed a greater advantage over the PCR technique in identifying animals that are carrying O. ovis., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Santa Ines, an indigenous Brazilian hair sheep, in comparison with European sheep breeds (e.g., Ile de France), show greater resistance against gastrointestinal nematode (GIN) infections, mainly to Haemonchus contortus. Here we bring novel findings that address some gaps regarding the resistance traits involved in the development of the immunity of young lambs in the first few weeks of life to H. contortus infection. This study aimed to compare parasitological, humoral, and local effector cell-mediated responses, also the histopathological differences in the abomasal mucosa of Santa Ines (SI) and Ile de France (IF) suckling lambs serially infected with H. contortus. Parasitological variables, local and circulating humoral immunity, and local cellular response were evaluated in naïve Santa Ines (n = 14) and Ile de France (n = 12) lambs, randomized into four groups: infected SI (n = 8), non-infected control SI (n = 6), infected IF (n = 8) and non-infected control IF (n = 4). Lambs from infected groups were first infected at 14 days old, and multiple infections were conducted every second day, until the age of 66 days old (52 days post first infection). In comparison with infected Ile de France, infected Santa Ines lambs had lower mean eggs per gram of faeces, lower total H. contortus worm burden, lower females' length, greater abomasal lymph node weight, greater mucosal thickness in the fundus, and also higher counts of eosinophils in the fundus, and mast cells and globule leukocytes in both fundic and pyloric mucosa of the abomasum. Intrabreed differences were observed into the infected Santa Ines group, with three of the eight lambs classified as highly resistant for displaying H. contortus burden ranging only from 1 to 42 worms. Overall, Santa Ines suckling lambs showed great resistance against H. contortus infection in comparison with Ile de France lambs, being able to mount a robust innate immune response at an early age, and before weaning., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Natural infection by intestinal nematodes was assessed in Santa Ines and Ile de France sheep breeds, over a five-month grazing period, with emphasis on the development of the immune response in lambs under two anthelmintic treatment regimens. Nineteen Santa Ines and 19 Ile de France male lambs were allocated into two treatment groups: animals under suppressive treatment with anthelmintics; and animals under targeted selective treatment (TST). The nematodes Trichostrongylus colubriformis and Strongyloides papillosus showed the highest infection intensity in the TST animals in both breeds. Parasite-specific immunoglobulin G levels were significantly higher and more rapidly induced in Santa Ines lambs. Additionally, these lambs had higher levels of parasite-specific immunoglobulin A in intestinal mucus. Animals under TST had higher means of mast cells and globule leukocytes in the intestinal mucosa due to longer and greater parasite challenge in comparison with the suppressive group. A breed effect (P < 0.05) was recorded for mast cells, with Santa Ines lambs presenting the highest counts. Although Santa Ines lambs had lower intestinal nematode counts than Ile de France lambs, they had shown a large variation in T. colubriformis burden, with most of them presenting a marked worm burden, despite showing an earlier and more intense immune response to intestinal nematode infection.

This study aimed to evaluate foraging distance (FD) from the dung, parasitological and physiological factors in 18 Crioula Lanada lambs naturally infected by nematodes with three infection levels (IL) in a Voisin Grazing System. In the pre-experimental phase animal feces collection, deworming, observer training, animal adaptation and dung demarcation were carried out; in the experimental phase, grazing distance, feces, pasture and blood sampling. An initial exploratory analysis was carried out (Kruskal-Wallis test). Fixed predictors were selected with a cumulative logit regression model; an ordinal logistic regression mixed model identified influencing factors of ordinal responses for (i) FD, (ii) infective larvae quantity (L3). Animals approached the dung when the radiation or temperature were more intense (P < 0.05). Paddock entry/exit, IgG and L3 influenced FD over time (P < 0.05). L3, in turn, was influenced by IL, FEC and corpuscular volume (CV). In the High IL group, FD varied between 60-100 cm. Greater L3 and FEC were found in the High and Low IL from the 4th week (P < 0.05). Naturally infected Crioula Lanada lambs increased the distance from the dung, which was not related to IL but to the dynamics of solar radiation and parasitological and immunological factors.

The sheep nasal bots Oestrus ovis is parasite of the nasal cavities and sinuses of small ruminants causing oestrosis, one of the most frequent parasitic diseases in sheep and goats. The widely use of ivermectin and closantel by the sheep breeders in the treatment and prophylaxis of gastrointestinal nematodes resulted in widespread cases of anthelmintic resistance. However, there is no report about cases of O. ovis with drug-resistance. In this study, we evaluated the prophylactics and therapeutic effects of both antiparasitics in sheep with O. ovis natural infestation. The trial was carried out from early December 2019 to March 2020, with 30 crossbred males lambs allocated into three groups of 10 animals each: control (without treatment), treated with ivermectin (0.2 mg/kg subcutaneously) and treated with closantel (10 mg/kg orally). The animals were kept together grazing the same pasture area. The treatment groups were drenched in two occasions 70 days apart: on 5th December 2019 and on 13th February 2020. On 19th March 2020, all lambs were slaughtered. The lamb heads were removed and sectioned along their longitudinal and sagittal axis to search for larvae. Recovered O. ovis larvae were counted and identified according to their developmental stage (L1, L2, and L3). Seven of the control lambs were infested with O. ovis larvae ranging from six to 17 larvae (11.6 mean infestation intensity). All recovered larvae from control group were intact and active. Three animals treated with ivermectin had O. ovis larvae (1-3 larvae), however they were dead and in degeneration. The animals treated with closantel did not have any larvae. The clinical suggestive signs of oestrosis were scarce over the experimental period. The averages of daily weight gain were similar ( p > 0.05) among groups. Closantel and ivermectin had high efficacy against oestrosis and O. ovis parasitism did not hinder the performance of lambs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Bello, Lins, Albuquerque, Ferreira, Amarante and Amarante.)

We estimated the losses caused by gastrointestinal nematode (GIN) infections in young Nellore cattle born in three consecutive calving seasons (Cycles I, II and III). Three groups of animals: Group 1 - free from GIN using 2.5 mg/kg of albendazole sulfoxide; Group II - free from Oesophagosomum radiatum using 0.2 mg/kg of ivermectin but infected with ivermectin-resistant Haemonchus placei and Cooperia spp.; and Group III - non-treated experimental control animals, infected with all GIN, were observed over a period of 12 months. Male and female calves were evaluated starting before weaning when the animals were an average of approximately four months of age. In Cycle I, only females were evaluated. All the animals continued to graze on pastures of Urochloa spp. (= Brachiaria). All the groups showed median faecal egg counts of fewer than 250 eggs per gram (EPG), and no clinical signs of parasitic gastroenteritis were observed. The blood variables were within the normal ranges, and no calf presented anaemia. In most of the samplings, the median EPGs were significantly lower (P < 0.05) in the group treated with albendazole. Throughout the experiment, the most prevalent parasite observed in the control group was Cooperia spp., followed in decreasing order by Haemonchus spp., O. radiatum and Trichostrongylus spp. All the groups of calves exhibited weight gain throughout the trials with control group displaying the lowest body weight gain. Body weight variables between the albendazole- and ivermectin-treated groups were not significantly different (P > 0.05). After weaning, females animals in albendazole and ivermectin treated groups exhibited higher body weights, 20.4% and 22.7%, respectively, than those of the control group. Likewise, males treated with albendazole and ivermectin exhibited 27.6% and 25.8%, respectively, more body weight gain than animals under control group. Because the main difference between the ivermectin and the control groups was the O. radiatum parasitism, most of the losses in the control group were possibly due to this nematode species. Nevertheless, the other nematodes species that occurred in relatively high intensities in control group could also have an additive effect in such losses., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Haemonchus contortus is the most important gastrointestinal nematode in the tropics and subtropics causing huge economic losses to the small ruminant industry. Vaccination is potentially a sustainable approach to control this parasite and the performance of Barbervax® a vaccine containing integral membrane glycoproteins from H. contortus intestinal cells, was evaluated in naturally infected grazing sheep during their development from sucking lambs to adults. The sheep were randomly assigned to two groups: Vaccine and Control. The Vaccine group were vaccinated 23 times over the course of this two-year trial at intervals of 3-6 weeks. They responded with anti-vaccine specific antibodies, had 80 % lower Haemonchus egg counts and were less anaemic compared with the controls. Packed cell volumes (PCV) were always greater than 25 % in the vaccinated sheep but averaged between 23 % and 24 % in the controls. Total plasma protein values were higher in the vaccinated group from the third vaccination until the end of the trial. Throughout the trial, 88 % of the control sheep were drenched (average of 3.1 drenches per treated animal) but only 57 % of vaccinates, needed a salvage anthelmintic treatment (average of 1.9 drenches per treated animal), however, between group no differences in body weight were observed. In summary, these results indicate that a continuous course of Barbervax® can provide lambs with substantial year-round protection against H. contortus until they reached adulthood., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study aimed at evaluating parasitological and blood variables in native breed Crioula Lanada sheep belonging to the same herd, to identify and compare susceptible or resistant individuals to gastrointestinal nematodes during gestation and lactation phases. For this purpose, 18 Crioula sheep were used within 2 years of evaluation, in which blood and feces samples and weight of the animals were taken from their 4th month of gestation until the weaning of their lambs, in the 4th month postpartum. Feces samples were used for counting eggs per gram of feces (EPG) and, thus, to identify 12 resistant ewes (EPG < 1,000) and 6 susceptible (EPG > 1,000) to gastrointestinal nematodes. The identification of infective larvae was also performed. Blood was used for analysis of packed cell volume (PCV), eosinophil count, total plasma protein, and immunoglobulin G level against Haemonchus contortus infective larvae. The Kruskal-Wallis non-parametric comparison test was used to evaluate the differences between days of parturition and linear mixed-effects model using package lme4 in R to evaluate the groups. The main parasite species observed in the feces samples were Haemonchus, Trichostrongylus, Oesophagostomum, Ostertagia , and Cooperia in similar proportions in both groups. Susceptible ewes presented peaks of EPG at the beginning of lactation and lower PCV values throughout the study. No difference between groups was observed concerning other blood variables or body weight, but some changes were observed along with the gestation and lactation phases within each group. The physiological response of sheep to nematode infection is a useful tool to identify the most susceptible individuals within the same breed and herd and to select the most genetically resistant individuals., (Copyright © 2020 Pereira, Longo, Castilho, Leme, Seugling, Bassetto, Amarante and Bricarello.)

The influence of shade in the silvopastoral system on the performance and degree of parasitism by endo- and ectoparasites in Angus x Nellore heifers was assessed. The experiment was conducted with 72 heifers, with an initial mean weight of 276 ± 5.67 kg and 9 months of age, in a total area of 25 ha, divided into 12 paddocks, with three treatments and four replicates. The experimental design was a randomized complete block design, to evaluate the conventional grazing system (CGS) without shade and silvopastoral systems with simple line (SPS1) or triple lines (SPS3) of eucalyptus. The evaluations were carried out every 28 days, from June 2017 to April 2018. During the trial, the number of eggs per gram of feces (EPG) presented a gradual increase in the three systems. Differences (P < 0.05) in the variables analyzed were recorded only in two occasions: the CGS group EPG mean ± standard error (1269 ± 105) was higher than the SPS1 group mean (402 ± 129 EPG) in March, and tick average of the CGS group (13 ± 2.7) was lower than the SPS3 (32 ± 5.3) in October. There were no significant differences (P ≥ 0.05) between groups in relation to horn fly counts, the numbers of infective nematode larvae on pasture, hematological variables, and weight gain. It was concluded that in comparison with the CGS system, the shading in the SPS1 and SPS3 systems did not have any major influence on the degree of parasitism or in the performance of the heifers.

The objectives of the present study were to evaluate the efficacy of three anthelmintic drugs, i.e. fenbendazole, ivermectin and moxidectin; to determine the genera and species of the most abundant strongyles; and to investigate parasite control measures used in herds of horses in the western region of the state of São Paulo, Brazil. This study was onducted between February and December 2013 on 10 farms in this region. Coprological evaluations were conducted for counting the numbers of eggs per gram of feces (EPG) and coprocultures were made in order to identify third-stage larvae (L3) of strongyles. Parasite control measures were investigated by surveying each farm's management and characteristics. A fecal egg count reduction test (FECRT) was used to determine the anthelmintic efficacy based on EPG count results before and after treatment. The FECRT showed that anthelmintic resistance to fenbendazole was present and that ivermectin and moxidectin had high effectiveness in the western region of the state of São Paulo. Identification of L3 revealed that there was high prevalence of cyathostomins among strongyle larvae. The highest prevalence of Strongylus vulgaris occurred on farms where the intervals between deworming were long. The questionnaire applied showed that ivermectin was the drug most used among these farmers., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The objective of this trial was to evaluate the period of spelling necessary for a pasture to become free of contamination by infective larvae of gastrointestinal nematodes (GIN) of sheep, in different seasons of the year, as well as to determine when the greatest pasture contamination occurs and how long it lasts. An area was divided into four paddocks, one for each season (spring, summer, autumn, and winter). In order to contaminate the paddocks with free living stages of GIN, eight ewes, naturally infected, grazed on each paddock for 14 consecutive days, starting on the following dates: autumn, on April 4, 2017; winter, on July 4, 2017; spring, on September 26, 2017; summer, on January 2, 2019. At the beginning and end of the grazing period, faecal samples were taken directly from the rectums of the ewes to count eggs per gram of faeces (EPG) and for faecal cultures. Every 14 days pasture samples were collected to assess the number of infective larvae (L3) per kilogram of dry matter. At the end of the 14 day ewe grazing period, 21 stakes were placed where there were faeces on the paddock. Subsequently, every 14 days, the faeces located at three of the stakes were collected and the L3 were recovered. After the exit of the ewes, monthly, two tracer lambs, free of helminth infection, were allocated into the paddock for 14 days. At the end of this period they were housed in covered stalls for 28 days. Faeces from the lambs were collected for individual EPG counting and faecal culture at 21 and 28 days after grazing. Infective larvae recuperation was observed from faeces and pasture in all seasons. In the autumn, spring, and summer, high EPG counts were observed in the first tracer lambs (8521, 4800, and 8064 EPG, respectively), while in winter, high infection (14132 EPG) of the animals was observed only from the second pair of tracer lambs. For a pasture to become "clean", 322 days, 350 days, 294 days, and 182 days following contamination were necessary, respectively, in the autumn, winter, spring, and summer. In autumn, spring, and summer, massive contamination of the pasture with L3 occurred soon after an area had been grazed by infected sheep, while in winter this took a little longer. The contamination persisted, approximately, from a minimum of six months post contamination in summer to up to almost one year post contamination in winter., 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 © 2020 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the dynamics of Haemonchus contortus and Haemonchus placei infections and hybridization between these species in grazing sheep without contact with cattle. On January 14, 2014, sixteen young sheep were infected with 4000 infective H. placei third-stage larvae L3; 11 days later, another group n = 16 was infected with 4000 H. contortus L3. The establishment rates of H. contortus and H. placei L3 were, on average, 61.6 % and 56.8 %, respectively, in the permanent sheep. After the establishment of patent infections, all permanent sheep were allocated together in the same clean pasture where they grazed for the next 12 months. Euthanasia of a sample of the permanent sheep was performed every three months: in May, August, November and February. Two weeks before the sheep were removed for euthanasia, 2 worm-free tracer sheep were introduced to the pasture to evaluate the larval population in the field. The tracer sheep grazed alongside the permanent sheep for 2 weeks. Then, they were housed indoors for 20 days; at the end of this period, they were euthanized. Parasites were recovered from the permanent and tracer sheep and identified using morphological and molecular techniques. A total of 432 worms (from permanent and tracer animals) were analyzed by PCR using species-specific primer pairs. Of these specimens, only two (0.46 %) male worms were identified as hybrids: one was recovered from a permanent animal euthanized in August and the other from a tracer sheep that grazed in May. The last detection of adult H. placei worms occurred in sheep euthanized in May (approximately 3.5 months after the beginning of the grazing period). The morphological evaluation of the L3 produced in fecal cultures showed that H. placei were progressively replaced by H. contortus populations starting in March. The last trace of H. placei L3 was found in August, when a small percentage (0.5 %) of infective larvae with H. placei morphology was identified in a fecal culture. In conclusion, hybridization between H. contortus and H. placei can occur in the field during coinfection. It was demonstrated that H. placei established successfully in artificially infected worm-free sheep; however, with concomitant natural reinfection with H. contortus, the H. placei population showed a rapid decrease and was eliminated within a few months in an environment without cattle., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Atomic force microscopy (AFM) was used in this work to investigate the ultrastructural and mechanical characteristics of Haemonchus contortus, the major gastrointestinal nematode that infects small ruminants worldwide. The biophysical characterization of this species is extremely important in order to reveal mechanisms of action of drugs and to classify its ultrastructure and biomechanical properties. High-resolution topographic images by AFM as well as data on biomechanical properties of cuticles were obtained at different stages of H. contortus. The results reveal details of the mechanical and structural properties of this nematode never observed before for nematodes parasite with other microscope techniques. Qualitative and quantitative reductions in the elasticity of the larvae stage egg were compared with those of the morulae stage, and the increased adhesion of unsheathed L 3 were compared with the same stage of sheathed larvae. The results presented here open possibilities for understanding the mechanisms of drug and biomolecular actions that can be used to control infections caused by H. contortus., Competing Interests: Declaration of Competing Interest The authors declare that there is conflict of interest regarding the publication of this article., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This paper describes the discovery of Lagochilascaris minor parasitizing a stray cat (Felis catus) in São Sebastião, a coastal city in the state of São Paulo, Brazil. The cat presented fistulas with purulent secretion in the ventral cervical region. A thorough evaluation revealed nematodes in the site. The parasites were removed, amounting to a total of about 100 specimens. The parasite species was identified based on measurements and morphological characteristics of adult worms. This report of parasitism by L. minor in domestic cats in the state of São Paulo reinforces the need for greater attention to this parasitosis, given the increase in cases among carnivores in Brazil., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Understanding the immunological basis of resistance to gastrointestinal nematode infections in livestock is important in order to develop novel methods of parasite control such as vaccination or genetic selection for parasite resistance. The present study aimed to investigate differences in immune response between parasite resistant Santa Ines and susceptible Ile de France sheep breeds to natural Haemonchus contortus infection. Parasitological parameters, humoral immunity, local and circulating cellular immune responses were evaluated in 19 Santa Ines and 19 Ile de France lambs undergoing different anthelmintic treatments regimens: suppressive treatments (SUP) or targeted selective treatments (TST) over a 5-month grazing period. Santa Ines lambs had significantly lower Haemonchus faecal egg count and worm burden compared to Ile de France regardless of treatment regime. In addition, circulating blood eosinophils count and parasite-specific IgG levels were significantly higher and more rapidly induced in Santa Ines lambs. Abomasal immune responses were generally greater in the resistant breed, which had significantly higher levels of parasite-specific IgA in mucus, and elevated number of globule leukocytes and CD3+ T cells within the abomasal mucosal. Furthermore, numbers of POU2F3+ epithelial cells, a tuft-cell specific transcription factor, were also elevated in the Santa Ines breed, suggesting that this breed is better able to initiate T-helper type 2 immune responses within the abomasum. In conclusion, the differential immunological responses detailed here are relevant to understanding resistance to gastrointestinal nematodes in other host breeds, as well as to resistance breeding as a sustainable control approach for parasitic infections.

This study aimed to evaluate the simultaneous infections of Haemonchus contortus and Haemonchus placei in sheep, as well as the production of hybrids. A parental group of lambs (n = 6) were mix-infected with 2000 infective larvae (L3) of H. placei and 2000 L3 of H. contortus. Faecal samples were taken from each of these six lambs to produce the first generation of L3 (F1-L3) in individual cultures. These F1-L3 were used to infect 12 lambs; six of them were euthanized at 42 days (Group F1-42) and six at 84 days (Group F1-84) post infection. Polymerase chain reaction (PCR) analysis, using species-specific primer pairs, was the gold standard method for identification of Haemonchus adult species and hybrids. The establishment rate of both species was similar in the parental group: 51.7% H. contortus and 48.3% H. placei. Of the 219 adult specimens from groups F1-42 and F1-84 analysed by PCR, eight (3.65%) were hybrids, 111 were H. contortus and 100 were H. placei. The morphological evaluation of the F1-L3 from the parental group showed a predominance of larvae with H. contortus size (51.5%) in comparison with H. placei (42.8%). In the second generation of L3 (F2-L3) produced by the F1-lambs, larvae with H. contortus morphology predominated, with 81.5% in the F1-42 group and 84.0% in the F1-84 group. In conclusion, an artificial mixed infection by H. contortus and H. placei was established in lambs and resulted in the production of a small number of hybrids among their offspring.

Background: Haemonchus contortus, a gastrointestinal nematode parasite of sheep, is mainly controlled by anthelmintics; the occurrence of anthelmintic resistance leads to treatment failures and increases economic burden. Because molecular mechanisms involved in drug resistance can be elucidated by genomic studies, an extreme quantitative trait locus (X-QTL) mapping approach was used to identify co-segregation of the resistance phenotype with genetic markers to detect the genome-wide variants associated with monepantel resistance in H. contortus., Methods: A cross between H. contortus isolates using parental susceptible (Par-S) males and monepantel resistant (Par-R) females resulted in SR progeny, while reciprocal cross resulted in RS progeny. Pools (n = 30,000) of infective larvae (L3) recovered from Par-R, and from SR and RS populations in the F3 generation, collected both before (unselected group) and 7 days after (selected group) selection with monepantel treatment in sheep hosts, were subjected to genome sequencing (Pool-Seq). Pairwise comparisons of allele frequencies between unselected and selected groups were performed for each population by Fisher's exact test (FET) and for both populations combined by a Cochran-Mantel-Haenszel (CMH) test., Results: Mapping rates varied from 80.29 to 81.77% at a 90.4X mean coverage of aligned reads. After correction for multiple testing, significant (P < 0.05) changes in allele frequencies were detected by FET for 6 and 57 single nucleotide polymorphisms (SNPs) in the SR and RS populations, respectively, and by the CMH test for 124 SNPs in both populations. The significant variants located on chromosome 2 generated a selection signal in a genomic region harboring the mptl-1, deg-3 and des-2 genes, previously reported as candidates for monepantel resistance. In addition, three new variants were identified in the mptl-1 gene., Conclusions: This study expands knowledge on genome-wide molecular events underlying H. contortus resistance to monepantel. The identification of a genome region harboring major genes previously associated with monepantel resistance supports the results of the employed X-QTL approach. In addition, a deletion in exon 11 of the mptl-1 gene should be further investigated as the putative causal mutation leading to monepantel resistance.

In this study we aimed to evaluate the effect of Trichostrongylus colubriformis infection on digesta passage rate, rumen fermentation and lamb performance. Eighteen three-month-old Santa Ines castrated male lambs (16.9 ± 1.43 kg of body weight) were randomly distributed in two experimental treatments: Infected with T. colubriformis (I, n = 9) and Uninfected (U, n = 9). The I lambs received a total of 45,000 L 3 larvae of T. colubriformis (5,000 infective larvae, three times per week for three weeks). Daily feed intake was assessed using the I lambs as a reference for their respective pairs on the U group (pair-fed). Weight, body condition score and faeces (stool) samples were obtained every 15 days for 75 days. In both treatments, faecal egg count (FEC), digesta passage rate, rumen fermentation parameters, protozoa count and short chain fatty acids (SCFA) were evaluated. The lambs presented moderate infection (FEC = 620). The retention time of the digesta in the rumen-reticulum segment was lower (P < 0.05) in I lambs. The I lambs presented no inappetence, however, lower concentrations of total SCFA and butyrate, while higher acetate concentration were observed in these lambs (P < 0.05). The present findings highlight that T. colubriformis infection decreased the retention time (solid and liquid content) of the digesta in the rumen-reticulum, as well as negatively affected lamb growth., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Gastrointestinal nematodes significantly affect the ovine industry, and Haemonchus contortus is considered the most pathogenic parasite in tropical regions. This situation is aggravated when the main strategy to control worms fails because of the genetic resistance that parasites acquire against anthelmintics. Aiming to anticipate the events involved in anthelmintic resistance, we induced monepantel resistance in H. contortus by in vivo subdosing of sheep hosts. Four successive passages of a monepantel-susceptible H. contortus isolate in Santa Ines or Ile de France sheep hosts resulted in three monepantel-resistant (efficacy varying from 0 to 58.5%) H. contortus isolates. Sheep hosts were treated from 0.075 mg/kg to the therapeutic dose of 2.5 mg/kg of monepantel in 19-26 rounds of selection for 112-133 weeks. Success in inducing H. contortus resistance to monepantel may have been affected by worm burden and by host-parasite interactions, including a possible effect of the breed of sheep hosts. We conclude that subdosing of sheep, although time-consuming, is an efficient in vivo strategy for the induction of monepantel resistance in H. contortus. The resistant parasites can be used in further studies to elucidate the genetic and biochemical events involved in the acquisition of anthelmintic resistance.

Molecules from natural sources, such as essential oils, have shown activity against parasites in vitro, but have not yet been explored extensively in vivo. Anethole and carvone (10% each), encapsulated with 80% of a solid matrix, referred to as EO (encapsulated oils), were tested in vivo in 2 experiments. In Experiment 1: Lambs were artificially infected with multidrug resistant Haemonchus contortus, or left uninfected, and treated (or not) with 50 mg/kg bw (body weight) of EO in a controlled environment. Thirty-two male lambs were kept in individual cages for a period of 45 days, after which animals were evaluated for parasitological, hematological, toxicological, and nutritional parameters. After 45 days of treatment, EO at 50 mg/kg bw provided a significant (P ≤ 0.05) reduction in fecal egg count (FEC). Although FEC was reduced, animals from both treatments had similar counts of total adult worms. The low FEC was caused probably by a significant reduction (P ≤ 0.05) in both male worm size and female fecundity. Dry matter intake of uninfected controls was significantly (P ≤ 0.05) reduced, although no toxicity was observed in treated animals. Thus, in Experiment 2, conducted for five months we used an EO dose of 20 mg/kg bw. Thirty-four weaned lambs, free of parasites, were divided in two groups and kept in collective pens. One group received EO at 20 mg/kg bw mixed with concentrate for 5 months and the other was kept as a control group (CTL). Parasitological and hematological parameters as well as body weight were evaluated. In the first 2.5 months, CTL and EO groups were confined, and both presented similar clinical parameters. Then, animals were allotted to graze on contaminated pastures to acquire natural infection for the next 2.5 months. The infection was patent after 25 days and both groups had similar decreases in weight gain, increases in FEC, and decreases in blood parameters. Coprocultures from CTL and EO groups established that parasite population was 90% Haemonchus sp. We concluded that the technology of encapsulation is safe and practical to deliver to lambs at the farm level and anethole and carvone at 50 mg/kg bw caused a significant decrease in FEC and, consequently, in pasture contamination by free living stages of H. contortus. However, EO at 20 mg/kg bw was not effective to prevent or treat sheep naturally-infected with gastrointestinal nematodes., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Haemonchus contortus is an economic problem in sheep farms worldwide, mainly in the tropics and subtropics. A vaccine against haemonchosis, called Barbervax ® , was evaluated in ewes under two nutritional status, naturally infected with gastrointestinal nematodes. Ewes were divided into four groups: Supplemented Diet - Vaccine; Supplemented Diet - No vaccine; Basal Diet - Vaccine and Basal Diet - No vaccine. Their lambs were divided in Vaccinated and No vaccine. Ewes were immunised six times starting about 1 month of pregnancy with the first three doses at 3 week intervals and the last three shots at 4 week intervals. Supplemented ewes had higher body weight, body score and packed cell volume compared with those fed a basal diet. Both groups of vaccinated ewes showed a similar response in circulating anti-vaccine antibodies but the vaccine had no discernible effect on either body weight, body score and packed cell volume. There was a marked group difference in the number of ewes that received precautionary treatments with anthelmintic. All 14 Basal Diet - No vaccine ewes required treatment. In contrast only 7 ewes, in the Supplemented Diet - Vaccine group required anthelmintic treatment. In the Basal Diet - Vaccine and in the Supplemented Diet - No Vaccine groups, 12 and 13 ewes needed anthelmintic treatment, respectively. Vaccinated lambs showed much higher antibody titres resulting in 80% less Haemonchus spp. egg counts comparing with no vaccine lambs. Taken together these results clearly suggest that in pregnant and lactating ewes a combined protective effect between vaccination and improved nutrition resulted in fewer precautionary anthelmintic treatments. Thus, it was possible to achieve a more sustainable level of control of the haemonchosis, less dependent on anthelmintic drugs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study examined the development of resistance to anthelmintics in Haemonchus contortus in lambs under suppressive or selective treatment regimens that included monepantel. Twenty Ile de France and 20 Santa Ines lambs were allocated to two anthelmintic treatment regimens, based on body weight and nematode faecal egg counts (FEC): targeted selective treatment (TST) or suppressive treatment, both with monepantel. Lambs of the TST group were treated individually when they presented with a packed cell volume (PCV) ≤20%. On 7 October 2016, the lambs were allocated to clean pastures, where they grazed in separated paddocks by group until late February 2017. The experimental area was contaminated with nematodes that were introduced with the experimental Ile de France and Santa Ines lambs, naturally infected with gastrointestinal nematodes. To maintain the grazing lambs in the suppressive treatment group and their pasture as free of worms as possible, these lambs were treated with anthelmintics before being allocated to their paddock and then were periodically treated with monepantel. However, the use of a suppressive treatment regimen that included monepantel over a period of 3 months resulted in the emergence of a population of resistant H. contortus. In the TST group, there was a rapid and progressive reduction in the efficacy of monepantel, which at the end of the experiment was only 76%. The Ile de France lambs were all treated one or more times during the experiment, whereas only two Santa Ines lambs in the TST required treatment. In conclusion, a population of H. contortus resistant to monepantel emerged quickly during the rainy season, even when sheep were submitted to selective treatment., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Haemonchus contortus and Haemonchus placei are among the major parasites of small ruminants and cattle. Although infection with these nematodes is host-specific, with H. placei predominating in cattle and H. contortus in sheep, cross-infections are observed in areas where both parasites are sympatric, and hybrid offspring can occur. Therefore, a fast and precise method is required for differentiating the parasites. Identification based on spicule morphometry is the most common technique for differentiating Haemonchus species. However, because these measurements overlap between species, morphological analysis is insufficient for differentiating between helminth species. In this work, we present a reliable, conventional polymerase chain reaction (PCR)-based method that uses two species-specific primer pairs to differentiate between H. contortus and H. placei specimens and their hybrids. Each primer pair produces one single and distinct amplification band for each species, which enables the detection of hybrid specimens. These primer pairs were validated by testing eight different populations of H. contortus, H. placei and hybrids.

Haemonchus contortus and Haemonchus placei are two closely related economically important parasites of ruminants. Their close morphological similarity, common occurrence as co-infections and ability to hybridize makes definitive diagnosis and epidemiological studies in field populations challenging. In this paper, we describe the development of a panel of microsatellite markers that can be used to discriminate and study the genetics of these two parasite species in co-infections and mixed field populations. We have identified two additional microsatellites (Hp52 and Hp53), in addition to three previously reported microsatellites (Hcms3561, Hcms53265 and Hcms36) that have a discrete set of alleles between the two species. Multilocus genotyping of worms with this 5 marker panel from 3 geographically diverse H. placei isolates and 4 geographically diverse H. contortus populations allows unambiguous species assignment of individual worms. This panel of markers should provide a valuable resource in studying the biology and epidemiology of these important ruminant parasite species in the field., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Helminths is a endoparasites that cause the major losses for profitable sheep production in Brazil. The increased development of resistant strains of endoparasites have enforced the search for sustainable alternatives. The aim of this paper was to provide information about endoparasites control with banana leaves in infected sheep as alternative control strategies and see its viability., Materials and Methods: In this study, we performed two trials to investigate the anthelmintic properties of banana leaves on endoparasites in sheep. In Trial 1, twelve sheep were artificially infected with Trichostrongylus colubriformis ; in Trial 2, eleven sheep were artificially infected with Haemonchus contortus . Clinical examinations, packed cell volume, total protein, faecal egg counts (FECs) and egg hatchability tests (EHTs) were performed. At the end of the trials, the sheep were humanely slaughtered, and total worm counts were performed., Results: In Trial 1 and 2, no significant FEC decreases were note but significant diference in EHTs were observed. Total worm counts, clinical and haematological parameters did not reveal significant changes between the treatment and control groups. These results suggest that feeding dried ground banana plant leaves to sheep may reduce the viability of Trichostrongylus colubriformis eggs, and this anthelmintic activity is potentially exploitable as part of an integrated parasite management programme., Conclusion: However, further investigation is needed to establish the optimal dosage, develop a convenient delivery form and confirm the economic feasibility of using banana plantation byproducts as feed for ruminant species. Abbreviations: Coproculture test (CT)., Faecal egg count (FEC)., Egg hatchability test (EHT).

The aim of this study was to determine the time required by different anthelmintic agents to reduce strongyle egg shedding in horses. Fifty horses were divided into five homogenous groups based on faecal egg counts (FECs). Treatment groups received either ivermectin; moxidectin; fenbendazole; piperazine; or no treatment (control group). Faecal examinations were performed 4, 8, 12, 18, 24, 36 and 48h after the anthelmintic treatment. After this period, faecal samples were taken every 24h over the next 12days and finally on alternate days (48-h intervals) for another 14days until the end of the experiment (28days post-treatment). The faecal egg count reduction (FECR) was calculated based on the post-treatment mean FECs in the controls and treated animals. Eggs were absent from the faecal examinations beginning at 72h and 4days, respectively, following treatment with moxidectin or ivermectin. Piperazine showed an FECR greater than 95% from 48h up to 9days post-treatment, with the highest FECR value recorded at 7days post-treatment (98.1%). However, the FECR was lower than 90% in the last two samplings (26 and 28days post-treatment). The febendazole group presented the lowest efficacy with FECR below 90% in all samplings. The faecal cultures showed that at the beginning of the trial, all of the groups presented with mixed infections and were predominantly composed of cyathostomins (92.8%), followed by Strongylus vulgaris (5.6%) and Triodontophorus serratus (1.6%). Only cyathostomin larvae were identified following treatment with fenbendazole or piperazine. In conclusion, horses in the present study had a segment of the cyathostomin population with resistance to fenbendazole and piperazine. The strongyle population was susceptible to macrocyclic lactones, with cessation in egg shedding three and four days after treatment with moxidectin and ivermectin, respectively., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Prevalence of allergic and autoimmune pathologies is clearly increasing in developed countries. This has been attributed to a decreased exposure to certain microorganisms and been referred as hygiene hypothesis. In this study we evaluated if a previous infection with Strongyloides venezuelensis would alter the progression of experimental autoimmune encephalomyelitis (EAE) in Lewis rats. Animals were initially infected with 4000 L3 infective larvae of S. venezuelensis by subcutaneous route. Encephalomyelitis was then induced during the acute phase of the infection by immunization with myelin basic protein emulsified with Complete Freund's Adjuvant plus Mycobacterium butyricum. Previous infection downmodulated cytokine production but did not change clinical and histopathological EAE manifestations. Cytometric analysis with antibodies specific for CD4+CD25+Foxp3+ regulatory T cells indicated that infection also did not alter the frequency of these cells in spleen and regional lymph nodes. This finding could partly explain the failure of this worm to avoid EAE progression. Altogether these results demonstrated that infection with S. venezuelensis was not able to modify EAE progression in Lewis rats. In the context of the hygiene hypothesis, these results reinforce the necessity of a comparative study among different helminth species to identify the ones with immunoregulatory competence.