Haemonchus Placei Research Articles

In vitro and In vivo Anthelmintic Activity of Nicotiana tabacum against Haemonchus placei in Cattle

In vitro and In vivo Anthelmintic Activity of Nicotiana tabacum against Haemonchus placei in Cattle

Toltrazuril + fenbendazole for cattle: Pharmacokinetics and efficacy against Eimeria spp. and gastrointestinal nematodes

The present work evaluated the pharmacokinetics and efficacy of the association of 15cmg/kg toltrazuril +5cmg/kg fenbendazole against Eimeria spp. and gastrointestinal nematodes (GINs) in calves of different regions of Brazil (Center-West, Southeast, and South). A total of seven experiments were carried out, five of which determined formulation efficacy against Eimeria spp., considering the following aspects: therapeutic, preventive, metaphylactic, and residual efficacy. Therapeutic efficacy experiments for GINs were carried out by parasitological necropsy. The toltrazuril + fenbendazole association demonstrated ≥95% efficacy against Eimeria spp. for 21 days post-treatment (DPT). When used preventively and metaphylatically, the same association demonstrated ≥97% efficacy against E. zuernii, E. ellipsoidalis, E. cylindrica, E. bovis, E. wyomingensis and E. auburnensis. Toltrazuril + fenbendazole administered seven days before challenge was 100% effective against all these Eimeria species. Results of therapeutic, preventive, metaphylactic and residual efficacies can be related to the pharmacokinetic results, especially considering toltrazuril sulfone, which was detected in animal plasma for a longer period than the parent compound. Toltrazuril + fenbendazole achieved 100% anthelminthic efficacy against the GINs Haemonchus placei (L4), Cooperia pectinata and Oesophagostomum radiatum; 99.94% against adult H. placei; and 99.98% against C. puntacta. The association of toltrazuril + fenbendazole, associated with other measures, is an important and suitable tool for the control and treatment of Eimeria spp. and GINs in young cattle.

Effect of aqueous extract of Azadirachta indica leaves on gastrointestinal nematodes in ruminants: an in vitro study

High rates of gastrointestinal parasite infestation lead to the frequent use of anthelmintics for their control, resulting in resistance in the main species of gastrointestinal nematodes. Natural plant extracts, particularly those from Azadirachta indica A. Juss leaves, have been described as having antiparasitic, antimicrobial, insecticidal, antioxidant, and anticorrosive activities, which are related to the diversity of their chemical composition. This research aimed to evaluate the effect of the aqueous extract of A. indica leaves on the control of gastrointestinal nematodes in ruminants. To achieve this, an in vitro test was performed using the larval development assay, with fecal samples obtained from Pelibuey sheep, Criolla goats in the development category, and Criolla calf cattle for the culture of gastrointestinal nematode larvae. Three concentrations (minimum, medium, and maximum) were evaluated for each animal species against Haemonchus placei and Cooperia sp larvae for cattle and Haemonchus contortus, Trichostrongylus sp, and Trichuris for sheep and goats. The survival percentage indicator was evaluated at 0, 24, 48, and 72 h. The research concluded that the minimum concentration was valid for sheep and goats but not for cattle, and the medium and maximum concentrations were effective against all three animal species, resulting in 100% larval mortality. Keywords: Azadirachta, drug effects, Ruminants, Trichostrongyloidea

Efficacy of a new fixed-dose combination injectable (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) in Australian cattle against artificial infections of gastrointestinal nematodes

We describe a new fixed-dose combination injectable (FDCI) formulated with doramectin and levamisole hydrochloride (HCl) to target broad and overlapping spectra of gastrointestinal nematodes (GINs) through two distinct modes of action. Here, we demonstrate the superior efficacy of the FDCI against mixed populations of cattle GINs in two dose confirmation studies conducted in Australia using artificially induced adult (Study 1) and immature (Study 2) GIN infections. Artificial infections consisted of Cooperia spp., Haemonchus placei, Ostertagia ostertagi, and Trichostrongylus axei. In both studies, cattle were inoculated with third-stage larvae and infections were confirmed by fecal egg count (FEC). Treatment groups in both studies were as follows: (1) negative control (saline, 0.9% sodium chloride), (2) positive control injectable endectocide (Study 1–0.2 mg/kg ivermectin; Study 2–0.2 mg/kg doramectin), (3) positive control injectable anthelmintic (7.5 mg/kg levamisole HCl), and (4) FDCI (0.2 mg/kg doramectin + 6.0 mg/kg levamisole HCl). Cattle were treated either 28 days (Study 1) or 6 days (Study 2) post-infection. On Days 14–16 (Study 1) or Days 20–21 (Study 2) post-treatment, cattle were euthanized and necropsied for the recovery, identification, and enumeration of worms. Treatment efficacy was calculated as reduction in worm burdens of treated cattle compared to saline-treated cattle, and treatments were considered effective if the geometric mean worm burden in the treatment group was reduced by ≥ 95% compared to the negative control group. In both studies, saline-treated cattle remained positive for GIN infections for the study duration. Ivermectin was less than 95% effective against Cooperia spp. (80.2%) and H. placei (24.8%) in Study 1, and levamisole HCl was less than 95% effective against Ostertagia spp. (47.1%) in Study 2. In contrast, the novel FDCI was 100% effective in treating adult and immature life stages of all cattle GINs included in the artificial infections, with no worms recovered at necropsy from doramectin + levamisole HCl-treated cattle. These data show a single administration of the FDCI provides broad-spectrum treatment of economically important cattle GINs.

Morphological and molecular characterization of Haemonchus contortus isolated from the small ruminants of south Gujarat, India.

The successful design of strategic control measures against the blood-sucking gastrointestinal nematode, Haemonchus contortus in small ruminants can be facilitated by revealing its general features from morphology to the molecular level. In the south Gujarat region of India, a total of 2408 H. contortus were collected from 84 slaughtered sheep's abomasum, consisting of 347 males and 2061 females (1:6 ratio) (p<0.05). Furthermore, 726 H. contortus were collected from 61 goats, comprising 145 males and 581 females (1:4 ratio) (p<0.05). The male worms were approximately 12±0.06 mm long, while female worms were about 20±0.09 mm long. The vulvar morphotypes of the female worms were found to be 17.7% linguiform, 76.6 % knobbed/button (p<0.05), and 5.7 % smooth type, demonstrating common features of H. contortus. The nucleotide sequences of the Internal Transcribed Spacer 1 (ITS-1) of 165 bp or ITS-2 plus of 256 bp were aligned, and it was found that the genotypes of male and female specimens of either sheep or goat origin were identical, with a 100 % match. The present isolates shared >95 % and >94 % homology with published sequences of ITS-1 and ITS-2 plus of H. contortus, respectively, with more nucleotide transitions than transversions in the aligned sequences. The reconstructed phylogram of either ITS-1 or ITS-2 plus revealed two major clades, one for H. contortus and another for other nematodes, with Haemonchus placei showing its proximity with the clade of H. contortus. The study established the role of morphological and molecular features in identifying and differentiating H. contortus parasite at the local level.

Refugia or reservoir? Feral goats and their role in the maintenance and circulation of benzimidazole-resistant gastrointestinal nematodes on shared pastures.

Gastrointestinal nematodes threaten the productivity of grazing livestock and anthelmintic resistance has emerged globally. It is broadly understood that wild ruminants living in sympatry with livestock act as a positive source of refugia for anthelmintic-susceptible nematodes. However, they might also act as reservoirs of anthelmintic-resistant nematodes, contributing to the spread of anthelmintic resistance at a regional scale. Here, we sampled managed sheep and cattle together with feral goats within the same property in New South Wales, Australia. Internal transcribed spacer 2 (ITS-2) nemabiome metabarcoding identified 12 gastrointestinal nematodes (Cooperia oncophora, Cooperia punctata, Haemonchus contortus, Haemonchus placei, Nematodirus spathiger, Ostertagia ostertagi, Teladorsagia circumcincta, Oesophagostomum radiatum, Oesophagostomum venulosum, Trichostrongylus axei, Trichostrongylus colubriformis and Trichostrongylus rugatus). Isotype-1 β-tubulin metabarcoding targeting benzimidazole resistance polymorphisms identified 6 of these nematode species (C. oncophora, C. punctata, H. contortus, H. placei, O. ostertagi and T. circumcincta), with the remaining 3 genera unable to be identified to the species level (Nematodirus, Oesophagostomum, Trichostrongylus). Both ITS-2 and β-tubulin metabarcoding showed the presence of a cryptic species of T. circumcincta, known from domestic goats in France. Of the gastrointestinal nematodes detected via β-tubulin metabarcoding, H. contortus, T. circumcincta, Nematodirus and Trichostrongylus exhibited the presence of at least one resistance genotype. We found that generalist gastrointestinal nematodes in untreated feral goats had a similarly high frequency of the benzimidazole-resistant F200Y polymorphism as those nematodes in sheep and cattle. This suggests cross-transmission and maintenance of the resistant genotype within the wild ruminant population, affirming that wild ruminants should be considered potential reservoirs of anthelmintic resistance.

Integration of ITS-2 rDNA nemabiome metabarcoding with Fecal Egg Count Reduction Testing (FECRT) reveals ivermectin resistance in multiple gastrointestinal nematode species, including hypobiotic Ostertagia ostertagi, in western Canadian beef cattle

A large-scale Fecal Egg Count Reduction Test (FECRT) was integrated with ITS-2 rDNA nemabiome metabarcoding to investigate anthelmintic resistance in gastrointestinal nematode (GIN) parasites in western Canadian beef cattle. The study was designed to detect anthelmintic resistance with the low fecal egg counts that typically occur in cattle in northern temperate regions. Two hundred and thirty-four auction market-derived, fall-weaned steer calves coming off pasture were randomized into three groups in feedlot pens: an untreated control group, an injectable ivermectin treatment group, and an injectable ivermectin/oral fenbendazole combination treatment group. Each group was divided into six replicate pens with 13 calves per pen. Individual fecal samples were taken pre-treatment, day 14 post-treatment, and at monthly intervals for six months for strongyle egg counting and metabarcoding. Ivermectin treatment resulted in an 82.4% mean strongyle-type fecal egg count reduction (95% CI 67.8–90.4) at 14 days post-treatment, while the combination treatment was 100% effective, confirming the existence of ivermectin-resistant GIN. Nemabiome metabarcoding of third-stage larvae from coprocultures revealed an increase in the relative abundance of Cooperia oncophora, Cooperia punctata, and Haemonchus placei at 14 days post-ivermectin treatment indicating ivermectin resistance in adult worms. In contrast, Ostertagia ostertagi third-stage larvae were almost completely absent from day 14 coprocultures, indicating that adult worms of this species were not ivermectin resistant. However, there was a recrudescence of O. ostertagi third stage larvae in coprocultures at three to six months post-ivermectin treatment, which indicated ivermectin resistance in hypobiotic larvae. The calves were recruited from the auction market and, therefore, derived from multiple sources in western Canada, suggesting that ivermectin-resistant parasites, including hypobiotic O. ostertagi larvae, are likely widespread in western Canadian beef herds. This work demonstrates the value of integrating ITS-2 rDNA metabarcoding with the FECRT to enhance anthelmintic resistance detection and provide GIN species- and stage-specific information.

Isotonic, aqueous-based media as simple and suitable test media for short-term Haemonchus placei adult worm motility assay

The solvating power of test media used in anthelmintic assays is critical to the validity of assay results, especially when evaluating plant extracts. High solutes in media lowers its solvating power, altering the range of concentrations available for investigation and assay performance. To identify simplified, well-tolerated media for adult Haemonchus placei with improved solvating power, we investigated the impact of varying solutions of pH (2.5–8.5), salinity (19–154 mM), and normal saline (NS) incorporating dissolution enhancers (acetone, propylene glycol, DMSO and Tween-80; 10–40% v/v) on the nematode over 3 h at room temperature. The performance of identified media, NS and 20% Tween-80 in NS, were evaluated by preparing sample extracts (acetone extract Sarcocephalus latifolius, AESL20&10; and chloroform extract Vernonia amygdalina, CEVA20&10) stock solutions (20 and 10 mg/mL) in them, assessed their apparent dissolution, and each highest stock solution that dissolves the extracts evaluated for anthelmintic activity against H. placei. We found isotonicity to be the critical-to-worm survival factor as H. placei survived 100% in pH solutions 3.5–8.5, and saline solutions 39–154 mM. The dissolution enhancers, at 40%, gave no survival. At 30% and 20%, only Tween-80 gave 92.5% and 100% survival, respectively. At 10%, Tween-80, acetone, DMSO and propylene glycol gave 100%, 100%, 87.5% and 0% survival, respectively. In 20% Tween-80 in NS, AESL20&10 and CEVA20&10 dissolved, furnishing wider concentration range (20-0 mg/mL); whereas only AESL10 dissolved in NS (narrower concentration range, 10-0 mg/mL). The LC50s (mg/mL) of 7.67 (AESL10, NS) and 7.48 (AESL20, Tween-80 in NS) were not significantly different (p > 0.05), while CEVA20 (Tween-80 in NS) gave 2.67. Our findings show that NS and 20% Tween-80 in NS, as isotonic, aqueous-based media, are suitable, and well-tolerated as test media for adult H. placei in a short-term motility assay. Up to 30% Tween-80 could be used to enhance dissolution where necessary.

Selection of Molecules with Immunological Potential from Excretory and Secretory Products from the Nematode Haemonchus placei by Cell Proliferation and Gene Expression Assays.

The nematodeHaemonchus placeiis a pathogenic parasite, the most seriously affecting ruminant's health and being responsible for enormous economic losses all over the world. The present protocol describes different in vitro techniques to select potential candidate antigens with immune-protective activity from excretory and secretory products (ESP) fromH. placeitransitory infective larvae (xL3). ESP from xL3were obtained from the in vitro infective larvae (L3) maintained in Hank's medium at 37 °C with 5% CO2for 48 h. Then, the presence of ESP proteins was confirmed by SDS-PAGE to be used in an in vitro proliferation assay with bovine peripheral blood mononuclear cells (PBMCs). The ESP were exposed to the PBMCs during two different periods (24 and 48 h). Genes associated with immune response against the nematode were analyzed using relative gene expression and bioinformatic tools. These are simple, economic, and helpful tools to identify potential immune-protective molecules under in vitro conditions for confirming the efficacy of future in vivo assays. Graphical overview.

In vitro effect of three tropical plants on adult Haemonchus placei, an haematophagous nematode from cattle

BackgroundVernonia amygdalina (leaf), Garcinia kola (seed), and Leucaena leucocephala (seed) are three well-known tropical plants used in African ethnomedicine to reduce parasitic worm burdens and are potential sources of alternative solution for controlling parasitic helminths infection in grazing livestock. This study investigated extracts from these plants for anthelmintic activity against adult Haemonchus placei, an haematophagous nematode from cattle abomasa. Powdered plant materials were macerated in acetone and the crude acetone extracts evaluated for anthelmintic activity using H. placei adult worm motility assay. Afterwards, fresh sample of V. amygdalina was macerated successively in chloroform and acetone and the extracts evaluated for anthelmintic activity. The chloroform extract was subjected to phytochemical and FT-IR analyses and fractionated by vacuum liquid chromatography. Anthelmintic data were fitted to a nonlinear regression equation (Log [extract or fraction] vs. lethality; variable slope) to produce best-fit sigmoidal curves and LC50 values computed with associated uncertainty.ResultsOf the three tropical plants, only V. amygdalina was active against adult H. placei with best-fit LC50 of 6.51 mg/mL (95% CI: 5.32–7.75). Evaluation of the two extracts obtained by successive maceration showed that chloroform extract (LC50, 2.46 mg/mL, 95% CI: 1.87–3.28) was 11 times as potent as acetone extract (LC50, 27.01 mg/mL, 95% CI: 21.32–48.57) (α < 0.0001). Chromatographic fractionation of the chloroform extract yielded four fractions (FA-FD) with FB (LC50, 2.38 mg/mL, 95% CI: 1.76–3.28) 2.19 times as potent as FC (LC50, 5.21 mg/mL, 95% CI: 4.40–5.79) against H. placei, while FA and FD were inactive. Phytochemical evaluation of the chloroform extract revealed the presence of saponins, steroids, terpenoids, cardiac glycosides, and the absence of tannins, flavonoids, alkaloids, and anthraquinones. FT-IR structural analysis of chloroform extract indicated the presence of key functional groups which are chemical fragments/ structural motifs known to be present in the two major classes of bioactive compounds (sesquiterpene lactones and steroid glucosides) reportedly to be found in V. amygdalina.ConclusionsThe findings showed that chloroform extract of V. amygdalina leaf possessed relatively good anthelmintic activity against adult H. placei. This could be indicative of its potential usefulness as an anthelmintic phytomedicine to control gastrointestinal nematodes infection in cattle.Key highlightsExtracts of three different plant materials (one leaf, two seeds) were tested against adult Haemonchus placei in vitro;Chloroform extract of Vernonia amygdalina was 11 times as potent as acetone extract;Fractionation of the chloroform extract yielded a bioactive fraction responsible for about 90% of the total lethal effect of the chloroform extract.Bioprocessing of V. amygdalina leaf could produce phytomedicines for organic livestock farming.Graphical abstract

Eprinomectin 5% w/v extended-release injection for the treatment of larval and/or adult Bunostomum phlebotomum, Haemonchus contortus, H. placei, Nematodirus helvetianus, Oesophagostomum radiatum, and Trichostrongylus colubriformis infections in cattle.

In an effort to generate data for regulatory purposes on the therapeutic efficacy of eprinomectin 5% w/v extended-release injection (Eprinomectin ERI) (LongRange®, Boehringer Ingelheim) against infections of immature and adult stages of some nematode species (or stages) which are generally less common or predominantly seen in younger cattle, nine studies (minimum two per parasite and stage) were conducted in the USA and Germany. A total of 218 young cattle were included in seven experimentally induced infection studies (180 animals) and two studies with naturally acquired nematode infections (38 animals), which were compliant with WAAVP and VICH guidelines. In each study, cattle were randomly assigned into groups which received 1mL per 50kg body weight of either saline (controls) or Eprinomectin ERI (equivalent to 1mg eprinomectin per kg body weight) via subcutaneous injection when the parasites were developing fourth-stage larval (L4) or adult nematodes. Following necropsy and parasite recovery, percentage efficacy was calculated based on the comparison of geometric mean nematode counts of the Eprinomectin ERI- vs. the saline-treated animals. Eprinomectin ERI-treated cattle had significantly (p < 0.05) lower counts of each species and stage of nematodes than the controls. Eprinomectin ERI treatment was demonstrated to be efficacious (> 90%) against L4 and adult Bunostomum phlebotomum and Nematodirus helvetianus; against L4 Haemonchus placei, Oesophagostomum radiatum, and Trichostrongylus colubriformis; and against adult H. contortus. Percentage efficacy against L4 H. contortus was variable (69.6 to 100%). All treatments were well accepted, and no treatment-related health problems were observed in any study.

Haemonchus placei

This datasheet on Haemonchus placei covers Identity, Distribution, Hosts/Species Affected, Pathology, Epidemiology, Impacts.

Mortalidade causada por nematódeos gastrintestinais em bovinos de corte submetidos a protocolo sanitário inadequado

ABSTRACT: The aim of this study was to describe the epidemiological, clinical, and pathological findings of two outbreaks of gastrointestinal nematode infections in beef cattle kept under an inadequate sanitary protocol. An outbreak of trichostrongylosis occurred in Amambai, Mato Grosso do Sul (MS), from May to July 2016. The herd consisted of 3,000 Nellore cows, and of these, fifteen died after showing emaciation and diarrhea and remaining in recumbency. At necropsy, the abomasum showed edema in the folds, white, raised areas, multifocal to coalescent, and small ulcers. Histopathology showed larvae compatible with trichostrongylid nematodes were present inside the abomasum glands. Trichostrongylus axei was identify in the abomasum. The hemonchosis outbreak began in October 2018, in a breeding and finishing property in Santa Rita do Pardo, MS. Of 4,000 cattle aged 8 to 18 months, 673 became ill and 117 died. Clinical signs were emaciation, weakness, dehydration, submandibular edema, and soft stools. At necropsy, large numbers of nematodes were found in the abomasum; they were morphologically classified as Haemonchus placei. Both outbreaks were caused by failures in the parasite control protocol. However, in Outbreak I, frost and immune stress caused by lack of food may have contributed to the deaths. In Outbreak I, the main failure in the devermination protocol was the use of anthelmintics without carrying out an efficacy test. In Outbreak II, there was no parasite control protocol in the rearing and finishing property, and in the breeding property, there was resistance to the anthelmintic used (ivermectin). The two outbreaks highlight the importance of gastrointestinal nematode control protocols in cattle and demonstrate that infections by T. axei and H. placei can be lethal for this species.

Isotonic, Aqueous-Based Media as Simple and Suitable Test Media for Short Term Haemonchus Placei Adult Worm Motility Assay

Isotonic, Aqueous-Based Media as Simple and Suitable Test Media for Short Term Haemonchus Placei Adult Worm Motility Assay

Investigation of fecal egg counts versus worm burden and helminth fauna in cattle treated or not with macrocyclic lactones in a tropical region

Gastrointestinal nematodes negatively impact the health and productivity of livestock. Macrocyclic lactones (MLs) are the most common class of chemicals used in helminth control programs, however, their intense use is accelerating the development of parasite resistance. For cattle, little is known regarding the relationship between fecal egg counts (FECs) and the helminth population inside the animal's gastrointestinal tract and so this study evaluated the correlation between FEC and worm burden in cattle treated or not with MLs (ivermectin 200 μg/kg, ivermectin 630 μg/kg, abamectin 200 μg/kg, doramectin 700 μg/kg and moxidectin 200 μg/kg). Animals were necropsied to determine if there were any modifications to the proportions of worm species due to chemical treatment. FECs of ML-treated and untreated animals showed significant (p ≤ 0.05 and R2 ≥0.70) positive linear correlations and substantial or perfect strength-of-agreement (LCCC ≥ 0.61) with total worms present, all of which were Haemonchus placei and Cooperia punctata. However, this correlation and strength-of-agreement did not occur when the efficacy of MLs was ≥80 % against these helminths. Among the active MLs tested, moxidectin had the weakest relationship between FEC and worm burden, except for H. placei. Analysis of the helminthological fauna found an increase of H. placei (35.6 % = 95 % CI 35.4–35.6) and a consequent decrease of C. punctata (52.5 % = 95 % CI 52.3–52.6) in untreated animals, when compared to studies carried out over 20–40 years ago, but this proportion was reversed for ML-treated animals (C. punctata 64.5 % = 95 % CI 64.4–64.8; H. placei 30.8 % = 95 % CI 30.6–30.8). It is possible that MLs mitigate this effect in the field and that C. punctata remained prevalent in cattle in different regions of Brazil.

Epidemiological, Morphological, and Morphometric Study on Haemonchus spp. Recovered From Goats in Egypt.

Goats can be infected by multiple groups of external and internal parasites. Haemonchus spp. are among abomasal parasites that can result in higher mortality and several considerable economic losses in goats. Early detection of parasites and better understanding of the major risk factors associated with infection are among the main strategies for controlling the infection. Considering this, information on hemonchosis in goats from Egypt, and the contribution of goats in the maintenance of the epidemiological foci of the disease is limited. This study investigated the prevalence of Haemonchus species among 240 abomasum samples collected during postmortem examination of goat carcasses from Assiut Governorate, Egypt. Moreover, the association of the major risk factors to describe the epidemiological pattern of the disease was explored. This study demonstrated that 16.66% of abomasa samples harbored Haemonchus species. Additionally, age, sex, and sampling season were the most significant risk factors associated with infection. Following the variable factors under study, goats aged 1 year or older were at higher risk, with an infection rate of 22.14% (31 of 140), than those younger than 1 year (9%) [p = 0.008; odds ratio (OR) = 2.87; 95% confidence interval (CI), 1.30–6.35]. The infection rate was 25% (19 of 76) in males and 12.8% (21 of 164) in females [p = 0.024; odds ratio (OR) = 2.26; 95% confidence interval (CI), 1.13–4.53]. Moreover, the exposure to infection was higher in summer (22.22%) than in winter (8.33%) (p = 0.007; odds ratio (OR) = 0.318; 95% confidence interval (CI), 0.139–0.725). More importantly, three species of the parasite—Haemonchus contortus, Haemonchus placei, and Haemonchus longistipes—were identified for the first time, and the confirmation of the identification and morphological characterization of the worms was performed using light microscopy and SEM. Collectively, this study reveals interesting epidemiological, morphological, and morphometric findings associated with the occurrence of hemonchosis among goats in Egypt. This study suggests further research for exploring the major circulating species of the parasite in Egypt, which is mandatory for controlling the disease.

Analysis of the immunomodulatory activity of excreted and secreted products from Haemonchus placei transition infective larvae (xL3)

The excretory/secretory (E/S) products released by infective transitory larvae (xL3) of Haemonchus placei have an important biological function in stimulating immune mechanisms during the invasive process. Our objective was to analyse the modulatory activity of 15 and 70 kDa E/S products from H. placei xL3. Both E/S products were collected from xL3in vitro cultures at 24 and 72 h. Proteins were confirmed by SDS-PAGE, and the corresponding spots were elicited by gel isoelectrofocusing (IEF) and characterised by mass spectrometry. Additionally, flow cytometry of CD4+/γδ+ T cells and immune gene expression were performed by proliferation assays using each E/S product to stimulate lymphocyte and peripheral blood mononuclear cells (PBMCs) from non-infected calves. The IEF results displayed two spots of 7.0 and 5.7 pI for the 15 and 70 kDa products, respectively. Additionally, 29 and 17 peptides from the 15 and 70 kDa E/S products, respectively, were identified with the hypothetical neurotransmitter and enzymatic functions necessary for larval development. The relative expression displayed upregulation of IL4, 5, 6, 8, 10, 13, IFNγ, and FCεR1A genes (from 2.0- to 17.6-fold, p < 0.05) stimulated by the 15 and 70 kDa proteins, indicating specific genes against haemonchosis. Although the percentage of median florescence intensity (MFI%) of CD4+/γδ+ T cells did not change for both E/S products compared to the negative control and concanavalin-A stimulated cells as the positive control (p > 0.05), the 15-kDa protein reduced the levels of both T cells, and the 70-kDa proteins increased the γδ+ cells slightly. Additionally, there was increased PBMCs proliferation by the 70 kDa proteins (p < 0.05), denoting the biological role of other immune cells. The 15 and 70 kDa protein E/S products from H. placei xL3 showed modulation of the immune response, and although more studies are required, they indicate important functions in the host/parasite interaction.

Anthelmintic activity and non-cytotoxicity of phaeophorbide-a isolated from the leaf of Spondias mombin L.

Ethnopharmacological relevanceHelminthosis (worm infection) is a disease of grazing livestock, with significant economic implications. Increasing resistance to existing synthetic anthelmintics used to control helminthosis and the unwanted presence of residues of the anthelmintics reported in meat and dairy products present a serious global health challenge. These challenges have necessitated the development of novel anthelmintics that could combat drug resistance and exhibit better safety profiles. Spondias mombin L. (Anacardiaceae) is a plant that has been used traditionally as a worm expeller. Aim of studyThe aim of the work reported herein was to isolate and characterise anthelmintic compound(s) from S. mombin leaf, establishing their bioactivity and safety profile. Materials and methodsAdult Haemonchus placei motility assay was used to assess anthelmintic bioactivity. Bioassay-guided chromatographic fractionation of acetone extract of S. mombin leaf was carried out on a silica gel stationary phase. The structure of the compound was elucidated using spectroscopy (1H and 13C NMR) and Liquid Chromatography-Mass Spectrometry (LC-ESI-MS). Screening to exclude potential cytotoxicity against mammalian cells (H460, Caco-2, MC3T3-E1) was done using alamar blue (AB) and CellTitreGlo (CTG) viability reagents. ResultsThe acetone extract yielded an active fraction 8 (Ethyl acetate: methanol 90:10; anthelmintic LC50: 3.97 mg/mL), which yielded an active sub-fraction (Ethyl acetate: Methanol 95:5; anthelmintic LC50: 53.8 μg/mL), from which active compound 1 was isolated and identified as phaeophorbide-a (LC50: 23.0 μg/mL or 38.8 μM). The compound was not toxic below 200 μM but weakly cytotoxic at 200 μM. ConclusionsPhaeophorbide-a (1) isolated from S. mombin leaf extract and reported in the plant for the first time in this species demonstrated anthelmintic activity. No significant toxicity to mammalian cells was observed. It therefore represents a novel anthelmintic pharmacophore as a potential lead for the development of novel anthelmintics.

Effect of antihelminths with contrasting efficacy against gastrointestinal nematodes on the live-weight gain of young Nellore cattle.

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 2.5mg/kg of albendazole sulfoxide; Group II - free from Oesophagosomum radiatum using 0.2mg/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 12months. 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.

Gastrointestinal parasites of a reintroduced semi-wild plains bison (Bison bison bison) herd: Examining effects of demographic variation, deworming treatments, and management strategy.

Bison (Bison spp) are being reintroduced into semi-wild, spatially constrained herds across North America and Europe. Herd managers are concerned about gastrointestinal (GI) nematode parasites as they care for the health of their bison. We examine how demographics, grazing location, herd management, and anthelmintic treatments affect the fecal egg counts (FECs) of GI nematodes within a reintroduced Plains bison (Bison bison bison) herd in the Great Plains. Our results suggest that younger bison (<2 years of age) experience higher GI parasite eggs/oocysts per gram (epg/opg) and that some taxa are more prevalent throughout different periods of a bison's early years. Demographic findings suggest that calf and yearling (0–2 yrs age) bison have the highest FECs and that these decline until reaching a low in peak adulthood and thereafter (x > 6 yrs of age). FECs of both Trichuris spp. and particularly Nematodirus spp. were much more abundant, relatively, during the first year of a bison's life. This pattern was also true of Moniezia spp. and Eimeria spp., however, strongyle-type spp. FECs appeared to peak in relative abundance during the second year of life. Our data also indicate that FECs are influenced by differences in land-use histories of pastures previously grazed by cattle or by the proportion of frequent flooding in different pastures. Treatment results suggest that fenbendazole may more effective than moxidectin at lowering FECs of bison over the long-term, and lasting effects of at least one administered anthelmintic treatment. Multiplex PCR assays revealed that American bison share GI nematodes with cattle including: Ostertagia spp., Haemonchus placei, Cooperia onchophora, and Oesophagostomum spp, but did not detect the presence Trichostrongylus columbriformis. Our results may have wider conservation implications for reintroduction efforts of American bison, as well as the endangered European bison (Bison bonasus).