The most challenging obstacles to testing products for their anthelmintic activity are: (1) establishing a suitable nematode in vitro assay that can evaluate potential product use against a parasitic nematode of interest and (2) preparation of extracts that can be redissolved in solvents that are miscible in the test medium and are at concentrations well tolerated by the nematode system used for screening. The use of parasitic nematodes as a screening system is hindered by the difficulty of keeping them alive for long periods outside their host and by the need to keep infected animals as sources of eggs or adults when needed. This method uses the free-living soil nematode Caenorhabditis elegans as a system to screen products for their potential anthelmintic effect against small ruminant gastrointestinal nematodes, including Haemonchus contortus. This modified method uses only liquid axenic medium, instead of agar plates inoculated with Escherichia coli, and two selective sieves to obtain adult nematodes. During screening, the use of either balanced salt solution (M-9) or distilled water resulted in averages of 99.7 (± 0.73)% and 96.36 (± 2.37)% motile adults, respectively. Adult worms tolerated DMSO, ethanol, methanol, and Tween 80 at 1% and 2%, while Labrasol (a bioenhancer with low toxicity to mammals) and Tween 20 were toxic to C. elegans at 1% and were avoided as solvents. The high availability, ease of culture, and rapid proliferation of C. elegans make it a useful screening system to test plant extracts and other phytochemical compounds to investigate their potential anthelmintic activity against parasitic nematodes.

Anthelmintic resistance is a worldwide concern in small ruminant industry and new plant-derived compounds are being studied for their potential use against gastrointestinal nematodes. Mentha piperita, Cymbopogon martinii and Cymbopogon schoenanthus essential oils were evaluated against developmental stages of trichostrongylids from sheep naturally infected (95% Haemonchus contortus and 5% Trichostrogylus spp.) through the egg hatch assay (EHA), larval development assay (LDA), larval feeding inhibition assay (LFIA), and the larval exsheathment assay (LEA). The major constituent of the essential oils, quantified by gas chromatography for M. piperita oil was menthol (42.5%), while for C. martinii and C. schoenanthus the main component was geraniol (81.4% and 62.5%, respectively). In all in vitro tests C. schoenanthus essential oil had the best activity against ovine trichostrongylids followed by C. martini, while M. piperita presented the least activity. Cymbopogon schoenanthus essential oil had LC(50) value of 0.045 mg/ml in EHA, 0.063 mg/ml in LDA, 0.009 mg/ml in LFIA, and 24.66 mg/ml in LEA. The anthelmintic activity of essential oils followed the same pattern in all in vitro tests, suggesting C. schoenanthus essential oil could be an interesting candidate for nematode control, although in vivo studies are necessary to validate the anthelmintic properties of this oil.

BackgroundTicks cause massive damage to livestock and vaccines are one sustainable substitute for the acaricides currently heavily used to control infestations. To guide antigen discovery for a vaccine that targets the gamut of parasitic strategies mediated by tick saliva and enables immunological memory, we exploited a transcriptome constructed from salivary glands from all stages of Rhipicephalus microplus ticks feeding on genetically tick-resistant and susceptible bovines.ResultsDifferent levels of host anti-tick immunity affected gene expression in tick salivary glands; we thus selected four proteins encoded by genes weakly expressed in ticks attempting to feed on resistant hosts or otherwise abundantly expressed in ticks fed on susceptible hosts; these sialoproteins mediate four functions of parasitism deployed by male ticks and that do not induce antibodies in naturally infected, susceptible bovines. We then evaluated in tick-susceptible heifers an alum-adjuvanted vaccine formulated with recombinant proteins. Parasite performance (i.e. weight and numbers of females finishing their parasitic cycle) and titres of antigen-specific antibodies were significantly reduced or increased, respectively, in vaccinated versus control heifers, conferring an efficacy of 73.2%; two of the antigens were strong immunogens, rich in predicted T-cell epitopes and challenge infestations boosted antibody responses against them.ConclusionMining sialotranscriptomes guided by the immunity of tick-resistant hosts selected important targets and infestations boosted immune memory against salivary antigens.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2136-2) contains supplementary material, which is available to authorized users.

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a b s t r a c tThis study's main goal was to evaluate the thermoregulatory responses velocity through the variation of rectal temperature (RT), related to the thermolytic pathways, respiratory rate (RR) and sweating rate (SR) among different sheep breeds. Ninety female sheep, eighteen of each breed: Santa Ines and Morada Nova (Brazilian hair breeds), Texel, Suffolk and Ile de France (wool breeds) were challenged during three non-consecutive summer days (22• 42 S, 47• 18 W, and 570 m of altitude, maximum air temperature of 33.5• C, average relative humidity of 52 ± 6.9%). The physiological variables were registered at 0800 h (T1), 1300 h (T2: after 2 h of shade rest), 1400 h (T3) (after one hour of sun exposure) and in the shade at 1415 h (T4), 1430 h (T5), 1445 h (T6) and 1500 h (T7) and a thermotolerance index (TCI) was calculated as (10-(T7 to T4)-T1). The statistical analysis was performed by a mathematical model including the fixed effects of breeds and time frames, and the interaction between these effects, besides random effects such as animal and day. The Santa Ines breed presented the lowest RT after sun exposure (39.3 ± 0.12• C; P < 0.05) and it was the only one to recover morning RT 60 min after heat stress (38.7 and 38.9 for 1300 h and 1500 h; P > 0.05). Hair breeds presented RR lower (P < 0.05) than wool breeds. Although thick wool or hair thickness differs among and within hair and wool breeds (P < 0.05), SR did not differ among breeds and time (227.7 ± 16.44 g m −2 h −1 ; P > 0.05). The thermotolerance index did not differ among breeds, but it showed similar response (P > 0.05) 45 min or 1 h of shade after sun exposure. One week post shearing is not enough to wool breeds present to show thermotolerance similar to hair breeds.

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