Responses of artificially reared cat fleas Ctenocephalides felis felis (Bouché, 1835) to different mammalian bloods.

The cat flea, Ctenocephalides felis felis (Bouche, 1835) (Siphonaptera: Pulicidae), which is found worldwide and which parasitizes many species of wild and domestic animal, is a vector and/or reservoir of bacteria, protozoa and helminths. To aid in the study of the physiology and behaviour of fleas and of their transmission of pathogens, it would be of value to improve the laboratory rearing of pathogen-free fleas. The conditions under which artificially reared fleas at the University of Bristol (U.K.) and the Rickettsial Diseases Institute (France) are maintained were studied, with different ratios of male to female fleas per chamber (25 : 50, 50 : 100, 100 : 100, 200 : 200). The fleas were fed with bovine, ovine, caprine, porcine or human blood containing the anticoagulants sodium citrate or EDTA. Egg production was highest when fleas were kept in chambers with a ratio of 25 males to 100 females. In addition, the use of EDTA as an anticoagulant rather than sodium citrate resulted in a large increase in the number of eggs produced per female; however, the low percentage of eggs developing through to adult fleas was lower with EDTA. The modifications described in our rearing methods will improve the rearing of cat fleas for research.

Wild deer as potential vectors of anthelmintic-resistant abomasal nematodes between cattle and sheep farms.

Gastrointestinal (GI) nematodes are among the most important causes of production loss in farmed ruminants, and anthelmintic resistance is emerging globally. We hypothesized that wild deer could potentially act as reservoirs of anthelmintic-resistant GI nematodes between livestock farms. Adult abomasal nematodes and faecal samples were collected from fallow (n = 24), red (n = 14) and roe deer (n = 10) from venison farms and areas of extensive or intensive livestock farming. Principal components analysis of abomasal nematode species composition revealed differences between wild roe deer grazing in the areas of intensive livestock farming, and fallow and red deer in all environments. Alleles for benzimidazole (BZ) resistance were identified in ß-tubulin of Haemonchus contortus of roe deer and phenotypic resistance confirmed in vitro by an egg hatch test (EC50 = 0.149 µg ml(-1) ± 0.13 µg ml(-1)) on H. contortus eggs from experimentally infected sheep. This BZ-resistant H. contortus isolate also infected a calf experimentally. We present the first account of in vitro BZ resistance in wild roe deer, but further experiments should firmly establish the presence of phenotypic BZ resistance in vivo. Comprehensive in-field studies should assess whether nematode cross-transmission between deer and livestock occurs and contributes, in any way, to the development of resistance on livestock farms.

Prevalence of nematode infection and faecal egg counts in free-range laying hens: relations to housing and husbandry.

1. Faecal samples from 19 commercial, 65 week old free-range egg laying flocks were examined to assess the prevalence and number of parasitic nematode eggs. Data were collected to characterise the housing, husbandry, behaviour and welfare of the flocks to examine possible relationships with the egg counts. 2. Eggs of at least one genus of nematode were present in the faeces of all 19 flocks. Heterakis eggs were detected in 17 (89%) flocks, Ascaridia in 16 (84%), Trichostrongylus in 9 (47%), and Syngamus in 6 (32%). Faecal egg counts (FEC) were greatest for Ascaridia and Heterakis. 3. For each nematode genus, there was no significant difference in FEC between organic (N = 9) and non-organic (N = 10) flocks, or between static (N = 8) and mobile (N = 11) flocks. 4. FEC were correlated with a range of housing, husbandry and management practices which varied between the nematode genus and included depth of the litter, percentage of hens using the range, and number of dead hens. Statistical analysis indicated relationships with FEC that included light intensity above the feeder, indoor and outdoor stocking density, fearfulness in the shed and on the range, distance to the nearest shelter, and swollen toes. 5. None of the FEC for any of the genera was correlated with weekly egg production or cumulative mortality. 6. Although nematode FEC were highly prevalent among the flocks, the overall lack of relation to other welfare and production measures suggests that these infections were not severe.

A survey of helminth control practices on sheep farms in Great Britain and Ireland.

A telephone survey of 600 farmers throughout Great Britain and Ireland was conducted in order to characterise helminth control practices, and identify factors correlated with perceived anthelmintic failure. Overall, 93% of surveyed farmers routinely treated their sheep against nematodes, 67% against liver fluke and 58% against tapeworms. Anthelmintic resistance in nematodes was perceived by farmers to be present on 10% of farms. Farmers who dosed ewes at mating were more likely to have observed anthelmintic failure, than those who were aware of national guidelines on parasite control. However, objective assessment of anthelmintic efficacy had only been undertaken on 19% of farms. Ewes were treated at mating and lambing on 63% and 62% of farms, respectively. On average, lambs were treated 3.6 times annually, depending on geographical region and on dates of lambing and finishing. Although 'quarantine' treatments were widely administered to bought-in stock, these were appropriately applied in only 3% of cases. This study provides baseline data against which the impact of future anthelmintic information campaigns can be assessed; it will facilitate the development of rational, farm-level mathematical models in support of sustainable parasite control, and will aid in the design of farm management practices that prolong the effective lifespan of novel classes of anthelmintic.

Calibration and diagnostic accuracy of simple flotation, McMaster and FLOTAC for parasite egg counts in sheep.

The present study was aimed at carrying out a calibration and a comparison of diagnostic accuracy of three faecal egg counts (FEC) techniques, simple flotation, McMaster and FLOTAC, in order to find the best flotation solution (FS) for Dicrocoelium dendriticum, Moniezia expansa and gastrointestinal (GI) strongyle eggs, and to evaluate the influence of faecal preservation methods combined with FS on egg counts. Simple flotation failed to give satisfactory results with any samples. Overall, FLOTAC resulted in similar or higher eggs per gram of faeces (EPG) and lower coefficient of variation (CV) than McMaster. The "gold standard" for D. dendriticum was obtained with FLOTAC when using FS7 (EPG=219, CV=3.9%) and FS8 (EPG=226, CV=5.2%) on fresh faeces. The "gold standard" for M. expansa was obtained with FLOTAC, using FS3 (EPG=122, CV=4.1%) on fresh faeces. The "gold standard" for GI strongyles was obtained with FLOTAC when using FS5 (EPG=320, CV=4%) and FS2 (EPG=298, CV=5%). As regard to faecal preservation methods, formalin 5% and 10% or freezing showed performance similar to fresh faeces for eggs of D. dendriticum and M. expansa. However, these methods of preservation were not as successful with GI strongyle eggs. Vacuum packing with storage at +4°C permitted storage of GI strongyle eggs for up to 21 days prior to counting. Where accurate egg counts are required in ovine samples the optimum method of counting is the use of FLOTAC. In addition, we suggest the use of two solutions that are easy and cheap to purchase and prepare, saturated sodium chloride (FS2) for nematoda and cestoda eggs and saturated zinc sulphate (FS7) for trematoda eggs and nematoda larvae.

Comparison of traps for the control of sheep blowfly in the U.K.

The ability of three commercially available trap types to catch Lucilia (Diptera: Calliphoridae) blowflies was assessed on three sheep farms in southwest England in 2008. The aim was to evaluate their relative value for the control of ovine cutaneous myiasis (sheep blowfly strike) on farms. There was a highly significant difference between the total number of female Lucilia caught per day by the traps, with an Agrilure Trap (Agrimin Ltd, Brigg, U.K.) catching more than the other trap types (Rescue Disposable Fly Trap, Sterling International, Spokane, U.S.A.; Redtop Trap, Miller Methods, Johannesburg, South Africa). However, there was no significant difference between the traps in the numbers of female Lucilia sericata (Meigen) caught. Nevertheless, consideration of the rate at which female L. sericata were caught over time showed that the Agrilure trap did not begin catching until about 30 days after its initial deployment. It subsequently caught L. sericata at a faster rate than the other two traps. The data suggest that the freeze-dried liver bait used in the Agrilure trap required a period of about 30 days to become fully rehydrated and decompose to the degree required to attract and catch L. sericata. Once the bait was attractive, however, the trap outperformed the other two traps in terms of the rate of L. sericata capture. The Agrilure trap would appear to be the most effective of the designs tested for use against sheep blowfly and blowfly strike in the U.K., but care would be needed to ensure that the traps were deployed in advance of the blowfly season so that the bait was suitably aged when trapping was required.

FLOTAC: an improved method for diagnosis of lungworm infections in sheep.

The FLOTAC techniques involve the spinning of faecal samples onto the surface of counting chambers to permit enumeration of parasitic elements (eggs, larvae, oocysts and cysts) to an accuracy of one parasitic element per gram of faeces. In the present study it is demonstrated that FLOTAC provides a rapid and very sensitive method for counting of lungworm larvae of sheep. The optimum flotation solution for lungworm larvae is zinc sulphate and mercury II iodide (s.g. 1.45) although zinc sulphate (s.g. 1.20 or 1.35) on its own also gave good results. Samples preserved in 5% formalin gave the highest counts but fresh, frozen and samples in 10% formalin also gave higher counts than McMaster and simple flotation. Larval counts of 307 field samples gave up to 1.27x more positives samples than use of Baermann funnels and up to 4.18x more larvae per sample. As FLOTAC is faster than Baermannisation of samples it offers a better method of counting larvae in ruminant faecal samples.

Susceptibility to praziquantel of male and female cercariae of praziquantel-resistant and susceptible isolates of Schistosoma mansoni.

Praziquantel (PZQ) is now widely used for the treatment of human schistosomiasis. However, in recent years, there has been a growing concern about the resistance of Schistosoma to PZQ. The mechanisms of PZQ action against Schistosoma and resistance of Schistosoma to PZQ are poorly understood. Here, we report differential susceptibilities to PZQ between male and female cercariae in the PZQ-susceptible and PZQ-resistant isolates of Schistosoma mansoni, using tail loss as a measurement of PZQ action. The miracidia were collected by hatching eggs collected from faeces of infected mice. Single-sex cercaria lines were made by infecting a single Biomphalaria glabrata snail with a single miracidium. The sex of each single-sex cercaria line was identified by a direct W1-specific polymerase chain reaction (PCR) technique. Single-sex cercariae of two isolates were exposed to four different concentrations of PZQ, respectively. The tail shedding of cercariae was observed under a dissecting microscope for five time points up to 100 min after adding PZQ. The results showed that male cercariae have higher tail-shedding rates than that of female cercariae when PZQ-susceptible isolates of S. mansoni are exposed to the same concentration of PZQ. But this phenomenon was not observed in the PZQ-resistant isolates. This sexual differential resistance phenomenon of S. mansoni suggests that resistance to PZQ is induced by decreasing the PZQ susceptibility of male worms. The experiment described here may also be useful for developing tests to detect PZQ resistance in the field.

Weight-based targeted selective treatment of gastrointestinal nematodes in a commercial sheep flock.

Targeted selective treatment (TST) strategies, in which a proportion of the flock or herd is left untreated so that anthelmintic-susceptible genotypes are preserved, are increasingly advocated as a means of prolonging the effective life of current anthelmintic drugs. The major limitation to this approach is a lack of efficient indicators for selection, which can be applied effectively on commercial farms to identify individuals that can be left untreated without fear of disease or production loss. With the advent of electronic identification and automated weighing technology, monitoring of short-term changes in weight gain shows promise as such an indicator, but its operation in the field as part of TST has yet to be evaluated. Widespread deployment of weight-based TST will be highly dependent on the likely production penalty from leaving the fastest growing animals untreated. On a commercial flock in south-west UK, the weight gain of 508 lambs of various breeds was tracked using an automated identification and weighing system, every one to ten weeks from June to December (one to four weeks in summer), and a variable proportion of the fastest growing individuals that also appeared to be in good condition with little breech soiling was left untreated during whole-flock dosing in June, July and August. In total, 51 lambs were selected for non-treatment on at least one occasion, while the other lambs were treated two or three times during the summer. Subsequent weight gain of untreated animals was not reduced relative to their peers in either the short-term or over the whole grazing season. Faecal egg counts from untreated individuals did not differ significantly from those of the rest of the flock, suggesting that animals left untreated on the basis of weight gain can contribute effectively to refugia. The application of TST in this case is cautious in its extent, but this is appropriate on a commercial farm with associated aversion to production loss. Results suggest that such losses can be avoided while leaving part of the flock untreated, and should encourage wider application of this approach to slow the development of anthelmintic resistance. Since the cost of investment in weighing and recording systems is likely to prove prohibitive to many farmers, other selective indicators should also be investigated. The co-ordination of TST with pasture use to maximise the benefit in terms of environmental refugia, and its integration with other control strategies, also requires further attention.

Mechanical recovery of inhibited cyathostomin larvae from equine intestinal tissue.

The Stomacher is very widely used in food and medical research for extracting tissues. To determine whether nematode larvae were disrupted by the Stomacher, L3 larvae of Haemonchus contortus were homogenised for up to 40 min at full power but no larval disruption occurred. Therefore, tissue from the mucosa and submucosa of the caecum of horses collected from a licenced abattoir was treated to determine whether inhibited cyathostomin larvae could be extracted. The optimum time on full power for a 10-g sample was 20 min, and in three out of five caecal samples from different horses, significantly more larvae were recovered than with 6 h pepsin HCl digestion. It is concluded that the Stomacher provides a simple fast method of extracting inhibited nematode larvae from gastrointestinal tissues in the horse that could replace digestion with pepsin HCl.

The role of targeted selective treatments in the development of refugia-based approaches to the control of gastrointestinal nematodes of small ruminants.

Anthelmintic resistance is recognised as a major problem affecting small ruminant production worldwide and now threatens the sustainability of many of these systems. One method that has been proposed to prolong the efficacy of our current anthelmintics is the maintenance of a parasite population in refugia (unexposed to a drug) which will maintain the genes for susceptibility within the parasite population. Management strategies that employ refugia-based methods include targeted or strategically timed whole flock treatments, targeted selective treatments (TST), whereby only a proportion of the flock is treated at any one time, and the dilution of resistant with susceptible parasites. The ability to effectively target anthelmintic use relies on the identification of those animals that will most benefit from treatment. This review explains the concept of refugia, describes the role of refugia-based approaches to the management of anthelmintic resistance and reviews the markers that have been studied as indicators for TSTs as well as the implementation of refugia-based strategies. Recent results suggest that targeting anthelmintic treatment on the basis of anaemia, milk production and liveweight gain may offer a means of reducing anthelmintic usage whilst still maintaining animal performance.

Impact of the South-to-North Water Diversion Project on the transmission of Schistosoma japonicum in China.

The South-to-North Water Diversion Project (SNWDP) is currently the key, national, water-conservation project in China, designed to optimise the use of water resources and relieve the water shortages in the north of the country. As one of the main water intakes for the project, that of the Eastern Route Scheme (ERS), is a breeding site for Oncomelania hupensis (the intermediate host of Schistosoma japonicum), there is concern that the snail may be carried far to the north, in the water passing through the project. To see if they could survive and breed to the north of their current range in China, O. hupensis were collected in marshland near Nanjing City and transferred to cages, on the banks of fish ponds, in the cities of Zhenjiang (in Jiangsu province, at 32 degrees 10'N), Xuzhou (in the same province but at a latitude of 34 degrees 23'N) and Jining (in Shandong province, at 35 degrees 23'N). Except over the first 6 months in Xuzhou, the snails moved north of their natural distribution did not survive and reproduce as well as those in Zhenjiang, and all those transferred to Jining died out within 1 year. Although the snail populations in Xuzhou survived for 7-8 years and retained their infectivity to S. japonicum, histological and histochemical studies revealed abnormalities in the reproductive organs of these snails. It is concluded that, unless global warming significantly increases the minimum winter temperatures in northern China, the SNWDP is unlikely to result in the northward spread of schistosomiasis japonica.

Control of head lice with a coconut-derived emulsion shampoo.

OBJECTIVE: To evaluate a novel coconut-derived emulsion (CDE) shampoo against head lice infestation in children. DESIGN: A school trial in which pupils were treated on days 0 and 7 and checked on days 8 and 15 and a family trial where product was applied by parents three times in 2 weeks or used as a cosmetic shampoo and checked on days 14 and days 70. SETTING: UK schools in Bristol and Western-super-Mare and families in Northern Somerset. MAIN OUTCOME MEASURE: Numbers of children free from infestation after treatment. RESULTS: In the school trial, percentage cures at day 8 were 14% (permethrin, n=7) and 61% (CDE, n=37). In the family trial where all family members were treated, cure rate was 96% (n=28), and if the shampoo was subsequently used as a cosmetic shampoo, only 1 of 12 children became re-infested after 10 weeks. CONCLUSION: CDE shampoo is a novel effective method of controlling head lice and used after treatment as a cosmetic shampoo can aid in the reduction of re-infestation.

An immunohistochemical assessment of the cutaneous immune response to louse infestation in cattle.

Skin samples were taken from 10 experimental cattle exposed naturally, during a period extending over two winters, to Bovicola bovis and Solenoptes capillatus, five becoming infested and five being protected from infestation by repeated treatment with ectoparasiticides. Skin sections were examined histopathologically and immunohistochemically for expression of the immune cell markers CD3, CD4, CD8 and class II antigens of the major histocompatibility complex (MHC). Louse-infested cattle had a mixed infiltration of the superficial dermis and perifollicular regions with eosinophils and mononuclear cells. The skin of infested cattle differed from that of non-infested cattle in showing significantly more cells expressing CD3, CD4 and MHC class II (P<0.05). Many of the MHC class II(+) cells had dendritic morphology, suggesting active antigen presentation within the lesions. Louse infestations have previously been thought to produce a type 1 hypersensitivity response, mediated by Th2 lymphocytes. However, the increased number of lymphocytes and antigen-presenting cells observed in the present study suggests that in chronic infestation there is activation of local cell-mediated (Th1) immunity.