Development of molecular detection methods of Bovicola ovis from sheep fleece.

The sheep body louse (Bovicola ovis) commonly referred to as sheep lice are small chewing ectoparasites of sheep. Infection results in significant economic costs to the Australian sheep industry due to reduced wool quality caused by chronic itching from sheep rubbing and biting fleece. Treatment relies on use of insecticides; however, resistance has developed against pyrethroid and other insect growth regulator lousicides. There is urgent need to develop cost-effective lice management to reduce the use of insecticides, with the application of insecticidal treatments only applied when an infestation is detected. However, the current detection method relies on fleece parting for detection of B. ovis which is highly dependent on the skill of the inspector, the number of sheep examined, and the prevalence and severity of the infestation. To improve B. ovis detection, a highly sensitive (5 × 10-8 ng/µL) and specific multiplex quantitative PCR which simultaneously detects sheep lice and sheep DNA was developed. In addition, a B. ovis loop-mediated isothermal amplification (LAMP) assay was developed for field use. The B. ovis LAMP (Bov-LAMP) assay was optimized to reliably detect B. ovis from wool samples down to 5 × 10-6 ng/µL, with time to positive (Tp) < 10 min. Both assays demonstrate high sensitivity and specificity, enabling rapid identification of B. ovis DNA from sheep fleece samples and have the capacity to be used for ongoing management and surveillance of B. ovis in Australian sheep flocks.

Bioactivity of tea tree oil from Melaleuca alternifolia against sheep lice (Bovicola ovis Schrank) in vitro.

Tea tree oil (TTO) from the Australian native plant Melaleuca alternifolia has wide ranging bio-active properties, including insecticidal and repellent activity against arthropods. Furthermore, composition of commercially available Australian TTO is specified under an International Organization for Standardization standard (ISO 4730), reducing the potential for variable effects often noted with botanical pesticides. The effect of TTO, meeting the ISO standard for terpinen-4-ol chemotype, was tested against sheep lice (Bovicola ovis Schrank) in a series of laboratory studies. Immersion of wool for 60s in formulations containing concentrations of 1% TTO and above caused 100% mortality of adult lice and eggs. Exposure to vapours from TTO, delivered as droplets in fumigation chambers and when applied to wool also caused high mortality in both lice and eggs. The main active component of TTO in the fumigant tests was terpinen-4-ol. Treated surface assays and tests with wool where the formulation was allowed to dry before exposure of lice indicated low persistence. These studies demonstrate that TTO is highly toxic to sheep lice and active at concentrations that suggest potential for the development of TTO-based ovine lousicides.

[Numerical characteristics of insect community structure in five stored export products along Yangtze River in Jiangsu Province].

This paper investigated the dominant species of insect communities in five stored export products along Yangtze River in Jiangsu Province. With the indexes species richness, ecological dominance, community diversity and evenness, the numerical characteristics of insect community structure in the stored products were analyzed, and the similarity of the communities was also analyzed. By the method of hierarchical cluster analysis, the numerical classification was made, and the communities were grouped into 4 groups. The insect communities in stored willow and feather products were in the same group, while those in other stored products were grouped into 3 groups. Except that in woodwork products, the insect community structures in other stored products, especially in leather products, were not rational.

A simple pump-assisted method for collecting live, undamaged Psoroptes ovis from sheep using circulating saline.

The availability of large numbers of undamaged sheep scab mites, Psoroptes ovis, would be beneficial for discovery screening and development trials. There are several reported procedures for removing scab mites from sheep but they have limitations. To overcome this, a simple but versatile method employing the use of pumped saline was developed to remove all stages of the P. ovis mite from sheep. The method takes no more than 2 min to remove mites from the selected site with relative ease and is not affected by the condition of the fleece or lesion. The number of mites removed with the new method was 5-10 times more than detected by visual examination. These mites were undamaged and survived off-host for up to 16 days. The robust, portable equipment is easy to use under field conditions, making this method suitable for use as a diagnostic tool for early detection and monitoring of scab mites thus providing opportunities for development of novel alternative control strategies.

Occupational rhinoconjunctivitis and asthma in a wool worker caused by Dermestidae spp.

BACKGROUND: The family Dermestidae belongs to the order Coleoptera. Occupational allergy has been described in museum personnel. A 31-year-old male wool worker presenting rhinoconjunctivitis and asthma episodes probably linked to exposure to Dermestidae-infected wool was investigated. METHODS: Extracts prepared either from insect bodies or from dust from parasitized wool were used for skin prick testing (SPT), conjunctival and bronchial provocation tests and in vitro determinations. RESULTS: SPT and provocation tests were positive to both extracts. PEFR measurement demonstrated the association between the patient's symptoms and occupational exposure to Dermestidae. Specific IgE to both extracts was detected and immunoblotting revealed several protein bands from 5 to 200 kDa that were reactive to IgE from the patient's serum. CONCLUSIONS: Dermestidae exposure in wool workers when handling parasitized wool can be a cause of IgE-mediated rhinoconjunctivitis and asthma.

Variation among Merino sheep in susceptibilty to lice (Bovicola ovis) and association with susceptibility to trichostrongylid gastrointestinal parasites.

Sheep of two bloodlines of Merino were artificially infested with equal numbers of lice (Bovicola ovis) and the resulting louse populations were monitored over the following 20 months. The sheep were shorn 6 and 17 months after infestation and, for analysis, the louse counts considered in 3 years separated by shearings. Nematode faecal egg counts (FECs) were assessed on faecal samples collected on five occasions, three times following natural challenge and twice after artificial challenge with 40,000 trichostrongyloid larvae (84% Trichostrongylus vitrinus). In addition, blood samples were collected and measured for B. ovis-specific immunoglobulins (predominantly IgG), B. ovis-specific IgE and serum total IgE. Bloodlines differed significantly in the size of louse populations at the end of year 2, FEC after both natural and artificial challenge and in serum levels of all three antibodies (p<0.05). There were also large variations in louse counts and FEC among sheep within bloodlines. Louse counts at inspections after louse populations had been allowed to build up were highly repeatable, both between and within years. However, correlations with counts at inspections soon after initial infestation and following shearing were lower. FEC after natural challenge was correlated with louse counts in year 2 (r=0.45, p<0.01) and year 3 (r=0.38, p<0.05), but the correlation with counts in year 1 was not significant (r=0.25, p>0.05). FEC following artificial challenge was significantly correlated with louse counts in year 3 (r=0.36, p<0.05), but not in year 2 (r=0.25, p>0.05) or year 1 (r=0.04, p>0.05). Louse counts in the 3 years were significantly correlated with anti-B. ovis antibody concentration (r=0.60, 0.48, 0.36), but not with levels of either anti-B. ovis or total serum IgE. These results suggest that sheep with greater resistance to gastrointestinal parasites also tend to be less susceptible to lice. Whether this is due to interaction of the effects of the parasites or to correlation in underlying resistance mechanisms requires clarification.

Sensitivity of two-stage sampling to detect sheep biting lice (Bovicola ovis) in infested flocks.

The sampling distribution of Bovicola ovis (Schrank) on sheep was examined in two flocks, one with a light and one with a heavy infestation of lice. The derived distributions were used to calculate the sensitivity of detecting lice on individual sheep and in flocks by fleece parting regimes that varied in number of parts per animal and number of sheep per flock, different scenarios of flock sizes, proportion infested and louse density were examined. Lice were aggregated among fleece partings in the heavily infested flock and described by a negative binomial distribution with k values between 0.3 and 1.92. The distribution was indistinguishable from Poisson in the lightly infested flock. The assumed distribution had little effect on sensitivity, except when only one fleece part per animal was examined. On individual sheep where louse density was 0.5 per 10 cm part or greater, there were only marginal gains from inspecting more than 10 parts per animal. Increasing the number of sheep inspected always increased sensitivity more than increasing number of parts per sheep by an equivalent amount. This advantage was greatest in situations where a low proportion of sheep in the flock were infested with a high density of lice, and less where a low proportion of sheep were infested with a low density of lice, or a high proportion of sheep were infested with a high density of lice.

Survival away from sheep and alternative methods of transmission of sheep lice (Bovicola ovis).

Transmission of sheep lice is thought to occur mainly by sheep to sheep contact although the possibility of other sources of infestation is often suggested. This study investigated the period of survival of Bovicola ovis after removal from sheep under varying conditions and assessed the likelihood of new infestations arising from contaminated facilities, wool caught on fences and shearers' footwear. In laboratory studies with lice held away from sheep at 4, 20, 25 and 36.5 degrees C, adults and nymphs survived longest at 25 degrees C (LT90 of 11.7 and 24.1 days for adults and large nymphs, respectively). Nymphs survived longer than adults and lice provided with raw wool survived longer than lice provided with wool that had been degreased. Nymphal lice survived for up to 29 days on unscoured wool at 36.5 degrees C, but the LT50 was less than 9 days in most experiments. In shearing sheds in winter and early spring lice survived for up to 14 and 16 days, respectively. These periods of survival are considerably longer than previously indicated for B. ovis. Most lice dropped out of wool staples attached to a fence within 1 h and only two of a total of 225 lice were still present after 24 h, suggesting that sheep are unlikely to become infested from wool caught on fences. Adult and nymphal lice readily transferred to shearers' moccasins and survived there for up to 10 days, indicating that transmission of lice on the footwear of shearers or other sheep handlers may be a cause of new infestations. Microwaving each moccasin for 5 min killed all lice and may provide a simple method of reducing the likelihood of transmission of B. ovis between properties.

Inspection of wool lots at sales as a diagnostic test for louse infestation.

The accuracy of visual inspection of wool lots for lice as a test for louse infestation was estimated using information provided by 178 woolgrowers in Queensland, Australia. The estimated sensitivity of inspection was 36% (95% confidence interval, 19-58%) and the specificity was 95% (95% CI, 88-98%). Accuracy was influenced by timing, after shearing, of pesticide application for louse control and by class of pesticide last applied after shearing. Visual inspection was less sensitive (29%) if pesticides were applied >3 months after shearing and less sensitive (21%) if an insect growth regulator was the class of pesticide last used after shearing. Based on 36% sensitivity, it was estimated that 16 inspections would have to be conducted to reduce the false negative test rate to <20% in the study population. We suggest that visual inspection of wool lots could be used to efficiently monitor the prevalence of louse infestations in Queensland sheep flocks. Positive inspection results are more likely to represent real louse infestations, rather than a false test result, in flocks grazed in the more extensive regions of Queensland.

Spatial distribution and spread of sheep biting lice, Bovicola ovis, from point infestations.

The spatial distribution of chewing lice (Bovicola ovis) on their hosts was examined in Polypay and Columbia ewes initially artificially infested on the midside or the neck. Densities of lice were determined at 69 body sites in eight body regions at approximately monthly intervals for 2 years. In the second year, half of the ewes were mated and lice were counted at 26 body sites on the resulting lambs. Polypay ewes had higher densities of lice than Columbias at most inspections but there was little effect of infestation point or mating on either numbers or the distribution of lice. During periods of high louse numbers densities were generally greatest on the sides or the back. Densities on the head were also high at times and peaked later than overall louse densities. Shearing markedly reduced density but increased the proportion of lice found on the neck, belly and lowleg sites. The distribution of lice on the lambs was similar to that on the ewes except that fewer lice were found on the head. Comparisons of lice per part with the numbers of lice extracted from clipped patches indicated that a sheep with wool bearing area of 1 m2 and a mean count of one louse per 10 cm fleece parting carried approximately 2000 lice. At most times of the year inspections for sheep lice should be concentrated on the sides and back, but in recently shorn sheep greater attention should be paid to the lower neck and ventral regions. Implications of the observed distributions of lice for the efficacy of chemical treatments are discussed.

Efficacy against sheep lice (Bovicola ovis) and fleece wetting of six shower dip preparations.

The relative efficacy of 6 shower dip chemicals most frequently used for the treatment of sheep lice (Bovicola ovis) in Western Australia was examined. Groups of 20 sheep infested with lice were treated with products containing either alphamethrin, cyhalothrin, diazinon or diazinon plus piperonyl butoxide and rotenone, formulated as emulsifiable concentrates, and with products containing either coumaphos or magnesium fluorosilicate, formulated as wettable powders. All treatments were applied through a shower dip (Sunbeam model SSD). Inspections for lice were conducted until 9 months after dipping. No lice were found on sheep treated with the 4 emulsifiable concentrate products. In contrast, treatment with the wettable powders, which contained either coumaphos or magnesium fluorosilicate as the active ingredient, did not eradicate the lice infestations. The degree to which the fleece was wetted was assessed 20 minutes after dipping and showed that the wettable powder dips penetrated the fleece less than the emulsifiable concentrate dips. Less fluid was retained by wool staples in an in-vitro test when dip wash was made with the wettable powders. It was concluded that the degree of wetting attained at dipping was an important factor in achieving eradication of sheep lice.

The prevalence of lice (Bovicola ovis) infested sheep flocks in Western Australia (1987-1993)

The proportion of wool bale brands with a positive test for sheep lice in baled wool decreased from 29.5% in 1987/88 to 23.2% in 1990/91 before increasing to 38.2% in 1992/93. Changes in the proportion of wool bale brands with a positive test for lice were highly correlated with changes in the Wool Market Price Indicator. The increase in the proportion of positive lice tests since 1990/91 was associated with an increase in failures to eradicate lice from flocks. These failures were partly a consequence of the reduced use of lousicidal treatments, the development of resistance to synthetic pyrethroid chemicals and an increase in the transmission of lice between flocks.

Persistence of the sheep body louse, Bovicola ovis, after treatment.

Lice were found in samples of baled wool from 198 of 464 flocks treated to eliminate an infestation in the period July 1988 to June 1990. In 287 flocks the insecticide was applied as a backline treatment and in 177 flocks a shower dip was used. Of these flocks, 41.5% and 44.6%, respectively, were found to be infested at the following shearing. After adjusting for the accuracy of the test, it was estimated that infestation in 2 consecutive 12-month periods occurred in 52.6% of flocks with a real incidence rate of 27.4%. Using these estimates, it was calculated that in 34.7% of infested flocks treatment did not kill all lice. There were no differences in the proportion of consecutive infestations between flocks treated with any of the 3 chemicals applied as backline treatments. Among flocks treated in a shower dip, 68.4% using coumaphos, 37.8% using diazinon and 41.5% using cyhalothrin had consecutive infestations (P < 0.05). The detection of lice in 63.2% of flocks that were treated with magnesium fluosilicate was, in part, attributed to the application of this chemical by one operator.

[One of the developmental dead ends or a method of dispersal of the pasture tick Dermacentor marginatus]. / Odin iz tupikov razvitiia ili sposob rasseleniia pastbishchnogo kleshcha Dermacentor marginatus.

Accumulation of pasture ticks Dermacentor marginatus has been discovered in the wool of sheep, 5-6 cm from the skin surface, the temperature in those sites being permanent 18-20 degrees C. The ticks died either in the course or following ovipositor. It may be assumed that this was followed by the appearance of larvae from oocysts. It is suggested that the above phenomenon is one of the deadlines in Dermacentor marginatus development or one of the unusual ways of their larvae habitation on pastures.