蚤蝇的昼夜活动节律及睡眠行为研究

蚤蝇是重要的法医昆虫,同时是实验室中遗传、发育和生物测定等研究的重要对象。然而,蚤蝇的昼夜活动节律和睡眠行为及其在脑部的神经网络目前还不清晰。本文通过捕获本地蚤蝇并对其进行分子鉴定,研究了蚤蝇的昼夜活动节律和睡眠行为,同时表征了蚤蝇脑部核心钟神经元和多巴胺神经元。结果表明:蚤蝇在12h光照∶12h黑暗(12L∶12D)条件下不存在对开灯前或关灯前的活动预期,其双峰活动模式是对开关灯的光反应行为。在全黑暗(DD)条件下蚤蝇内源活动周期接近24h。黑腹果蝇神经肽PDF抗体免疫显示蚤蝇脑部核心钟神经元4~5个,不像黑腹果蝇一样存在明显的神经轴突。在睡眠行为上,蚤蝇雄虫和雌虫在整体活动强度、睡眠节律模式、总睡眠上均没有明显差异。相反,雄虫总睡眠次数和晚上睡眠次数低于雌虫,而总睡眠持续时间、晚上睡眠持续时间、总入睡时间和晚上入睡时间高于雌虫。此外,影响睡眠的重要多巴胺神经元在蚤蝇脑部的分布与黑腹果蝇类似。     

Role of Rusa deer Cervus timorensis russa in the cycle of the cattle tick Boophilus microplus in New Caledonia

Two field experiments were conducted to evaluate the efficiency of Rusa deer in the development of the cattle tick Boophilus microplus in comparison with that of steers in the same pastures and under the same conditions of infestation. No difference was noted between a mixed steer/deer herd and a pure steer or pure deer herd in the infestation pattern of each host, suggesting that attachment to the alternative host is mechanical and not affected by the simultaneous presence of the primary host on the pasture. Deer are capable of producing engorged viable females, with weight and reproductive performances similar to or even better than females fed on steers. For moderate levels (1 million larvae per hectare) and high levels (32 million larvae per hectare) of pasture infestation, tick burdens on steers were not very different (e.g. average 1911 and 2681ticks per m² skin, respectively, on day 24). This may be because of saturation of steer skin sites at the moderate larval dose. Deer harboured 2.7–33 times fewer ticks than steers and produce no engorged females at the moderate larval level and 32 times fewer engorged females than steers at the high larval level. Infestation of deer was dosedependent with averages of 12 and 399ticks per m² skin on day 25 at the moderate and high larval levels, respectively. At a high infestation level of the environment, Rusa deer may contribute, but to a limited extent, to infestation of pastures and, consequently, of cattle. However, their role in sustaining a viable tick population requires further investigation.     

Major ectoparasites of cattle in and around Mekelle, northern Ethiopia

Studies on the identification of major ectoparasites of cattle in and around Mekelle in northern Ethiopia were carried out from November 2006 to June 2007. Ectoparasite prevalence, infestation, and the possible risks associated with ectoparasites were investigated. Data collected were analyzed using the χ²‐test. A total of 849 cattle were examined: 40.2% (341), 53.2% (452) and 5.9% (50) were infested with ticks, lice and Demodex, respectively. A positive correlation was observed between prevalence and production system and breed of cattle. The results also revealed widespread occurrence of ectoparasites that have affected the general health and productivity of cattle in the study area. Seven different species of ticks were found on the cattle, with some capable of transmitting pathogens harmful to both cattle and humans.     

Sequence Variations in the Boophilus Microplus Bm86 Locus and Implications for Immunoprotection in Cattle Vaccinated with this Antigen

Cattle tick infestations constitute a major problem for the cattle industry in tropical and subtropical regions of the world. Traditional control methods have been only partially successful, hampered by the selection of chemical-resistant tick populations. The Boophilus microplus Bm86 protein was isolated from tick gut epithelial cells and shown to induce a protective response against tick infestations in vaccinated cattle. Vaccine preparations including the recombinant Bm86 are used to control cattle tick infestations in the field as an alternative measure to reduce the losses produced by this ectoparasite. The principle for the immunological control of tick infestations relies on a polyclonal antibody response against the target antigen and, therefore, should be difficult to select for tick-resistant populations. However, sequence variations in the Bm86 locus, among other factors, could affect the effectiveness of Bm86-containing vaccines. In the present study we have addressed this issue, employing data obtained with B. microplus strains from Australia, Mexico, Cuba, Argentina and Venezuela. The results showed a tendency in the inverse correlation between the efficacy of the vaccination with Bm86 and the sequence variations in the Bm86 locus (R² = 0.7). The mutation fixation index in the Bm86 locus was calculated and shown to be between 0.02 and 0.1 amino acids per year. Possible implications of these findings for the immunoprotection of cattle against tick infestations employing the Bm86 antigen are discussed.     

Efficiency and cost of strategic use of acaricide for tick control in N'Dama cattle in The Gambia

The efficiency of strategic and strategic/selective applications of flumethrin spray formulation for controlling ticks were assessed, respectively, in two groups of fourteen N'Dama cattle (Group S and Group S/S) by comparison of the number of feeding ticks with thirteen untreated N'Dama cattle (Group U) over a period of 11 months (June 1996 to April 1997). During the expected peak of tick abundance, acaricide was applied fortnightly on the whole body in animals in Group S and only on the most infested body areas in cattle in Group S/S. Weight changes and skin lesions, directly associated with tick attachment, were recorded in cattle in the three groups. The costs of the two tick control schemes were estimated. Maximum level of ticks, all species together, feeding on cattle was observed in the rainy season. Both in Group S/S and Group S, cattle carried a lower (P < 0.001) number of feeding ticks than animals in Group U over the whole study period. Percentage of tick control, over the entire period of tick investigation, was satisfactory in both acaricide-treated groups, reaching 61.2 and 75.2% in Groups S/S and S, respectively. However, the proportion of control varied according to tick species or genus. Significantly lower prevalence of skin lesions was observed on the ano-genital and udder region in cattle in Group S/S (P < 0.05) and Group S (P < 0.01) in comparison with cattle in Group U. Mean amount of acaricide solution used and relative estimated cost of treatment in cattle in Group S/S were, respectively, 25- and 14-fold lower than those in cattle in Group S. At the end of the study, animals in Groups S/S and S were, respectively, 7.2 and 15.9 kg heavier than animals in Group U. The difference was statistically significant (P < 0.02) only between Groups S and U. However, the efficiency, low cost and derived benefits of the strategic/selective acaricide application scheme indicated that it might be the most cost effective.     

Detecting Resistance to Organophosphates and Carbamates in the Cattle Tick Boophilus Microplus, with a Propoxur-based Biochemical Test

Rapid and sensitive detection of resistance to insecticides in arthropods is needed. In the cattle tick, Boophilus microplus, resistance to a variety of acaricides is widespread. The most commonly used assay for resistance, the larval packet test, takes at least two, but generally six weeks for a one-host tick like B. microplus to complete and may take up to three months to complete for three-host ticks. Here we describe a test for resistance to organophosphate acaricides that can be used on larvae and adult ticks which takes less than 24 hours. The test measures the difference in acetylcholinesterase (AChE) activity in homogenates of ticks in the presence and absence of propoxur, a carbamate acaricide. We found clear discrimination of organophosphate-susceptible and organophosphate-resistant adults with 100 M propoxur. AChE from susceptible ticks had almost no activity at this concentration of propoxur whereas AChE from resistant ticks had 67% of its potential activity. AChE from heterozygote ticks could also be distinguished from AChE from homozygous-susceptible and homozygous-resistant ticks. This is the first biochemical test for resistance to an acaricide. Rapid, sensitive tests like ours will allow resistance to organophosphates to be detected soon after it develops in the field, thus, the spread of resistance might be slowed and the useful life of acaricides extended.     

Development of enzyme linked immunosorbent assays to measure Bm86 antigen of Boophilus microplus (cattle tick) and to detect anti-Bm86 antibodies in serum samples

The immunization of cattle with the Boophilus microplus Bm86 antigen has been successful for the control of cattle tick infestations. To monitor the Bm86 production process and to measure the anti-Bm86 antibody titers in vaccinated cattle, mAb-based ELISA were developed and validated. The development of both immunological methods is essential to obtain a product with high quality and immunogenic properties and to monitor the immunological protection induced in vaccinated cattle against B. microplus.     

Integrated Control of Acaricide-Resistant Boophilus Microplus Populations on Grazing Cattle in Mexico Using Vaccination with Gavac™ and Amidine Treatments

Throughout most of the twentieth century, tick infestations on cattle have been controlled with chemical acaricides, typically administered by dipping or spraying. This approach can cause environmental and residue problems and has created a high incidence of acaricide resistance within tick populations in the field. Recently we developed a vaccine against Boophilus microplus employing a recombinant Bm86 antigen preparation (Gavac), (Heber Biotec S.A., Havana, Cuba) which has been shown to induce a protective response in vaccinated animals. Here we show for the first time under field conditions a near 100% control of B. microplus populations resistant to pyrethroids and organophosphates, by an integrated system employing vaccination with Gavac and amidine treatments. This method effectively controls tick infestations while reducing the number of chemical acaricide treatments and consequently the rise of B. microplus populations resistant to chemical acaricides.     

Ixodid Tick Infestation in Cattle and Wild Animals in Maswa and Iringa,Tanzania

Ticks and tick-borne diseases are important in human and livestock health worldwide. In November 2012, ixodid ticks were collected and identified morphologically from cattle and wild animals in the Maswa district and Iringa urban, Tanzania. Amblyomma gemma, A. lepidum, and A. variegatum were identified from Maswa cattle, and A. variegatum was the predominant species. A. marmoreum, Hyalomma impeltatum, and Rhipicephalus pulchellus were identified from Iringa cattle in addition to the above 3 Amblyomma species, and A. gemma was the most abundant species. Total 4 Amblyomma and 6 Rhipicephalus species were identified from wild animals of the 2 areas. A. lepidum was predominant in Maswa buffaloes, whereas A. gemma was predominant in Iringa buffaloes. Overall, A. variegatum in cattle was predominant in the Maswa district and A. gemma was predominant in Iringa, Tanzania.     

Seasonal prevalence of ticks and tick-transmitted haemoparasites in traditionally managed N'Dama cattle with reference to strategic tick control in the Gambia

Abstract A survey of tick spatial and seasonal distribution in traditional managed N' Dama cattle over 1 year old was carried out in The Gambia over 16 months. Presence of Anaplasma marginale and Babesia spp. in the blood smears and their antibodies in the serum of same animals were also examined. Tick species, in decreasing order of abundance, were: Boophilus geigy, Rhipicephalus senegalensis, B.decoloratus, Hyalomma truncatum, H. marginatum rufipes and Amblyomma variegatum. All tick species peaked during the rainy season. Additional B.geigy peaks occurred towards the end of the dry season. The most infested animal body areas were anogenital, udder and abdomen. A.variegatum, Hyalomma spp. and, to a lesser extent, Boophilus spp. showed strong preferences for these anatomical regions. Overall prevalence of A.marginale, B.bigemina and B.bovis haemoparasites were 3.2%, 0.9% and 0.1 %, respectively. Peaks of A.marginale and B.bigemina occurred at the end of the rainy season-beginning of the dry season; an additional A.marginale peak was observed at mid-end dry season. Overall antibody seroprevalence were 29.6% for A.marginale , 44.7% for B.bigemina and 5.2% for B.bovis; monthly fluctuations in percentage of seroreactors were limited. Endemic stability for A.marginale and B.bigemina is postulated. Implications of the outcomes are discussed in relation to tick-control strategy in The Gambia.     

A competitive enzyme linked immunosorbent assay for recombinant Bm86 produced in Pichia pastoris

A competitive enzyme linked immonosorbent assay (ELISA) was developed for quantification of recombinant Bm86 protein (rBm86), antigen of the vaccine GavacTM against the cattle tick Boophilus microplus. Monoclonal antibody (Mab) SSBm1 showed identical recognition to folded and denatured antigen and was used in the competition assay. This ELISA was sensitive enough to detect 60 ng/ml of rBm86. Within-assay coefficient of variation was 5.7 to 6.2 % and between-assay variation was 7.7 to 11.6 %. The level of expression of recombinant Bm86 in Pichia pastoris reached about 2.7 g per liter of culture after 80 hour of fermentation. © Rapid Science Ltd. 1998     

The effects of the brown ear-tick, Rhipicephalus appendiculatus, on milk production of Sanga cattle

Abstract. Lactating Sanga cows of the Mashona breed from Zimbabwe, receiving either a low or a high level of nutritional supplement, were exposed to two levels of infestation of adults of the brown ear-tick, Rhipicephalus appendiculatus in the highveld of Zimbabwe. The effect of the ticks on the milk yield was measured over an 11-week period during the rainy season from January to April 1986. A technique in which calves were weighed before and after suckling was used to estimate milk yield. There were significant treatment effects of ticks (P < 0.05) on milk production but no significant differences in liveweight gain between calves from tick-free and tick-infested dams. The loss in milk production was poorly related to the number of female ticks that engorged, being 9 g (SEM 4) per tick. A Friesian x Hereford (Bos taurus) reference group of cattle carried 50% more ticks than the Mashona cows, illustrating a difference in resistance between the breeds. Thirteen screw-worm (Chrysomya bezziana) strikes were recorded amongst the thirty-two Mashona cows compared with twenty-one amongst the ten Friesian × Hereford animals between January and the end of March.     

The effects of the brown ear-tick, Rhipicephalus appendiculatus, on milk production in dairy cattle

Abstract. The effect is reported of artificially controlled levels of infestation with adults of Rhipicephalus appendiculatus on the milk yield of twenty commercial Bos taurus dairy cattle on a high plane of nutrition and eighteen crossbed B.taurus × Sanga cattle on a lower plane of nutrition in the highveld of Zimbabwe. The results showed no significant effect on milk yield of infestations averaging twenty engorging ticks per animal per day, despite severe ear damage in some animals. They indicate that milk production of dairy cattle under commercial management is not sensitive to infestation with R.appendiculatus. The results have important implications for management of ticks in Africa, but need to be interpreted within the context of the control of tick-borne diseases.     

A comparison of Ixodes rubicundus (Acari: Ixodidae) infestations on Friesian and Bonsmara cattle in South Africa

This study was conducted to compare the relative resistance of crossbred Bos indicus X B. taurus Bonsmara and B. taurus Friesian cattle to Ixodes rubicundus (Karoo paralysis tick) infestations. During periods of peak abundance of the ticks, Friesian oxen harboured almost twice or more than twice as many ticks as either Bonsmara oxen or cows. During periods of low tick abundance tick burdens on both cattle breeds were closely similar. It is envisaged that cattle can play an important role in an integrated control strategy against the Karro paralysis tick.