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1.
A rapid method is described for preparing and staining salivary glands of Rhipicephalus appendiculatus ticks infected with Theileria parva. The technique, involving the use of a modified methyl green pyronin stained minimizes the risk of losing material and allows examination of stained glands within minutes of preparation. The technique was applied in a series of studies in which ticks were either infected with T. parva under different conditions, or maturation of parasites in adult ticks was stimulated by different means. When nymphal ticks were fed on the ears of cattle the subsequent infection rate of the adult ticks showed no correlation with the parasitaemia of the cattle at the time of nymphal engorgement. There was no difference in infection rates between adult ticks in which parasite maturation had been stimulated either by incubation at 37 degree C or by feeding on rabbits. However, parasite maturation took about 1 day longer in incubated ticks than in rabbit-fed ticks. Female ticks were consistently more highly infected than males, both in terms of the percentage of ticks infected and the mean number of infected acini/tick. Ticks were infected with T. parva by injection of nymphs with parasitaemic bovine blood, but the resultant adult infection was lower than that in ticks which had been infected naturally by feeding on cattle.  相似文献   

2.
Nymphs of the brown ear tick, Rhipicephalus appendiculatus, were fed on heparinised bovine blood infected with Theileria parva parasites in an in vitro feeding system consisting of rabbit skin membranes. The main feeding and development parameters such as the mean attachment rate, feeding duration and engorgement weights of membrane-fed ticks were not significantly different from nymphs fed on cattle. The moulting rate was also comparable although a slight significant difference was observed. Assessment of infection prevalence and abundance with T. parva in adults indicated that the membrane-fed ticks acquired infection to the same level as those fed on cattle. Stabilates prepared from both the membrane- and cattle-fed adult ticks were found to be infective and caused severe reactions in susceptible cattle. When the immunised cattle were challenged with a lethal homologous dose of T. parva (Marikebuni), they were found to be immune.  相似文献   

3.
Theileria parva is an intracellular protozoan parasite transmitted by ticks that causes a fatal lymphoproliferative disease of cattle known as East Coast Fever. Vaccination against the disease currently relies on inoculation of the infective sporozoite stage of the parasite and simultaneous treatment with long-acting formulations of oxytetracycline. Sporozoites are maintained as frozen stabilates of triturated infected ticks and the method requires accurate titration of stabilates to determine appropriate dose rates. Titration has traditionally been undertaken in cattle and requires large numbers of animals because of individual variation in susceptibility to infection. An alternative tissue culture-based method is laborious and time consuming. We have developed a flow cytometric method for quantifying the infectivity of sporozoite stabilates in vitro based on the detection of intracellular parasite antigen. The method allows clear identification of parasitized cells with a high degree of sensitivity and specificity. Analysis of infected cells between 48 and 72 h post-infection clearly defines the potential transforming capability of different stabilates.  相似文献   

4.
The variation in Theileria parva infection rates of experimental batches of adult Rhipicephalus appendiculatus ticks, used during the course of several years, was examined. It was found that considerable variation occurred, but that this could not always be correlated with the piroplasm parasitaemia in the cattle on which the ticks engorged as nymphs. Statistical analysis showed that the infection rate of ticks fed on cattle with a parasitaemia of 41–50 per cent was significantly higher than that of ticks fed on cattle with lower parasitaemias. A number of experiments were then carried out in which one or several factors of this aspect of the host-parasite relationship remained constant whilst others were altered. None of these factors was seen to play a major part in the variation. Finally, randomly selected groups of 10 ticks which had dropped engorged as nymphs from the same animal on the same day were examined. The variation observed even in these groups was so great that it was concluded that the infection rate could depend on a factor such as the juxtaposition of possibly-infected gut epithelial cells and developing salivary glands during the nymphal moult.  相似文献   

5.
The effect of Anaplasma marginale antibodies ingested with the tick blood meal was tested on infected male ticks that were allowed to feed on cattle immunized with the erythrocytic stage of A. marginale. The experiments were done in two trials. Trial 1 was done using splenectomized calves (two calves per treated and control groups) while ticks in trial 2 were fed on intact yearling cattle (four cattle per treated and control groups). The cattle were immunized with purified outer membrane proteins of erythrocyte-derived A. marginale using saponin (trial 1) or monophosphoryl lipid-A-trehalose dicorynomycolate adjuvant (trial 2). The corresponding control cattle received adjuvant only. All cattle were challenged using Dermacentor andersoni males infected as adults that were allowed to feed for 7 days. In trial 1, the ticks were allowed to feed a second time on susceptible calves to test whether exposure of ticks to immunized cattle affected their ability to transmit anaplasmosis. Infections in fed ticks were monitored by determining the infection rates in salivary glands with an A. marginale-specific RNA probe and light microscopy. Vaccine-derived antibodies ingested with the tick blood meal did not appear to affect the development of A. marginale in previously infected ticks. The infection rates in the salivary glands were not significantly different among ticks fed on immunized versus adjuvant control cattle. When the vaccine-exposed ticks in trial 1 were allowed to feed a second time on susceptible calves, the resulting clinical symptoms of anaplasmosis were similar to those of the controls. There was no statistically significant effect of tick exposure to the anti-erythrocytic stage antibody on the development of salivary gland infection or transmission of A. marginale by ticks.  相似文献   

6.
Ticks are important disease vectors that can cause considerable economic losses by affecting animal health and productivity, especially in tropical and subtropical regions. In this study, we investigated the prevalence and diversity of bacterial and protozoan tick-borne pathogens in ticks collected from the vegetation and cattle in Nigeria by PCR. The infection rates of questing ticks were 3.1% for Rickettsia species, 0.1% for Coxiella burnetii and 0.4% for Borrelia species. Other pathogens, such as Babesia, Theileria, Anaplasma, and Ehrlichia species, were not detected in ticks from the vegetation. Feeding ticks collected from cattle displayed infection rates of 12.5% for Rickettsia species, 14% for Coxiella burnetii, 5.9% for Anaplasma species, 5.1% for Ehrlichia species, and 2.9% for Theileria mutans. Babesia and Borrelia species were not detected in ticks collected from cattle. Mixed infections were found only in feeding ticks and mainly Rickettsia species and Coxiella burnetii were involved. The diversity of tick-borne pathogens in Nigeria was higher in feeding than in questing ticks, suggesting that cattle serve as reservoirs for at least some of the pathogens studied, in particular C. burnetii. The total estimated herd infection rates of 20.6% for a Rickettsia africae-like species, 27% for Coxiella burnetii, and 8.5% for Anaplasma marginale/centrale suggest that these pathogens may have considerable implications for human and animal health.  相似文献   

7.
Three experiments were carried out in which Theileria parva was irradiated in its tick vector, Rhipicephalus appendiculatus. In the first experiment, infected unfed adult ticks were irradiated at doubling doses from 4 to 32 krad. Some of the ticks were then fed for 4, 5, 6, 7 and 8 days on rabbits, and the parasites in their salivary glands examined. Five male and 5 female ticks from each irradiation dose were put onto each of a pair of susceptible cattle, whose reactions were recorded. Increasing doses of irradiation resulted in progressive destruction of the parasites. All cattle receiving ticks irradiated at doses up to and including 16 krad died of East Coast fever (ECF), and one of the cattle receiving ticks irradiated at 32 krad died.In the second experiment, recently engorged nymphs were irradiated at 1, 2 or 4 krad, and moulting nymphs at doses of 2, 4, 8, 12, 16 or 32 krad. The salivary glands of the resultant adult ticks were examined after the ticks had fed for 4, 5, 6, 7 or 8 days on rabbits. Engorged nymphs irradiated at 4 krad failed to moult, whilst moulting nymphs irradiated at 32 krad moulted but failed to attach to rabbits. Doses of irradiation survived by the ticks had no apparent morphological effect on the parasites they contained.In the third experiment, infected unfed adult ticks were irradiated at 0, 5, 10, 15, 20 25 30, 35, 40, 45, 50 or 60 krad. The ticks were fed on rabbits for 5, 6 or 7 days. Some of them were then examined morphologically, whilst others were ground in MEM/BPA and aliquots of the supernatant used to inoculate groups of 5 cattle. The reaction of these cattle, together with the morphological examination of the parasites, suggested that increasing doses of irradiation destroyed increasing numbers of parasites.  相似文献   

8.
We report on the detection of Theileria annulata in infected Hyalomma ticks by the PCR using primers derived from the gene encoding the 30 kDa major merozoite surface antigen (Tams1–1). No inhibition of the PCR was observed and as little as 0.1 pg of parasite DNA, corresponding to 12 sporozoites, could be detected in non-infected tick DNA samples, spiked with T. annulata genomic DNA. Hyalomma dromedarii ticks, fed on a calf experimentally infected with T. annulata, were used to validate the PCR further. The infection rate in the adult ticks, fed as nymphs during the febrile reaction, was high (62%), dropped to zero for 1 day in tick batches that engorged after treatment with ButalexTM and increased to 30% 2 days later and 38% of the ticks acquired the infection after feeding as nymphs during a carrier state piroplasm parasitaemia of less than 0.1%. As an internal control, 16S tick rDNA sequences could be amplified from T. annulata-negative tick samples. Finally, 202 adult ticks from Mauritania, collected from zebu cattle carrying low levels of Theileria piroplasms, were tested by the PCR. Thirty-eight out of 52 (73%) and 17 out of 30 (57%) H. dromedarii from the Gorgol and Trarza regions, respectively and two out of 30 (7%) Hyalomma marginatum rufipes from the Gorgol region were positive. Hyalomma marginatum rufipes, Rhipicephalus evertsi evertsi and Rhipicephalus guilhoni from the Trarza region were negative. These findings confirm that H. dromedarii is the main vector of T. annulata in Mauritania and that the PCR is a useful method of determining the infection rates in ticks collected from cattle carrying low levels of T. annulata piroplasms.  相似文献   

9.
Watt, D. M., Walker, A. R., Lamza, K. A., and Ambrose, N. C. 2001. Tick-Theileria interactions in response to immune activation of the vector. Experimental Parasitology 97, 89-94. Immune mechanisms towards the haemoprotozoan parasite Theileria parva were investigated in their tick vector, Rhipicephalus appendiculatus. The exoskeletons of adult ticks were initially pierced with bacteria-coated, saline-coated, or sterile dry glass needles. Haemolymph was extracted from the ticks at 6, 24, 48, and 72 h postinjection and applied to bacterial plates to measure the growth inhibition effects. The inhibition zones were larger with all the injected groups compared to uninjected controls. The largest inhibition zones were seen 24 h after injection with bacteria-coated needles. An experiment was carried out to investigate whether antibacterial immune responses were relevant to the parasite/tick relationship and, if so, which parasite form was most vulnerable. R. appendiculatus nymphs were infected with T. parva by feeding on an infected calf and were then injected with needles on days 7, 13, 15, and 17 throughout their moult in an attempt to induce tick immune responses at the same time as different lifecycle forms of T. parva would be present. Salivary glands from the moulted adult ticks in the control and different treatment groups were dissected out and examined for the presence of T. parva sporoblasts. No difference in infection levels was seen in any of the treatment groups compared with the controls, suggesting that immune responses in R. appendiculatus, induced by bacterial injection, do not affect T. parva infections. The fecundity of injected ticks was compared with that of uninjected controls to ensure that the injection procedure itself was not detrimental to the ticks. Injected females had higher engorgement masses than controls but reduced levels of egg hatching.  相似文献   

10.
A monoclonal antibody specific for the Theileria parva sporozoite, which recognizes a determinant on the surface coat and blocks sporozoite infectivity, was used to investigate the presence of the determinant on other stages of the parasite lifecycle. Immunofluorescence techniques did not demonstrate this determinant on the kinete, schizont, merozoite, or piroplasm stages of the parasite. Immunoautoradiography, using a tritiated form of the monoclonal antibody, on sections of infected salivary glands collected from ticks that had fed for 0, 1, 2, 3, or 4 days revealed that the determinant recognized was synthesized predominantly during sporogony, between 2 to 3 days after the tick started feeding. Immunoelectron microscopy was performed on ultrathin frozen sections of infected tick salivary glands incubated with the monoclonal antibody followed by Protein-A--colloidal gold. The antigen or its precursor could be detected in the developing parasite. In ticks fed 2 days, the sporoblast was labeled, both in the cytoplasm and on parasite membranes, often including the nuclear envelope. In sections from ticks fed 4 days, the sporozoite surface membrane was labeled, as were membrane-bounded sporozoite organelles identified as micronemes. Observation by immunofluorescence, on sporozoites incubated with bovine peripheral blood lymphocytes, suggested that the antigen recognized by the monoclonal antibody does not enter the lymphocyte during sporozoite endocytosis. We conclude that synthesis of the antigen or its precursor(s) occurs during sporogony in the feeding tick, at the time of maximal parasite proliferation, and precedes the formation of morphologically mature sporozoites; the antigen's role in the parasite life cycle also appears to be limited to events associated with the sporozoite entry process.  相似文献   

11.
Cryopreservation of infective particles of Theileria parva   总被引:7,自引:0,他引:7  
Cryopreservation of infective particles of Theileria parva. International Journal for Parasitology3: 583–587. Infective particles of Theileria parva, the causative organism of East Coast fever of cattle, were obtained from infected Rhipicephalus appendiculatus ticks, either by using an in vitro feeding technique or by grinding the ticks in a suitable medium. If foetal calf serum containing 15% glycerol (v/v) was added to the infective material and it was then distributed either to glass capillary tubes (in vitro tick feed) or glass tubes (ground tick supernate) it could be slowly frozen to either ?80°C or ?196°C without loss of viability. Stabilates, tested by rapid thawing and inoculation into ECF-susceptible cattle, remained viable for up to a year at these temperatures.  相似文献   

12.
We describe the generation of an auto-annotated index of genes that are expressed in the salivary glands of four-day fed female adult Rhipicephalus appendiculatus ticks. A total of 9162 EST sequences were derived from an uninfected tick cDNA library and 9844 ESTs were from a cDNA library from ticks infected with Theileria parva, which develop in type III salivary gland acini. There were no major differences between abundantly expressed ESTs from the two cDNA libraries, although there was evidence for an up-regulation in the expression of some glycine-rich proteins in infected salivary glands. Gene ontology terms were also assigned to sequences in the index and those with potential enzyme function were linked to the Kyoto encyclopedia of genes and genomes database, allowing reconstruction of metabolic pathways. Several genes code for previously characterized tick proteins such as receptors for myokinin or ecdysteroid and an immunosuppressive protein. cDNAs coding for homologs of heme-lipoproteins which are major components of tick hemolymph were identified by searching the database with published N-terminal peptide sequence data derived from biochemically purified Boophilus microplus proteins. The EST data will be a useful resource for construction of microarrays to probe vector biology, vector-host and vector-pathogen interactions and to underpin gene identification via proteomics approaches.  相似文献   

13.
One hundred fifty Amblyomma americanum were examined between March and September 1986 from Cookson Hills Wildlife Refuge in eastern Oklahoma (USA). Of these ticks, 11% (17 of 150) were infected with Theileria cervi. Field-collected nymphal ticks had an 8% (3 of 37) prevalence of infection averaging 1.0 infected acini/nymph. Female ticks had a 16% prevalence of infection averaging 1.6 infected acini/female; T. cervi was not observed in salivary glands of field-collected male ticks. When laboratory reared A. americanum nymphs were allowed to feed on experimental white-tailed deer (Odocoileus virginianus) with varying T. cervi parasitemias (less than 1, 2, 6 and greater than 20%), only ticks which fed on deer with parasitemias greater than 1% became infected. Although prevalence and intensity of infection varied in the infected ticks, there was no significant difference in prevalence of infection between males and females. However, females did acquire significantly greater intensities than males. The data from these studies confirm that T. cervi overwinters in A. americanum and suggests that the prevalence, intensity and abundance of infection of T. cervi in ticks is influenced by the parasitemia of the deer host. Furthermore, fawns may play a more important role in the epidemiology of T. cervi transmission than do adult deer because of the coordination between tick activity patterns and deer fawning.  相似文献   

14.
Questing ticks from various districts and agro-ecological zones (AEZ) in the Tanga Region of Tanzania were studied for a two-year period between September 1999 and July 2001. Collections of both nymphal and adult ticks occurred at 29 sites using a blanket or white cloth dragging technique. The species recorded in the order of decreasing abundance were Rhipicephalus appendiculatus, Boophilus spp., and Ambylomma variegatum. Rhipicephalus appendiculatus field infestation levels varied across administrative districts and seasons, with Amani and the cool-to-dry season having lower tick counts ( = -2.09, SE = 0.71, P < 0.001 and = -1.54, SE = 0.56, P = 0.006 for Amani and cool to dry season, respectively). Based on the polymerase chain reaction technique, Theileria parva infection prevalence in adult R. appendiculatus was estimated to be 2.6%. Implications of these findings in light of the spatial and temporal distribution of ticks are discussed for the accurate diagnosis of multiple tick-borne diseases, the infected vector tick, the endemic status of T. parva in the region, and the implemention of control strategies.  相似文献   

15.
Theileria annulata, a protozoan parasite of cattle and domestic buffaloes, is transmitted by ticks of the genus Hyalomma, and causes a disease named Mediterranean or tropical theileriosis. In this research 50 cattle naturally infected with Theileria annulata were treated with the extract of the plant Peganum harmala. The treatment was continued for 5 days, the dose of the extract being 5 mg/kg per day. After the treatment, 39 cattle responded to the treatment and recovered, but 11 did not respond to the treatment and died. The recovery rate of animals treated with the extract of the plant Peganum harmala was 78%.  相似文献   

16.
We examined the relationship between body size and the phenology of the tick complex Rhipicephalus appendiculatus/Rhipicephalus zambeziensis. These ticks transmit Theileria parva in cattle. In Africa, the body size of R. appendiculatus increases with latitude while the body size of the morphologically similar Rhipicephalus zambeziensis is constant at two different latitudes. A larger body size is necessary once survival becomes a constraint. The most plausible explanation for the smaller R. appendiculatus in equatorial Africa is the cost to produce a larger egg. The consequences of these findings for the introduction of R. appendiculatus in new environments are discussed. New field observations from southern Zambia indicate that R. appendiculatus body size does not vary seasonally as compared to eastern Zambia. This is an additional indication of the presence of a single diapausing population of larger ticks.  相似文献   

17.
Sera and blood from cattle and sheep were examined for the presence of Babesia and Theileria spp by microscopy and serology at the Parasitology Department of the Istituto Zooprofilattico Sperimentale of Abruzzo and Molise (IZSAM). Of the 47 bovine herds (323 animals) tested, 15 were found positive for Babesia bigemina and 1 for Babesia bovis. Two outbreaks occurred, one caused by B. bigemina and one by B. bovis. The B. bigemina outbreak occurred in Abruzzo and has been followed for two years. The isolate of B. bigemina was very pathogenic leading to the death of two cows out of 57. The vector responsible of the transmission appeared to be Rhipicephalus bursa. Parasites were observed in the erythrocytes for 30 days whereas sera were positive to indirect immunofluorescence (IFA) for at least one year. The B. bovis outbreak occurred in the province of Mantova (Northern Italy) in a group of 70 beef cattle imported from France. The infection resulted in the death of 5 animals and severe illness in another 6. In contrast with what occurred for Babesia infection, no clinical cases were recorded in cattle when species of Theileria were detected by microscopy. Of the 24 bovine herds (252 animals) tested for Theileria, 21 were found positive for the T. "sergenti"/buffeli/orientalis group. Single and mixed infection of T. "sergenti" and T. buffeli/orientalis were detected in herds of cross-bred cattle from Abruzzo and Marche. The parasites were identified by using a polymerase chain reaction which amplified DNA encoding p32/34. Most of the collected ticks (90%) were adults of R. bursa whereas the others were adults of Hyalomma detritum. During the period the animals have been observed (18 months), no clinical cases have been recorded and no associations have been found between blood abnormalities and animals found infected with Theileria. Prevalences of subclinically infected carriers increased from February till December (95.4%) even if the animals were indoors and no ticks were present. The prevalence then dropped dramatically six months later (76.7%). In calves less than 1 year old, the prevalence of infection significantly (p<0.05) increased with age, however intraerythrocytic stages of Theileria were found in the blood of three newborn calves (<7 days of age). Of the 18 ovine flocks tested for Babesia spp. (150 animals examined), 1 was positive for B. ovis and 2 for B. motasi. B. motasi infection was not associated with symptoms, while an outbreak of babesiosis caused by B. ovis occurred in Abruzzo. The infection resulted in the death of 3 animals (0.75% of the flock), two rams (20% of the total number) and a ewe, and severe illness in another 5 ewes (2% of the flock). Specimens of R. bursa and R. turanicus were collected from the infected animals. Of the 18 flocks (150 animals) examined, 12 were microscopically positive for Theileria spp. No clinical cases were recorded and identification at species level was not possible on the basis of morphological criteria. The prevalence distribution of infected herds and infected animals within herds and flocks have been calculated by a Monte Carlo simulation model, running 10,000 iterations. The most likely levels of prevalence of infected herds and infected animals within herds found for the species observed were as follows: 20% for B. bigemina with a prevalence within herd of 27%, 11% for B. bovis (18% within herd), 10% for Babesia ovis (19% within herd), 10% for B. motasi (17.5% within herd), 63% for Theileria in cattle (66% within herd) and 51% for Theileria in sheep (55% within herd).  相似文献   

18.
SYNOPSIS. Infective particles (IPs) of Theileria parva , the causative organism of East Coast Fever of cattle, were harvested from the tick vector, Rhipicephalus appendiculatus , using an in vitro feeding technic. In a ranging experiment, pairs of cattle were inoculated with aliquots of suspensions of IPs irradiated at doses of 4–137.6 krad. Doses of irradiation in excess of 8 krad appeared to destroy the parasite. In the 2nd and 3rd experiments, groups of 5 cattle were inoculated with aliquots of suspensions containing low and high concentrations of IPs respectively, irradiated at doses of 4–32 krad. In the 2nd experiment, doses of irradiation in excess of 10 krad appeared to destroy the parasite. In the 3rd experiment, at least 1 animal became infected when inoculated with an aliquot of a suspension irradiated at 16 krad. In all experiments, it appeared that increasing doses of irradiation destroyed increasing numbers of IPs. There was no conclusive evidence that IPs which survived irradiation were attenuated, and it appears that vaccination of cattle against ECF is unlikely to be achieved by inoculation of irradiated IPs using the methods described.  相似文献   

19.
Abstract. Groups of cattle of four different cross-breeds (Ankole × Friesian, Ankole × Brown-Swiss, Ankole × Guernsey, Ankole × Sahiwal) plus a group of pure Ankole cattle were immunized against tick-borne diseases: East Coast Fever, using a Theileria parva trivalent vaccine, and anaplasmosis, babesiosis and, subsequently, heartwater. With the exception of two small subgroups of Ankole and Ankole × Sahiwal which were treated by weekly spraying using Delnav, all the groups were exposed to natural field challenge of ticks to quantify the development of host resistance to ticks. The results indicate resistance to the most abundant species, Rhipicephalus appendiculatus , as follows: Ankole > Ankole × Sahiwal > Ankole × Brown Swiss > Ankole × Friesian > Ankole × Guernsey. In both Amblyomma variegatum and Rhipicephalus evertsi evertsi , the manifestation of different levels of resistance between breeds is inapparent. Correlations between total ticks and standard female ticks were good for R. appendiculatus (r = 0.73) and R. evertsi (r - 0.51) but poor for A. variegatum (r = 0.31). Correlations between species were reasonably consistent (r = 0.43-0.59). There was no significant correlation between tick burdens and daily liveweight gain (DLWG) over the. whole period of the study. Of the cross-bred groups, Ankole × Sahiwal, which had the highest level of resistance, had the highest DLWG. In contrast, the Ankole × Guernsey which developed the lowest level of resistance had only marginally lower DLWG, the difference being only 8 g/day. During the first period of exposure to ticks, however, the untreated Ankole × Sahiwal group had markedly lower DLWG, which may indicate that the development and manifestation of resistance is at the expense of productivity.  相似文献   

20.
M Samish  E Pipano 《Parasitology》1978,77(3):375-379
The time-course for the development of infectivity was studied in Hyalomma detritum ticks fed as pre-imagoes on calves infected with Theileria annulata. Unfed adults derived from infected nymphs were non-infectious when inoculated into susceptible calves, whereas ticks of both sexes that had fed for 2-3 days or longer on calves or rabbits were always infectious. Some adults fed for only 1-2 days were infectious, whereas others were not. As few as 2 ticks were capable of infecting susceptible calves.  相似文献   

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