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1.
The development of Anaplasma marginale was studied in Dermacentor andersoni nymphs after they had fed on a calf with ascending Anaplasma infection. Gut tissues were collected on day 4 of tick feeding, from newly replete (fed) nymphs and on postfeeding days (PFD) 5, 10, 15, 20, and were processed for light and electron microscopy to determine density of A marginale colonies. Homogenates of gut tissues were prepared from nymphs collected on the same days and inoculated into susceptible, splenectomized calves to test for infectivity. Anaplasma colonies were detected in gut cells on PFD 5, 10, 15, and 20. Although colony density appeared to be higher on PFD 10 and 15, differences were not significant. Nymphal type-1 colonies were detected in highest numbers on PFD 5 and 10, transitional colonies were seen in highest numbers at PFD 10 and 15, and nymphal type-2 colonies were observed only on PFD 20. Gut homogenates that were collected from ticks at 4 days of feeding, when newly replete, and on PFD 20 caused anaplasmosis when injected into susceptible calves, but homogenates made from ticks collected on PFD 5, 10, and 15 were not infective. The data indicate that of the colony types of A marginale that develop in replete nymphs, nymphal type-1 and transitional colonies may contain organisms that are not infective for cattle.  相似文献   

2.
Colonies of Anaplasma marginale in midgut epithelial cells of experimentally infected Dermacentor andersoni were studied in adult ticks 1, 3, and 6 months old. Longevity of the parasite in ticks was assessed by evaluating its infectivity for splenectomized calves; calves were exposed by feeding ticks and by inoculation of tick gut homogenates. Longevity was also evaluated by determining size, type, and density of colonies in male and female ticks. The effect of incubation (2.5 days at 37 C) on colony density was also examined for ticks at each age period. All methods used to assess longevity of A marginale in ticks (tick transmission, calf inoculation, and histologic studies) indicated a decrease of the numbers of organisms in 6-month-old ticks. Furthermore, when tick gut homogenates from 6-month-old nonincubated ticks were not infectious for susceptible calves, incubation of ticks before dissection restored infectivity of homogenates. Colonies of A marginale were detected in gut tissues of 6-month-old ticks that were not infective; therefore, infectivity of ticks could not be confirmed merely by presence of A marginale colonies.  相似文献   

3.
Anaplasma marginale is a tick-borne hemoparasite of cattle worldwide. The Virginia isolate of A. marginale was propagated previously in a cell line derived from embryos of the tick, Ixodes scapularis. The cultured Anaplasma (VA-tc) was passaged continuously for over 4 years and retained its infectivity for cattle and antigenic stability. We report herein the continuous in vitro cultivation of a second isolate of A. marginale derived from a naturally infected cow in Oklahoma (OK-tc). Blood from the infected cow was subinoculated into a splenectomized calf and blood collected at peak parasitemia was frozen, thawed and used as inoculum on confluent tick cell monolayers. Colonies of Anaplasma were apparent in low numbers at 9 days post exposure (PE) and infection in monolayers reached 100% by 4-5 weeks PE. Cultures were passaged by placing supernatant onto fresh tick cell monolayers at a dilution of 1:5 or 1:10. By the third passage development of the OK-tc was similar to that of the VA-tc and a 1:5 dilution resulted in 100% infection in 10-12 days. Inoculation of OK-tc into a splenectomized calf caused clinical anaplasmosis and Dermacentor ticks that fed on this calf transmitted the organism to a second susceptible calf. Major surface proteins (MSPs) 1-5 of the OK-tc were compared with homologous proteins present on VA-tc and the erythrocytic stage of the Oklahoma isolate. The MSPs 1, 2, 4, 5 were conserved on the OK-tc but there was evidence for structural variation in MSP3 between the cultured and erythrocytic stage of Anaplasma. MSP2 and MSP3 were the major proteins recognized by serum from infected cattle. Two-dimensional gels also identified positional differences between VA-tc and OK-tc in MSP2 and MSP3. The OK-tc may have potential to be used as antigen for development of an improved vaccine for anaplasmosis in the South Central United States.  相似文献   

4.
Infectivity and antigenicity of Anaplasma marginale from tick cell culture   总被引:1,自引:0,他引:1  
The infectivity and immunogenicity of Anaplasma marginale grown in a tick cell culture from embryonic Dermacentor variabilis ticks were assessed in splenectomized and intact calves, respectively. Culture 1 consisted of the cell line inoculated with midguts of adult ticks infected with the Mississippi isolate of A marginale and dissected 5 to 10 days after repletion and detachment from an experimentally infected calf. Cultures 2 and 3 consisted of the cell line inoculated with midguts of ticks infected with the Virginia isolate of the organism. Inoculum for culture 2 was derived from nymphal ticks dissected 5 to 10 days after repletion and detachment from the infected calf; inoculum for culture 3 was midguts from adult ticks that were fed as nymphs, allowed to molt in the laboratory and dissected 21 to 24 days after molting. In trial 1, cultures 1, 2, and 3 were maintained at pH 6.9 and incubated at 28 C; in trial 2, cultures 1 and 3 were maintained at pH 7.4 and incubated at either 28 C or 37 C. Cultures 1, 2, and 3 failed to induce infection when injected IV and SC into 6 calves in 2 separate trials. Pre-challenge sera from these calves reacted with 2 purified Anaplasma antigens in the ELISA, but failed to react in the complement-fixation test. Results of a trial to use cultures 1 and 3 in combination with an oil-in-water adjuvant to immunize intact calves against A marginale were inconclusive. However, pre-challenge sera from immunized calves reacted with the 2 purified Anaplasma initial body antigens in the ELISA but failed to react in the complement-fixation text.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

5.
Rhipicephalus evertsi evertsi larvae were fed on the ears of rabbits. Seven days after larval infestation, unfed, newly moulted nymphae were manually removed to infest a splenectomized donkey showing a patent Babesia caballi infection. Engorged nymphae were collected from the donkey and the ensuing adult ticks were placed on a susceptible horse. The horse contracted a B. caballi infection showing a prepatent period of 19 days after tick infestation. A very low parasitaemia, (highest score 2), which was patent for only 10 days, was recorded. The lowest packed cell volume recorded was 16%.  相似文献   

6.
Isolate of Anaplasma marginale not transmitted by ticks   总被引:3,自引:0,他引:3  
The tick-borne transmissibility of 2 isolates of Anaplasma marginale was compared. Dermacentor variabilis were exposed to A marginale as nymphs by feeding on 1 of 4 splenectomized calves during periods of ascending parasitemia (maximum 49% to 81% parasitized erythrocytes) induced by injection of a stabilate. Tick-borne transmission was attempted, using 26 to 224 adult ticks within 30 to 220 days after molting. Adult D variabilis did not transmit an Illinois isolate of A marginale in 7 tick-borne transmission experiments (P = 0.0047), including 2 experiments in which calves were inoculated IV with homogenates of adult ticks. In contrast, a Virginia isolate of A marginale was readily transmitted by the same tick colony. Thus, previously reported morphologic and immunologic differences among A marginale isolates may extend to tick-borne transmissibility. The Virginia and Illinois A marginale isolates had an inclusion appendage that was not a marker for tick transmissibility.  相似文献   

7.
Hyalomma anatolicum anatolicum engorged nymphs and flat adults were collected from two areas in northern Sudan. Various developing stages of Trypanosoma theileri-like flagellates were observed in the engorged nymphs, freshly moulted adults and mature adults partially engorged on rabbits. When these ticks were applied to two calves, one calf became infected with the trypanosome. The parasites were observed for one day in the enlarged lymph node nearest to the tick-feeding site 5 days after the tick application. Subsequently the trypanosomes were re-isolated in vitro from the infected calf. Inoculation of a ground-up tick supernatant suspension from the infected batch of ticks containing 10(4) trypanosomes into a calf did not produce a patent infection.  相似文献   

8.
Since the 1983 summer outbreak of anaplasmosis in southern Saskatchewan, the role of the tick, Dermacentor andersoni as an overwintering reservoir for Anaplasma marginale has been questioned. The purpose of this study was to determine if spring-collected ticks carried virulent A. marginale. Sixteen splenectomized calves were assigned randomly to two groups of 14 principals and two controls. Adult D. andersoni, collected in April from areas having high transmission rates of A. marginale, were confined to the ears of the principals by special bags and allowed to feed for eight days. The two control calves were subsequently challenged intravenously with blood from a calf infected with the Virginia strain of A. marginale. Principals and controls were monitored for 60 and 50 days postexposure respectively for signs of infection by clinical, hematological and serological procedures. None of the principals developed anaplasmosis but both control calves developed signs of disease.  相似文献   

9.
1986年从病牛身上摘下饱血雌蜱,经实验室饲养,孵化为第二代成蜱,人工越冬。于1987年将上述蜱100只接种于已去脾23天的一岁龄健康水牛,以后每天从该牛身上取蜱查其唾液腺。结果在吸血期间,始终查到梨形虫体,其平均大小为1.3×0.68微米。接种后第4天,牛血中开始出现巴贝斯虫,至第19天,红细胞感染率高达9.6%,单梨形虫体大小为0.84~2.64×0.42~1.22微米,双梨形虫体大小为0.64~1.64×0.37~0.97微米,以后逐渐下阵。该牛出现体温升高)41.6℃)、严重贫血(血红蛋白量2克,红细胞压积14%,红细胞总数177万/毫米~3)、黄疸及精神萎顿等症状。上述结果表明,该牛感染了巴贝斯虫病,由此证实镰形扇头蜱是经卵传递巴贝斯虫,其感染阶段是成蜱。  相似文献   

10.
Two Bos taurus calves were made resistant to tick infestation by exposing them to approximately 500 rabbit-reared nymphs of Hyalomma anatolicum anatolicum twice at a 2-week interval. These two calves, together with a tick-susceptible control calf, were inoculated with a stabilate of Theileria annulata (Ankara). Patent infection resulted in all three calves. Seven-hundred and fifty gerbil-reared nymphs were then applied on each of these calves as well as another tick-susceptible calf that was Theileria free. This infestation was carried out on Day 8 post-inoculation. Ticks that dropped on Day 13 post-inoculation were examined to note the development of T. annulata in them and the histological changes that occurred in the gut and salivary glands. During the second phase of feeding, the gut epithelia of the ticks from the tick-resistant calves were less active. There were no notable differences in the characteristics of the developmental stages of T. annulata between the ticks from the tick-resistant calves and those from the susceptible calf. However, ticks from one calf that acquired a higher level of tick resistance were significantly less susceptible to infection by T. annulata. Bovine tick resistance therefore compromises the vector capacity of H. a. anatolicum and this may be of epidemiological significance in the endemic areas of tropical theileriosis.  相似文献   

11.
Laboratory-reared Dermacentor andersoni ticks experimentally infected as nymphs with Anaplasma marginale were allowed to feed as adults from 1 to 9 days on susceptible, splenectomized calves to determine when, during feeding, the hematozoan was transmitted from ticks to cattle. In experiment 1, ticks were allowed to feed on calves for 1, 2, 3, 4, 5, or 6 days and anaplasmosis did not result. The same calves were used for experiment 2, and ticks were allowed to feed for 1, 3, 6, 7, 8, or 9 days and anaplasmosis occurred in all calves on which ticks fed for greater than or equal to 6 days. In 2 trials in experiment 3, ticks were allowed to feed on calves for 1 to 9 days. Anaplasmosis developed only in calves on which ticks fed for 7, 8, or 9 days. The prepatent periods shortened with longer tick feeding, and linear regression analysis of combined prepatent periods of both trials of experiment 3 indicated a significant (P = 0.05) slope with an estimated daily decrease of 7.75 days from day 7 to 9 of feeding. There was no apparent correlation between length of tick feeding and severity of clinical signs in those calves that developed anaplasmosis. Seemingly, A marginale can be transmitted to cattle by adult D andersoni ticks no earlier than the 6th or 7th day of feeding.  相似文献   

12.
Studies on the pathogenesis of epizootic bovine abortion   总被引:2,自引:0,他引:2  
Ornithodoros coriaceus Koch ticks were fed on 37 pregnant cows. The fetuses were obtained from the cows at 23 to 126 days after maternal tick exposure. Characteristic lesions of epizootic bovine abortion were observed only in those fetuses recovered 100 days more or after maternal tick exposure. Fetuses collected between 50 and 100 days showed mild-to-moderate lymphoid and mononuclear cell hyperplasia. Reaction in fetuses studied less than 50 days after maternal tick exposure were mild. Lesions could not be seen in 2 of the youngest fetuses. Increases in serum immunoglobulin concentrations were present only in those fetuses examined 80 days or more after their dams had been exposed to ticks. The specificity of the immunoglobulins could not be determined. Sera from 12 fetuses tested failed to fix complement in tests for group-specific chlamydial antibodies. A wide variety of microbiological cultivation attempts were made to recover the causative agent of epizootic bovine abortion from these fetuses; however, no agent was recovered regularly, and chlamydial organisms were not recovered from any. The significance of 2 recovered agents, apparently viral, is still to be determined. Fetal tissues, both frozen and fresh, collected from fetuses of dams exposed to a feeding of ticks were capable of reproducing the disease after inoculation into pregnant cows or directly into fetuses.  相似文献   

13.
Several conclusions of importance to studies on the epidemiology of heartwater were drawn from an investigation in which the numbers of adult Amblyomma hebraeum ticks, to which a closed herd of Hereford cattle were exposed over a period of 6 1/2 years, were manipulated. With a tick Cowdria ruminantium infection rate of 3-5%, an endemically stable situation was created by dipping the herd only when an average of 10 adult male and female A. hebraeum ticks were counted on 10 animals. When the average was increased to 15 during the calving period, 97% of calves acquired a tick-mediated immunity at the age of 6 months. Because only adult ticks confined to the hindquarters are counted, this procedure is recommended as a feasible and practical guideline to stock owners wishing to determine a dipping programme that would ensure endemic stability. The indirect fluorescent antibody test gave a true reflection of the infection rate through ticks in calves 3-6 months old, but not in older animals that had been re-infected more than once. This is because on one hand antibody may persist for 2 years after withdrawal from tick exposure and on the other the artificial re-infection of cattle with a tick acquired immunity is not always followed by a rise in antibody titres and may even result in seronegativity. Four cows infected and re-infected through ticks, remained immune to challenge for 2 years after withdrawal from tick exposure. Within the confines of one farm 3 isolates of C. ruminantium that differed in pathogenicity and immunogenicity were recovered from ticks. One of these isolates was almost non-pathogenic to cattle.  相似文献   

14.
A crossbred calf (3 months old) obtained from a farm where regular control of ticks was practised and found to be free of blood parasites was inoculated with 20 ml pooled blood collected from four field cattle which had very low Trypanosoma theileri parasitaemias (one parasite per 70 microl blood as determined by the haematocrit centrifugation technique). Trypanosoma theileri was present in the blood 6 days after injection and a peak parasitaemia of 42 parasites per 70 microl blood was recorded by day 12. Hyalomma anatolicum anatolicum nymphs were applied on the ears of the calf on day 8 and they dropped engorged by days 13 and 14. The resulting adult ticks were examined for the presence of T. theileri by severing a leg and making a smear of the clear haemolymph which exuded from the wound. The smear was fixed in methanol and stained with Giemsa stain. The infection rate with T. theileri in the ticks was 43.3% (26 out of 60). The intensity of infection was very high and various developmental stages of the flagellates were observed (epimastigotes, sphaeromastigotes, trypomastigotes and other intermediate stages). The haemolymph from 12 ticks was also collected in tissue culture medium and the trypanosomes survived for 25 weeks before eventually dying. The results demonstrated unequivocally the high vectorial capacity of the tick H. a. anatolicum for T. theileri.  相似文献   

15.
Antibodies against the 24 kDa Rhipicephalus sanguineus (Rs24p) protein were detected by ELISA to evaluate the relationship between antibodies and tick infestation. The mean titer of 3 dogs that underwent 2 experimental infestations with adult ticks was transiently increased after the second infestation. There was a significant difference in mean titers between positive control dogs naturally infested with ticks and tick-naive dogs. These results suggested that anti-Rs24p antibodies detected by ELISA are a marker of tick exposure. There was no significant difference in mean titers between tick-naive dogs and seropositive dogs to Ehrlichia canis. Some dogs positive for E. canis antibodies showed, however, higher titers than most tick-naive dogs. R. sanguineus may be related to the E. canis infection in Japan.  相似文献   

16.
Seven specific-pathogen-free (SPF) ponies, 1-5 years old, were exposed to Borrelia burgdorferi-infected adult ticks while being treated with dexamethasone over 5 consecutive days. One SPF pony (pony No. 178) was first exposed to laboratory-reared nymphs without B. burgdorferi infection and 3 weeks later was exposed to B. burgdorferi-infected adult ticks with concurrent dexamethasone treatment for 5 consecutive days. Four uninfected ponies treated with dexamethasone, exposed to laboratory-reared ticks without B. burgdorferi infection served as uninfected controls. Clinical signs, bacteriologic culture, polymerase chain reaction (PCR) for bacterial DNA, immunologic responses, and gross lesions and histopathologic changes were investigated during the experiment or at necropsy 9 months after tick exposure. In all of the seven challenged ponies, infection with B. burgdorferi was detected from monthly skin biopsies and various tissues at postmortem examination by culture and by PCR. However, pony No. 178 exposed to laboratory-reared nymphs (without B. burgdorferi infection) and challenged with B. burgdorferi-infected adult ticks 2 months later did not develop a B. burgdorferi infection. All of the infected ponies seroconverted. Control ponies and pony No. 178 were negative by culture, PCR, and serology. Except for skin lesions, we failed to induce any significant histopathologic changes in this study. This is the first report of successful tick-induced experimental infection in ponies by exposure to B. burgdorferi-infected ticks. This Lyme disease model will be very useful to evaluate efficacy of vaccines against the Lyme agent and the effect of antibiotic therapy on horses infected with B. burgdorferi.  相似文献   

17.
An aqueous emulsion of amitraz (Bovitraz, Bayer AH), prepared and applied according to the manufacturer's recommendations, was evaluated for its activity against cattle ticks on a dairy farm in Ethiopia. Eight crossbred heifer calves aged between 6 and 8 months and heavily infested with ticks were selected and divided into equal treatment and control groups. The calves in the treatment group were hand-sprayed with the amitraz emulsion while the control group was left untreated. Each calf was restrained and ticks were counted and identified in situ. One hundred percent tick control was achieved on Day 3 after acaricide application, and this was maintained for a further 18 days. The residual effect was long, and protected the animals from re-infestation for 21 days. There was a significant difference between the mean number of ticks on the control group compared to the treated group (P < 0.05). An important finding in this trial was the rapid detachment of all tick instars from animals treated with the test acaricide.  相似文献   

18.
Blood from 2 buffalo harbouring Theileria organisms was inoculated into a splenectomized Ayrshire calf. The calf developed an infection which extended over a long period. The infection was transmitted to two cattle with Amblyomma gemma by transstadial transmission between the larvae and nymphs. Severe anaemia developed in these cattle and correlated with the parasitaemia. Schizonts morphologically characteristic of T. mutans were detected for short periods in the lymphoid cells of cattle infected by the ticks. The antigens and sera prepared from the cattle reacted with T. mutans sera and antigens in the indirect fluorescent antibody test. After recovery from the primary parasitaemia, the cattle had detectable organisms and antibodies to T. mutans for more than two years.  相似文献   

19.
A novel spotted fever group rickettsia has recently been isolated from Amblyomma hebraeum ticks in Zimbabwe. In a survey of 172 goat sera collected throughout Zimbabwe the highest prevalence of antibodies reactive with this rickettsia was in the south of the country, the area where A hebraeum is most commonly found. Nine goats were infected using male and female A hebraeum taken from a tick line shown to be infected with the novel rickettsia. By week 3 after infection, seroconversion occurred in all nine goats but no clinical signs of disease could be detected. A leucocytosis due to a mature neutrophilia one to two weeks after infection was the only abnormality. Rickettsaemia was detected only on day 3 after exposure to infected ticks. Immunosuppression failed to induce recrudescence of the rickettsaemia.  相似文献   

20.
Dermacentor variabilis were infected as nymphs with Anaplasma marginale by allowing the ticks to feed on a single infected donor calf. Two weeks after molting to the adult stage, the ticks were allotted into 1 of 3 groups and were allowed to overwinter at room temperature (25 C) in the laboratory (group 1), cold storage (4.5 C) in the laboratory (group 2), or outdoors in leaf litter (group 3). Persistence of A marginale was assessed by determining density of colonies (number of colonies/0.1 mm2 of gut tissue examined) in tick gut specimens at 3, 5, 7, 9, and 12 months after molting to the adult stage. Colonies of A marginale were found in all groups at every density evaluation period. Highest colony densities were observed uniformly in specimens collected at month 7 (May); densities decreased at month 9 and were lowest at month 12. Statistical analysis indicated that ticks subjected to cold storage and to outdoor conditions had similar colony densities of A marginale; the density curve in these 2 groups indicated significant quadratic effects over time, with peak densities in May. Mean colony density in ticks kept at room temperature fit a different quadratic equation. The morphologic data indicated that A marginale overwinters in Dermacentor variabilis, and that increasing numbers of organisms are found from January to May.  相似文献   

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