Diagnosis of heartwater at post-mortem in ruminants and the confirmation of Cowdria ruminantium in mice

Criteria that are required to confirm heartwater in ruminants and mice are discussed. In ruminants it entails the macroscopical and microscopical lesions as well as the identification of Cowdria ruminantium in brain smears or histological sections. Macroscopical lesions in the majority of animals that die of the disease include effusion of body cavities, hydropericardium, oedema of the lungs, brain, mediastinum, and its associated lymph nodes, and splenomegaly. The effect of specific chemotherapy on the morphology of heartwater organisms in vivo is outlined. A severe nephrosis in heartwater-infected Angora goats, treated after the first day of the febrile reaction, is described.     

Histologic and immunochemical study of the pathogenesis of heartwater (Cowdria ruminantium infection) in goats and mice

Eleven adult goats and 32 adult outbred mice were inoculated IV with Cowdria ruminantium-infected blood (Kwanyanga isolate), monitored clinically, then serially euthanatized. Predominant clinical signs of disease in goats were depression, head tremors, seizures, and dyspnea. In mice, dyspnea and depression were the only clinical signs of disease noticed. Tissues were examined histologically and immunohistochemically for C ruminantium colonies or antigen. In goats, C ruminantium was detected only in endothelial cells of the brain, even though gross and microscopic lesions were confined to the thorax. In mice, C ruminantium was detected only in endothelial cells of the heart and lungs.     

Specific immunity in mice to heartwater

Mice develop a specific immune response following infection with the mice strains of heartwater. In the case of the Kümm strain the agent can persist in some tissues for up to 365 days. Transfer of spleen cells from immune mice confers protection against homologous challenge in recipient mice showing that cell mediated immunity is important. A comparison with immune mechanisms occurring in other Rickettsia is discussed.     

Resistance of leopard tortoises and helmeted guineafowl to Cowdria ruminantium infection (heartwater).

Experimental infection trials were conducted to investigate susceptibility of leopard tortoises (Geochelone pardalis) and helmeted guineafowl (Numida meleagris) to infection with Cowdria ruminantium, the causative agent of heartwater, a tickborne disease of domestic and wild ruminants. Ten guineafowl were inoculated intravenously with a virulent dose of C. ruminantium derived from bovine endothelial cell cultures, and four leopard tortoises were exposed to C. ruminantium infection by the feeding of infected Amblyomma hebraeum ticks. Uninfected A. hebraeum ticks (on both tortoises and guineafowl) and Amblyomma marmoreum ticks (on tortoises only) were fed on the animals during weeks 2 and 3 post-exposure in an attempt to detect infection. These ticks were analyzed for C. ruminantium infection by xenodiagnosis and with the C. ruminantium-specific pCS20 polymerase chain reaction (PCR) assay. Attempts to detect infection in ticks fed on either species were negative by both tests. These results suggest that leopard tortoises and helmeted guineafowl are refractory to C. ruminantium infection and, therefore, are unlikely to be capable of introducing heartwater directly into new areas. However, leopard tortoises are efficient hosts of A. marmoreum and A. hebraeum and are likely to be important epidemiologically in the transport and maintenance of these tick vector species.     

12株中国伊氏锥虫对小鼠的毒力试验

为了确定中国伊氏锥虫各株的毒力强弱,对中国伊氏锥虫:安徽水牛株(AHB)、广东阳江水牛株(GDB_1)、广东水牛株(GDB_2)、广东马株(GDH)、广西骡株(GXM)、湖北骡株(HBM)、湖南水牛株(HNB)、江苏高邮水牛株(JSB_1)、江苏盱眙水牛株(JSB_2)、新疆骆驼株(XJCA)、云南水牛株(YNB)、浙江水牛株(ZJB)进行小鼠的毒力试验。以各组鼠死亡率和平均存活天数作为毒力强弱的主要依据。结果表明最强致死率为100%,最弱30%;致死所需时间平均为7.5 d～25 d。对小鼠的致病力强弱依次是:AHB>YNB>GDB_2>XJCA>HBM>HNC>JSB_1>GXM>GDB_1>GDH>JSB_2>ZJB。提示不同株的毒力差异显著,测定结果可为伊氏锥虫相关科学研究选株提供依据。     

The pathology of heartwater. I. A study of mice infected with the Welgevonden strain of Cowdria ruminantium

Gross and microscopical lesions in mice intravenously infected with the Welgevonden strain of Cowdria ruminantium closely resembled the lesions described in cattle, sheep and goats. A high concentration of organisms was present in alveolar endothelial cells. Cytopathic changes in parasitized and non-parasitized endothelial cells and the morphology of the organisms are described and compared with the Ball3 strain of C. ruminantium. Possible mechanisms in the development of the lung oedema are considered and the role of mice as animal model is discussed.     

Possible death of a buffalo calf (Syncercus caffer) due to suspected heartwater (Ehrlichia ruminantium)

Rickettsial organisms resembling Ehrlichia ruminantium (the causative organism of heartwater) were demonstrated in brain smears and formalin-fixed brain sections derived from a buffalo calf that died on a private game reserve in northern KwaZulu-Natal. The possibility that the tick-free environment of a quarantine boma may have affected the calf's immunity, is discussed. These findings suggest that monitoring heartwater in wild ruminants and making brain smears as a routine during post mortem evaluations of wild ruminants, should be encouraged.     

Protection of goats against Caribbean and African heartwater isolates by the Ball 3 heartwater vaccine

Summary Two groups of castrated male adult goats (three goats/group) were infected intravenously with the Ball 3 vaccine strain ofCowdria ruminantium and treated with long-acting oxytetracycline at the onset of clinical disease 10 days later. Five weeks post-vaccination one group was challenged with a Caribbean isolate (Gardel) and the other group with a West African isolate (Mali) ofC. ruminantium. Non-vaccinated controls infected with either the Gardel or Mali isolate died. All of the vaccinated animals challenged with the Gardel strain reacted and survived. The three vaccinated animals challenged with the Mali isolate also reacted and two died of heartwater.

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Proteccion De Cabras Con La Vacuna Ball 3 Contra Las Cepas Caribeña Y Africana Causantes De Hidropericardio

Resumen Se infectaron tres grupos de machos cabríos castrados vía intravenosa (tres cabras/grupo), con la vacuna Ball 3 deCowdria ruminantium. Los animales se trataron posteriormente con tetraciclina LA, diez días después al comienzo de la enfermedad clínica. Cinco semanas después de la vacunación, un grupo se descargó con la cepa Caribeña (Gardel) y el otro grupo, con la cepa Africana Occidental (Malí) deC. ruminantium. Los controles no vacunados, infectados con las cepas Gardel y Malí, murieron. Todos los animales vacunados que recibieron la descarga con la cepa-Gardel, sobrevivieron. Los tres animales que recibieron la descarga con la cepa Malí reaccionaron también, muriendo dos de ellos.

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Protection Apportee Aux Chevres Vis-A-Vis Des Souches antillaises Et Africaines De Heartwater Par Le Vaccin Heartwater Ball 3

Résumé Deux groupes de chèvres adultes (mâles castrés, 3 animaux par groupe) ont été infectés par voie intraveineuse avec la souche vaccinale Ball 3 deCowdria ruminantium, et traités avec une oxytetracycline à longue action lors de l'application des signes cliniques dix jours plus tard. Cinq semaines après la vaccination, un groupe a été éprouvé avec une souche antillaise (Gardel) et l'autre groupe avec une souche ouest-africaine (Mali) deC. ruminantium. Les animaux contrôlés, non vaccinés, infectés soit par la souche Gardel, soit par la souche Mali, sont morts. Les animaux vaccinés, éprouvés avec la souche Gardel ont réagi mais ont survécu. Les trois animaux éprouvés avec la souche Mali ont aussi réagi et deux sont morts de heartwater.

     

Isolation and transmission ofCowdria ruminantium (causal agent of heartwater disease) in Blue Nile Province,Sudan

Heartwater was diagnosed in sheep at the Umm Banein Livestock Research Station, Blue Nile Province, Sudan.Cowdria ruminantium was isolated from sheep by subinoculation of blood into a goat. AdultAmblyomma lepidum ticks fed as nymphs on this goat transmitted heartwater to a Friesian (Bos taurus) calf. Brain stabilates from this calf, either snapfrozen into liquid nitrogen or frozen slowly in a deepfreezer at –70°C, with DMSD as cryoprotectant, were infective to cattle when inoculated subcutaneously.     

The infection and treatment method of vaccination against heartwater

The history of vaccination techniques against heartwater and the development of the infection and treatment method are reviewed briefly. Present techniques and their shortcomings are defined and possible areas of improvement discussed.     

An assessment of the economic impact of heartwater (Cowdria ruminantium infection) and its control in Zimbabwe.

Heartwater, caused by the rickettsial organism Cowdria ruminantium, is a serious constraint to livestock development in much of sub-Saharan Africa. Traditionally, the disease has been controlled by the use of chemical acaricides to control the vector tick. The University of Florida/USAID-supported heartwater research project (based in Zimbabwe) is developing a new inactivated vaccine to control the disease. In order that the vaccine is used effectively, the project has been studying the epidemiology of the disease in different livestock production systems of Zimbabwe, and evaluating the economic impact of the disease and of its future control using a vaccine such as the one under development. Initially, field studies were conducted to characterise the communal and commercial livestock-productions systems at risk from heartwater and to understand the epidemiology of the disease. The data from these studies were then applied to an infection-dynamics model of heartwater, which was used to provide estimates of disease incidence and impact under various scenarios over a period of 10 yr. Two principal outputs of the epidemiological model (cumulative annual heartwater incidence and infection-fatality proportion) were key inputs into an economics model. The estimated total annual national losses amount to Z$ 61.3 million (US$ 5.6 million) in discounted value terms over 10 yr. Annual economic losses per animal in the commercial production system (Z$ 56 discounted values) are 25 times greater than the losses in the communal system (Z$ 2.2). The greatest component of economic loss is acaricide cost (76%), followed by milk loss (18%) and treatment cost (5%). Losses in outputs other than milk (beef, traction and manure) appear to be minimal. A new vaccine has the promise of a benefit: cost ratio of about 2.4:1 in the communal and 7.6:1 in the commercial system. A control strategy based on a new vaccine would yield additional non-financial benefits to farmers and the government resulting from reductions in the use of chemical acaricides.     

Variations in innate resistance to experimental Toxocara canis infection in two strains of mice

Strain-, sex- and age-based resistance of mice to Toxocara canis infection was demonstrated. Infection doses of similar numbers resulted in significantly (P less than 0.05) greater worm establishment in CD1 mice than in NIH mice, as well as a greater heterogeneity of responses between individuals. Male mice had significantly (P less than 0.05) fewer larvae in the brain and more larvae in the liver than female mice, when mice of greater than or equal to 3 months old were used. Appreciably more larvae were retained in the liver of male CD1 mice than in NIH mice. Orchidectomized mice had higher levels of brain infestation and a lower larval recovery from the liver compared with intact male mice. Infections in young mice of both strains showed more diversity in establishment and migration pattern between individuals than in older mice. Despite these differences in distribution of larvae and in the numbers recovered from experimental infections, the acute and chronic clinical signs observed in the two strains were very similar.     

Detection of Cowdria ruminantium antigen and antibody during the course of heartwater disease in sheep by means of an enzyme-linked immunosorbent assay

An enzyme-linked immunosorbent assay was used to detect Cowdria ruminantium antibodies during the course of heartwater disease. IgM antibodies reached a maximum on the 4th day after infection and disappeared on the 7th day. IgG antibodies first appeared on the 8th day and continued to increase during the remainder of the observation period of 28 days. The presence of C. ruminantium in the blood fractions of diseased animals was demonstrated by an enzyme-linked immunosorbent assay. The earliest day of C. ruminantium antigen detection was in plasma and serum on the 4th day after inoculation. Of all the blood fractions investigated, the red blood cells showed the highest concentration, and this reached a maximum on the 12th day after infection.     

The role of males of the bont tick (Amblyomma hebraeum) in the transmission of Cowdria ruminantium (heartwater)

The role of males of the bont tick (Amblyomma hebraeum) in the transmission of Cowdria ruminantium (heartwater) was investigated. Transstadial (nymph to adult) and intrastadial transmission were demonstrated. Males transferred from live or dead hosts to live hosts were shown to transmit C. ruminantium repeatedly. It was concluded that male transmission is of importance in the epidemiology of heartwater.     

The use of electro-encephalography and brain biopsy in the clinical diagnosis of heartwater

Both electro-encephalography and brain biopsy were examined as possible aids in the confirmation of a clinical diagnosis of heartwater in field cases. Results showed that electro-encephalographic changes associated with heartwater were non-specific and probably caused by the associated brain oedema. Brain biopsy on the other hand appears to be a reliable technique for the confirmation of heartwater, and the technique if properly performed holds little risk for the animal.     

The heterogenicity of Cowdria ruminantium stocks: cross-immunity and serology in sheep and pathogenicity to mice

Ten stocks of Cowdria ruminantium (Ball 3, Breed, Comoro, Germishuys, Kümm, Kwanyanga, Mali, Mara, Nonile and Welgevonden) were compared from a cross-immunity, serological and mouse pathogenicity point of view. They were found to differ in varying degrees. Except for the Ball 3, Comoro and Germishuys stocks that were similar but not identical, there was no pattern in the antigenic diversity of the 10 stocks. The Welgevonden stock emerged as the stock that elicits an immunity against most of the South African stocks. The inability of the reference Ball 3 stock to protect sheep against no fewer than 6 other stocks questions the advisability of retaining this stock as the vaccine stock. The antigenic diversity of the 10 stocks could not be correlated with the antibody levels detected with the indirect fluorescent antibody test, since the sera against all 10 stocks reacted positively to the Kümm stock antigen and the variation in titres was not stock-related.     

Protection of cattle against heartwater in Botswana: comparative efficacy of different methods against natural and blood-derived challenges

Five groups of Tswana-cross castrated male cattle between 20 and 30 months of age (a total of 158 animals) were transported from a ranch in a heartwater-free area of south Botswana to a feedlot near Gaborone in the east of Botswana where heartwater is endemic. On arrival, one group was vaccinated intravenously with the Onderstepoort sheep blood heartwater vaccine, one group was vaccinated intravenously with the new Onderstepoort tick-derived heartwater vaccine and a third group was vaccinated subcutaneously with this tick-derived vaccine. Vaccine reactions were blocked with long acting oxytetracycline on the first day of fever. A fourth group had a series of injections of long acting oxytetracycline on days 0, 7, 14 and 21 after arrival, and a fifth served as untreated controls. The animals remained at the feedlot for 65 days during which time they faced a low level of challenge by Amblyomma hebraeum ticks. None contracted heartwater and so they were then challenged, together with a further group of control cattle, with a dose of the sheep blood vaccine. Some animals in all groups had severe heartwater reactions and died despite therapy, but 76.7 per cent, 64.5 per cent and 74.3 per cent of the cattle in the blood vaccine, intravenous tick vaccine and long acting oxytetracycline groups respectively were resistant to challenge, compared with 48.3 per cent of the subcutaneous tick vaccine group and 36.4 per cent of the controls. It was concluded that intravenous vaccination of susceptible adult cattle with either the blood or the tick-derived vaccine needs careful monitoring in the month after vaccination and does not necessarily result in immune animals.(ABSTRACT TRUNCATED AT 250 WORDS)