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Heartwater (Ehrlichia Ruminantium)

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Heartwater (Ehrlichia Ruminantium) Clinical communication — Kliniese mededeling Possible death of a buffalo calf (Syncercus caffer) due to suspected heartwater (Ehrlichia ruminantium) S Pfitzera*, R Lastb and D T De Waalc northern KwaZulu-Natal. It was the 2nd ABSTRACT buffalo calf found dead within 2 months Rickettsial organisms resembling Ehrlichia ruminantium (the causative organism of heart- in this reserve. In addition, 1 blue wilde- water) were demonstrated in brain smears and formalin-fixed brain sections derived from a beest and one impala ram (Aepyceros buffalo calf that died on a private game reserve in northern KwaZulu-Natal. The possibility melampus) had died of unknown causes. that the tick-free environment of a quarantine boma may have affected the calf’s immunity, The dam of this buffalo calf came from the is discussed. These findings suggest that monitoring heartwater in wild ruminants and tuberculosis (TB) eradication programme making brain smears as a routine during post mortem evaluations of wild ruminants, should carried out by KwaZulu-Natal Conserva- be encouraged. tion Service (KZNCS) in quarantine Key words: buffalo, Cowdria ruminantium, cowdriosis, Ehrlichia ruminantium, heartwater, bomas at Phinda Resource Reserve. South Africa. The dam had initially originated from the Pfitzer S, Last R, De Waal D T Possible death of a buffalo calf (Syncercus caffer) due to sus- Hluhluwe–Umfolozi Game Reserve and pected heartwater (Ehrlichia ruminantium). Journal of the South African Veterinary Association had been introduced onto the property (2004) 75(1): 54–57 (En.). Pongola Animal Clinic, PO Box 682, Pongola, 3170 South Africa. about 1 year before this investigation. The calf was born in the private game reserve, which had been free of domestic rumi- INTRODUCTION deaths due to infection with E. ruminan- nants for more than 10 years. Cowdria ruminantium,4 the causative tium have been reported. To date, no organism of heartwater,has recently been deaths due to heartwater have been Pathology reclassified as Ehrlichia ruminantium based documented in the African buffalo. How- on genetic analysis as well as biological ever, African buffalo2,24 as well as eland, Post mortem and protein characteristics.6 Ehrlichia kudu (Tragephalus strepsiceros strepsiceros), The calf died during the morning of ruminantium is transmitted by 3 host ticks blue wildebeest, giraffe (Giraffa camelo- 5 June 2000. The carcass was kept cool belonging to the genus Amblyomma5. pardalis)15, sable (Hippotragus equinus) and until the post mortem could be carried out Amblyomma hebraeum, the vector of E. tsessebe (Damaliscus lunatus)16 are known at 15:00 that afternoon. ruminantium in South Africa, occurs to seroconvert after artificial infection and Gross post mortem findings revealed the mainly in the Limpopo, KwaZulu-Natal become carriers of the parasite. One animal to be in good condition with a low and in the Eastern Cape provinces of African buffalo showed signs of heart- tick burden. Although ticks were not South Africa23. water after artificial infection with E. specifically identified, some Amblyomma Heartwater is one of the most important ruminatium, but it recovered spontane- were present. Rigor mortis was still tick-borne diseases of domestic rumi- ously24. Different strains of E. ruminan- apparent. No signs of struggling or inju- nants in South Africa18. Mortality rates are tium have the ability to grow in ries were evident. The peripheral blood high in susceptible cattle, sheep and goats endothelial cell lines of sable antelope, smear was negative for anthrax and blood introduced into endemic regions and buffalo and eland20. These findings parasites. The rumen contained some peracute, acute, subacute and clinically suggest the possibility that wild rumi- roughage; milk was found in the aboma- inapparent forms of heartwater have nants play an important role in the sum. Mild splenomegaly and hepato- been reported in these domestic rumi- epidemiology and spread of heartwater. megaly was present, as well as ascites. nants21. In wild ruminants, such as the This hypothesis is strengthened by the Whether hydrothorax was present, could blue wildebeest (Connochaetes taurinus), results of a study in the Kruger National not be determined, as the pericard was springbok (Antidorcas marsupialis), Park of South Africa where it was shown damaged when skinners opened the bushbuck (Tragelaphus scriptus)13, black that a wildlife cycle can maintain the thorax. Lungs revealed a foamy, frothy wildebeest (Connochaetes gnu), blesbok transmission of E. ruminantium inde- material in the major bronchi and distal (Damaliscus albifrons)11, steenbok (Raphi- pendent of domestic ruminants17. 3rd of the trachea. A few lobules in the cerus campestris)8, sitatunga (Tragelaphus Numerous E. ruminantium isolates have cranial lobe appeared hepatized. Accen- spekei)14 and eland (Taurotragus oryx)25 been made, and named after the farm or tuation of lobulation due to interstitial a country of origin. These strains differ in oedema and emphysema in the lung was Pongola Animal Clinic, PO Box 682, Pongola, 3170 3,7 South Africa. E-mail: [email protected] their pathogenicity for ruminants . visible. Atrial petechiae were observed bVetdiagnostix – Veterinary Pathology Services, PO Box on the heart. In the brain blood vessels 13624, Cascades, 3202 South Africa. appeared engorged. A brain smear was cParasitology Division, ARC-Onderstepoort Veterinary CASE REPORT Institute, Private Bag X05, Onderstepoort, 0110 South performed and stained with Kyro-Quick Africa. Present address: Department of Veterinary Micro- 19 biology and Parasitology, Faculty of Veterinary Medicine, History stain (Kyron Laboratories) . Numerous University College Dublin, Belfield, Dublin 4, Ireland. On 5 June 2000, a dead buffalo calf of rickettsial-like organisms were observed E-mail: [email protected] approximately 5 months of age was in the endothelial cells of the capillary *Author for correspondence. Received: July 2003. Accepted: January 2004. discovered on a private game reserve in blood vessels, which was also seen in 54 0038-2809 Tydskr.S.Afr.vet.Ver. (2004) 75(1): 54–57 Giemsa-stained brain smears (Fig. 1). Un- stained brain smears as well as samples of frozen brain were taken. Additional samples of brain, bone marrow, heart, kidney, liver, spleen, lymphonodes and lung were preserved in 10 % formalin for further histopathological investigation. Microscopic examination Brain. Severe congestion of meningeal and parenchymal vessels was evident. Prominent vasogenic oedema with widening of Virchow-Robin spaces, peri- neural spaces and dilatation of myelin sheaths in the central white matter neu- trophils were present as well as diffuse gliosis with scant perivascular infiltrates of individual mononuclear cells. Distinct intracytoplasmic colonies of rickett- sial-like organisms resembling E. rumi- Fig. 1: Many large colonies of small Ehrlichia ruminantium-like organisms, intra- nantium were identified within the endo- cytoplasmically within endothelial cells of brain capillaries. Giemsa; ×1000. thelial cells (Fig. 2). Spleen. Diffuse congestion of splenic rhages were scattered throughout the double-unit membrane were found in sinuses was present while lymphoid myocard. Interstitial oedema was present capillary endothelial cells, features that follicles had a washed-out appearance with teasing apart of myofibres. Individ- are characteristic of the Ehrlichia group. with depletion of lymphocytes from the ual myofibres appeared amorphous and germinal centres (lympholysis). eosinophilic (myofibre hyalinization). DNA probes Lung. Severe diffuse congestion was Frozen brain tissue was also subjected evident with engorgement of bronchial Immunohistochemistry to PCR and DNA probe examination in an vasculature and alveolar wall capillaries. Immunohistochemical (immunoperox- attempt to confirm the identity of the Effusion of fibrinous fluid had resulted idase) stains for heartwater were per- organism. The pCS20 probe22 is known to in interstitial distension (interstitial formed on formalin-fixed brain sections discriminate between E. ruminantium and oedema). There was scant interstitial infil- and revealed positive-staining granules other Ehrlichial organisms1. Unfortu- trates of mixed mononuclear cells and in many endothelial cells of parenchymal nately, the sample was in a too-advanced individual leukocytes. In individual capillaries. Confirmed cases of Coxiella stage of decomposition and the results lobules a severe inflammatory reaction burnetti and Chlamydia psittaci were used were inconclusive. was visible; bronchi were plugged with as negative controls and showed no posi- necrotic plugs consisting of fibrin, tive staining. Diagnosis necrotic cell debris and infiltrating leuko- The cause of death was suspected heart- cytes (necrotic bronchitis). Peribronchial Transmission electron microscopy water with complicating acute bacterial and intra-alveolar infiltrates contained in- On transmission electron microscopic bronchopneumonia. numerable neutrophils accompanied by (TEM) examination, pleomorphic, coc- proliferating alveolar macrophages coid, ovoid, filamentous, horseshoe- DISCUSSION (alveolar pneumonia). Large bacterial shaped and polygonal organisms, 0.49– Demonstration of rickettsial organisms, colonies consisting of bipolar, Gram-neg- 2.7 µm in diameter and enclosed by a resembling E. ruminantium, in brain ative organisms resembling Pasteurella were observed in many of these necrotic areas. Liver. The liver showed diffuse conges- tion with engorgement
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