Theileria annae (syn. Babesia microti-like) infection in dogs in NW Spain detected using direct and indirect diagnostic techniques: clinical report of 75 cases.

BACKGROUND: In north-western Spain, piroplamosis caused by Theileria annae is now recognized as a serious problem because veterinarians, despite being aware of the clinical signs of piroplasmosis, lack the necessary information on its epidemiology or specific diagnostic tools for its management. This, along with the fact that T. annae infection is also refractory to current piroplamosis treatments, prompted this study designed to assess the clinical presentation and diagnosis of this largely unknown parasitic disease in dogs. METHODS: One hundred and twenty dogs in NW Spain suspected clinically of having piroplasmosis were examined and piroplasm species detected by light microscopy (LM) observation of Giemsa-stained blood smears, immunofluorescent antibody test (IFAT), and PCR plus sequencing. RESULTS: Seventy five of the sick dogs were confirmed to be infected with T. annae by PCR (designated "true infection cases"). Intraerythrocytic ring-shaped bodies morphologically compatible with small piroplasms were observed by LM in 59 (57 true infections) of the 120 blood samples. Anti-Babesia antibodies were detected by IFAT in 59 of the 120 sera (55 of which were "true infections"). Using PCR as the reference method, moderate agreement was observed between positive LM vs PCR and IFAT vs PCR results (kappa values: 0.6680 and 0.6017, respectively). Microscopy examination and IFAT were moderately sensitive in detecting the pathogen (76% and 73.3%, respectively). In the 75 cases of "true infection", the most common clinical signs observed were pale mucous membranes, anorexia and apathy. Blood cell counts consistently revealed severe regenerative anaemia and thrombocytopenia in dogs with piroplasmosis due to T. annae. Young dogs (≤3 year) (p = 0.0001) were more susceptible to the disease. CONCLUSION: Microscopy showed moderate diagnostic sensitivity for acute T. annae infection while IFAT-determined antibody titres were low (1/64 to 1/128). The infecting species should be therefore confirmed by molecular tests. Our results suggest that the disease affects dogs in regions of Spain bordering the endemic Galicia area where this piroplasm has not been previously reported (Asturias, northern Spain). Further epidemiological surveys based on serological and molecular methods are required to establish the current geographical range of T. annae infection.

Theileria gilberti n. sp. (Apicomplexa: Theileriidae) in the Gilbert's potoroo (Potorous gilbertii).

The morphology and genetic characterisation of a new species of piroplasm identified in the blood of the Gilbert's potoroo (Potorous gilbertii) from the Two Peoples Bay Nature Reserve near Albany, Western Australia, is described from blood and tissue samples from 16 Gilbert's potoroos. Microscopy of blood showed these parasites are highly pleomorphic with a mean length of 1.8 mum and mean width of 0.85 mum. Phylogenetic analysis of 18S rRNA sequence data identified the piroplasm as a new species of Theileria that is closely related to other Australian marsupial piroplasm species. Based on biological and molecular data, it is proposed that the parasite from Gilbert's potoroo be given the name Theileria gilberti n. sp.

Theileria electrophorin.sp., a parasite of the electric eel Electrophorus electricus (Osteichthyes: Cypriniformes: Gymnotidae) from Amazonian Brazil.

The name Theileria electrophori n.sp. is proposed for a small parasite described in the erythrocytes of the electric eel, Electrophorus electricus, from Amazonian Brazil. Division of the organism in the erythrocyte produces only four bacilliform daughter cells which become scattered in the host cell, without a cruciform or rosette-shaped disposition. Exoerythrocytic meronts producing a large number of merozoites were encountered in Giemsa-stained impression smears of the internal organs, principally in the liver, and are presumably the source of the intraerythrocytic forms of the parasite. This developmental pattern is characteristic of piroplasms within the family Theileriidae, where the author considers the parasite of E. electricus to most appropriately belong. It effectively distinguishes the organism from the dactylosomatid parasites Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbé, 1894 also found in fishes. This appears to be the second report of Theileria Bettencourt, Franca and Borges, 1907 in a fish.

Theileria electrophorin.sp., a parasite of the electric eel Electrophorus electricus (Osteichthyes: Cypriniformes: Gymnotidae) from Amazonian Brazil

The name Theileria electrophori n.sp. is proposed for a small parasite described in the erythrocytes of the electric eel, Electrophorus electricus, from Amazonian Brazil. Division of the organism in the erythrocyte produces only four bacilliform daughter cells which become scattered in the host cell, without a cruciform or rosette-shaped disposition. Exoerythrocytic meronts producing a large number of merozoites were encountered in Giemsa-stained impression smears of the internal organs, principally in the liver, and are presumably the source of the intraerythrocytic forms of the parasite. This developmental pattern is characteristic of piroplasms within the family Theileriidae, where the author considers the parasite of E. electricus to most appropriately belong. It effectively distinguishes the organism from the dactylosomatid parasites Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbé, 1894 also found in fishes. This appears to be the second report of Theileria Bettencourt, Franca and Borges, 1907 in a fish.

Redescription of Babesia equi Laveran, 1901 as Theileria equi Mehlhorn, Schein 1998.

The horse-parasitizing species Babesia equi Laveran, 1901 was redescribed as Theileria equi Mehlhorn, Schein 1998 and, thus, transferred from one valid genus to another. This transfer was needed since it turned out that this horse parasite showed the relevant characteristics of theilerians with regard to biological data, morphological features, biochemical properties, and molecular biological relationships.

The same but different: the biology of Theileria sporozoite entry into bovine cells.

Theileria are important tick-transmitted protozoan parasites that infect wild Bovidae and domestic animals throughout much of the world. Much of our understanding of Theileria sporozoite invasion of bovine cells is based on work on T. parva, the causative agent of East Coast fever in cattle throughout east, central and southern Africa. Sporozoite entry involves a defined series of sequential but separable steps that differ in important details from the invasion process in other apicomplexans such as Plasmodium and Toxoplasma. While the morphological features of invasion are fairly well documented, the detailed biology of the individual steps is only now becoming clear. This review summarizes much of this recent work on the biology of sporozoite entry. In particular, recent studies on the role of Ca2+ and cell activation processes in sporozoite entry suggest that the initial sporozoite binding event triggers the mobilization of intrasporozoite Ca2+ and the activation of both kinase and G-protein associated signalling processes in the parasite. These processes in turn regulate the invasive capacity of the sporozoite although the identity of these parasite molecules and how they contribute to the invasion process remain to be determined.

Theileriosis of sheep and goats in China.

Theileriosis is an important disease of sheep and goats in West China. Its main distribution includes Qinghai, Gansu, Ningxia, Inner Mongolia, Shaanxi and Sichuan. The epidemic period is from late March to July with April-May being the peak months. This is the period of most intensive tick attack by Haemaphysalis qinghaiensis (77.2-99.24%) during the year. It has been proved that the nymphs and adults, which develop from larvae and nymphs engorged on infected sheep or goats can transmit the pathogen. Experimental infections revealed the incubation period, clinical signs and the pathogen's morphological characteristics. The disease was more serious in lambs and exotic adult animals than native adult animals. The sickness rates of lambs, exotic and native adult animals were 78-85%, 41% and 9% respectively; death rates were 81.41%, 62.5% and 65% respectively. Clinical prophylaxis, chemical therapy and destruction of ticks have been carried out with antiparasitic compounds to control the disease since 1982. Experiments demonstrated that the curative rate was up to 76-86%, but no satisfactory prophylactic methods were found.

An unidentified species of Theileria infective for cattle discovered in China.

Infection experiments, morphological observations and transmission experiments were conducted with an unidentified Theileria sp. isolated from a naturally infected ox. The results showed that the protozoa could multiply extensively in a splenectomized ox and the parasitaemia could reach 52.69%. The Theileria sp. was polymorphic: being pear-shaped, circular, elliptical, rod-like, comma-shaped, three-leafed- or cross-shaped and having many other irregular-shapes which were seldom detected. In erythrocytes, the anaplasma-like protozoa grew, producing protoplasm which could extend and clump together, and developed into many polymorphic protozoa. Some of the protozoa propagated themselves by budding. The protozoa could not be transmitted by Haemaphysalis longicornis or Hyalomma detritum. The pathogenicity, vector ticks and life cycle of this protozoan are unknown.

Light and electron-microscopical observations of the early developmental stages of Theileria sergenti in the gut of Haemaphysalis longicornis.

Morphological characteristics of the early developmental stages of Theileria sergenti in the vector tick were observed by light and electron-microscopies and compared with those of other Theileria species. Just before the repletion of nymphs, various stages of the parasite were observed in gut contents of tick by light microscopy: the spherical intra-erythrocytic stage, the ring-form stage, the spindle-shaped stage and the round-form stage. Electron microscopy revealed that the spherical intra-erythrocytic stages were 1-2 microns in diameter and were marginally located in host erythrocytes. The organisms liberated from host erythrocytes into the tick gut lumen had a large nucleus, an endoplasmic reticulum, electron dense-organelles and a double-walled, mitochondrion-like organelle. In addition ticks had ray-bodies in the gut lumen. These bodies had short protrusions containing 1-3 microtubules and a labyrinthine structure containing highly electron-dense organelles, so they were considered as the developing microgamont of T. sergenti.

Histological observations on the schizonts in cattle infected with Japanese Theileria sergenti.

Histological observations were performed on the schizonts of Japanese Theileria sergenti in three calves necropsied 8 and 10 days after application of nymphal Haemaphysalis longicornis infected with T. sergenti. In all the three calves, schizonts were observed in the cytoplasm of huge cells formed in the drainage lymph nodes, liver and spleen. The huge cells were 50 to 200 microns in diameter. Schizonts had granular appearance and an irregular shape and were 1 to 7 microns in diameter. Ultrastructurally, schizonts had more than one nuclei and were formed in the unit enlarged cell. Schizonts showed a specific reaction against anti-T. sergenti anti serum, therefore, it was concluded that the schizonts were those of T. sergenti histologically.