Control of tropical theileriosis with attenuated schizont vaccine in China.

Tropical theileriosis, a tick borne disease of cattle caused by the protozoan Theileria annulata, was widely distributed in northern China causing great economical losses before the 1980's. In the 1960's blood passaging, irradiation and in vitro culture of parasite were used for the development of attenuated vaccine. Two lines of the schizont stage were obtained which both provided sufficient protection against tick infestation or blood challenge in experimental and field trials. A gelatin protection technique allowed storage at 4 degrees C, significantly prolonged shelf life and permitted transport without a cold chain. The vaccines were commercialized and widely administered in epidemic areas, leading to control of T. annulata infection in China.

Molecular phylogenetic studies on an unnamed bovine Babesia sp. based on small subunit ribosomal RNA gene sequences.

The 18S small subunit ribosomal RNA (18S rRNA) gene of an unnamed Babesia species (designated B. U sp.) was sequenced and analyzed in an attempt to distinguish it from other Babesia species in China. The target DNA segment was amplified by polymerase chain reaction (PCR). The PCR product was ligated to the pGEM-T Easy vector for sequencing. It was found that the length of the 18S rRNA gene of all B. U sp. Kashi 1 and B. U sp. Kashi 2 was 1699 bp and 1689 bp. Two phylogenetic trees were, respectively, inferred based on 18S rRNA sequence of the Chinese bovine Babesia isolates and all of Babesia species available in GenBank. The first tree showed that B. U sp. was situated in the branch between B. major Yili and B. bovis Shannxian, and the second tree revealed that B. U sp. was confined to the same group as B. caballi. The percent identity of B. U sp. with other Chinese Babesia species was between 74.2 and 91.8, while the percent identity between two B. U sp. isolates was 99.7. These results demonstrated that this B. U sp. is different from other Babesia species, but that two B. U sp. isolates obtained with nymphal and adultal Hyalomma anatolicum anatolicum tick belong to the same species.

A comparison of small-subunit ribosomal RNA gene sequences of bovine Babesia species transmitted by Haemaphysalis spp. in China.

The ribosomal small-subunit RNA gene sequences of six Chinese Babesia stocks infective to cattle, including a Babesia bigemina isolate, a B. bovis isolate, two B. ovata isolates, a Babesia sp. Wenchuan isolate and a B. major isolate, were compared and analyzed. The target DNA segment was amplified by polymerase chain reaction and the product ligated into the pGEM-T Easy vector for sequencing. The length of the 18S rRNA gene of all Babesia species involved in this study varied between 1,653 and 1,693 bp. The phylogenetic trees were inferred based on the 18S rRNA sequence of the Chinese isolates as well as other species of Babesia available in GenBank. The results showed that the B. ovata transmitted by Haemaphysalis longicornis and Babesia sp. Wenchuan isolate were confined to the same group as B. ovata Korea, with an identity among them of >96.5%, while B. major transmitted by H. punctata was situated in another branch, and identity with other bovine Babesia species was less than 92.5%. B. ovata should, therefore, be a valid species, differing from B. major according to the 18S rRNA gene sequence.

Efficacy of different dosages of ivermectin injectable against the Hypoderma spp. in yaks.

A chemotherapy trial was conducted to determine the lowest dosage of injectable preparation of ivermectin against Hypoderma spp. infestation in yaks in Tibetan areas in Tianzhu county, Gansu province, in northwest of China. One hundred and sixty yaks were randomly divided into four groups of 40 yaks for the trial. The first three groups were treated by subcutaneous injection in the neck with 0.1% ivermectin (respectively, 1, 5, 10 microg/kg body weight). The fourth group was not treated and considered as control group. All the experiments were performed in November 2000 and the animals were examined for the presence of warbles in the next March and May. The results indicated that there was no warbles found on the back of treated animal while third stage larvae were palpated on back of some of the yaks in control group. It is concluded that dosage of 1 microg/kg ivermectin injectable was sufficient to kill or stop development of larvae of Hypoderma spp. in naturally infected yaks if administrated in November.

Experimental transmission of an unnamed bovine Babesia by Hyalomma spp., Haemaphysalis longicornis and Boophilus microplus.

Experiments were undertaken to determine the mode of transmission to cattle of an unnamed Babesia sp. by Hyalomma anatolicum anatolicum, Hyalomma detritum, Hy. rufipes koch, Haemaphysalis longicornis and Boophilus microplus. The unnamed Babesia species designated Babesia U sp. was isolated by infesting cattle with nymphs from female Hy. a. anatolicum ticks collected from Xinjiang province. Adults of laboratory reared Hy. a. anatolicum, Hy. detritum and Hy. rufipes koch were infected with Babesia U sp. by feeding on infected cattle, isolated with nymphal ticks of Hy. a. anatolicum derived from females collected from field. The experiments revealed that Hy. a. anatolicum was capable of transmitting Babesia U sp. transovarially in larval (2 of 4 calves), nymphal (6 of 6 calves) and adult (3 of 8 calves) stages, with prepatent periods of 16, 12, and 8 days, respectively, and that this Babesia was also transovarially transmitted by both the nymphal and adult stages of Hy. detritum and Hy. rufipes. Attempts to transmit this Babesia U sp. transovarially with Hae. longicornis and B. microplus, and transstadially with Hyalomma spp., were carried out, and the results proved to be negative.

Experiments on transmission of an unidentified Theileria sp. to small ruminants with Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum.

Experiments on the transmission of an unidentified Theileria sp. infective for small ruminants by Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum were carried out. Three Theileria-free batches of adult, larvae, and nymphs of laboratory reared H. qinghaiensis and Hy. a. anatolicum ticks were infected by feeding them on sheep infected with Theileria sp. The Theileria sp. was originally isolated from adult ticks of H. qinghaiensis, by inoculation of blood stabilates or tick transmission. H. qinghaiensis has been shown to be capable of transmitting the Theileria sp. infective for small ruminants transstadially to sheep and goats. The nymphs developed from the larvae engorged on the sheep infected with the parasite transmitted the pathogen to splenectomized sheep with prepatent periods of 30, 31 days, respectively; but the subsequent adult ticks of H. qinghaiensis derived from the nymphs did not transmit the pathogen to sheep. However, adults developed from the nymphs engorged on the sheep infected with the parasite transmitted the pathogen to sheep with prepatent periods of 24-27 days. The larvae, nymphs and adult ticks derived from female H. qinghaiensis ticks engorged on infected sheep were not able to transmit the parasite transovarially. The same experiments were done with Hy. a. anatolicum, but examination for presence of piroplasma of Theileria sp. from all animals were negative, demonstrating that Hy. a. anatolicum could not transmit the organism to sheep or goats.

Description of a new Babesia sp. infective for cattle in China.

A Babesia species has been identified and shown to be transmitted by Hyalomma anatolicum anatolicum in China. When larvae, nymphs and adults developed from engorged females H. a. anatolicum collected from cattle in Xinjiang province were infested onto the Babesia-free calves, piroplasms of Babesia sp. were seen in blood smears from cattle infested with nymphs (2 calves), but not from the calves infested with larvae (1 calf or adult ticks (2 calves). This Babesia sp. proved to be of low virulence, causing 3% parasites which lasted 3-4 days.

Transmission of an unidentified Theileria species to small ruminants by Haemaphysalis qinghaiensis ticks collected in the field.

The transmission of a recently identified Theileria species pathogenic for sheep and goats in northern China is described. Haemaphysalis qinghaiensis nymphs which had been collected from grass in epidemic areas were able to transmit this Theileria species to sheep. The pathogen was also transmitted to sheep and goats by three batches of adult ticks collected from grass, ticks collected when moving about on sheep and ticks which were found partially engorged on sheep or goats.