Spread of parasites transported with their hosts: case study of two species of cattle tick.

Like all parasites, ticks can be spread easily along with their hosts. Ticks are obligate parasites of vertebrates, to which they attach themselves for varying periods of time, and are well-adapted to this mode of transport. Once the transport stage is complete and they have detached at destination, they are also able to wait several months for the arrival of a new host on which they will continue their life cycle. This leads to the establishment of a secondary tick population. Two tropical cattle tick species, Rhipicephalus microplus and Amblyomma variegatum, have perfected this strategy of colonisation and occupation of favourable zones. Rhipicephalus microplus, which originated from South and Southeast Asia, is highly specific for ungulates, and thanks to cattle movements it has spread throughout the tropical belt, apart from the remotest areas. Amblyomma variegatum, which originated in Africa, was transported to Madagascar and the Mascarene Islands, as well as to the West Indies, during the time of the Atlantic triangular trade. These two ticks are vectors of particularly serious cattle diseases: babesiosis and anaplasmosis in the case of R. microplus, and heartwater (cowdriosis) in the case of A. variegatum. Anticipated climate changes are likely to modify the potential geographical range of these two parasite species and numerous others. Even now there are still many areas of the Americas, Asia and Oceania into which A. variegatum has not yet spread, but which it would find favourable. It could be spread not only by the transport of cattle, but also by the migration of some of its other hosts, such as birds. Surveillance--and know-how--is needed to identify these parasites when they first appear and to rapidly contain new outbreaks. Efforts should be made to raise the awareness of livestock professionals about the risks of transporting cattle. Regulations should be implemented and precautions taken to avoid such artificial expansion of the range of ticks and the diseases they transmit.

Tick reference collection of the late Dr. P.C. Morel: a tool for tick taxonomists and veterinarians.

Host preference of ticks is an important, but still controversial, subject. Recent developments in molecular biology provide new opportunities to test some hypotheses about host preference in a given environment if appropriate specimens are available. Since the unique collection gathered by Dr. P.C. Morel could help achieve this goal, we present an overview of the samples available in his collection.

Serotypes in Cowdria ruminantium and their relationship with Ehrlichia phagocytophila determined by immunofluorescence.

Two tick-borne rickettsial pathogens of ruminants, Cowdria ruminantium (causative agent of heartwater disease) and Ehrlichia phagocytophila (causative agent of tick-borne fever), were successfully cultivated in caprine or ovine neutrophilic granulocytes. Infected cultures were subsequently used as antigens in the indirect fluorescent antibody test. Low-level bilateral serological cross-reactions could be detected between Cowdria and Ehrlichia. In addition, comparison of five Cowdria stocks using immunofluorescence demonstrated the existence of distinct serotypes within the genus of Cowdria. It is concluded that the occurrence of these serotypes will considerably complicate the current serodiagnosis of heartwater.

International collaborative research: significance of tick-borne hemoparasitic diseases to world animal health.

A general review is given of the tick-borne hemoparasitic diseases of greatest economic importance in ruminants, babesiosis, anaplasmosis, theileriosis and cowdriosis, each caused by one or more species of hemoparasites. Most affected are cattle and small ruminants, buffalo are more resistant and little is known regarding camels. The situation varies from one continent or region to another. Innate and breed susceptibility to these diseases are of tremendous importance. Disease in the field cannot be considered separated from the whole complex of tick-borne diseases and from the ticks themselves, particularly if the aim is to attain endemic stability. International coordination is needed now that research funds are scarce. An appendix contains tables with hemoparasites of various domestic animals and notes with background details.

Piroplasms of domestic animals in the Macedonia region of Greece. 1. Serological cross-reactions.

During a serological survey on haemoparasites in Macedonia, serum samples were collected from cattle, sheep and goats. All sera were tested by the indirect immunofluorescence test (IFAT); the cattle sera against Theileria orientalis, T. annulata, Babesia bigemina, B. bovis, B. divergens and B. major antigens; the sheep and goat sera against T. ovis, B. ovis, B. motasi and B. crassa antigens. Parallel tests of negative and positive control sera against all the antigens showed the existence of cross-reactions of different degrees between species of the same genus. In cattle, the most important cross-reactions were obtained against B. bigemina antigen, especially with the anti-B. bovis serum, in small ruminants against B. motasi with the anti-B. crassa serum. In the field sera, there was a high correlation between the antibody titres of B. bigemina and B. bovis, and also between the titres of these two Babesia spp. and B. divergens. A high correlation was also found between B. motasi and B. crassa, and lower ones between these two and B. ovis. The correlations of the sera titres were due to mixed infections or to cross-reactions. Therefore, the use of the IFAT is not always satisfactory for diagnosing infections in regions where animals are infected with different piroplasms.

Relationships between Cytauxzoon felis and African piroplasmids.

A cat which had recovered from Cytauxzoon felis infection following treatment with the anti-theilerial drug, parvaquone, showed an increase in piroplasm parasitemia after splenectomy and its blood was suitable for the preparation of antigen smears for the indirect fluorescent antibody test. High levels of antibodies were found in this cat after recovery, in two other cats sub-lethally infected with piroplasms and in sera of naturally infected bobcats. Cats recovered from piroplasm infection died from cytauxzoonosis when challenged with organ material containing C. felis schizonts. Tests with piroplasm antigens and positive sera of C. felis, South African Babesia felis and African Theileria taurotragi showed no significant serological relationship between C. felis and the African parasites. C. felis was not shown to be infective for splenectomized sheep.

Immunization of cattle against Theileria parva bovis and their exposure to natural challenge.

Theileria parva bovis isolates were tested for their immunizing capacity under natural field challenge on Willsbridge Farm in the highveld of Zimbabwe. Fifteen susceptible Sussex yearlings were immunized with the Boleni stock and 15 with a mixture of three isolates from the farm, using tick-derived sporozoite stabilates. No chemoprophylaxis was used. A dose of 0.1 ml of stabilate appeared to be safe in preliminary laboratory experiments, but the reactions were severe in the Sussex cattle and one died despite treatment. Twenty-nine immunized animals and 10 controls first experienced a mild infection, starting about 15 days after their arrival at the farm. Ten of the immunized animals and four controls had schizonts in peripheral lymph nodes for variable periods; one third of those had pyrexia. Nymphal Rhipicephalus appendiculatus ticks applied to three of the reacting immunized calves transmitted Theileria taurotragi to two animals and T. parva to a third. A second Theileria infection, due to T. parva bovis, was detected shortly after the first one. Schizonts were detected in seven out of 10 controls. Pyrexia was more severe and prolonged. Two of the controls died of theileriosis. At the same time schizonts were seen in three immune animals and eight of them had short periods of pyrexia. Intercurrent infections with Babesia bigemina, Borrelia theileri and Eperythrozoon were detected and may have contributed to the fever. Tick infestations were low during the exposure. In the second year of exposure, four out of eight new control animals had severe reactions, and one died. None of the immunized animals became ill, but one animal from the first year control group, which had not reacted previously, had clinical theileriosis. It is concluded that immunization provided an effective protection against field challenge.

Skin nodules in East Coast fever.

A skin eruption developed in five calves with advanced experimental East Coast fever (ECF) (Theileria parva infection). In three cases the eruption was due to intradermal nodules containing numerous theilerial schizonts, in one animal the nodules contained abscesses filled with Demodex mites, and in the fifth calf both types of nodules were present. No nodules were seen in 15 other experimental cases of ECF, which generally died or were killed in an earlier stage of the disease. It is postulated that demodicosis may become apparent in healthy carriers of the mite in the course of ECF, because of immuno-depression caused by the latter disease.

Is Mukherjeella caprae Bandaranayake and Neitz, 1961 (Rickettsiales) a microorganism?

Longitudinal structures in red cells in Giemsa-stained blood smears of goats in the Netherlands were in distinguishable from what has been described as Mukherjeella caprae Bandaranayake and Neitz, 1961 (Rickettsiales). Electron microscopic studies and animal experiments showed, however, that these structures are not parasitic in nature, but are fibres consisting of bundles of microfilaments embedded in the erythrocytic cytoplasm, apparently identical with the longitudinal fibres in goat erythrocytes which are considered to be a haemoglobin abnormality. The condition appears to be genetic. M caprae should be considered as a nomen dubium.

Cowdria ruminantium (Rickettsiales) in primary goat kidney cell cultures.

Fresh or cryopreserved kidney tissue from goats suffering from heart water (Cowdria ruminantium infection) was infective when inoculated intravenously into susceptible goats. Primary kidney cell cultures were established from 14 goats reacting to C ruminantium infection; they were tested for infectivity by intravenous injection into susceptible goats after periods varying from five to 31 days. Three cultures, five, 12 and 13 days old, induced heartwater in recipients. The other 11 cultures, varying in age from six to 31 days, did not cause any reaction and all 11 recipient goats died from heartwater on challenge. C ruminantium could not be detected microscopically in the cell cultures.

Observations on mouse-infective stocks of Cowdria ruminantium: attempts to demonstrate antigenic changes of the Kwanyanga stock in mice.

Starting from a stabilate of caprine blood infected with the Kwanyanga stock of Cowdria ruminantium, eight serial passages were made in groups of mice, and eight parallel serial passages in goats. Cross-immunity tests in goats and mice failed to demonstrate any difference between stabilates made after the eighth passage. The Kwanyanga stock was of exceptionally low virulence for Dutch goats.

Theileria mutans in Nigeria.

Two strains of bovine Theileria from northern Nigeria were shown to be identical to Theileria mutans in the indirect immunofluorescent antibody test. One of the strains was transmitted by the tick Amblyomma variegatum; large macroschizonts, typical for T mutans, could be demonstrated in infected cattle. It is concluded from these experiments and from the literature that there is reliable evidence so far for the occurrence in Nigeria of only two bovine Tehileria species, T mutans and T velifera.

Antigenic diversity of Cowdria ruminantium isolates determined by cross-immunity.

Antigenic diversity in five stocks of the tick-borne rickettsia Cowdria ruminantium, the causal agent of heartwater disease of ruminants, was studied by cross-immunity trials in goats and sheep. Complete absence of cross-protection was found only between the Kümm and Kwanyanga stocks, and in all other combinations there were various degrees of cross-immunity. Immunological strain differences were more pronounced in goats than in sheep.

Artificial infection of Rhipicephalus appendiculatus with Theileria parva by percutaneous injection.

Rhipicephalus appendiculatus nymphs were inoculated with fresh or cryopreserved blood containing Theileria parva piroplasms, or with cell culture grown stages of T parva. The use of fresh blood was successful. Cryopreserved blood containing dimethylsulphoxide (DMSO), killed most nymphs after inoculation: DMSO could be removed by slow dialysis, without destroying the infectivity of the blood. Attempts to infect ticks by inoculating cell culture grown stages of T parva failed, even when large numbers of merozoites were present in the inoculum.

Theileria orientalis occurs in Central Africa.

An outbreak of bovine theileriosis in Burundi was investigated and the occurrence of Theileria orientalis in Central Africa was confirmed for the first time. Its identity was established on morphological and serological grounds. The parasite was associated with clinical signs and further complicates the disease picture of bovine theileriosis in Africa. Amblyomma variegatum is suspected as being the local vector.

Observations on mouse-infective stocks of Cowdria ruminantium: microscopical demonstration of the Kwanyanga stock in mouse tissue and the carrier-status of the Senegal stock in mice.

The behaviour of two different stocks of Cowdria ruminantium was investigated in mice. The mouse-pathogenic Kwanyanga stock of C ruminantium was microscopically demonstrated in mice in capillary endothelial cells of the lung, spleen, kidney, liver and brain. Mice of the ninth passage of the Senegal stock, which is infective but not pathogenic to mice, were kept alive for a year. Their blood and homogenised spleens, inoculated intravenously, caused fatal heartwater in a goat. However, the Senegal stock could not be demonstrated microscopically in mice. These results indicate the possible role of rodents in the epidemiology of heartwater.

Theileria orientalis, a cosmopolitan blood parasite of cattle: demonstration of the schizont stage.

Serological and morphological comparison of Theileria orientalis stocks from Australia, Britain, Iran, Japan and the USA with a more pathogenic stock from Korea, corresponding to T sergenti of Russian literature, showed that they all belong to one species, for which the name T orientalis is recommended. T orientalis is now known to occur on all continents. Macroschizonts and microschizonts, found in some of the calves infected with the Korean stock, are described and illustrated. Infections with the Korean stock were associated with early hyperthermia during the period schizonts are found, and commonly with high parasitaemias and anaemia even in unsplenectomised calves. The higher pathogenicity of this stock may be related to a faster rate of division. T orientalis may cause latent infection in sheep. With the exception of the stock from the USA, all stocks could be transstadially transmitted by Haemaphysalis longicornis and, or, H punctata.

Erythrocytic forms of Theileria velifera.

The enigmatic veil associated with the cattle parasite Haematoxenus veliferus has been shown to be crystallike in ultrastructure and to stain with benzidine like haemoglobin. The ultrastructure of the organism is similar to that of other theilerial parasites. As less distinct veil-like structures are known to occur with some other Theileria species, and in one case, have also been shown to be crystalline in structure, it is proposed to sink the genus Haematoxenus as a synonym of Theileria.

Observations on mouse-infective stocks of Cowdria ruminantium: attempts to demonstrate persistence of the organism in mice immune to the Kwanyanga stock.

Mice immunised against the Kwanyanga stock of Cowdria ruminantium by infection and treated with oxytetracycline proved immune to challenge on day 40 and also to a second challenge on day 125 after infection. Treatment with the experimental dithiosemicarbazone gloxazone on days 59 and 73 did not abolish immunity to challenge on day 125. No persistence of the organism in immune mice that had been challenged on day 40 could be demonstrated by subinoculating blood and liver homogenate on day 126. These results are different from findings reported elsewhere with the mouse-infective Kumm stock.