[Eperythrozoon parvum (Rickettsiales) in swine in the Netherland].
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Anaplasmataceae Infections↗
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Experimental infections and natural outbreaks of eperythrozoonosis in pigs identified by PCR-DNA hybridizations.
Eperythrozoon-specific DNA amplification reactions and subsequent hybridizations with an eperythrozoon DNA probe (KSU-2) were used in experimental infection studies to identify Eperythrozoon suis DNA in the blood of splenectomized and nonsplenectomized pigs. The results indicate that E. suis DNA is present in nonsplenectomized pigs at levels that can be amplified by polymerase chain reaction (PCR) and identified in DNA hybridizations within 24 hours after infection. The ability of the E. suis PCR/hybridization assay to detect eperythrozoonosis was further demonstrated in blood samples collected from pigs in 2 separate natural outbreaks in Oklahoma. Results from these initial samplings indicate that pigs infected with E. suis from geographically distinct locations can be identified using the eperythrozoon-specific PCR/hybridization assay, which offers many advantages over conventional laboratory procedures for diagnosing eperythrozoonosis in pigs.
Animals↗
On molecular taxonomy: what is in a name?
Gene sequences of small portions of the genome are often used for premature detailed taxonomic changes, neglecting polyphasic taxonomy, which should also consider phenotypical characteristics. Three examples are given: (i) Recently, members of the genera Eperythrozoon and Haemobartonella have been moved, correctly so, from the Rickettsiales to the Mycoplasmatales, but were assigned to the genus Mycoplasma, mostly on the basis of 16S rRNA sequence analysis. Not only is the 16S rRNA sequence similarity between 'classical' Mycoplasma and these species of Eperythrozoon and Haemobartonella less than that between some other well-recognised bacterial genera, but their biological differences amply justify their classification in different genera of the Mycoplasmatales. Furthermore, the move creates considerable confusion, as it necessitates new names for some species, with more confusion likely to come when the 16S rRNA sequences of the type species of Eperythrozoon, a name which has priority over Mycoplasma, will be analysed. (ii) In the Rickettsiales, members of the genera Anaplasma, Ehrlichia, Cowdria, Neorickettsia and Wolhbachia are so closely related phylogenetically on the basis of 16S rRNA sequences, and for some also of groESL operon sequences, that they have recently been fused, correctly so, into one family, the Anaplasmataceae, while the tribes Ehrlichieae and Wolbachieae have been abolished. Sequence diversity within the 'classical' genus Ehrlichia has led to classifying E. phagocytophila (including E. equi and the agent of human granulocytic ehrlichiosis), E. platys and E. bovis in the genus Anaplasma, while others have been retained in Ehrlichia, which also includes Cowdria ruminantium. E. sennetsu and E. risticii have been transferred to the genus Neorickettsia. 16S rRNA and GroEL sequences of 'classical' Anaplasma and some members of 'classical' Ehrlichia do show a close relationship, but differences in citrate synthase gene sequences, the GC content of this gene, and sequences of the gene encoding the beta-subunit of RNA polymerase, not to speak of the phenotypical differences, do not justify the fusion into one genus. Because of the phylogenetical diversity in Ehrlichia it is recommended that a new genus name be created for the E. phagocytophila genogroup (and E. platys and E. bovis). (iii) One of the conclusions of studies on the phylogeny of ticks of the subfamilies Rhipicephalinae and Hyalomminae, based on nucleotide sequences from 12S rRNA, cytochrome c oxidase I, the internal transcribed spacer 2, 18S rRNA, as well as morphological characters, is that Boophilus should be considered as a subgenus of Rhipicephalus. While Boophilus and Rhipicephalus are undoubtedly close, the obviously important morphological and biological differences between the genera Rhipicephalus and Boophilus are thus overruled by similarities in the sequences of a number of genes and this leads to considerable confusion. Polyphasic taxonomy amply justifies maintaining Boophilus as a separate genus, phylogenetically near to Rhipicephalus. This note is a plea for a cautious and balanced approach to taxonomy, taking into account molecular genotypical information, as far as is possible from different genes, as well as phenotypical characteristics.
Anaplasmataceae↗
[An epidemic investigation of eperythrozoonsis in Shandong province].
OBJECTIVE: To learn the status of infection of eperythrozoonsis in Shandong province. METHODS: Seven hundred and seventy-six human blood samples, 198 domestic animal blood samples and 114 wild rat blood samples were collected. Eperythrozoon was detected in the human and animal blood stain slides under microscope. RESULTS: Overall infection rate of Eperythrozoon in human population was 7.3%. Rates were significantly different (P < 0.01) in different districts but no significant difference was found between men and women (P > 0.05). The rate among adults was higher than that of teenagers and elderly. It was also noticed that the rates of people affected with chronic diseases as hepatitis, diabetes and arthritis were higher than those without. The infection rates of goat, cows and rats were 15.6%, 3.2% and 0, respectively. CONCLUSION: Data showed that Eperythrozoon infection was intensively identified in the people and animals in Shandong province.
Adolescent↗
[Disseminated intravascular coagulation in eperythrozoonosis of swine].
Investigations were carried out on the influence of latent and clinically manifest Eperythrozoon suis infection upon haemostasis in swine. The study was carried out with 14 German Landrace pigs. Latent eperythrozoonosis was induced in 7 animals by experimental infection. Splenectomy of these 7 animals and 2 spontaneously infected pigs led to clinical manifestation of eperythrozoonosis. Five clinically healthy pigs were splenectomized and served as controls. In healthy pigs splenectomy was followed by a transient rise in fibrinogen and platelet count. Latent infection with Eperythrozoon suis did not cause an impairment of haemostasis. Acute eperythrozoonosis was associated with increased haemorrhagic tendency considered to be a consequence of intravascular coagulation and subsequent consumption coagulopathy. There was a prolongation of partial thromboplastin time and prothrombin time (Quick) and a decrease of platelet count. Thrombelastography showed prolongation of reaction and clot building time and a short-term decrease of maximum amplitude. Deviation from normal values was proportional to the number of red blood cells infected with Eperythrozoon suis. Anti-rickettsial therapy led to quick normalization of haemostasis. Various aspects of the cause and the consequences of the haemostatic defect are discussed with special regard to the underlying disease.
Anaplasmataceae Infections↗
Studies on the blood parasites of pigs in Ibadan, Nigeria.
The blood parasites in the blood of local pigs kept under free range, and exotic pigs kept under intensive management of Ibadan were investigated. The species found were Babesia trautmanni, B. perroncitoi, Eperythrozoon suis and E. parvum. Eperythrozoon suis was the most predominant in both breeds. Most of the local pigs carried mixed infections, while the same number of the exotic pigs carried single and mixed infections. Mixed infection of individual pigs with all four blood parasites was common. The parasitaemia was generally low in pigs of both breeds.
Anaplasmataceae Infections↗
Comparison of blood smear and indirect fluorescent antibody techniques in detection of haemoparasite infections in trade cattle in Nigeria.
The incidence of blood parasites in trade cattle was surveyed with emphasis on tick-borne parasites, using blood smears and immunofluorescent antibody (IFA) techniques. With the blood smear method, about 9 and 8.9% of cattle examined were found positive for Babesia bigemina and Anaplasma marginale, respectively. Percentage infections with other parasites were 3.33, 1.92, 0.75, 0.75 and 0.58, respectively, for Babesia bovis, Trypanosoma brucei, Anaplasma centrale, Eperythrozoon and Theileria species as well as Trypanosoma congolense. The incidence of A. marginale infection was at its peak during the rainy season while B. bigemina was most prevalent during the dry season. There were mixed infections of Anaplasma and Babesia (1.42%); Babesia and trypanosomes (1.00%); Babesia and Eperythrozoon (0.75%) and Babesia and Theileria (0.75%). Using the indirect fluorescent antibody test, 93, 55 and 68% of cattle sera examined were found to be positive for B. bigemina, B. bovis and A. marginale, respectively. Forty-nine percent of the positive sera of B. bigemina had highest titres. The importance of using serological means for determining the endemic levels of tick-borne diseases in cattle in Nigeria is discussed.
Anaplasmosis↗
Development of a recombinant Anaplasma marginale DNA probe.
An Anaplasma marginale DNA probe has been developed by using an improved method for the isolation of genomic DNA. Purified genomic A. marginale DNA from the St. Croix isolate was partially digested with Sau 3A1 into fragments (greater than or equal to 5.0 kb). The restriction fragments were cloned using standard techniques in the pBR322 vector and used to transform E. coli (DH5) host cells. The recombinant A. marginale DNA library was screened by the colony lifting procedure. Colonies containing plasmids with A. marginale DNA inserts were identified by hybridization with a genomic A. marginale DNA radiolabeled probe (32P). Seven recombinant A. marginale DNA probes were evaluated by dot-blot in vitro hybridization assays to identify candidates as diagnostic tools in bovine anaplasmosis studies. Specificity and sensitivity experiments were carried out by using heterologous and homologous DNAs. The heterologous panel contained bovine DNA (WBC) and blood parasites DNA from Babesia bovis (Bb), Babesia bigemina (Bbi), Eperythrozoon suis (Es) and Eperythrozoon wenyoni (Ew). The homologous DNA panel included A. marginale DNAs of 12 different isolates which were isolated in the Caribbean, Mexico, and the U.S.A. The selected diagnostic probe was identified as pSt. Croix A1, and labeled with 32P by using in vitro nick translation and random primer techniques. The pSt. Croix A1 probe demonstrated 100% specificity and high sensitivity by hybridization in dot blotting and Southern blotting. The probe can detect 500-1000 infected erythrocytes per microliters which corresponds to a parasitemia of less than 0.01%. The A. marginale DNA insert was approximately 6.4 kb in size and a partial restriction map has been constructed.
Anaplasma↗
Tick-borne diseases of sheep and goats and their related vectors in Iran.
Haemoparasitic diseases have long been considered as a major problem to efficient sheep and goats production in Iran. Theileriosis due to Theileria hirci and babesiosis due to Babesia ovis and B. motasi are the most pathogenic protozoa. B. crassa, Anaplasma ovis and Eperythrozoon ovis are usually non-pathogenic and do not cause any apparent problem. The major tick genera found on sheep and goats are Hyalomma, Rhipicephalus, Haemaphysalis, Ixodes and rarely Dermacentor distributed in all part of Iran. Our studies indicate that only Hyalomma ticks transmit Theileria species but the others transmit Babesia, Anaplasma and Eperythrozoon. The two latter ones can also be transmitted through some biting flies and mechanical means. Control methods presently available in Iran rely almost entirely on: (i) vaccination of sheep and goats with cell culture vaccine for theileriosis; (ii) chemotherapeutic treatment for babesiosis and anaplasmosis; (iii) acaricides for control of tick vectors.
Anaplasmosis↗
Theileria sergenti and T. buffeli: polymerase chain reaction-based marker system for differentiating the parasite species from infected cattle blood and infected tick salivary gland.
Benign Theileria species in cattle. Theileria sergenti and T. buffeli, are morphologically indistinguishable. The polymerase chain reaction (PCR) was used to amplify the genes encoding the 33- and 34-kDa major piroplasm antigens (p33/34) of T. sergenti and T. buffeli from cattle blood infected with these parasites and tick salivary gland infected with T. sergenti. Following amplification, the p33 gene from T. sergenti and the p34 gene from T. buffeli were clearly differentiated using the restriction enzyme sites that were not shared between them. The oligonucleotide primer set, designed from the p33/34 genes, was specific for these Theileria species, since no amplification was detected with DNA from Babesia ovata, B. bovis, Anaplasma marginale, A. centrale, Eperythrozoon wenyoni, bovine white blood cells, and uninfected tick salivary glands. One tenth vol of the template prepared from either 25 microliters of blood with 0.5% parasitemia or individual tick salivary glands with six infected acini allowed sufficient amplification for differentiation of the two parasite species by restriction enzyme digestion. In addition, this system could be used to demonstrate the simultaneous, experimentally induced infection of cattle with T. sergenti and T. buffeli. The PCR-based marker system therefore provides a means to differentiate T. sergenti from T. buffeli in infected cattle blood and infected tick salivary glands. This system may also be useful for the characterization of other benign Theileria species in cattle.
Animals↗
Enzyme-linked immunosorbent assay using solubilised antigen for detection of antibodies to Anaplasma marginale.
An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against Anaplasma marginale. A marginale bodies were separated from parasitised erythrocytes by a modified nitrogen decompression method, sonicated, then solubilised with Triton X-100 and used as the ELISA antigen. In this ELISA system the required amount of antigen protein was 16.2 ng for each well. In the course of experimental infections, of calves, significant antibody levels were detected by ELISA and the complement fixation test (CFT) at almost the same time. Antibodies against A. marginale were detectable for longer periods using the ELISA than using the CFT. Sera from calves infected with Babesia bigemina, B. bovis, B. ovata, Theileria sergenti and Eperythrozoon wenyoni gave no reaction; however, antisera against A. centrale did react with the A. marginale ELISA antigen.
Anaplasma↗
The use of tick transmission by Boophilus microplus to isolate pure strains of Babesia bovis, Babesia bigemina and Anaplasma marginale from cattle with mixed infections.
Pure strains of Babesia bovis, Babesia bigemina and Anaplasma marginale were isolated from cattle infected with all 3 species as well as Theileria sp. and Eperythrozoon teganodes, using only transmission by the tick, Boophilus microplus. Unengorged adult ticks transferred to susceptible cattle transmitted A. marginale, but not Babesia. Engorged adults gave rise to progeny that transmitted Babesia, B. bovis by larvae and B. bigemina by male ticks. The Theileria and E. teganodes were not transmitted by the ticks and thus did not appear in calves used for isolating the pure strains of Babesia and A. marginale.
Anaplasma↗
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.
Animals↗
Incidence and prevalence of tick-borne haemoparasites in domestic ruminants in Ghana.
Giemsa-stained thin blood smears prepared monthly from cattle, sheep and goats in the Greater Accra region of Ghana between May 1994 and December 1996 were examined for presence of tick-borne haemoparasites. The majority of animals were less than 2 months old at the start of the survey. Monthly and cumulative incidences are presented of Anaplasma sp., Babesia bigemina, Borrelia sp., Eperythrozoon sp., Theileria mutans and Theileria velifera in cattle, Anaplasma sp., Borrelia sp., and Theileria sp. in sheep, and Anaplasma sp. in goats. T. mutans was the commonest parasite in cattle, with 100% incidence in calves by 10 months of age, and Anaplasma was commonest in small ruminants. The relative prevalence of these haemoparasites in blood smears from cattle, sheep and goats sampled on a single occasion at sites in all 10 regions of Ghana was found to be similar, though actual infection rates were lower. Packed cell volume (PCV) measurements from the sampled animals are also presented; no seasonal trends were evident in the PCV of the cattle, sheep and goats sampled monthly. In animals sampled on a single occasion, mean PCV was significantly higher in cattle and sheep without detectable haemoparasite infection, and in cattle was lowest in animals positive for both Babesia and Anaplasma, while there was no difference in mean PCV levels between parasitised and non-parasitised goats.
Anaplasmosis↗
The isolation and characterization of a Babesia from red deer (Cervus elaphus).
On three occasions, antibody positive blood from wild red deer produced overt infections with Babesia when inoculated into splenectomized red deer. One of the deer also became infected with Eperythrozoon sp. Babesia divergens, B. capreoli and the Babesia of red deer are morphologically similar and the marginal position of the parasites in the host cell is characteristic. Babesia were not seen and no antibody was formed in five out of six splenectomized bovine calves which were injected with parasitaemic red deer blood. Two of these calves when challenged with B. divergens were fully susceptible. A transient infection with the deer Babesia may have occurred in the sixth calf since antibody was detected and the animal resisted challenge with B. divergens. In indirect fluorescent antibody tests there was little or no difference in the titre of sera from naturally or experimentally infected cattle and deer when reacted with B. divergens or the red deer Babesia antigens. Despite their similarities, specific status for B. divergens and the red deer Babesia is probably justified; at present there is insufficient evidence to justify separation of the red deer Babesia from B. capreoli.
Animals↗
Investigations on a Babesia isolated from Scottish sheep.
The pathognomic, serologic and morphometric identity of a Babesia sp. recently isolated from sheep in Argyllshire was studied. Initially, problems were encountered due to the occurrence of Eperythrozoon ovis as a contaminant of the isolate and treatment with neoarsphenamine failed to remove it. Finally, a combination of oxytetracycline therapy and rapid passage allowed isolation of the Babesia sp. and its preservation in the form of a glycerolized, infected blood stabilate. Inoculation of the stabilate into splenectomized sheep produced a mile non-pyrogenic disease. Serologically the parasite was seen to have no affinity with either of the two Babesia recognized as parasitizing sheep, but some affinity with the cattle parasite B. divergens. Morphometrically it was seen to have affinity with the deer parasite B. capreoli and with the Babesia sp. isolated from red deer in Scotland, and the low number of budding forms of the parasite seen suggested that the parasite might be reproducing slowly in an alien host. It was concluded that the parasite was likely to be B. capreoli derived from red deer via the tick Ixodes ricinus.
Animals↗
The indirect fluorescent antibody test for bovine anaplasmosis.
An indirect fluorescent antibody test was used successfully for the serodiagnosis of experimental Anaplasma infections in cattle. Specific antibodies were detected three to ten days after anaplasma bodies were found in the blood, and persisted at least 15 weeks post-infection. An American and an African stock of A. marginale were used to prepare antigens, and gave comparable results when tested on sera positive to either of these stocks, as well as to an A. central-like stock from Korea. There were no cross-reactions with several Theileria, Babesia, Trypanosoma and Eperythrozoon species.
Anaplasma↗
Acute hepatitis associated with mouse leukemia. V. The neurotropic properties of the causal virus.
Observations on the behavior of MHV (Pr) in the cerebral tissue of Princeton and Swiss weanling mice indicated a limited neurotropism. The virus migrated to the brain on intraperitoneal injection and was established there by cranial passage, though with difficulty in Swiss mice. Intracerebral multiplication was rarely followed by outward signs of nervous disorder. A slight pathologic reaction occurred in the brains of intracerebrally injected Princeton mice, but it was negligible compared with that of the ensuing hepatitis. In Swiss mice, injected intracerebrally with a mixture of MHV (Pr) and Eperythrozoon coccoides, a related virus with restricted pathogenicity and host range, possibly a mutant, was isolated from the liver and brain. MHV (C), an actively hepatotropic virus recovered from leukemic Balb C mice, was much more neurotropic than MHV (Pr). Intracerebral injection of Balb C and Swiss weanling mice was attended by marked leptomeningeal and encephalitic lesions. Paralysis of the extremities occurred in some of the animals. The virus was essentially inactive in Princeton mice. During the intracerebral passage of MHV (C) in Swiss mice a pleuropneumonia-like organism was isolated from the brain. In conjunction with the virus this organism produced a vigorous leukocytic reaction.
Animals↗