Bovine babesiosis (Babesia bovis infection) in Zambia

Summary

Two cases of Babesia bovis, a parasite associated with the tick Boophilus microplus, are reported for the first time from the central part of Zambia. It is concluded that infected B. microplus ticks are occasionally introduced into central Zambia by tick‐infested cattle from the north‐eastern part of the country where B. bovis is endemic. The spread of B. microplus in Southern Africa in a westward direction is discussed and related to the epidemiology of bovine babesiosis in Zambia.     

Use of cerebellar brain smears in the diagnosis of babesiosis (Babesia bovis) in cattle

Summary Cerebral and cerebellar smears were made from 4 animals acutely reacting toBabesia bovis and 94 animals free from clinical babesiosis. The brain smears were stained by the Giemsa method and examined for the presence ofB. bovis parasites. In animals showing clinical babesiosis capillaries congested with parasitised erythrocytes were abundant in cerebral and cerebellar smears. Results obtained from both types of brain smears in animals free from clinical babesiosis agreed closely (83% conformity) as to the presence or absence of parasites. A third group of 39 animals from which cerebral and cerebellar smears were taken was also examined serologically by the indirect immunofluorescent antibody test (IFAT); about 69% of the IFAT positive and doubtful animals showed parasites in the cerebellar brain smears. The existence of false negatives in the IFAT test has been shown and discussed. It has been concluded that cerebellar samples obtained through the foramen occipitale can be used for the microscopic detection ofB. bovis parasites in latently infected bovines. This method can also be used in field cases suspected of cerebral babesiosis permitting brain sampling without resorting to the opening of the skull. Such an approach might prove particularly useful in areas where rabies occurs and the animal's head has to be sent to a diagnostic centre.

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El Empleo De Frotis De Cerebelo En El Diagnostico De Babesiosis (Babesia Bovis) En Bovinos

Resumen Se hicieron frotis de cerebro y cerebelo de 4 animales con síntomas agudos de babesiosis porBabesia bovis y de 94 animales clínicamente sanos. Los frotis se tiñeron con Giemsa y se examinaron por la presencia deB. bovis. En animales enfermos, los capilares del cerebro y cerebelo se encontraron congestionados y repletos de eritrocitos parasitados. Los resultados obtenidos en ambos tipos de frotis en animales libres de la enfermedad clínica, estuvieron de acuerdo (83% de conformidad) en cuanto a la presencia o ausencia de parásitos. Un tercer grupo de 39 animales de los cuales se tomaron frotis de cerebro y cerebelo, se examinaron también serológicamente mediante la técnica indirecta de anticuerpos fluorescentes (TIAF); cerca del 69% de animales positivos y dudosos por immunofluorescencia revelaron parásitos en los frotis de cerebelo. La existencia de falsos positivos se demostró mediante TIAF. Se concluye que los frotis de cerebelo tomados a través del agujero occipital pueden usarse para detectarB. bovis microscópicamente en animales con infecciones latentes. Este método se puede utilizar en el campo para no tener que abrir el craneo, sobre todo en áreas en donde existe rabia y las cabezas se envían al laboratorio.

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Examen De Frottis De Matière Cervelette Pour Le Diagnostic De La Babésiose (babesia bovis) Du Bétail

Résumé Des frottis de cerveau et de cervelet ont été effectués à partir de 4 animaux atteints de babésiose aiguë et de 94 autres n'ayant présenté aucun symptôme de la maladie. Les frottis ont été colorés au Giemsa et examinés pour mettre en évidence la présence deB. bovis. Chez les animaux atteints de babésiose clinique, les capillaires étaient congestionnés avec présence abondante d'érythrocytes parasités. Il existe une excellente concordance (83 p.100) entre les résultats des examens des deux types de frottis, qu'il s'agisse d'absence ou de présence des parasites. Un troisième groupe de 39 animaux dont les frottis de cerveau et de cervelet ont été faits ont également fait l'objet d'une étude sérologique pour déceler des anticorps par le test de l'immunofluorescence indirecte (I.F.A.T.); environ 69 p.100 des animaux reconnus positifs et douteux par l'I.F.A.T. ont montré des parasites dans les frottis du cervelet. L'existence de cas négatifs faux dans le test IFAT a été montrée et discutée. Il en a été conclu que les échantillons de cervelet obtenus à travers le foramen occipital peuvent être utilisés pour la détection microscopique deB. bovis chez des animaux en état d'infection latente. Cette méthode peut être également utilisée en brousse lors de suspicion de babésiose cérébrale car elle permet d'obtenir les prélèvements de tissus cérébraux nécessaires sans ouverture de la boite cranienne. Une telle approche peut être particulièrement utile dans les régions où sévit la rage et où il est nécessaire d'expédier la tête des animaux suspects à un centre de diagnostic.

     

Bovine babesiosis: severity and reproducibility of Babesia bovis infections induced by Boophilus microplus under laboratory conditions

A total of 61 intact Holstein-Friesian calves were exposed to Babesia bovis (= B argentina) by the injection of infected blood or the application of infected Boophilus microplus larvae. Tick-induced infections were uniformly severe, even when induced by relatively small numbers of infected ticks. In contrast, calves infected with carrier blood experienced mild, subclinical reactions despite detectable parasitaemia. The greater severity of tick-induced reactions appeared to be due to the large number of infective doses injected by each infected tick rather than greater virulence of tick versus blood-origin babesiae. The severity of tick-induced babesiosis was related to the age of the experimental calves, with more severe reactions and high mortality occurring among older animals. The mean daily temperature rise increased with age and was highest in 15 animals dying of babesiosis. No relationship could be found between peripheral blood parasitaemia, packed cell volume reduction and mortality in tick-induced infections. One thousand larvae from infected colonies induced babesiosis in all of 29 animals exposed. Although larval transmission of B bovis is difficult to quantitate, there would seem to be no objection to using a controlled tick-borne challenge in the laboratory for assessment of susceptibility.     

用重组的牛巴贝斯虫棒状体蛋白1作为ELISA诊断抗原进行牛群中牛巴贝斯虫病的血清流行病学调查

用重组的牛巴贝斯虫棒状体蛋白1（Bc—RAP-1）作为ELISA诊断抗原,对采自青海省湟中县的120份奶牛血清样品,进行抗Bc—RAP-1特异性抗体的检测。结果检出7份阳性,阳性率为5．83％,说明该地区牛群存在牛巴贝斯虫的感染。     

Immune control of Babesia bovis infection

Babesia bovis causes an acute and often fatal infection in adult cattle, which if resolved, leads to a state of persistent infection in otherwise clinically healthy cattle. Persistently infected cattle are generally resistant to reinfection with related parasite strains, and this resistance in the face of infection is termed concomitant immunity. Young animals are generally more resistant than adults to B. bovis infection, which is dependent on the spleen. Despite the discovery of B. bovis over a century ago, there are still no safe and effective vaccines that protect cattle against this most virulent of babesial pathogens. Immunodominant antigens identified by serological reactivity and dominant T-cell antigens have failed to protect cattle against challenge. This review describes the innate and acquired immune mechanisms that define resistance in young calves and correlate with the development of concomitant immunity in older cattle following recovery from clinical disease. The first sections will discuss the innate immune responses by peripheral blood- and spleen-derived macrophages in cattle induced by B. bovis merozoites and their products that limit parasite replication, and comparison of natural killer cell responses in the spleens of young (resistant) and adult (susceptible) cattle. Later sections will describe a proteomic approach to discover novel antigens, especially those recognized by immune CD4+ T lymphocytes. Because immunodominant antigens have failed to stimulate protective immunity, identification of subdominant antigens may prove to be important for effective vaccines. Identification of CD4+ T-cell immunogenic proteins and their epitopes, together with the MHC class II restricting elements, now makes possible the development of MHC class II tetramers and application of this technology to both quantify antigen-specific lymphocytes during infection and discover novel antigenic epitopes. Finally, with the imminent completion of the B. bovis genome-sequencing project, strategies using combined genomic and proteomic approaches to identify novel vaccine candidates will be reviewed. The availability of an annotated B. bovis genome will, for the first time, enable identification of non-immunodominant proteins that may stimulate protective immunity.     

Experimental Babesia bovis infection in Holstein calves

One intact and two splenectomized Holstein calves were infected intravenously with a Mexican strain of Babesia bovis and killed following the onset of severe clinical disease. A light and electron microscopic study was conducted on selected tissues to examine the relationship between parasitized erythrocytes and microvascular endothelial cells. The pattern and degree of specific organ sequestration of parasitized erythrocytes was assessed and correlated to lesions. Red blood cells infected with Babesia bovis exhibited stellate membrane protrusions. This morphological change appeared to mediate erythrocyte sequestration in the microvascular and capillary beds of the brain, kidney, and adrenal gland by an as yet unknown mechanism(s).     

Heterologous strain immunity in bovine babesiosis using a culture-derived soluble Babesia bovis immunogen

The cross-protective capacity of culture-derived soluble immunogens against heterologous Babesia bovis strains from different geographical locations of Latin America was examined. Susceptible yearling cattle were either immunized with immunogens derived from Venezuelan or Mexican strains, or were administered a multi-component immunogen containing antigens of the Australian, Mexican and Venezuelan strains. Cattle were challenged with virulent B. bovis organisms of the Argentinian, Colombian, Ecuadorean, Mexican and Venezuelan strains. The major parameters used to evaluate cross-protection were the following: presence, level and duration of parasitemia; maximal PCV reduction; level and duration of fever; determination of fibrinogen and cryofibrinogen; homologous and heterologous antibody levels; and net gains in body weight. Results showed good protection with a Venezuelan B. bovis immunogen after homologous and heterologous challenge exposures. A low degree of cross-immunity was observed when cattle vaccinated with the Mexican immunogen were challenged with each of the heterologous strains.     

Genotypic diversity in field isolates of Babesia bovis from cattle with babesiosis after vaccination

Objective To determine whether particular genotypes of Babesia bovis were common to field isolates obtained from cattle properties in Queensland where the B bovis vaccine had apparently failed.
Design A comparative study of polymerase chain reaction genotypes in different populations of B bovis .
Procedure Two polymerase chain reaction assays were applied to analyse DNA extracts of B bovis vaccine (K, T and Dixie strains) and 27 field isolates from 24 properties where disease outbreaks had occurred despite the use of the vaccine. To evaluate the stability of the genotypes identified, 11 of the field isolates were inoculated into experimental cattle that had either been previously vaccinated with T strain or not vaccinated.
Results No particular genotype of B bovis was responsible for the problems observed in previously vaccinated herds. None of the isolates had genotypes identical to the vaccine strains used. No geographic trends among the genotypes were observed. Isolates that originated from the same property also had different genotypes. Blood passage of the 11 field isolates in either previously vaccinated or nonvaccinated cattle did not alter the original genotype.
Conclusion No particular genotypes identified by the Bv80 and BvVA1 polymerase chain reaction assays could be associated with vaccine failures.     

Bovine babesiosis in Colombia

Babesiosis is a tick-borne disease of cattle which occurs in many tropical and subtropical areas of the world. Despite the extensive investigations which have been carried out since the discovery of the organism (Babes, 1888) many problems of major importance remain to be solved in Babesia spp. — host complex. In Colombia (South America) the experiments were carried out to identify the existing Babesia spp. by morphologic and immunoserologic methods. The immunoserologic relationship of Babesia spp. were studied by several serologic techniques. Attempts were made to develop a sensitive and practical serologic test for diagnosis of latent Babesia spp. infections. Several groups of intact and splenectomized calves were inoculated with various antigens isolated from Babesia spp. infections and the response to vaccination, premunition and tick-borne challenge were studied. The second part of this investigation was mainly concerned with evaluating the system of chemoprophylaxis against Babesia spp. infections under actual field conditions.     

Proteomic profiling of Rhipicephalus (Boophilus) microplus midgut responses to infection with Babesia bovis

Differences in protein expression in midgut tissue of uninfected and Babesia bovis-infected southern cattle ticks, Rhipicephalus (Boophilus) microplus, were investigated in an effort to establish a proteome database containing proteins involved in successful pathogen transmission. The electrophoretic separation of midgut membrane proteins was greatly improved by using liquid-phase isoelectric focusing combined with one-dimensional or two-dimensional (2-D) gel electrophoresis. A selection of differentially expressed proteins were subjected to analysis by capillary-HPLC-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). Among the identified Babesia-affected tick midgut proteins were six proteins that are implicated in signaling processes, including three Ca(2+)-binding proteins, a guanine nucleotide-binding protein, a protein with signal peptide activity and a translocon-associated receptor protein. Up-regulation of five metabolic enzymes indicated parasite-induced changes in electron and proton transport, protein processing and retinoic acid metabolism. Among the down-regulated proteins were a molecular chaperone, a cytoskeletal protein and a multifunctional protein of the prohibitin family. Identification of these proteins may provide new insights into the molecular interactions between B. bovis and its tick vector, and could lead to identification of anti-tick and transmission-blocking vaccine candidates.     

Differential Bos taurus cattle response to Babesia bovis infection

Bovine babesiosis is a tick-borne disease caused by Babesia spp. haemoprotozoans. The disease is of great importance at tick enzootic unstable areas and hampers cattle production in several developing countries. The available immunisation alternatives are pre-immunition and attenuated vaccines. Despite being efficient and protective, they are unsafe as they use cattle blood cells as inoculum and may potentially spread other diseases. Another alternative to help in babesiosis control would be the identification of genetically resistant cattle to Babesia bovis infection. The objective of this work was to phenotype cattle based on primary response against B. bovis infection. Two-hundred and forty half-sib Hereford and Aberdeen Angus heifers (120 animals from each breed), 12-18-month-old na?ve cattle, originated from a tick-free area in Southern Brazil, were used in the experiment. Animals were monitored following an inoculation with 1x10(7)B. bovis parasitised erythrocytes. Results showed three different phenotypes: 1-'susceptible', animals with babesiosis clinical signs that received treatment to avoid death; 2-'intermediate', animals with clinical signs: parasitaemia, >or=21.5% reduction in packed cell volume (PCV) and increase in body temperature when compared to their pre-challenge physiological parameters, no specific treatment was needed as animals self recovered from the disease, and 3-'resistant', animals without clinical signs that showed B. bovis presence in blood smears, <21.5% PCV reductions, with little or no increase in body temperature and no need for babesiosis treatment. The frequencies of each phenotype were: 45.4, 26.7, and 27.9%, respectively, demonstrating the existence of phenotypic variation for B. bovis in Bos taurus cattle.     

Evaluations of buparvaquone as a treatment for equine babesiosis (Babesia equi)

We evaluated the efficacy of buparvaquone in eliminating Babesia equi of European origin in carrier horses and in experimentally infected splenectomized ponies. When administered at the rate of 2.5 mg/kg of body weight, IM, 4 times at 96-hour intervals, buparvaquone was effective in eliminating B equi carrier infection in 1 horse. Such results could not be repeated at the same dosage or at 3.5 or 5 mg/kg, IM. Buparvaquone given at the rate of 4 to 6 mg/kg IV and/or IM was therapeutically effective in 4 of 5 acute B equi infections in splenectomized ponies. The treated ponies became carriers.     

Bovine babesiosis in the European community

There are apparently three species of bovineBabesia within the European Community-B. divergens,B. major andB. bovis. The former is the principal cause of disease, but lack of information about the other two, particularly about the ability of the tickIxodes ricinus to act as their vector, makes further investigation essential. The potential hazard of indiscriminate movement of cattle within the E.C. is discussed, and a plea is made for the introduction of routine serological screening of cattle before their export.

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Kurzfassung In der Europäischen Gemeinschaft scheint es drei Arten vonRinderbabesia zu geben,B. divergens,B. major undB. bovis. Die erste Art ist die Hauptkrankheitsursache, aber mangelnde Kenntnisse über die anderen beiden, besonders über die Fähigkeit der ZeckeIxodes ricinus als übertrager zu dienen, lässt weitere Untersuchungen wesentlich erscheinen. Die potentiellen Gefahren durch uneingeschränkte Viehbewegungen innerhalb der EG werden erörtert, und es wird für die Einführung routinemässiger serologischer Untersuchungen der Tiere vor ihrer Ausfuhr plädiert.

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Resume Il existe apparemment trois sortes debabésia bovine dans la Communauté Européenne-B. divergens,B. major etB. bovis. La première est la cause principale de la maladie, mais le manque d'informations concernant les deux autres, particulièrement la capacité de la tiqueIxodes ricinus d'agir en tant que vecteur de ces maladies, rend des recherches complémentaires absolument nécessaires. Le danger virtuel que constituent les mouvements désordonnés de bétail à l'intérieur de la Communauté Européenne fait l'objet d'un discussion, et il est préconisé l'analyse systématique du sérum du bétail avant son exportation.

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Riassunto Nella Comunità europea pare esistano tre specie diBabesia bovina cioè-B. divergens,B. major eB. bovis. Principale causa di malattia è la prima, ma la mancanza di sufficienti informazioni sulle altre due, soprattutto riguardo al ruolo della zeccaIxodes ricinus come vettore, richiede lo svolgimento di ulteriori ricerche. I rischi potenziali inerenti al movimento indiscriminato dei bovini nella C.E. sono oggetto di discussione e si suggerisce l'istituzione di regolari controlli sierologici dei bovini prima che gli stessi vengano esportati.

     

Involvement of a host erythrocyte sialic acid content in Babesia bovis infection

Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant alpha2-3-linked SA residues but not alpha2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of alpha2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining with the alpha2-3-linked SA-specific lectin: a selective invasion of B. bovis was found only in the intact RBCs bearing the SAs, but not in the desialylated RBCs. Furthermore, a significant reduction of the parasitized RBCs was also observed in the culture supplemented with exogenous 3'-sialyllactose containing the alpha2-3-linked SAs. However, the complete inhibition of parasite proliferation was not achieved in the culture. These findings indicate that while the alpha2-3-linked SA-dependent pathway is needed for highly efficient invasion of host RBCs by B. bovis, there might also be other potential alternative pathways.     

Epidemiology of bovine babesiosis and anaplasmosis in Zambia

The serological prevalence of bovine babesiosis and anaplasmosis in the traditional farming sector of six provinces of Zambia was determined using the indirect fluorescent antibody test (IFAT) for babesiosis and the card agglutination test (CAT) for anaplasmosis. Antibodies to Babesia bigemina occurred throughout the country whereas the prevalence of B. bovis followed the distribution of its tick vector Boophilus microplus which is limited to the north-eastern part of the country. Low numbers of B. bovis serologically positive cattle were demonstrated in central and southern Province. Anaplasma spp. occurred throughout Zambia but the overall percentages of positive sera were low ranging between 14.7% and 38.6% using the CAT. Two hundred sera were retested for anaplasmosis using an enzyme-linked immunosorbent assay (ELISA). Sero-prevalence rates were 1.5 to 2.3-fold greater with the ELISA than with the card agglutination test.