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Biomedical subjects

M Ristic

Publications and source records attributed to M Ristic.

At least 37 records · Page 2Linked to original sources

Unexplained febrile illnesses after exposure to ticks. Infection with an Ehrlichia?

The Ehrlichia are tick-borne rickettsial organisms that cause disease in animals throughout the world but that have been previously recognized as human pathogens only in Asia. We have identified six patients with serological evidence of recent infection with an Ehrlichia: a fourfold or greater rise or fall in titer to Ehrlichia canis. All of the patients reported recent tick bites. Rigors, myalgia, headache, nausea, and anorexia were each reported by five patients. Fever was present in all patients and was accompanied by relative bradycardia and leukopenia in five patients, thrombocytopenia and abnormal liver function test results in four, and anemia in three. Five of the six patients were treated with tetracycline hydrochloride, and all recovered. Infection with Ehrlichia should be considered in patients with unexplained febrile illnesses after tick exposure.

Adult

Bovine babesiosis: induction of protective immunity with culture-derived Babesia bovis and Babesia bigemina immunogens.

The comparative protective efficacy of culture-derived Babesia bovis and B. bigemina immunogens against challenge exposure of susceptible crossbred cattle to heterologous strains was examined and correlated with the antigenic threshold requirements for induction of protective immunity. Strong protection was observed in animals vaccinated with 10 ml-equivalent doses of soluble, B. bovis exoantigen-containing supernatant fluids. Similar protective responses to B. bigemina exoantigens were evident even at 1 ml-equivalent dosages. In addition, the efficacy of a combined B. bovis-B. bigemina immunogen was assessed with a dose-response analysis in highly susceptible, purebred cattle. Vaccinated animals were protected against clinical babesiosis, and significant weight gains were recorded after challenge infection with virulent parasites.

Animals

Antigenic relationship between Plasmodium falciparum and Babesia bovis: reactivity with antibodies to culture-derived soluble exoantigens.

Antigenic similarities between Plasmodium and Babesia parasites of the phylum Apicomplexa have been previously demonstrated primarily by the serological cross reactivity observed in the indirect fluorescent antibody (IFA) test. We have now studied the antigenic relationship between the human malaria parasite, Plasmodium falciparum, and the hemoparasitic agent of cattle, Babesia bovis, using rabbit monospecific antibodies produced against individual culture-derived P. falciparum polypeptides and bovine polyspecific antibodies to B. bovis exoantigens. These respective antibodies were found to be distinctly cross reactive in the IFA test using infected erythrocytes (squirrel monkey--P. falciparum; bovine--B. bovis) as antigen substrates. Immunofluorescence was shown to be highly specific for parasite surfaces. Additionally, the degree of reactivity with soluble exoantigens contained in Plasmodium and Babesia culture supernatants was monitored by a two-site enzyme immunoassay employing the cross-reactive antibodies. Further evidence for antigenic cross reactivity between P. falciparum and B. bovis parasites was shown with the in vitro inhibition assay. Antibodies to P. falciparum and B. bovis were found to be highly inhibitory for the in vitro growth of P. falciparum in human erythrocytes.

Animals

Detection of culture-derived Babesia bovis exoantigen using a two-site enzyme immunoassay.

Soluble exoantigens in the supernatants of Babesia bovis cultures have been shown to be efficient immunogens against bovine babesiosis. We used a two-site enzyme immunoassay to monitor the release of these antigens during in vitro cultivation. Bovine immunoglobulin G was isolated from serum of an adult cow previously immunized with culture-derived B. bovis exoantigens and challenged via needle with virulent parasites. The specific immunoglobulin G was used as a capture antibody and as an enzyme-conjugated recognizing antibody. The optimal protein concentration of capture antibody was 10 micrograms/ml. The 24-h cultures showed the greatest antigen concentration. The test was sensitive for detection of differences in species-specific antigenic activity among B. bovis isolates, for determining loss of antigenicity during storage and formalinization, and for monitoring the kinetics of exoantigen release during in vitro cultivation. Antigens cross-reactive with the other major Babesia species of cattle, Babesia bigemina, were also detected with this assay. The high specificity, sensitivity, and reproducibility of this technique should facilitate detection and quantitation of Babesia antigens during purification and in standardization of candidate immunogens.

Animals

Development of Dot-enzyme immunoassay for diagnosis of canine babesiosis.

A reagent conservative Dot-enzyme immunoassay (Dot-EIA) was developed primarily for canine babesiosis caused by Babesia canis. The technique is simple, specific, and sensitive. All steps were carried out at room temperature. Strong agreement was observed between Dot-EIA and the conventionally used indirect immunofluorescence test. The procedure is adaptable to other protozoal disease, e.g., bovine babesiosis and human malaria.

Animals

In vitro and in vivo adaptation of the Geneve/SGE-1 strain of Plasmodium falciparum to growth in a squirrel monkey (Saimiri sciureus) model.

Human erythrocytic culture-adapted parasites of the Geneve/SGE-1 strain of Plasmodium falciparum were successfully adapted to grow in an in vitro culture system containing squirrel monkey erythrocytes and serum. These monkey culture-adapted organisms were then used to produce a patent infection in a splenectomized squirrel monkey. Fresh infected blood from this animal was introduced into another splenectomized monkey and was subsequently serially passed through seven intact squirrel monkeys. High level parasitemias (greater than 10%) were obtained in the animals from the last two passes following inoculation of moderate numbers of parasites. It is anticipated that this squirrel monkey-adapted Geneve/SGE-1 strain of P. falciparum will continue to produce high level parasitemias in intact Bolivian Saimiri, and consequently will be suitable for challenge of these monkeys.

Animals

Pathologic changes associated with fatal Plasmodium falciparum infection in the Bolivian squirrel monkey (Saimiri sciureus boliviensis).

Fatal cases of experimental Plasmodium falciparum (Indochina I) in Bolivian squirrel monkeys (Saimiri sciureus boliviensis) were examined by histologic and ultrastructural methods. Gross lesions were characterized by hepatosplenomegaly and interstitial pulmonary changes. Histologically, there was marked diffuse reticuloendothelial hyperplasia, pulmonary alveolar septal thickening, mesangioproliferative glomerulonephropathy, sequestration of parasitized erythrocytes in deep vascular beds, degenerative parenchymal changes in the liver and myocardium, and in one case retinal and cerebral hemorrhage. These data indicate that the Bolivian squirrel monkey is a good model for studying pathologic changes associated with human falciparum malaria.

Animals

Correlative clinical biochemistry and hematological profiles of laboratory-bred Bolivian squirrel monkeys (Saimiri sciureus).

Twenty-five clinical and biochemical parameters were determined on 17 normal, male, laboratory-bred Bolivian squirrel monkeys (Saimiri sciureus). Parallel hematologic parameters were conducted. The correlation of biochemical and hematological observations and the distribution characteristics, range, and standard error of the mean were determined. The purpose of the present communication is to establish baseline biochemical values for laboratory-bred squirrel monkeys and to present a correlative comparison between selected cellular elements and major blood electrolytes.

Animals

Diagnosis of equine monocytic ehrlichiosis (Potomac horse fever) by indirect immunofluorescence.

The recent establishment of a system for the continuous in vitro propagation of Ehrlichia risticii, the causative agent of equine monocytic ehrlichiosis (EME; synonym, Potomac horse fever), has facilitated the development of an indirect fluorescent antibody test for the diagnosis of this disease under laboratory and field conditions. The field diagnostic application of the test has aided in the recognition of the disease in 16 states of the United States and in 1 province of Canada. A limited epidemiologic study conducted between January and September 1985, in an area where the disease is known to be enzootic, revealed that conversion from seronegative to seropositive status is not always accompanied by clinical manifestations of the disease. Confirmatory findings in experimentally inoculated horses suggest the existence of clinically undetectable infections.

Animals

Isolation, experimental transmission, and characterization of causative agent of Potomac horse fever.

Potomac horse fever, a disease characterized by fever, anorexia, leukopenia, and occasional diarrhea, is fatal in approximately 30 percent of affected animals. The seasonal occurrence of the disease (June to October) and evidence of antibodies to the rickettsia Ehrlichia sennetsu in the serum of convalescing horses suggested that a related rickettsia might be the causative agent. Such an agent was isolated in cultured blood monocytes from an experimentally infected pony. This intracytoplasmic organism was adapted to growth in primary cultures of canine blood monocytes. A healthy pony inoculated with these infected monocytes also developed the disease. The organism was reisolated from this animal which, at autopsy, had pathological manifestations typical of Potomac horse fever. Cross serologic reactions between the newly isolated agent and antisera to 15 rickettsiae revealed that it is related to certain members of the genus Ehrlichia, particularly to Ehrlichia sennetsu. Since the disease occurs in other parts of the United States as well as in the vicinity of the Potomac River, and since it has also been reported in Europe, the name equine monocytic ehrlichiosis is proposed as being more descriptive.

Animals

Seroepidemiology of bovine anaplasmosis and babesiosis in Venezuela.

The serological prevalence of bovine anaplasmosis and babesiosis in the Centro-Occidental region of Venezuela was determined using the indirect fluorescent antibody (IFA) and latex agglutination (LAT) tests. Anaplasma marginale IFA and LAT prevalence rates were 57.7 and 48.6% respectively for cattle of all ages and breeds. The respective prevalence of IFA activity to B. bigemina and B. bovis was 78.2 and 38.8%. The LAT test specific only for Babesia spp. revealed that 61.4% of the animals tested were serologically positive for Babesia parasites. Anaplasma marginale and B. bigemina organisms were detected in peripheral blood smears of 21.1 and 7.5% respectively of cattle surveyed. A longitudinal survey of antibody activity and incidence of parasitaemia for Anaplasma and Babesia infections was conducted in two groups of calves from their birth to seven months of age. The animals generally experienced infections between three and four months of age the period subsequent to decline in colostral antibody levels. Seroepidemiological data are considered in the formulation of proposed vaccination regimens for anaplasmosis and babesiosis.

Anaplasmosis

Purification and characterization of culture-derived exoantigens of Plasmodium falciparum.

An 83 kDa glycoprotein and a 100 kDa glycoprotein have been purified from the supernatant fluid of in vitro cultures of Plasmodium falciparum by conventional cation-exchange liquid chromatography, size exclusion high performance liquid chromatography, and anion-exchange high performance liquid chromatography. Both proteins exist as dimers in the native state and have been identified as parasite antigens by Western immunoblotting and by their specific reactivity in the indirect enzyme-linked immunosorbent assay. The N-terminal amino acid sequence of these two proteins has been determined and they are at least 90% homologous. The use of monospecific rabbit antisera raised against the individual pure proteins confirm their cross-reactivity. We postulate that the 83 kDa protein is a specific processing product of the larger 100 kDa protein. The presence of these proteins in the culture supernatant suggests they could both be derived from the merozoite surface coat and are potential protective antigens.

Amino Acid Sequence

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.

Animals