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Biomedical subjects

M Ristic

Publications and source records attributed to M Ristic.

At least 19 recordsLinked to original sources

Hemophagocytic syndrome: a cause of pancytopenia in human ehrlichiosis.

A 67-year-old white man with human ehrlichiosis infection complicated by pancytopenia, hemophagocytic syndrome, disseminated intravascular coagulopathy and septic shock is presented. The patient had been on a three-week camping trip to California, Colorado, Utah, and New Mexico. The diagnosis of human ehrlichiosis was confirmed by sixteen-fold rise in antibody titer to Ehrlichia canis, and supported by the characteristic cytoplasmic inclusions. Human ehrlichiosis should be considered in the differential diagnosis in patients with fever and cytopenia associated with hemophagocytosis. Pancytopenia associated with ehrlichiosis is transient; however, it may be severe, and appears to be associated with destruction of normal blood elements.

Aged

An overview of research on ehrlichiosis.

Ehrlichiosis is a disease caused by a group of rickettsiae, known as ehrlichiae, which parasitize circulating leukocytes of man and a variety of domestic and wild animals. A characteristic morphologic feature of ehrlichiae is their occurrence in membrane-bound vacuoles in the cytoplasm of leukocytes, forming inclusions that contain variable numbers of organisms. The tick is the apparent vector of ehrlichiae, but such evidence is not available for all species.

Animals

Goat serum, a substitute of bovine serum in cultivation of Babesia bovis.

Babesia bovis (a Mexican isolate) was cultivated in MASP culture system using goat serum in various concentrations as substitute of bovine serum. It was observed that 20% goat serum + 20% bovine serum + 60% Parker's medium 199 supported the growth of the parasite, which was maintained in this medium through 8 subcultures. The soluble exoantigen (vaccine) present in the culture supernatant is to be quantified and tested in vitro. Goat serum from slaughterhouses may be utilized for in vitro cultivation of the parasite and, expectedly, production of vaccine. This study may prove to be useful in reducing the cost of vaccine at least in tropical countries.

Animals

Immunogenicity and protective efficacy of affinity-purified Plasmodium falciparum exoantigens in Aotus nancymai monkeys.

Soluble Plasmodium falciparum polypeptides, affinity-purified from culture supernatant fluids using sequential immunoadsorptions employing both monoclonal and polyclonal antibodies, induced protective immunity against experimental falciparum malaria in Peruvian Aotus nancymai monkeys. Susceptible monkeys were vaccinated with polypeptides affinity-purified from supernatant fluids of P. falciparum Indochina I/CDC cultures. Eighteen animals (6 immunized with purified antigens plus adjuvants, 6 injected with only the adjuvant preparation, and 6 untreated) were challenged with whole blood containing monkey-adapted virulent organisms of the Indochina I/CDC strain. Selected hematologic, serologic and parasitologic profiles served as potential indicators of protection. This immunogen, when fortified with an aluminum hydroxide/Quil-A saponin adjuvant combination, elicited good antibody responses to major P. falciparum antigens. Protection in vaccinated animals was evidenced by a significantly limited reduction in hematocrit and hemoglobin levels and a relatively moderate course of infection after homologous needle-challenge with Aotus monkey-adapted P. falciparum parasites.

Animals

Canine ehrlichiosis in Connecticut.

The first case of canine ehrlichiosis in Connecticut is reported. A female Brittany spaniel from Milford presented with lethargy, anorexia, fever, petechiae, splenomegaly, thrombocytopenia, anemia, elevated serum alkaline phosphatase, lymphopenia, and hypoalbuminemia. Serologic analysis revealed antibodies to Ehrlichia canis (titer, 1:2,560). This documents a more northern geographic distribution in the United States for this infectious agent than had previously been suspected.

Animals

Use of the dot enzyme-linked immunosorbent assay with isolated Anaplasma marginale initial bodies for serodiagnosis of anaplasmosis in cattle.

Isolated Anaplasma marginale initial bodies were successfully used in a dot ELISA for rapid detection of antibodies to Anaplasma organisms. The enzyme immunoassay used only 25 ng of antigen dotted onto nitrocellulose disks. Antigen-antibody complexes were detected by use of alkaline phosphatase-conjugated protein A, and reactions were read visually after addition of a precipitable, chromogenic substrate. The test allowed the processing of multiple sera, either for screening or for titer determination, in less than 3 hours and was found to be as sensitive as the indirect fluorescent antibody test. The overall performance of the dot ELISA, using isolated A marginale initial bodies, for 580 bovine serum samples was as follows: sensitivity, 93%; specificity, 96%; and predictive value, 95%. Cross-reactivity was not observed with sera positive to Babesia bovis and B bigemina, Trypanosoma vivax, or common bacteria or viruses infecting cattle. The antigen dotted onto nitrocellulose disks was stable when stored at -20, 4, or 25 C. Compared with the indirect fluorescent antibody test, the dot ELISA allowed easier, faster, and more objective interpretation of results. Its simplicity and low cost combined with high sensitivity and specificity indicate that this assay could effectively replace serologic assays currently used for diagnosis of anaplasmosis in cattle.

Anaplasma

Monthly prevalence (in 1986) of antibody titers against equine monocytic ehrlichiosis in apparently healthy horses in Illinois.

The seroprevalence and seasonal trend of antibody titers against equine monocytic ehrlichiosis (Potomac horse fever) were determined in apparently healthy horses in selected areas of Illinois in 1986. Sera from 1,367 horses (6 months to 29 years old) were evaluated for the presence of antibodies against Ehrlichia risticii with indirect immunofluorescence. The majority (88%) of the horses were Thoroughbred or Standardbred racehorses. The number of horses with antibodies against E risticii was 229/1,367 (16.75%). The titers in these horses ranged from 1:10 to 1:640. As the year progressed, the number of seropositive horses (titers greater than or equal to 1:10) and the magnitude of the titers increased significantly, both reaching a maximum in July and August, respectively (P less than 0.05). A relationship between seropositivity and gender was not detected. In the year prior to sampling, 56.8% of the seropositive horses had not been ill, whereas 0.8% had diarrhea, an episode of acute abdominal pain, or laminitis. It was concluded that a large number of horses in Illinois are exposed to E risticii, that maximal exposure occurs in July, and that the most common form of the disease in Illinois is not associated with clinical signs.

Animals

Glomerulopathy in squirrel monkeys with acute Plasmodium falciparum infection.

The renal pathology of 9 squirrel monkeys (Saimiri sciureus) with acute Plasmodium falciparum infection was studied by light and electron microscopy. Endocapillary proliferative glomerulonephritis was the major pathological change observed. The peroxidase anti-peroxidase method demonstrated the presence of IgG, IgM, and P. falciparum antigens in the mesangium and basement membrane. These findings were consistent with those seen in humans with acute P. falciparum infection and indicates that squirrel monkeys are likely to be a good model for the study of renal pathology in malaria research.

Acute Disease

Susceptibility of cats to infection with Ehrlichia risticii, causative agent of equine monocytic ehrlichiosis.

Eight adult cats were inoculated IV (n = 6) or SC (n = 2) with Ehrlichia risticii-infected P388D1 (continuous murine macrophage) cells or with E risticii released from P388D1 cells. Three additional cats were inoculated with organism-free P388D1 cultured monocytes, and 1 cat, which served as a medium control was inoculated with balanced salt solution. Clinical signs of illness were observed in the IV inoculated cats from which E risticii was isolated. One cat developed intermittent diarrhea between postinoculation days (PID) 8 and 18, and the other cat developed lymphadenopathy, acute depression, and anorexia between PID 20 and 24. Ehrlichia risticii was isolated in cultures from 2 of 6 IV inoculated cats on PID 6, 10, and 17. Both cats were inoculated with E risticii released from the P388D1 cells. Ehrlichia risticii was not isolated from SC inoculated cats or from control cats. All 8 cats inoculated with E risticii seroconverted between PID 10 and 23. A pony inoculated with E risticii isolated from 1 of the inoculated cats developed clinical signs of equine monocytic ehrlichiosis including fever, anorexia, depression, and mild colic. Ehrlichia risticii was isolated from the blood of this pony on PID 7, 9, 11, and 16.

Animals

Susceptibility of dogs to infection with Ehrlichia risticii, causative agent of equine monocytic ehrlichiosis (Potomac horse fever).

Adult dogs 1 to 5 were inoculated IV and/or SC with 3, 5, or 6 ml of a suspension containing 1.2 x 10(4) Ehrlichia risticii-infected cells (derived from primary canine monocyte cell cultures)/ml. Dogs 6 to 8 were inoculated IV and/or SC with 3 or 6 ml of 1.2 x 10(5) organism-free cultured canine monocytes/ml. Ehrlichia risticii was isolated in cultures from inoculated dogs 3, 4, and 5 on postinoculation days (PID) 10 to 16, but not from dogs 6 to 8. Dogs inoculated with E risticii seroconverted between PID 6 and 12. Clinical signs of illness were not observed in these 5 E risticii-inoculated dogs. A pony, inoculated with E risticii isolated from inoculated dog 5, developed clinical signs of equine monocytic ehrlichiosis, including fever, anorexia, depression, and diarrhea, and E risticii was isolated from the pony's blood. This E risticii isolate was then inoculated into susceptible dog 9, and E risticii was repeatedly isolated from dog 9 during PID 6 to 17. Dogs were susceptible to infection with E risticii and may serve as a reservoir of the organism in the field.

Animals

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