Comparable cytological diagnostic of blood smears on babesia infection

Abstract

In last time Babesiosis as a tick-borne hemoprotozoans human disease have a very important role in differentil diagnostics of modern infectious medicine. It caused by protozon of the genus Babesia, which invade and destory erythrocytes. Babesiosis olso has been called tick fever. So, Babesia has been known by other genus names, including Nuttallia, Microbabesia, Babesialla, and Gonderia. Because all Babesia species are piroplasms, a more inclusive term for anthropozoonotic infections caused by these organisms would be piroplasmosis.They detective complicacy are bild that, tick-borne disease agents from prolongate life cycles involving arthropod and vertebrate host. The complexity is enhanced by the diversity of hosts in different biotopes, which depends on factors life type of vegetation, climate and/or human influence, such as restoration of former industrial sites, which leads to the development of new biotopes. So, on the one hand, new habitats for plants and animals including ticks, and nature are created. About the first case of babesiosis infection was reported as a cause of human sickness in1969 in northeastern United State. Several hundred cases are now reported from this region each year. The disease is characterized by a grandual oncet of malaise with anorexia, fever, headaches, myalgia, and other vague symptoms, which may persist for long period. Occasionally dangerous fulminating infections occur particularly in immunocompromised or aged individuals. The purpose of the present research was to study of the cytological diagnostic of blood smears from object’s with the Babesia infection. Materials and methods. Blood smears (by Romanovsky- Gimze (standart), Wright’s standart and staining, the author’s modification, 2014) of domestic dogs (n = 31) of both sexes with Babesia infection at the age from 3 months to 6 years served as the material for the study. The preparations were fixed during 1-2 seconds with 96 % ethyl alcohol. Then warmed (t = 36.0 ± 2.0)°С commercial matrix solutions of eosin, azure and methylene blue were applied one by one. The smears were rinsed (1-2 seconds) in distilled water and dehydrated. The procedure ended with short-term drying in a diffused stream of warm dry air (Samsung house fan, power 220 W). The results were compared with intact control. Smears were contrasted and analysed under a microscope LOMU (LOMO, Russia): x 300; x400; x1000; x1350 and photographed with a digital camera “Canon EOS-3000”. Results. Blood samples infected with Babesia species were collected (may-october) from naturally (promenade in forest-park) tick-borne infected dogs (Canis familiaris) in all Kharkov region and sity. All (experimental) animals were monitored twice daily by veterinary doctors for clinical signs and had rectal temperatures taken (authors have a greate thankness for the cooperation and consolidation Chif -Mr. Yu. V. Al’okhin and veterinary personal of Kharkov Center of Clinical Veterinary). Blood was drawn daily for hematocrit determination and peripheral blood smear were made from ear vien blood to determine parasitemia status. As result of the analysis of blood smears it was found out that against a background of orange erythrocyte cytoplasm the preparation area easily revealed crimson- and red-lilac pyriform (n = 8-12 in the field of vision of the preparation), annular (n = 9-16 in the field of vision), amoebiform haemoparasites and those with other shapes (Σ=13), thereby indicating a high level of infection (81.8 %). Owing to their own chromatophilic feature, protozoan cells looked geometrically marked and clearly contrasted against a background of the saturated red-violet colour of nuclei. The developed technique of staining facilitated: a more qualitative analysis of ontogenetic staging (III) of Babesia (trophozoites, merozoites, sporozoites); improvement of differential diagnosis of the haemoparasites with blood platelets (the latter were distinguished from cells of the causative agent by the presence of marked ovaloid azurophilic granules in the cytoplasm of young forms (Σ = 1-3 in the field of vision) and azurophilic granularity in mature forms; better differential diagnosis with intracellular inclusions (intraerythrocytic Cabot rings, Howell–Jolly bodies); improved differential diagnosis with solid elements of sediments of used stains (the above artifacts were in a saturated dark blue or black colour and observed very seldom). Conclusions. By the results of the cytological diagnostic of blood smears it was revealed that domestic dogs with clinically detected Babesia infection had a high level of contaminaiton with parasites. In our studies this level was 81.8 %. Has been established that using of the blood smears by Romanovsky- Gimze in Wright’s the author’s modification (2014) are very effective to extrimal medecine and perspective for next clinical investigation.