Characterization of Camel Leukocytes by Flow Cytometry and Microscopic Evaluation of Granulocyte Phagocytosis of Fluorescent Bacteria

Abstract

Peripheral blood was obtained from four lactating camels (Camelus bactrianus) and flow cytometry was used to characterize cell populations. The ability of the granulocytes to engulf fluorescent bacteria was studied in vitro using fluorescence microscopy.Three clusters of blood cells (mononuclear cells, neutrophils and eosinophils) were identified by flow cytometry and fluorescent microscopy. Milk, on the other hand, was dominated by cell fragments and no distinct cluster formation was found. The mean yield of blood granulocytes and monocytes isolated on Ficoll gradient was 92.2 5 5.4 % and 7.9 f 5.7 YO, respectively. Cell viability was 95 %. The mean percentage of phagocytic cells was 71.8 f 5.9 % at 10 min and increased to 97.3 2 0.5 '70 at 60 min when observation was terminated. The average number of bacteria per phagocyte was 8.7 * 2.1 and 13.1 k 0.9 at 10 and 60 min incubation time. BUSCHMANN, 1987b) and flow cytometry (SAAD and HAGELTORN, 1985; FOERSTER and WOLF, 1990; JENSEN-WAERN et al., 1994). T h e cells in mammary gland secretions and their relationships with various physiological and pathological states of the udder have been recognized in many animal species (see JAIN, 1976). Unlike other dairy animals, little is known regarding physiological and pathological states of the mammary gland of the camel. The aim of U.S. Copyright Clearance Center Code Statement: 0931 -184X/96/4302-0111$11.50/0 112 ABDURAHMAN and SAADthe present s t u d y was to characterize leukocyte populations from peripheral blood and milk of camels by means of flow cytometry and to evaluate phagocytic activity of granulocytes from blood using fluorescence microscopy. Materials and Methods AnimalsFour clinically healthy camels at Kilmirden park were used. The camels were of different ages and at different stages of lactation.

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“…Phagocytosis flow cytometry is a high throughput, quantitative assay, which assesses leukocytes' capacity to phagocytose fluorescently labelled bacteria [25,27]. It has numerous advantages for use in comparative immunology and has proven to be a rapid, reliable tool to evaluate phagocytosis in different species ranging from teleosts [28], mice and macaques [27] to salamanders [29], seals [30], camels [31], sea turtles [22,32], beluga whales [33] and Tasmanian devils [34]. The ex vivo nature of the phagocytosis flow cytometry assay also has advantages over in vivo immunologic tests such as allograft rejection, which may be ethically challenging to carry out in critically endangered species.…”