A rapid high-precision flow cytometry based technique for total white blood cell counting in chickens

Seliger, Christian; Schaerer, Beatrice; Kohn, Marina; Pendl, Helene; Weigend, Steffen; Kaspers, Bernd; Härtle, Sonja

doi:10.1016/j.vetimm.2011.10.010

Cited by 109 publications

(113 citation statements)

References 33 publications

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“…In this regard, it can be stated that the comparison of values of leukogram found by the traditional manual enumeration based on morphological characteristics using a microscope with their values, which were determined by the automatic determination using an analyzer, can be sometimes greatly misleading. Such a comparison can affect negatively Boever et al 2010;Seliger et al 2012;Bílková et al 2017). Besides, our results of the differential leukocyte counts in Cherry Valley hybrid ducks showed that LYM are the most abundant type of leukocytes, which is in accordance with the previous findings of Li et al (2012) and Eze et al (2014).…”

Section: Discussionsupporting

confidence: 92%

Haematological indicators in hybrid mallard ducks (Anas platyrhynchos) with regard to the use of meal from whole white lupin seeds in their diet

et al. 2017

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The objective of our study was to assess the effect of replacing soybean meal with the meal from whole white lupin seeds (Lupinus albus) of the Zulika variety in diets on selected haematological indicators in 40-day-old fattened hybrid mallard ducks. A total of 180 Cherry Valley ducks were divided into three groups (E1, E2, and control). The control group was fed a diet containing soybean meal. Soybean meal replaced with 50% and 100% meal of white lupin seeds were used in group E1 and group E2, respectively. At the end of the fattening, 12 ducks (6 males and 6 females) were randomly selected from each group for a haematological examination. From the result of this study, it is clear that the effect of the diet was found only on the slightly varying number of white blood cells and on the proportion of monocytes. Ducks of group E2 showed a slight increase in the total number of leukocytes which was accompanied by a decrease in the percentage share of monocytes (P < 0.05). Based on the results, it can be claimed that the replacement of soybean meal with meal from the Zulika variety of whole white lupin seeds in the diet did not have a negative effect on the determined blood indicators. Therefore, whole white lupin seeds were successfully used as the important protein component of the diet for fattening hybrid mallard ducks.

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Section: Discussionsupporting

confidence: 92%

Haematological indicators in hybrid mallard ducks (Anas platyrhynchos) with regard to the use of meal from whole white lupin seeds in their diet

et al. 2017

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“…When this calculation is applied to the in vivo model, where 1.5ml of injected yolk was cleared from the body cavity in 24h, a minimum of 2.5×10 10 macrophages would be required to clear all yolk in one round of yolk uptake. Assuming a 0.75 blood volume to body mass ratio (Kiepper, 2009) and an average blood monocyte count of 1300cellsμl -1 blood (Seliger et al, 2012), the average 500g chicken used in this experiment had ~5×10 7 total monocytes available for recruitment from the blood into the body cavity, or three orders of magnitude fewer monocytes than would be required for effective yolk clearance. Conversely, if every available blood monocyte were to participate in yolk clearance then each cell would have to perform at least 1000 rounds of yolk uptake and efflux.…”

Section: Discussionmentioning

confidence: 99%

Mobilization of ectopic yolk in Gallus domesticus: a novel reverse lipid transport process

Cornax

Walzem

Larner

et al. 2013

Journal of Experimental Biology

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SUMMARYIn many oviparous animals, bursting type atresia of ovarian follicles occurs during the reproductive cycle, resulting in the escape of yolk into the extracellular compartment. In birds, this ectopic yolk is rapidly cleared by an unknown process that involves the appearance of yolk-engorged macrophage-like cells. To study this unique type of lipid transport, we injected young male chickens intra-abdominally with egg yolk. Absorption of egg yolk from the body cavity markedly increased the triacylglyceride-rich fraction (TRL) of plasma lipoproteins and was coincident with increased levels of plasma triacylglycerides (TAGs) but not non-esterified fatty acids (NEFAs). Thus, the transport of yolk lipids from the abdominal cavity appears to occur in lipoproteins and be more similar to the transport of hepatic TAGs to the periphery via lipoproteins than to transport of adipose TAGs to the periphery via NEFAs released by the action of lipases. When macrophages were exposed to yolk in vitro, they quickly phagocytized yolk; however, it is unclear whether this level of phagocytosis contributes significantly to total yolk clearance. Instead, the chicken macrophage may function more as a facilitator of yolk clearance through the modification of yolk lipoproteins and the regulation of the local and systemic immune response to ectopic yolk. Yolk appears to be anti-inflammatory in nature. Yolk did not increase levels of the inflammatory cytokines IL-1, IL-6 and IFNγ either in vivo or in vitro; in fact, yolk dampened many inflammatory changes caused by lipopolysaccharide (LPS). Conversely, LPS-induced inflammation retarded yolk clearance from the abdominal cavity and plasma TAG levels.

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“…The additional dextran separation step significantly reduced the number of cells obtained (ficoll alone: 119×10 6 ± 37.1×10 6 /mL of blood v. dextran/ficoll: 10×10 6 ± 1.8×10 6 /mL of blood; P = 0.005), but enough purified cells for immunophenotyping and functional assays could be obtained from only 1 mL of whole blood of very young chickens. The presence of abundant nucleated thrombocytes in PBMCs interferes with quantification of avian leukocyte populations, and requires an additional staining (anti K1 specific and anti K55) for exclusion of thrombocytes (SELIGER et al, 2012;BOHLS et al, 2006). Thus, our combined dextran/ficoll gradient separation could improve immunophenotyping simply by increasing the percentage of lymphocytes within PBMCs, which could be of great importance for rare subpopulations of lymphocytes (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Functional T cell assays require efficient isolation of peripheral blood mononuclear cells (PBMCs), but this is not necessary if phenotyping of lymphocytes and monocytes is done without downstream functional assays (SELIGER et al, 2012). Isolation of PBMCs in avian species is complicated by significant contamination with nucleated thrombocytes.…”

mentioning

confidence: 99%

“…In comparison to lymphocytes in peripheral blood (PBLs), thrombocytes express lower levels of antigen CD45 (De BOEVER et al, 2010), and a lack of CD3 surface antigen (CD45 +low CD3 -). On the other hand, they express the K1 marker, just like monocytes and macrophages, but they lack a monocyte/macrophage Kul-01 marker (k1 + Kul-01 -) (SELIGER et al, 2012;BOHLS et al, 2006). Therefore, cell sorting could be considered as the method of choice to remove thrombocyte contamination from PBLs, but it is expensive and requires additional equipment.…”

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confidence: 99%

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Combined dextran and ficoll separation yields pure populations of chicken peripheral blood mononuclear cells - short communicationia

et al. 2017

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The ficoll density gradient method, the most widely used method for isolating avian peripheral blood mononuclear cells (PBMCs), results in significant contamination with nucleated thrombocytes. This thrombocyte contamination can affect both immunophenotyping and functional assays of T cells. We used full blood samples from ten vaccinated male layer hybrid chickens for isolating PBMCs, and have developed a low cost, combined dextran and ficoll density gradient separation method which, in comparison to the standard ficoll density gradient separation, markedly reduces thrombocyte contamination of the isolated PBMCs. An ex vivo T cell proliferation assay showed an overall reduced background proliferation in unstimulated and stimulated samples after combined dextran and ficoll separation of PBMCs, resulting in a statistically significant difference upon in vitro NDV challenge.

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A rapid high-precision flow cytometry based technique for total white blood cell counting in chickens

Cited by 109 publications

References 33 publications

Haematological indicators in hybrid mallard ducks (Anas platyrhynchos) with regard to the use of meal from whole white lupin seeds in their diet

Haematological indicators in hybrid mallard ducks (Anas platyrhynchos) with regard to the use of meal from whole white lupin seeds in their diet

Mobilization of ectopic yolk in Gallus domesticus: a novel reverse lipid transport process

Combined dextran and ficoll separation yields pure populations of chicken peripheral blood mononuclear cells - short communicationia

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