A practical quantification method for Heinz bodies in birds applicable to rapid response field scenarios

Abstract

Abstract-Oil-induced oxidative injury to red blood cells results in Heinz body hemolytic anemia. Here, we evaluated three Heinz body staining techniques in brown pelican (Pelecanus occidentalis) blood. Using a range of in vitro acetylphenylhydrazine incubations, we validated a field-adapted technique against laboratory wet-mounts and verified the stability of this technique for one month following preparation. Employing this technique during petrochemical spill responses allows for delays between sample collection and analysis. Environ. Toxicol. Chem. 2013;32:401-405. # 2012 SETAC

Hematological indices of injury to lightly oiled birds from the Deepwater Horizon oil spill

“…These repercussions of anemia have obvious implications for individual fitness and suggest that sublethal physiological injury associated with small amounts of oil exposure may have important negative long‐term repercussions for individuals. In addition, the present study demonstrates that proper new methylene blue staining to enumerate reticulocytes and erythrocytic inclusions consistent with Heinz bodies is an important technique that can be applied under field conditions during petroleum‐associated wildlife incidents .…”

“…Exposure to PAHs and their metabolites can result in oxidative injury to cytoplasmic hemoglobin, and aggregates of this damaged hemoglobin can be highlighted by cell staining techniques (e.g., new methylene blue vital staining) that reveal inclusion bodies within erythrocytes . These aggregations of denatured hemoglobin, called Heinz body inclusions (also, Heinz‐Ehrlich bodies), are pathognomonic for oxidative damage to erythrocytes.…”

Haematological and immunological characteristics of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) infected and co-infected with endo- and ectoparasites

“…These slides were stained with either Azure–Xanthene stain (duplicate slides; Diff-Stain kit; IMEB, Inc.) or Wright–Giemsa stain (triplicate slides; Camco Quik Stain II) for visualizing WBCs and trypanosomes, respectively. In addition, we also prepared duplicate slides in the field using a vital stain to reveal changes in RBC morphology ( Fallon et al ., 2013 ). To do this, we aliquoted 20 µl of whole blood into a 0.5 ml Eppendorf vial containing 20 µl of new methylene blue stain (Ricca Chemical Co., Arlington TX, USA).…”

“…We examined slides prepared with new methylene blue stain at ×400 magnification to characterize red blood cell morphology further, to identify vital-staining inclusion bodies and to distinguish immature RBCs (reticulocytes). In other vertebrates, vital stains such as new methylene blue can be useful for evaluating regenerative responses to anaemia by highlighting immature erythrocytes, termed reticulocytes, and for identifying oxidative injury to RBCs ( Campbell and Ellis, 2007 ; Fallon et al ., 2013 ). New methylene blue has also been used to highlight trypanosome organisms in humans ( Ferreira et al ., 2006 ).…”

Dietary intake of Deepwater Horizon oil-injected live food fish by double-crested cormorants resulted in oxidative stress

“…These findings thereby suggest that exposure to oil during the DWH oil spill resulted in oxidative stress that resulted in hemoglobin damage and led to the development of hemolytic anemia in these birds. Anemia is a particularly important oxidative outcome because it causes reduced availability of oxygen to tissues (Fallon et al, 2013). Birds suffering from hemolytic anemia are often fatigued and have reduced energy available for metabolic processes, which could have profound impacts on exercising birds that rely on high energy to sustain energy intensive flights.…”