Preserving whole blood in formalin extends the specimen stability period for manual cell counts for fish

Arnold, Jill; Matsche, Mark A.; Rosemary, Kevin

doi:10.1111/vcp.12214

Cited by 25 publications

(19 citation statements)

References 10 publications

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“…14 However, there is also typically a high degree of imprecision (>9% CV) in manual RBC counts for fish. 25 Leukocytes reported in this study include small and large lymphocytes (most abundant), neutrophils, monocytes, and heterophils (least abundant). In teleosts, lymphocytes generally account for the majority of leukocytes in blood, and nonneutrophilic granulocytes are generally either absent or infrequent.…”

Section: Hematologymentioning

confidence: 69%

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A comparison of hematology, plasma chemistry, and injuries in Hickory shad (Alosa mediocris) captured by electrofishing or angling during a spawning run

Matsche

Rosemary

Stence

2017

Veterinary Clinical Pathol

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Section: Hematologymentioning

confidence: 69%

“…The range of RBC counts among teleosts is variable (1–5 × 10 12 /L), with a general trend to higher values in more active species . However, there is also typically a high degree of imprecision (>9% CV) in manual RBC counts for fish …”

Section: Discussionmentioning

confidence: 99%

A comparison of hematology, plasma chemistry, and injuries in Hickory shad (Alosa mediocris) captured by electrofishing or angling during a spawning run

Matsche

Rosemary

Stence

2017

Veterinary Clinical Pathol

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“…Coelomic fluid was placed in both lithium heparin and ethylenediaminetetraacetic acid (EDTA) micro tubes (Sarstedt Inc., Princeton, NJ). Formalin preparations of 50 µl coelomic fluid in 200 µl 10% neutral buffered formalin and 50 µl EDTA coelomic fluid in 200 µl 10% neutral buffered formalin were prepared for coelomocyte counts for comparison (34). Sediment smears were prepared after centrifugation of 200 µl of coelomic fluid at 605 relative centrifugal force (RCF) for 10 min, drawing off the supernatant, re-suspending the pellet in 50 µl supernatant, and preparation of smears from this concentrated cellular material for dry mount cytology.…”

Section: Methodsmentioning

confidence: 99%

Coelomic Fluid Evaluation in Pisaster ochraceus Affected by Sea Star Wasting Syndrome: Evidence of Osmodysregulation, Calcium Homeostasis Derangement, and Coelomocyte Responses

et al. 2020

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Sea Star Wasting Syndrome (SSWS) is one of the largest marine wildlife die-offs ever recorded, killing millions of sea stars from more than 20 Asteroid species from Alaska to Mexico from 2013 to 2015 from yet undetermined cause(s). Coelomic fluid surrounds the sea star's organs, playing critical roles in numerous systemic processes, including nutrient transportation and immune functions. Coelomocytes, which are cellular components of coelomic fluid and considered functionally equivalent to vertebrate leukocytes, are responsible for innate cell-mediated immunity. The objectives of this study were to (1) evaluate changes in coelomic fluid chemistry, coelomocyte counts, and cytology from ochre sea stars (Pisaster ochraceus) (n = 55) with clinical signs consistent with SSWS at varying intensity (SSWS score 1: n = 4, score 2: n = 2, score 3: n = 3, score 4: n = 18, score 5: n = 26) in comparison to coelomic fluid from clinically normal sea stars (n = 26) and to (2) correlate SSWS score with cellular and biochemical analytes. SSWS-affected sea stars had wider ranges of all electrolytes, except calcium; statistically significantly higher chloride, osmolality, and total protein; lower calcium; and higher coelomocyte counts when compared to clinically normal sea stars maintained under identical environmental conditions. Free and/or phagocytized bacteria were noted in 29% (16 of 55) coelomic fluid samples from SSWS-affected sea stars but were absent in clinically normal sea stars. SSWS score correlated significantly with increasing chloride concentration, osmolality, and coelomocyte counts. These chemistry and cytological findings in coelomic fluid of SSWS-affected sea stars provide insight into the pathophysiology of SSWS as these results suggest osmo-and calcium dysregulation, coelomocyte responses, and presumptive opportunistic bacterial Wahltinez et al. Wasting Sea Star Coelomic Fluid infection in SSWS-affected sea stars. This information provides potential future research applications for the development of treatment strategies for sea stars in managed care and for understanding the complexity of various biochemical and cellular pathophysiological mechanisms involved in sea star wasting.

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“…Formalin preparations containing 50 μl of coelomic fluid in 200 μl of 10% neutral buffered formalin and 50 μl of EDTA coelomic fluid in 200 μl of 10% neutral buffered formalin were prepared for coelomocyte counts (Arnold et al. ). Sediment smears were prepared after centrifugation of 200 μl of coelomic fluid at 3,000 RPM for 10 minutes.…”

Section: Methodsmentioning

confidence: 99%

“…Manual coelomocyte counts were performed on native (non‐EDTA preserved) preparations and EDTA‐anticoagulated formalin‐fixed preparations as described in Arnold et al. () for lobster hemolymph and fish blood. Direct and sediment dry mount smears were treated with a Wright‐Giemsa stain for cytologic evaluation.…”

Section: Methodsmentioning

confidence: 99%

Coelomic Fluid Evaluation in Clinically Normal Ochre Sea Stars Pisaster ochraceus: Cell Counts, Cytology, and Biochemistry Reference Intervals

et al. 2019

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Coelomic fluid sampling is a noninvasive technique that is used to access the body fluid of sea stars for diagnostics and research. Given recent mortality events including sea star wasting disease, which has killed millions of sea stars along the Pacific coast since 2013, there is a need for validated diagnostic tests to evaluate sea star health. The objectives of this study were to establish coelomic fluid reference intervals for clinically normal ochre sea stars Pisaster ochraceus in an open system aquarium, to describe the cytologic findings, and to compare the chemistries of coelomic fluid with open system tank water. Coelomic fluid from 26 clinically normal sea stars was sampled for coelomocyte counts, cytologic evaluation, and biochemical analysis including magnesium, sodium, potassium, chloride, calcium, and total protein. The number of coelomocytes and total protein did not fit normal distribution and were excluded from analyses. Reference intervals were established for other chemistry analytes. There was no statistical difference in biochemistries between sea star coelomic fluid and water from five open system tanks, which supports previous evidence that sea stars are osmoconformers. Very low numbers of coelomocytes were observed cytologically. These results provide a useful baseline and diagnostic tool for health assessments of sea stars.

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Preserving whole blood in formalin extends the specimen stability period for manual cell counts for fish

Cited by 25 publications

References 10 publications

A comparison of hematology, plasma chemistry, and injuries in Hickory shad (Alosa mediocris) captured by electrofishing or angling during a spawning run

A comparison of hematology, plasma chemistry, and injuries in Hickory shad (Alosa mediocris) captured by electrofishing or angling during a spawning run

Coelomic Fluid Evaluation in Pisaster ochraceus Affected by Sea Star Wasting Syndrome: Evidence of Osmodysregulation, Calcium Homeostasis Derangement, and Coelomocyte Responses

Coelomic Fluid Evaluation in Clinically Normal Ochre Sea Stars Pisaster ochraceus: Cell Counts, Cytology, and Biochemistry Reference Intervals

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