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Abstract

Objective

To evaluate light microscopic, cytochemical, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes.

Animals

10 healthy snakes.

Procedure

Various stains, including Wright-Giemsa, benzidine peroxidase, Sudan black B, chloroacetate esterase, α-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff with diastase, and toluidine blue, were used to stain leukocytes differentially on multiple blood smears. Electron microscopy also was performed.

Results

Lymphocytes were the most commonly observed leukocyte and could be distinguished from thrombocytes, using periodic acid-Schiff stain with diastase. Azurophils also were commonly observed; their granules stained with peroxidase. Eosinophils were not identified; however, 2 morphologic variations of heterophils were seen in the blood of all snakes and were considered the same cell type at different stages of cytoplasmic granule development. Heterophil granules were better preserved, using a one-step Wright-Giemsa method that did not require alcohol fixation prior to staining. Degranulated heterophils were observed in all preparations.

Conclusions

Most leukocytes of eastern diamond-back rattlesnakes can be identified easily on Wright-Giemsa-stained preparations. However, hematologic stains that do not require alcohol fixing prior to staining may be preferred for leukocyte evaluation in certain reptiles. A limited degree of heterophil maturation may continue in the blood of healthy snakes. This, along with degranulation of heterophils, may result in a variable staining pattern in this cell type, regardless of the stain used.

Clinical Relevance

Results provide baseline data for use in hematologic testing in diagnosis of disease and monitoring of treatment of sick or injured snakes. (Am J Vet Res 1999;60:507-514)

Free access
in American Journal of Veterinary Research

Abstract

Objectives—To characterize protein composition of shell scute of desert tortoises and to determine whether detectable differences could be used to identify healthy tortoises from tortoises with certain illnesses.

Animals—20 desert tortoises.

Procedures—Complete postmortem examinations were performed on all tortoises. Plastron scute proteins were solubilized, scute proteins were separated by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and proteins were analyzed, using densitometry. Two-dimensional immobilized pH gradient-PAGE (2D IPG-PAGE) and immunoblot analysis, using polyclonal antisera to chicken-feather β keratin and to alligator-scale β keratin, were conducted on representative samples. The 14-kd proteins were analyzed for amino acid composition.

Results—The SDS-PAGE and densitometry revealed 7 distinct bands, each with a mean relative protein concentration of > 1%, ranging from 8 to 47 kd, and a major protein component of approximately 14 kd that constituted up to 75% of the scute protein. The 2D IPG-PAGE revealed additional distinct 62- and 68- kd protein bands. On immunoblot analysis, the 14-, 32-, and 45-kd proteins reacted with both antisera. The 14-kd proteins had an amino acid composition similar to that of chicken β keratins. There was a substantial difference in the percentage of the major 14- kd proteins from scute of ill tortoises with normal appearing shells, compared with 14-kd proteins of healthy tortoises.

Conclusions and Clinical Relevance—The major protein components of shell scute of desert tortoises have amino acid composition and antigenic features of β keratins. Scute protein composition may be altered in tortoises with certain systemic illnesses. ( Am J Vet Res 2001;62:104–110)

Full access
in American Journal of Veterinary Research