COMPARISON OF SUBCARAPACIAL SINUS AND BRACHIAL VEIN PHLEBOTOMY SITES FOR BLOOD COLLECTION IN FREE-RANGING GOPHER TORTOISES (GOPHERUS POLYPHEMUS).

Paired blood samples were collected from the brachial vein and subcarapacial sinus of 42 wild gopher tortoises (Gopherus polyphemus) for comparison of hematological and plasma biochemical, vitamin, mineral, and protein electrophoresis parameters with general linear models with repeated measures designs. Comparisons revealed higher brachial vein mean values for 38 of 42 parameters with significance (P ≤ 0.05) for 21 parameters. Significantly lower brachial vein means were noted for two parameters. Coefficients of variation for brachial vein samples were lower for 30 out of 42 parameters with significantly lower values for 11 parameters. Of the 12 parameters with lower coefficients of variation for subcarapacial sinus samples, significant differences were noted for four parameters. Both the lower coefficient of variation and the higher mean values for most parameters from brachial vein samples are attributed to greater hemodilution of subcarpacial samples with lymph compared to the paired brachial vein samples. To address the clinical relevance and reliability of the parameter ranges obtained from both paired phlebotomy sites, we compared these values to those obtained from other selected investigations of gopher tortoise and desert tortoise (Gopherus agassizii) clinicopathology. Although statistically significant differences were detected, available data demonstrated that most mean ± standard deviation values and/or ranges for both phlebotomy sites of our study are similar to those reported for the selected studies. Based on these results, utilization of either set of our study values in the assessment of a gopher tortoise population or individual gopher tortoises would be acceptable for clinical or management decisions. It is also advisable that investigators develop institutional blood parameter ranges for healthy individuals by phlebotomy site when possible in addition to maintaining standardized sample collection and handling protocols.

A modified diet to support conservation of the Atala hairstreak butterfly (Eumaeus atala Poey).

Raising insects in a laboratory for release into the wild is a common conservation practice, but maintaining breeding colonies year-round can be limited by seasonal food availability. Food availability is particularly challenging for insects which depend on specific host plants. For example, our early efforts to rear the imperiled Atala hairstreak butterfly (Eumaeus atala Poey) resulted in colony failure during winter due to lack of food. To overcome this barrier, we developed a modified freeze-dried host plant diet to support the colony. The diet consisted of reconstituted freeze-dried leaves and stems from fresh-growth coontie (Zamia integrifolia), the host plant for the Atala butterflies. We fed larvae less than 9 mm on this freeze-dried diet and transferred them to live coontie plants after they were more than 9 mm. We reared a colony of Atala butterflies using these methods for 859 days, resulting in more than 3400 animals released into the wild. Comparing colony counts during that time period to the 548 days we reared them without modified freeze-dried diet showed a clear benefit in using freeze-dried diet. A growth trial (N = 40) of larvae fed on only freeze-dried diet compared to larvae fed on fresh coontie cuttings found no significant difference in larval or pupal development between groups (p = 0.71 and p = 0.47, respectively). We, therefore, conclude that the freeze-dried diet provided an appropriate alternative for Atala colonies when fresh growth from the host plant is unavailable, and we recommend use of this technique for raising other host plant-dependent insect species of conservation concern.

Nutrition and health in amphibian husbandry.

Amphibian biology is intricate, and there are many inter-related factors that need to be understood before establishing successful Conservation Breeding Programs (CBPs). Nutritional needs of amphibians are highly integrated with disease and their husbandry needs, and the diversity of developmental stages, natural habitats, and feeding strategies result in many different recommendations for proper care and feeding. This review identifies several areas where there is substantial room for improvement in maintaining healthy ex situ amphibian populations specifically in the areas of obtaining and utilizing natural history data for both amphibians and their dietary items, achieving more appropriate environmental parameters, understanding stress and hormone production, and promoting better physical and population health. Using a scientific or research framework to answer questions about disease, nutrition, husbandry, genetics, and endocrinology of ex situ amphibians will improve specialists' understanding of the needs of these species. In general, there is a lack of baseline data and comparative information for most basic aspects of amphibian biology as well as standardized laboratory approaches. Instituting a formalized research approach in multiple scientific disciplines will be beneficial not only to the management of current ex situ populations, but also in moving forward with future conservation and reintroduction projects. This overview of gaps in knowledge concerning ex situ amphibian care should serve as a foundation for much needed future research in these areas.