Collection and characterization of semen from Chilean rose tarantulas (Grammostola rosea)

Kate E. Archibald College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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Larry J. Minter College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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Gregory A. Lewbart College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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C. Scott Bailey College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27607.

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Abstract

Objective—To establish a nonterminal semen collection method for use in captive Chilean rose tarantulas (Grammostola rosea) and to evaluate tools for investigating morphology and viability of spermatozoa.

Animals—7 mature male Chilean rose tarantulas.

Procedures—Each tarantula was anesthetized in a 500-mL induction chamber containing a cotton ball infused with 2 mL of isoflurane. Semen collection was performed by applying direct pressure to the palpal bulbs (sperm storage organs) located on the distal segment of the palpal limbs. Morphology of spermatozoa was examined by light microscopy and transmission and scanning electron microscopy. Propidium iodide and a fluorescent membrane-permeant nucleic acid dye were used to evaluate cell viability.

Results—Semen was collected successfully from all 7 tarantulas. Microscopic examination of semen samples revealed coenospermia (spherical capsules [mean ± SD diameter, 10.3 ± 1.6 μm] containing many nonmotile sperm cells [mean number of sperm cells/capsule, 18.5 ± 3.8]). Individual spermatozoa were characterized by a spiral-shaped cell body (mean length, 16.7 ± 1.4 μm; mean anterior diameter, 1.5 ± 0.14 μm). Each spermatozoon had no apparent flagellar structure. The fluorescent stains identified some viable sperm cells in the semen samples.

Conclusions and Clinical Relevance—The described technique allowed simple and repeatable collection of semen from Chilean rose tarantulas. Semen from this species was characterized by numerous spherical capsules containing many nonmotile spermatozoa in an apparently quiescent state. Fluorescent staining to distinguish live from dead spermatozoa appeared to be a useful tool for semen evaluation in this species.

Abstract

Objective—To establish a nonterminal semen collection method for use in captive Chilean rose tarantulas (Grammostola rosea) and to evaluate tools for investigating morphology and viability of spermatozoa.

Animals—7 mature male Chilean rose tarantulas.

Procedures—Each tarantula was anesthetized in a 500-mL induction chamber containing a cotton ball infused with 2 mL of isoflurane. Semen collection was performed by applying direct pressure to the palpal bulbs (sperm storage organs) located on the distal segment of the palpal limbs. Morphology of spermatozoa was examined by light microscopy and transmission and scanning electron microscopy. Propidium iodide and a fluorescent membrane-permeant nucleic acid dye were used to evaluate cell viability.

Results—Semen was collected successfully from all 7 tarantulas. Microscopic examination of semen samples revealed coenospermia (spherical capsules [mean ± SD diameter, 10.3 ± 1.6 μm] containing many nonmotile sperm cells [mean number of sperm cells/capsule, 18.5 ± 3.8]). Individual spermatozoa were characterized by a spiral-shaped cell body (mean length, 16.7 ± 1.4 μm; mean anterior diameter, 1.5 ± 0.14 μm). Each spermatozoon had no apparent flagellar structure. The fluorescent stains identified some viable sperm cells in the semen samples.

Conclusions and Clinical Relevance—The described technique allowed simple and repeatable collection of semen from Chilean rose tarantulas. Semen from this species was characterized by numerous spherical capsules containing many nonmotile spermatozoa in an apparently quiescent state. Fluorescent staining to distinguish live from dead spermatozoa appeared to be a useful tool for semen evaluation in this species.

Contributor Notes

Dr. Minter's present address is Great Plains Zoo and Delbridge Museum of Natural History, 805 S Kiwanis Ave, Sioux Falls, SD 57104.

Supported by The Triangle Community Foundation George H. Hitchings New Investigator Award in Health Research.

Presented in part as an abstract in the North Carolina State University CVM Annual Research Forum and Litwack Lecture, Raleigh 2013.

The authors thank Dr. Michael Dykstra, Dr. Jeanette Shipley-Phillips, Katelyn Cordle, and Hendrick Smock for technical assistance.

Address correspondence to Dr. Bailey (scott_bailey@ncsu.edu).
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