Antiretroviral efficacy of a 98% solution of glycerol or ethylene oxide for inactivation of feline leukemia virus in bone

George S. Coronado Jr Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
Present address is Ocean State Veterinary Specialists, 1480 County Trail, East Greenwich, RI 02818.

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 DVM, MS
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Cheryl L. Swenson Department of Pathobiology and Diagnostic Investigation and Laboratory for Comparative Virology, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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 DVM, PhD

Abstract

Objective—To determine whether infectious retrovirus was inactivated in bones from FeLV-infected cats after ethylene oxide (ETO) sterilization or preservation in a 98% solution of glycerol in an in vitro cell culture system.

Sample Population—Metatarsal bones obtained from 5 FeLV-infected cats and cultured with feline fibroblast cells.

Procedure—Metatarsal bones were treated with 100% ETO, a 98% solution of glycerol, or left untreated. Twenty-five flasks of feline fibroblast cells were assigned to 5 groups: negative control, positive control, ETO-treated bone, glycerol-treated bone, and untreated bone with 5 replicates/group for 4 passages. Media and cell samples were harvested from every flask at each passage to measure FeLV p27 antigen and the number of copies of provirus per 100 ng of DNA, respectively.

Results—All negative control and ETO-treated group replicates were negative for FeLV p27 antigen and provirus throughout the study. All positive control group replicates were positive for FeLV p27 antigen and provirus at passages 1 to 4. Untreated bone group replicates were positive for FeLV p27 antigen at passages 3 and 4 and provirus beginning at passage 2. Glycerol-treated group replicates had delayed cell replication and were negative for FeLV p27 antigen and provirus at passages 1 to 4 and 2 to 4, respectively.

Conclusions and Clinical Relevance—Ethylene oxide sterilization of bone from FeLV-infected cats appeared to abrogate transmission of infectious retrovirus and effectively sterilized bone allografts.

Impact for Human Medicine—Additional studies to confirm effectiveness of ETO treatment of allograft tissues for prevention of pathogen transmission via transplantation are warranted. (Am J Vet Res 2004;65:436–439)

Abstract

Objective—To determine whether infectious retrovirus was inactivated in bones from FeLV-infected cats after ethylene oxide (ETO) sterilization or preservation in a 98% solution of glycerol in an in vitro cell culture system.

Sample Population—Metatarsal bones obtained from 5 FeLV-infected cats and cultured with feline fibroblast cells.

Procedure—Metatarsal bones were treated with 100% ETO, a 98% solution of glycerol, or left untreated. Twenty-five flasks of feline fibroblast cells were assigned to 5 groups: negative control, positive control, ETO-treated bone, glycerol-treated bone, and untreated bone with 5 replicates/group for 4 passages. Media and cell samples were harvested from every flask at each passage to measure FeLV p27 antigen and the number of copies of provirus per 100 ng of DNA, respectively.

Results—All negative control and ETO-treated group replicates were negative for FeLV p27 antigen and provirus throughout the study. All positive control group replicates were positive for FeLV p27 antigen and provirus at passages 1 to 4. Untreated bone group replicates were positive for FeLV p27 antigen at passages 3 and 4 and provirus beginning at passage 2. Glycerol-treated group replicates had delayed cell replication and were negative for FeLV p27 antigen and provirus at passages 1 to 4 and 2 to 4, respectively.

Conclusions and Clinical Relevance—Ethylene oxide sterilization of bone from FeLV-infected cats appeared to abrogate transmission of infectious retrovirus and effectively sterilized bone allografts.

Impact for Human Medicine—Additional studies to confirm effectiveness of ETO treatment of allograft tissues for prevention of pathogen transmission via transplantation are warranted. (Am J Vet Res 2004;65:436–439)

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