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Applied and Environmental Microbiology, October 2008, p. 6077-6084, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.00317-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Effects of Hypercapnic Hypoxia on Inactivation and Elimination of Vibrio campbellii in the Eastern Oyster, Crassostrea virginica{triangledown}

Brett M. Macey,* Ikenna O. Achilihu, Karen G. Burnett, and Louis E. Burnett

Grice Marine Laboratory, College of Charleston, and Hollings Marine Laboratory, Charleston, South Carolina

Received 6 February 2008/ Accepted 28 July 2008

The Eastern oyster, Crassostrea virginica, inhabits shallow coastal waters that frequently experience periods of low dissolved oxygen (hypoxia) and elevated CO2 (hypercapnia) levels. Bacteria are extremely abundant in these environments and accumulate in large numbers in filter-feeding oysters, which can act as passive carriers of human pathogens. Although hypercapnic hypoxia (HH) can affect certain specific immune mechanisms, its direct effect on the inactivation, degradation and elimination of bacteria in oysters is unknown. This research was conducted to determine whether exposure to HH reduces the ability of C. virginica to inactivate and eliminate Vibrio campbellii following its injection into the adductor muscle. Oysters were held in fully air-saturated (normoxic; partial O2 pressure [PO2] = 20.7 kPa, CO2 < 0.06 kPa, pH 7.8 to 8.0) or HH (PO2 = 4 kPa, CO2 = 1.8 kPa, pH 6.5 to 6.8) seawater at 25°C for 4 h before being injected in the adductor muscle with 105 live Vibrio campbellii bacteria and remained under these conditions for the remainder of the experiment (up to 24 h postinjection). Real-time PCR was used to quantify the number of intact V. campbellii bacteria, while selective plating was used to quantify the number of injected bacteria remaining culturable in whole-oyster tissues, seawater, and feces/pseudofeces at 0, 1, 4, and 24 h postinjection. We found that oysters maintained under normoxic conditions were very efficient at inactivating and degrading large numbers of injected bacteria within their tissues. Moreover, a small percentage (~10%) of injected bacteria were passed into the surrounding seawater, while less than 1% were recovered in the feces/pseudofeces. In contrast, HH increased the percentage of culturable bacteria recovered from the tissues of oysters, suggesting an overall decrease in bacteriostasis. We suggest that poor water quality may increase the risk that oysters will harbor and transmit bacterial pathogens hazardous to human and ecosystem health.

* Corresponding author. Present address: Department of Environmental Affairs and Tourism, Marine and Coastal Management, Private Bag X2, Roggebaai 8012, South Africa. Phone: (27 21) 430-7009. Fax: (27 21) 434-2144. E-mail: bmacey{at}deat.gov.za

{triangledown} Published ahead of print on 1 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6077-6084, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.00317-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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