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Applied and Environmental Microbiology, April 2002, p. 1955-1961, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1955-1961.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Yeast Species Associated with Orange Juice: Evaluation of Different Identification Methods{dagger}

Covadonga R. Arias, Jacqueline K. Burns, Lorrie M. Friedrich, Renee M. Goodrich, and Mickey E. Parish*

Citrus Research and Education Center, University of Florida, IFAS, Lake Alfred, Florida 33850

Received 1 October 2001/ Accepted 14 January 2002

Five different methods were used to identify yeast isolates from a variety of citrus juice sources. A total of 99 strains, including reference strains, were identified using a partial sequence of the 26S rRNA gene, restriction pattern analysis of the internal transcribed spacer region (5.8S-ITS), classical methodology, the RapID Yeast Plus system, and API 20C AUX. Twenty-three different species were identified representing 11 different genera. Distribution of the species was considerably different depending on the type of sample. Fourteen different species were identified from pasteurized single-strength orange juice that had been contaminated after pasteurization (PSOJ), while only six species were isolated from fresh-squeezed, unpasteurized orange juice (FSOJ). Among PSOJ isolates, Candida intermedia and Candida parapsilosis were the predominant species. Hanseniaspora occidentalis and Hanseniaspora uvarum represented up to 73% of total FSOJ isolates. Partial sequence of the 26S rRNA gene yielded the best results in terms of correct identification, followed by classical techniques and 5.8S-ITS analysis. The commercial identification kits RapID Yeast Plus system and API 20C AUX were able to correctly identify only 35 and 13% of the isolates, respectively. Six new 5.8S-ITS profiles were described, corresponding to Clavispora lusitaniae, Geotrichum citri-aurantii, H. occidentalis, H. vineae, Pichia fermentans, and Saccharomycopsis crataegensis. With the addition of these new profiles to the existing database, the use of 5.8S-ITS sequence became the best tool for rapid and accurate identification of yeast isolates from orange juice.

* Corresponding author. Mailing address: Citrus Research and Education Center, University of Florida, IFAS, 700 Experiment Station Rd., Lake Alfred, FL 33850. Phone: (863) 956-1151. Fax: (863) 956-4631. E-mail: mparish{at}lal.ufl.edu.

{dagger} This study was approved for publication as Journal Series no. R-08477 of the Florida Agricultural Experiment Station.

Applied and Environmental Microbiology, April 2002, p. 1955-1961, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1955-1961.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wenning, M., Seiler, H., Scherer, S. (2002). Fourier-Transform Infrared Microspectroscopy, a Novel and Rapid Tool for Identification of Yeasts. Appl. Environ. Microbiol. 68: 4717-4721 [Abstract] [Full Text]  


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