This article was downloaded by: On: 25 August 2010 Access details: Access Details: Free Access Publisher Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37- 41 Mortimer Street, London W1T 3JH, UK New Zealand Journal of Marine and Freshwater Research Publication details, including instructions for authors and subscription information: http://www.informaworld.com/smpp/title~content=t918959567 Pfiesteria shumwayae (Pfiesteriaceae) in New Zealand Lesley L. Rhodesa; Joann M. Burkholderb; Howard B. Glasgowb; Parke A. Rubleec; Coy Allenb; Janet E. Adamsona a Cawthron Institute, Nelson, New Zealand b Center for Applied Aquatic Ecology, North Carolina State University, Raleigh, NC, United States c University of North Carolina at Greensboro, Greensboro, NC, United States Online publication date: 30 March 2010 To cite this Article Rhodes, Lesley L. , Burkholder, Joann M. , Glasgow, Howard B. , Rublee, Parke A. , Allen, Coy and Adamson, Janet E.(2002) 'Pfiesteria shumwayae (Pfiesteriaceae) in New Zealand', New Zealand Journal of Marine and Freshwater Research, 36: 3, 621 — 630 To link to this Article: DOI: 10.1080/00288330.2002.9517117 URL: http://dx.doi.org/10.1080/00288330.2002.9517117 PLEASE SCROLL DOWN FOR ARTICLE Full terms and conditions of use: http://www.informaworld.com/terms-and-conditions-of-access.pdf This article may be used for research, teaching and private study purposes. Any substantial or systematic reproduction, re-distribution, re-selling, loan or sub-licensing, systematic supply or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material. New Zealand Journal of Marine and Freshwater Research, 2002, Vol. 36: 621-630 621 0028-8330/02/3603-0621 $7.00 © The Royal Society of New Zealand 2002 Short communication Pfiesteria shumwayae (Pfiesteriaceae) in New Zealand LESLEY L.RHODES1 Keywords Pfiesteria; dinoflagellate; estuarine; 2 fish kills; molecular probes JOANN M. BURKHOLDER HOWARD B. GLASGOW2 PARKE A. RUBLEE3 INTRODUCTION COY ALLEN2 Pfiesteria Steidinger et Burkholder (Dinamoebales; JANET E. ADAMSON1 Pfiesteriaceae) is a heterotrophic dinoflagellate with 1 Cawthron Institute planktonic zoospore populations sourced from seed Private Bag 2 beds of cysts and amoebae in sediments (Steidinger Nelson, New Zealand et al. 1996; Burkholder et al. 1992; Burkholder & 2Center for Applied Aquatic Ecology Glasgow 1997). Pfiesteria exhibits "ambush- North Carolina State University predator" behaviour in the presence of fish 620 Hutton St., Suite 104 (Burkholder et al. 1998), which may lead to non- Raleigh focal as well as deep-focal, ulcerous lesions and NC 27606, United States death, as demonstrated in laboratory experiments (Burkholder et al. 1992, 1995, 2001a; Noga et al. 3University of North Carolina at Greensboro 1996; Marshall et al. 2000). The fish attacks are non- P.O. Box 26 174 specific and more than 20 native and exotic species Greensboro tested in the United States have proved vulnerable NC 27412, United States (Burkholder et al. 1997). The United States has experienced massive fish Downloaded At: 19:40 25 August 2010 kills, clearly attributed to Pfiesteria, in certain estuarine systems along its eastern seaboard (Burkholder et al. 1992; Burkholder & Glasgow Abstract Pfiesteria shumwayae Steidinger et 1997; Samet et al. 2001; Brownie et al. in press) and, Burkholder is now known to be present in New in the Albemarle-Pamlico Estuarine System of North Zealand and occurs in estuaries around the country. Carolina, >1 × 109 fish were killed during Pfiesteria The presence of Pfiesteria was initially determined outbreaks over the last decade (Burkholder et al. by a polymerase chain reaction (PCR)-based 2001a; Glasgow et al. 2001a). Pfiesteria piscicida detection assay, using oligonucleotide primers Steidinger et Burkholder (Steidinger et al. 1996) has targeted at ribosomal DNA extracted from estuarine been linked to measurable neurotoxic effects in water and sediments. Presence was confirmed by humans, including central nervous system isolation from fresh sediments in the presence offish impairment (for example, mostly reversible short- (Oreochromis mossambicus), followed by term memory loss that can last for weeks to months; identification by scanning electron microscopy. The Glasgow et al. 1995; Grattan et al. 1998), as well as New Zealand isolates of P. shumwayae were autonomic and peripheral nervous system dys- ichthyotoxic in bioassays, but there is no historic function, skin lesions, and other effects (Glasgow et evidence offish kills in New Zealand associated with al. 1995; Schmechel & Koltai 2001). The economic the dinoflagellate. costs related to Pfiesteria have been high; in Maryland the presence of the dinoflagellate in association with the death of c. 50 000 Atlantic menhaden (Brevoortia tyrannus Latrobe) was M02013; published 17 September 2002 Received 20 February 2002; accepted 30 May 2002 responsible for an estimated cost of US$65 million 622 New Zealand Journal of Marine and Freshwater Research, 2002, Vol. 36 (Epstein 1998; Lipton 1998). This was due to indirect In the United States, Pfiesteria has been found market effects based on provocative media reports from New York State to Texas (Rublee et al. 1999, and public perception rather than particularly heavy 2001). The geographic spread and toxicity of fish losses (Anderson et al. 2000; Committee on Pfiesteria spp. indicate a tolerance to a wide range Environment and Natural Resources (CENR) 2000; of salinities and temperatures (0-35 and 12-33°C Burkholder & Glasgow 2002). respectively; Burkholder et al. 2001a; Glasgow et al. The Pfiesteria species complex comprises the two 2001a,b, 2002). It should be noted that Pfiesteria described species, P. piscicida (Steidinger et al. strains have been TOX-B at most locations where it 1996) and P. shumwayae Glasgow et Burkholder has been detected, with toxicity requiring induction, (Glasgow et al. 2001b). Both species have complex and toxic events (mainly massive fish kills) taking life cycles and may occur in three functional types place at sites of nutrient enrichment (Burkholder et (toxicity status) as: (1) highly toxic or TOX-A al. 1997, 2001b; Glasgow et al. 2001a; Samet et al. (actively toxic, requiring the presence of live finfish 2001; Burkholder & Glasgow 2002). or their fresh tissues and excreta); (2) temporarily DNA-based diagnostics are available in several non-toxic in the absence of fish or TOX-B; and (3) formats for the detection of Pfiesteria spp. in water, non-inducible, apparently (based on present know- sediments, and fish (Rublee et al. 1999,2001; Oldach ledge) unable to produce toxin in response to fish et al. 2000; Bowers et al. 2000). In this study (Burkholder et al. 2001b). Other toxic algae, molecular probe technology was used to determine including dinoflagellates, are known to have benign the distribution of Pfiesteria in New Zealand. or non-inducible as well as toxic strains (Gentien & Arzul 1990; Anderson 1991; Bates et al. 1998; Edvardsen & Paasche 1998). METHODS Pfiesteria spp. can consume small, photosynthetic microalgae, and retain their chloroplasts in an active Collection, isolation, and culture state, presumably utilising their photosynthetic Water samples (1 litre) for dinoflagellate analysis products (Glasgow et al. 1998,2001b; Lewitus et al. were collected from sites from just above sediments 1999), and Pfiesteria can also consume fresh shellfish which had been lightly disturbed by hand. A total of tissue, preying on the pediveliger stage (Springer 26 estuarine localities and embayments were 2000). P. piscicida can survive passage through the gut selected from around New Zealand (Fig. 1) where of adult shellfish in cyst form, ultimately excysting and flushing rates were low, i.e., up to 5 days residence then capable of further predation (Springer et al. 2002). time. Sediment cores (3 cm diam.) were also There is no evidence to date that human illnesses collected from the top 5 cm at those sites, for analysis Downloaded At: 19:40 25 August 2010 result from eating fish or shellfish that have been for cysts, with 5-10 ml of seawater added to prevent exposed to a Pfiesteria event. Recent research by sediments drying out. All samples were maintained Springer et al. (2002), however, has demonstrated that in insulated containers at c. 15°C. Temperature (°C) toxic Pfiesteria cells can be concentrated by some and salinity (Practical Salinity Scale; Orion 140 shellfish (for example, the subadult eastern oyster salinity meter) were recorded in situ whenever Crassostrea virginica Gmelin). Thus, risks to humans possible. from seafood consumption cannot be ruled out until Aliquots (50-100 ml) of water samples were the Pfiesteria toxin(s) can be identified, so that their filtered (Whatman Ltd., GF/C glass microfibre), and presence/absence in fish tissues can be conclusively the filter immersed in CTAB (cetyltrimethylam- determined and quantified (Fairey et al. 1999; Kimm- monium bromide) buffer (1 ml) in 1.5 ml vials; Brinson et al. 2001;