Baseline / Marine Pollution Bulletin 50 (2005) 208–236 211 Acknowledgment Muir, D., Braune, B., DeMarch, B., Norstrom, R., Wagemann, R., Lockhart, L., Hargrave, B., Bright, D., Addison, R., Payne, J., The authors wish to thank Hector de Haro for the Reimer, K., 1999. Spatial and temporal trends and effects of contaminants in the Canadian Arctic marine ecosystem: a review. sample preparations and the staff of the Natal Sharks Science of the Total Environment 230, 83–144. Board who made the tissue sampling possible. Serrano, R., Ferna´ndez, M.A., Herna´ndez, L.M., Herna´ndez, M., Pascual, P., Rabanal, R.M., Gonza´lez, M.J., 1997. Coplanar polychlorinated biphenyl congeners in shark livers from the North- References Western African Atlantic Ocean. Bulletin of Environmental Contamination and Toxicology 58, 150–157. Agency for Toxic Substances and Diseases Registry (ATSDR)/US Storelli, M.M., Marcotrigiano, G.O., 2001. Persistent organochlorine Public Health Service, 1994. 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SETAC Press, Pensacola, FL, pp. 197– white shark, Carcharodon carcharias from the East Coast of South 217. Africa. Fisheries Bulletin 97, 153–169. 0025-326X/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.marpolbul.2004.11.032 Comparative paralytic shellfish toxin profiles in the strains of Gymnodinium catenatum Graham from the Gulf of California, Mexico I. Ga´rate-Liza´rraga a,*, J.J. Bustillos-Guzma´n b, L. Morquecho b, C.J. Band-Schmidt a, R. Alonso-Rodrı´guez b, K. Erler c, B. Luckas c, A. Reyes-Salinas b, D.T. Go´ngora-Gonza´lez b a Departamento de Plancton y Ecologı´a Marina, Centro Interdisciplinario de Ciencias Marinas (CICIMAR-IPN), Apdo. Postal 592, La Paz, B.C.S. 23000, Me´xico b Centro de Investigaciones Biolo´gicas del Noroeste (CIBNOR), Apdo. Postal 128, La Paz, B.C.S. 23000, Me´xico c Faculty of Biology and Pharmacy, Department of Food Chemistry, Friedrich-Schiller University, Dornburgerstraße 25, 07743 Jena, Germany Gymnodinium catenatum was described for the first time in the Gulf of California, Mexico by Graham * Corresponding author. Tel.: +52 112 25344/+52 612 1230350x82434; fax: +52 112 25322/+52 612 122 5322. (1943). The known global distribution of this species E-mail addresses: [email protected], [email protected] has increased dramatically in the past few decades, (I. Ga´rate-Liza´rraga). and it is now reported in every continent, its vegetative 212 Baseline / Marine Pollution Bulletin 50 (2005) 208–236 forms having been recorded in Argentina, Alexandria, Australia, Brazil, Cuba, Italy, Japan, Korea, Philip- pines, Portugal, Spain, Singapore, Uruguay, and Vene- zuela (Hallegraeff and Fraga, 1996; Negri et al., 2001; La Barbera-Sa´nchez and Gamboa-Maruez, 2001; Holmes et al., 2002; Go´mez, 2003; Leal et al., 2003; Park et al., 2004). G. catenatum is the only known gymnodi- niod dinoflagellate that produces paralytic shellfish poi- soning (PSP) (Hallegraeff and Fraga, 1996), and up to 20 saxitoxin analogues have been reported (Negri et al., 2001, 2003). G. catenatum is an important compo- nent of the phytoplankton along the Mexican Pacific coastline (Graham, 1943; Mee et al., 1986; Corte´s-Alta- mirano et al., 1999; Figueroa-Torres and Zepeda-Esqui- vel, 2001; Herrera-Galindo, 2002; Ga´rate-Liza´rraga et al., 2004a,b; Morquecho and Lechuga-Deve´ze, 2004; Band-Schmidt et al., 2005). High concentrations have been reported with few human fatalities, and extensive fish mortalities (Mee et al., 1986; Corte´s-Altamirano and Alonso-Rodrı´guez, 1997). Recently, massive mor- tality events of nauplii and adult shrimp related to G. catenatum blooms have been documented along the Fig. 1. Sampling stations ( ) and historical records (d) of distribution Sinaloa shoreline (Alonso-Rodrı´guez and Pa´ez-Osuna, of Gymnodinium catenatum along the Gulf of California (Graham, ´ 1943; Gilbert and Allen, 1943; Mee et al., 1986; Manrique and 2003; Alonso-Rodrıguez et al., 2004). Molina, 1997; Corte´s-Altamirano and Alonso-Rodrı´guez, 1997; Corte´s- Paralytic shellfish toxin (PST) profiles have been Altamirano et al., 1999; Morquecho and Lechuga-Deve´ze, 2004; determined in several strains of G. catenatum isolated Ga´rate-Liza´rraga et al., 2001; Ga´rate-Liza´rraga et al., 2004a, 2004b; from Bahı´a Concepcio´n, and nine saxitoxin analogues Alonso-Rodrı´guez et al., 2004; Band-Schmidt et al., 2004; this study). were determined (Band-Schmidt et al., 2005). In wild phytoplankton samples containing G. catenatum from Bahı´a Concepcio´n, Bahı´a de Mazatla´n and Bahı´ade riched with GSe (Doblin et al., 2000). Vegetative cells La Paz, also nine saxitoxin analogues were determined; were isolated from the enrichment medium with micro- SXT and neoSXT were observed only in the Bahı´adeLa pipettes using an inverted microscope (Carl Zeiss, Axio- Paz, and Bahı´a Concepcio´n samples, respectively vert 100). Cells were transferred individually to 24-well (Ga´rate-Liza´rraga et al., 2004a,b). Differences found in plates, previously filled with modified GSe medium the toxin profiles of three G. catenatum populations (Blackburn et al., 1989), and maintained at 21 ± 1 °C, led us to compare the toxicity of Mexican strains of L:D, 12:12, and 65 lEmÀ2 sÀ1 light intensity. Culture G. catenatum isolated from three bays of the Gulf of media were prepared using aged Bahı´a Concepcio´n California in culture conditions to determine if they and Bahı´a de La Paz seawater (salinity 35). Both sea- have a typical or similar PST profiles. water and nutrients were sterilized through a 0.22-lm G. catenatum strains were isolated from three embay- filter. Cultures from wells were transferred into 75-mL ments from the Gulf of California (Fig. 1). Four strains culture flasks for experimental and identification from Bahı´a Concepcio´n (BACO) were isolated from purposes. Nine clonal cultures from vegetative cells vegetative cells collected by vertical tows of a 20 lm and one strain from cyst germination of G. catenatum net during April 2001. A fifth strain was obtained from were established (Table 1). Batch cultures (50 mL) were resting cysts found in sediment samples collected with a grown in tissue culture flasks at 20 ± 1 °C with overhead gravity corer (5–20 cm length, 1.3 cm in diameter) dur- illumination of 150 lEmÀ2 sÀ1 (12:12 h L:D cycle) and ing June 2000. The cores were wrapped in aluminum foil harvested during exponential growth phase after inocu- and stored in darkness at 4 °C. Strains from Bahı´ade lation with G. catenatum strains. Cultures were har- Mazatla´n were isolated from water samples taken with vested by filtration through GF/F glass fiber filters and a Van Dorn bottle during a red tide of G. catenatum immediately frozen at À20 °C. These filters were used during April 2003. Strains from Bahı´a de La Paz were for toxin analysis. also isolated from phytoplankton net (20 lm) samples Paralytic shellfish toxins were extracted by adding collected during January 2003. 2 mL acetic acid (0.03 N) to each G. catenatum contain- Cell concentrates were sieved through a 60-lm mesh ing filter, sonicated at 35 kHz for 60 s in an ice bath, cen- to eliminate larger organisms and to inoculate 250-mL trifuged at 3000 rpm for 5 min, and the supernatant culture container, previously filled with seawater en- filtered with a single-use syringe-filter (0.45 lm).