870 



The occurrence of the provisional 

 Brazilian subspecies of spiny lobster 

 (Panuiirus argus westonii) in Florida waters* 



Shane K. Sarver 



Department of Biology 

 Black Hills State University 

 Spearfish, South Dakota 57799 

 E-mail address: ssarverfgjmystic bhsu edu 



D. Wilson Freshwater 



Center for Marine Science Research 

 7205 Wnghtsville Avenue 

 Wilmington, North Carolina 28403 



Patrick J. Walsh 



Division of Marine Biology and Fishenes 

 Rosenstiel School of Marine and Atmosphenc Science 

 University of Miami 

 Miami, Florida 33149 



The Caribbean spiny lobster, Panuii- 

 rus ar-gus, is distributed from Brazil 

 throughout the Caribbean and the Gulf 

 of Mexico to approximately North Car- 

 olina and Bermuda (Holthius, 1991). 

 It supports major commercial fisheries 

 in Florida, the Caribbean and Brazil. 

 Commercially, P. argus is especially 

 important to the state of Florida, where 

 the spiny lobster fishery ranks second 

 only to shrimp in terms of economic 

 value. There is also a significant rec- 

 reational fishery for P. argus, particu- 

 larly in Florida. A number of studies 

 have been initiated to gather informa- 

 tion to manage the lobster resource 

 carefully. 



Several studies have used genetic 

 techniques to examine population-level 

 patterns of differentiation to delineate 

 reproductively isolated stocks of P. 

 argus. Evidence of the stock structure 

 of spiny lobster populations could then 

 be used to implement more effective 

 fishery management plans. These stud- 

 ies have provided somewhat ambiguous 

 results. Using allozymes, Menzies and 

 Kerrigan (1979) and Menzies (1981) 

 provided some evidence for genetic dif- 

 ferences among populations, but more 

 recent studies using restriction frag- 

 ment length polymorphism (RFLP) 

 analysis of mitochondrial DNA have 



suggested little evidence for genetic 

 differences among Caribbean popula- 

 tions of P. argus (Silberman et al., 

 1994a. 1994b). Analysis of mtDNA 

 RFLPs have revealed suprisingly high 

 levels of diversity among individuals, 

 and several individuals were diverged 

 at levels not usually seen within a spe- 

 cies. Because previous studies did 

 not include populations from Brazil, 

 Sarx'er et al. ( 1998) compared P. a>-gus 

 from Caribbean locations with P. argus 

 from Brazil. Using DNA sequence anal- 

 ysis of two mitochondrial genes, Sarver 

 et al. (1998) found high levels of 

 sequence divergence between Carib- 

 bean and Brazilian P. argus. The levels 

 of nucleotide sequence divergence were 

 greater than that seen between rec- 

 ognized species of Panuiirus. In addi- 

 tion, there are striking color differences 

 between Caribbean and Brazilian P. 

 argus. As a result of these findings 

 Sarver et al. (1998) suggested provi- 

 sional recognition of two subspecies of 

 P. argus {P. argus argus in the Carib- 

 bean and P. argus westonii in Brazil) 

 until a formal taxonomic revision could 

 be done. 



The results from these studies, which 

 suggest that P. argus in Brazil are 

 genetically and taxonomically distinct 

 from their (Caribbean counterparts. 



have raised questions about the status 

 of the three genetically distinct spiny 

 lobsters reported by Silberman et 

 al. (1994a) in their original survey 

 of mtDNA diversity of Caribbean 

 P. aj-gus. This is significant because 

 two of these spiny lobsters found in 

 Silberman et al. (1994a) were caught 

 off the coast of Miami, Florida. Our 

 study uses DNA sequence analysis to 

 identify the three genetically distinct 

 spiny lobsters observed by Silberman 

 et al. (1994a) as the Brazilian form of 

 P. argus (provisionally recognized as P. 

 argus westonii, in Sarver et al., 1998). 



Materials and methods 



DNA samples were obtained from the 

 samples examined by Silberman et al. 

 (1994a). Caribbean Panuiirus argus 

 samples were randomly selected from 

 the samples examined by Silberman 

 et al. (1994a). Samples from Brazil 

 were collected near the Rio Grande do 

 Norte region of Brazil. Tissue samples 

 of Panuiirus argus from Brazil were 

 frozen prior to DNA isolation. Stan- 

 dard phenol or chloroform DNA extrac- 

 tion techniques were used for DNA 

 isolation (Hillis et al., 1990). A region 

 of the mitochondrial 16S rRNA gene 

 was amplified by the polymerase chain 

 reaction (PCR) by using primers 16Sar 

 and 16Sbr given in Palumbi et al.' 

 PCR products were then purified and 

 used as templates for DNA sequenc- 

 ing by using the JTaq cycle sequencing 

 kit (U.S. Biochemical Corp., Cleveland, 

 OH). Cycle sequencing reactions were 

 performed by using y-^'P-dATP end- 



Contribution 221 of the Center for Marine 

 Science Research, Wilmington. NC 2840.3. 

 Palumbi. S. R., A Martin, S. Romano, W. O. 

 McMillan, L. Stice, and G. Grabowski. 

 1991. A simple fools guide to PCR, vers. 

 2.0. Special publication of the Univer- 

 sity of Hawaii Department of Zoology and 

 Kcwalo Marine Laboratory. 46 p. Depart- 

 ment of Zoology and Kewalo Marine Labo- 

 ratory, Univ. Hawaii, Honolulu. HI 96822. 



Manuscript accepted 1 June 2000. 

 Fish. Bull. 98:870-873 (20001. 



