{"refrec":{"BRefID":131636,"RR":"<b>Vasileiadou, K.; Papakostas, S.; Triantafyllidis, A.; Kappas, I.; Abatzopoulos, T.J.</b> (2009). A multiplex PCR method for rapid identification of <i>Brachionus</i> rotifers. <i>Mar. Biotechnol. 11(1)</i>: 53-61. <a href=\"https://dx.doi.org/10.1007/s10126-008-9119-4\" target=\"_blank\">https://dx.doi.org/10.1007/s10126-008-9119-4</a>","BEntID":125682,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":0,"RefStringPartII":". <i>Mar. Biotechnol. 11(1)</i>: 53-61. <a href=\"https://dx.doi.org/10.1007/s10126-008-9119-4\" target=\"_blank\">https://dx.doi.org/10.1007/s10126-008-9119-4</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Vasileiadou, K.; Papakostas, S.; Triantafyllidis, A.; Kappas, I.; Abatzopoulos, T.J.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Vasileiadou, K. <i>et al.</i>","Englishabstract":"Cryptic species are increasingly being recognized in many organisms. In <i>Brachionus</i> rotifers, many morphologically similar yet genetically distinct species/biotypes have been described. A number of <i>Brachionus</i> cryptic species have been recognized among hatchery strains. In this study, we present a simple, one-step genetic method to detect the presence of those <i>Brachionus</i> sp. rotifers that have been found in hatcheries. With the proposed technique, each of the <i>B. plicatilis</i> sensu stricto, <i>B. ibericus, Brachionus</i> sp. Nevada, <i>Brachionus</i> sp. Austria, <i>Brachionus</i> sp. Manjavacas, and <i>Brachionus</i> sp. Cayman species and/or biotypes can be identified with polymerase chain reaction (PCR) analysis. Based on 233 cytochrome c oxidase subunit I sequences, we reviewed all the available cryptic <i>Brachionus</i> sp. genetic polymorphisms, and we designed six nested primers. With these primers, a specific amplicon of distinct size is produced for every one of the involved species/biotypes. Two highly sensitive protocols were developed for using the primers. Many of the primers can be combined in the same PCR. The proposed method has been found to be an effective and practical tool to investigate the presence of the above six cryptic species/biotypes in both individual and communal (bulk) rotifer deoxyribonucleic acid extractions from hatcheries. With this technique, hatchery managers could easily determine their rotifer composition at the level of cryptic species and monitor their cultures more efficiently. 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