Complex of primary and secondary parasitoids (Hymenoptera: Encyrtidae and Signiphoridae) of Hypogeococcus spp. mealybugs (Hemiptera: Pseudococcidae) in the New World Author Triapitsyn, Serguei V. Department of Entomology, University of California, Riverside, California, 92521, USA; E-mails: serguei. triapitsyn @ ucr. edu (S. V. T.); paul. rugman-jones @ ucr. edu (P. F. R. J.) serguei.triapitsyn@ucr.edu Author Aguirre, María B. FuEDEI, Simón Bolívar 1559, Hurlingham, Buenos Aires, Argentina; E-mails: glogarzo @ fuedei. org (G. A. L.); redbell _ @ hotmail. com (M. B. A.) glogarzo@fuedei.org Author Logarzo, Guillermo A. FuEDEI, Simón Bolívar 1559, Hurlingham, Buenos Aires, Argentina; E-mails: glogarzo @ fuedei. org (G. A. L.); redbell _ @ hotmail. com (M. B. A.) glogarzo@fuedei.org Author Hight, Stephen D. USDA-ARS, 6383 Mahan Dr., Tallahassee, Florida, 32308, USA; E-mail: Stephen. Hight @ ARS. USDA. GOV (S. D. H.) Author Ciomperlik, Matthew A. USDA-APHIS-PPQ-CPHST, Mission Laboratory, 22675 N. Moorefield Rd., Edinburg, Texas, 78541 - 9398, USA; E-mail: Matt. A. Ciomperlik @ aphis. usda. gov (M. A. C.) Matt.A.Ciomperlik@aphis.usda.gov Author Rugman-Jones, Paul F. Department of Entomology, University of California, Riverside, California, 92521, USA; E-mails: serguei. triapitsyn @ ucr. edu (S. V. T.); paul. rugman-jones @ ucr. edu (P. F. R. J.) serguei.triapitsyn@ucr.edu Author Verle Rodrigues, Jose C. Center for Excellence in Quarantine & Invasive Species, Agricultural Experimental Station-Río Piedras, Crops and Agro-Environmental Sciences Department, University of Puerto Rico-Mayagüez, 1193 Calle Guayacán, San Juan, Puerto Rico 00926, USA; E-mail: jose _ carlos @ mac. com (J. C. V. R.) jose_carlos@mac.com text Florida Entomologist 2018 2018-09-30 101 3 411 434 http://www.bioone.org/doi/10.1653/024.101.0320 journal article 10.1653/024.101.0320 1938-5102 13260321 Anagyrus tanystis De Santis, 1964 ( Figs. 37–41 ) Anagyrus tanystis De Santis 1964: 61−63 . Type locality: Autonomous City of Buenos Aires , Argentina . Anagyrus tanystis De Santis : Triapitsyn et al. 2014b: 221 (key). Type Material Examined . Holotype female [ MACN ] on slide ( Fig. 37 ) labeled: 1. “CAPITAL FEDERAL Col: A. Zotta I-II/1913”; 2. [partially printed in red] “ Anagyrus tanystis Det. De Santis HOLOTIPO MUSEO DE LA PLATA ”; 3. [ MACN type number] “A 89”. Distribution . Autonomous City of Buenos Aires , Argentina ( De Santis 1964 ). Hosts . Unknown. Comments . The holotype female of A. tanystis is dissected under 1 large square coverslip ( Fig. 37 ); it lacks almost the entire flagellum of 1 of the antennae except for about a half of F1. Illustrated here, to facilitate its recognition, are its features ( Figs. 38–41 ). This species is known only from a single specimen, which was not examined by Triapitsyn et al. (2014b) during preparation of the key to the described species of Anagyrus in Argentina . GENETIC ANALYSIS DNA sequences of all 3 loci (28S, ITS2 and COI) were successfully obtained for 29 of 30 specimens . The only “shortfall” was the COI of a single specimen ( PR17-254 ) from Argentina ,which failed to amplify de- spite multiple attempts. Direct-sequencing of the COI locus also failed for a second specimen from Argentina ( PR 17-256) and 3 from Brazil ( PR 17-247 through 249) due to co-amplification of a NUMT (GenBank accessions MG748829-832 ). However, a valid COI sequence was obtained for each of these specimens by amplicon cloning . ML analyses of the concatenated 28S (MG731466-495) and COI (MG731496-524) sequences recovered 3 distinct clades ( Fig. 42 ).These 3 clades corresponded to the species Leptomastidea hypogeococci , Anagyrus cachamai , and Anagyrus quilmes . Although discarded for the purpose of phylogenetic inference, sequences of ITS2 (MG731525- 554) identified the same 3 clades. Leptomastidea hypogeococci – DNA sequences of the rRNA loci 28S and ITS2 were identical across specimens from Puerto Rico , Argentina , and Brazil , with the slight exception that the 28S sequence of 3 specimens ( PR 17-253 and PR 17-257 from Puerto Rico , and PR17-254 from Argentina ) harbored a heterozygous peak at position 140 (G or A, rather than just a G). The Puerto Rican specimens shared an identical COI haplotype, but on average this differed from the COI of Brazilian and Argentinian specimens (obtained from cloned amplicons) by 3.7% (average pairwise p-distance = 0.037). Variation in COI among Brazilian and Argentinian specimens was 10-fold lower (average p-distance = 0.003) . Anagyrus cachamai – DNA sequences of 28S were more or less identical across specimens from Argentina and Paraguay , with just a separate single base insertion at positon 447 and 451 in specimens PR17-246 ( Paraguay ) and PR17-252 ( Argentina ), respectively. ITS2 was more variable, with 26 single nucleotide substitutions, insertions or deletions across 5 specimens and 695 aligned bases. In addition, 2 specimens (PR14-212 and PR14-213; MG731548-548 ) differed from the other 3 ( MG731534-536 ) with a 29bp deletion (positions 148-176). These 2 specimens also shared a COI haplotype ( MG731518-519 ) that differed from that of the other 3 specimens ( MG731504-506 ) by over 5% (average pairwise p-distance = 0.054). Variation among the latter 3 specimens was 4-fold lower (average p-distance = 0.013) . Figs. 34–36. Prochiloneurus spp. males (34–35, P. argentinensis , Loreto , Misiones, Argentina [photographs courtesy of D. A. Aquino]; 36, P. narendrani , Mona Island, Puerto Rico): 34, habitus; 35, antennae; 36, habitus. Anagyrus quilmes – DNA sequences of 28S and ITS2 were identical across all specimens from Puerto Rico , Argentina , Brazil , and Paraguay . Eight COI haplotypes were detected, and there was a strong phylogeographic signal in the distribution of these haplotypes ( Fig. 43 ). Pairwise divergence between 7 of these haplotypes was typically low (average pairwise p-distance = 0.005), but the remaining haplotype, harbored by PR14-214, differed from all others by over 5% (average pairwise p-distance = 0.054). BIOLOGICAL CONTROL OF THE HARRISIA CACTUS MEALYBUG IN PUERTO RICO The biological control program for Harrisia cactus mealybug in Puerto Rico was advanced with 2 shipments of 2 parasitoid species from South America to the quarantine facility at the University of Puerto Rico Center for Excellence in Quarantine & Invasive Species. The species transported to the quarantine facility were A. cachamai and A. lapachosus . Both parasitoid species accepted the Puerto Rico Harrisia cactus mealybug,and host range tests were initiated. Quarantine host range tests are expected to be completed by spring 2018, and a request for field release should be submitted before summer 2018. In addition, studies on natural enemy population dynamics and parasitoid biological parameters are planned for the 2 quarantined parasitoid colonies from South America, and a newly described parasitoid species already established and attacking the Harrisia cactus mealybug in Puerto Rico . This project is poised to successfully de- velop the only method that challenges the Harrisia cactus mealybug pest before we experience the devastation of cactus biodiversity in Puerto Rico .