KlebsiellaBacteria Isolated from the Genital Chamber ofPhyllophaga obsoleta

Attraction of Adults of Cyclocephala lunulata and Cyclocephala barrerai (Coleoptera: Scarabaeoidea: Melolonthidae) towards Bacteria Volatiles Isolated from Their Genital Chambers

“…This is the first study of the activity of VOCsB in members of Melolonthidae from Mexico in terms of the isolation of bacteria from the genital chamber, identification of VOCsB emitted from it, and the confirmation of biological activity. The isolation and identification of microorganisms in these species of Cyclocephala are the first recorded for this genus and the third report for Melolonthidae [ 16 , 17 ]; thus, this work provides more information on the diversity of microorganisms associated with the reproductive system of these beetles. The bacterial diversity of the genital chamber of C. lunulata and C. barrerai are like each other and to that reported in other insect species [ 29 , 34 , 42 ].…”

“…The genome of K. oxytoca suggests that this microorganism may emit short-chain alcohols [ 45 ]. The emission of aromatic alcohols is common in bacteria of the family Enterobacteriaceae [ 18 ] to which Cyclocephala bacteria belong, as well as the bacteria isolated from other Melolonthidae previously studied [ 16 , 17 ]. The emission of 3-methyl butane-1-ol in K. oxytoca is the first report with an amino acid-rich culture medium [ 24 ].…”

“…Based on the technique proposed by Rosete-Enríquez and Romero-López [ 17 ], bacteria were extracted from the genital chamber of 10 females of C. lunulata and C. barrerai . The colony-forming units of each bacterial strain were cultivated to describe their colonial morphology and identifying bacterial morphology using Gram staining.…”

“…Genomic DNA was extracted and purified according to the Wizard ® Genomic DNA Purification Kit (Promega, Madison, WI, USA). The amplification of two segments of the 16S rRNA gene was performed by a polymerase chain reaction (PCR) based on the protocol proposed by Rosete-Enríquez and Romero-López [ 17 ]. Briefly, two reactions carried out using 10 ng of genomic DNA, 200-μM deoxynucleotide solution mix (dNTPs), 1 μM of each pair of upper and lower primers, 2 or 1.5-mM MgCl 2 , 1.25 units of PlatimunTM Taq DNA polymerase, 1X PCR buffer (Invitrogen, Carlsbad, CA, USA), and DNase free water to make 20 μL.…”

“…These microorganisms produce volatile organic compounds from bacteria (VOCsB) such as phenol, which has been reported as a component of the sex pheromone of C. zealandica [ 15 , 16 ]. The presence of the enterobacteria Klebsiella oxytoca and Klebsiella michiganensis have also been recorded in the genital chamber of Phyllophaga obsoleta , where a similar relationship may occur [ 17 ]. These relationships have not been studied in Cyclocephala (Coleoptera: Scarabaeoidea: Melolonthidae) [ 3 ].…”