In Vitro Culture and Phylogenetic Analysis of "Candidatus Arsenophonus triatominarum," an Intracellular Bacterium from the Triatomine Bug, Triatoma infestans

Abstract

An intracellular symbiotic bacterium was isolated from the hemolymph of Triatorna infestans and cultured in an Aedes albopictus cell line. 16s ribosomal DNA sequence analysis revealed that the bacterium was a member of the y-3 subgroup of the class Proteobacteria, having 96.2% sequence identity with the most closely related bacterium, Arsenophonus nasoniae, the causative agent of the son-killer trait in the parasitoid wasp Nasonia vitripennis. These bacteria share morphological features and a common tissue distribution and transmission mode. The A. nasoniae-T. infestans symbiont branch represents a lineage of insect symbionts which may be capable of horizontal transmission between phylogenetically distant host insects. We propose that the intracellular symbiont from T. infestans be classified as "Candidatus Arsenophonus triatominarum." The bacterium found in the hemocytes of T. infestans is designated the type strain of this species.

Thorsellia anophelis is the dominant bacterium in a Kenyan population of adult Anopheles gambiae mosquitoes

“…Artificial culture has been achieved only on insect cell lines (Hypsa and Dale, 1997;Dale et al, 2006). Different species of Arsenophonus from different hosts share 499% 16S rRNA gene sequence similarity, suggesting recent host acquisition (Dale et al, 2006).…”

Endosymbiotic Bacteria in Insects: Their Diversity and Culturability

“…Meanwhile, the molecular mechanisms underlying insect-microbe interactions are much less well understood, because most of these insect mutualistic symbionts are neither culturable nor genetically manipulatable. Recent studies have succeeded in the isolation of several facultative symbionts by using insect cell lines or axenic media (27,29,32,57,65,87,92), revolutionizing studies of insect endosymbiosis. The present article reviews the amazing diversity of bacterial endosymbiosis in insects, focusing on several model systems with culturable endosymbionts, which provides a new perspective regarding how intimate symbiotic associations may have evolved and how they are maintained within insects.…”

Diferenciación de especies de Rhodococcusmediante una prueba de PCR-RFLP basada en los genes codificantes para la subunidad 16S ribosomal

“…Electroforesis en gel de agarosa al 2% coloreado con bromuro de etidio conteniendo 15 ml del producto de digestión. E. coli sin digerir (1), perfil de restricción con la enzima PstI para: E. coli (2), R. rhodnii, (3), R. equi (4) y A. baumani (5), perfil de restricción con la enzima HindIII para: E. coli (6), R. rhodnii, (7), R. equi (8) y A. baumani (9), perfil de restricción con la enzima BstEII para: E. coli (10), R. rhodnii, (11), R. equi (12) y A. baumani (13). Los tamaños de los fragmentos se indican en pb.…”

“…Condiciones de la reacción de PCR: el ADN bacteriano fue amplificado con los iniciadores: 16F (5'-GCTTAACACATGCAAG-3') y 16R (5'-ACGGGCAGTGTGTACAAGACC-3'), correspondientes a las posiciones 45 a 61 y 1406 a 1386, respectivamente, del gen codificante para el ARN ribosomal 16S de E. coli (13). La PCR se realizó en un volumen final de 50 ml, según lo descrito por Hypsa y Dale, (1997) y el producto de amplificación fue visualizado mediante electroforesis horizontal en geles de agarosa al 1% coloreados con bromuro de etidio (12).…”