Molecular evidence for multiple infections as revealed by typing of Asaia bacterial symbionts of four mosquito species.
The recent increased detection of acetic acid bacteria (AAB) of the genus Asaia as symbionts of mosquitoes, such as Anopheles spp. and Aedes spp., prompted us to investigate the diversity of these symbionts and their relationships in different mosquito species and populations. Following cultivation-dependent and -independent techniques, we investigated the microbiota associated with four mosquito species, Anopheles stephensi, Anopheles gambiae, Aedes aegypti, and Aedes albopictus, which are important vectors of human and/or animal pathogens. Denaturing gradient gel electrophoresis (DGGE) analysis based on the 16S rRNA gene revealed the presence of several bacterial taxa, among which Asaia sequences were among the dominant in most of the samples. A collection of 281 Asaia isolates in cell-free media was established from individuals belonging to the four species. The isolates were typed by internal transcribed spacer (ITS)-PCR, tRNA-PCR, BOX-PCR, and randomly amplified polymorphic DNA (RAPD)-PCR, revealing that different Asaia strains are present in different mosquito populations, and even in single individuals.
Chouaia
Bessem
B
Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria, Università degli Studi di Milano, Via Celoria 10, 20133 Milan, Italy. claudio.bandi@unimi.it
Rossi
Paolo
P
Montagna
Matteo
M
Ricci
Irene
I
Crotti
Elena
E
Damiani
Claudia
C
Epis
Sara
S
Faye
Ingrid
I
Sagnon
N'fale
N
Alma
Alberto
A
Favia
Guido
G
Daffonchio
Daniele
D
Bandi
Claudio
C
D016947:Acetobacteraceae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D000330:Aedes / Q000382:microbiology* / Q000502:physiology; D000818:Animals; D000852:Anopheles / Q000382:microbiology* / Q000502:physiology; D015373:Bacterial Typing Techniques; D044822:Biodiversity; D016000:Cluster Analysis; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA; D013559:Symbiosis; D014481:United States
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012343:RNA, Transfer