Evaluation of the presence of Paenibacillus larvae in commercial bee pollen using PCR amplification of the gene for tRNA Cys

• Published in: Brazilian journal of microbiology Vol. 50; no. 2; p. 471
• Main Authors: Andrade, Vicente Daniel Moreno, Flores, José Luis Hernández, López, Miguel Angel Ramos, Hernández, Andrés Cruz, Gómez, Sergio Romero, Medina, Rosa Paulina Calvillo, Calvillo, Rosa Paulina Medina, Martínez, Ana Gabriel Estrada, Pérez, Juan Caballero, Hernández, Iván Arvizu, Hidalgo, Erika Álvarez, Osuna, Claudia Álvarado, Jones, George H, Guillén, Juan Campos
• Format: Journal Article
• Language: English
• Published: Brazil 01-04-2019
• Subjects: Animals; Bacillus - genetics; Bees - microbiology; Nucleic Acid Amplification Techniques; Paenibacillus larvae - genetics; Paenibacillus larvae - isolation & purification; Pollen - microbiology; Polymerase Chain Reaction; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; RNA, Transfer, Cys - genetics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; United States; United States; Amplicon-16S rRNA; MALDI-TOF MS; Paenibacillus larvae; tRNACys-PCR
• ISSN: 1678-4405

American foulbrood (AFB) caused by Paenibacillus larvae is the most destructive honeybee bacterial disease and its dissemination via commercial bee pollen is an important mechanism for the spread of this bacterium. Because Mexico imports bee pollen from several countries, we developed a tRNA
-PCR strategy and complemented that strategy with MALDI-TOF MS and amplicon-16S rRNA gene analysis to evaluate the presence of P. larvae in pollen samples. P. larvae was not detected when the tRNA
-PCR approach was applied to spore-forming bacterial colonies obtained from three different locations and this result was validated by bacterial identification via MALDI-TOF MS. The genera identified in the latter analysis were Bacillus (fourteen species) and Paenibacillus (six) species. However, amplicon-16S rRNA gene analysis for taxonomic composition revealed a low presence of Paenibacillaceae with 0.3 to 16.2% of relative abundance in the commercial pollen samples analyzed. Within this family, P. larvae accounted for 0.01% of the bacterial species present in one sample. Our results indicate that the tRNA
-PCR, combined with other molecular tools, will be a useful approach for identifying P. larvae in pollen samples and will assist in controlling the spread of the pathogen.