Dataset Information

Evaluation of the presence of Paenibacillus larvae in commercial bee pollen using PCR amplification of the gene for tRNACys.

ABSTRACT: 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 tRNACys-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 tRNACys-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 tRNACys-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.

SUBMITTER: Andrade VDM

PROVIDER: S-EPMC6863256 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Evaluation of the presence of Paenibacillus larvae in commercial bee pollen using PCR amplification of the gene for tRNA<sup>Cys</sup>.

Andrade Vicente Daniel Moreno VDM Flores José Luis Hernández JLH López Miguel Angel Ramos MAR Hernández Andrés Cruz AC Gómez Sergio Romero SR Medina Rosa Paulina Calvillo Calvillo Rosa Paulina Medina RPM Martínez Ana Gabriel Estrada AGE Pérez Juan Caballero JC Hernández Iván Arvizu IA Hidalgo Erika Álvarez EÁ Osuna Claudia Álvarado CÁ Jones George H GH Guillén Juan Campos JC

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] 20190121 2

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<sup>Cys</sup>-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 tR ...[more]

PMID: 30666531

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Project description:American foulbrood disease of honey bees is caused by the bacterium Paenibacillus larvae. Infection occurs per os in larvae and systemic infection requires a breaching of the host peritrophic matrix and midgut epithelium. Genetic variation exists for both bacterial virulence and host resistance, and a general immunity is achieved by larvae as they age, the basis of which has not been identified. To quickly identify a pool of candidate genes responsive to P. larvae infection, we sequenced transcripts from larvae inoculated with P. larvae at 12 hours post-emergence and incubated for 72 hours, and compared expression levels to a control cohort. We identified 75 genes with significantly higher expression and six genes with significantly lower expression. In addition to several antimicrobial peptides, two genes encoding peritrophic-matrix domains were also up-regulated. Extracellular matrix proteins, proteases/protease inhibitors, and members of the Osiris gene family were prevalent among differentially regulated genes. However, analysis of Drosophila homologs of differentially expressed genes revealed spatial and temporal patterns consistent with developmental asynchrony as a likely confounder of our results. We therefore used qPCR to measure the consistency of gene expression changes for a subset of differentially expressed genes. A replicate experiment sampled at both 48 and 72 hours post infection allowed further discrimination of genes likely to be involved in host response. The consistently responsive genes in our test set included a hymenopteran-specific protein tyrosine kinase, a hymenopteran specific serine endopeptidase, a cytochrome P450 (CYP9Q1), and a homolog of trynity, a zona pellucida domain protein. Of the known honey bee antimicrobial peptides, apidaecin was responsive at both time-points studied whereas hymenoptaecin was more consistent in its level of change between biological replicates and had the greatest increase in expression by RNA-seq analysis.

Dataset Information

Evaluation of the presence of Paenibacillus larvae in commercial bee pollen using PCR amplification of the gene for tRNACys.

Publications

Evaluation of the presence of Paenibacillus larvae in commercial bee pollen using PCR amplification of the gene for tRNA<sup>Cys</sup>.

Similar Datasets

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