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Data from: Pyrosequencing-Based Analysis of the Microbiome Associated with the Horn Fly, Haematobia irritans

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posted on 2024-02-08, 19:22 authored by Azhahianambi Palavesam, Felix D. Guerrero, Andrew M. Heekin, Ju Wang, Scot E. Dowd, Yan Sun, Lane D. Foil, Adalberto A. Pérez de León

The bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) method was used to carry out the classification analysis of bacterial flora in adult female and male horn flies and horn fly eggs.

The bTEFAP method identified 16S rDNA sequences in our samples which allowed the identification of various prokaryotic taxa associated with the life stage examined. This is the first comprehensive report of bacterial flora associated with the horn fly using a culture-independent method. Several rumen, environmental, symbiotic and pathogenic bacteria associated with the horn fly were identified and quantified. This is the first report of the presence of Wolbachia in horn flies of USA origin and is the first report of the presence of Rikenella in an obligatory blood feeding insect.

Adult horn flies were collected on a single date from pastured cattle at the Louisiana State University Agricultural Center, St. Gabriel Research Station using aerial nets. Within 1 h after collection the flies were transferred to large sterile Erlenmeyer flasks and maintained in total darkness for 1.5 h and 30°C to allow flies to oviposit on the flask bottom [73]. Adult flies were released from the flasks into a cage and eggs were collected by rinsing with distilled water onto a filter paper. Both the eggs and adult flies were frozen at −80°C. To preserve nucleic acid integrity, adults were sexed on dry ice prior to freezing. Each sample used for DNA extraction and pyrosequencing consisted of 5 adult males, 5 adult females or 50 eggs pooled together and homogenized. Three replicates of adult male, adult female and eggs were analyzed.

The horn fly, Haematobia irritans, is one of the most economically important pests of cattle. Insecticides have been a major element of horn fly management programs. Growing concerns with insecticide resistance, insecticide residues on farm products, and non-availability of new generation insecticides, are serious issues for the livestock industry. Alternative horn fly control methods offer the promise to decrease the use of insecticides and reduce the amount of insecticide residues on livestock products and give an impetus to the organic livestock farming segment. The horn fly, an obligatory blood feeder, requires the help of microflora to supply additional nutrients and metabolize the blood meal. Recent advancements in DNA sequencing methodologies enable researchers to examine the microflora diversity independent of culture methods.


Resources in this dataset:

  • Resource Title: Supporting Information: Table S1 (CSV).

    File Name: journal.pone_.0044390.s001.csv

    Resource Description: The file contains bacterial species load in the adult female horn flies, the adult male horn flies and horn fly eggs. Bacterial load was determined by dividing the 16S rDNA numbers representing the tally for each species by the total tally detected in each life stage. We report the average from the three replicates in the text.

    http://dx.doi.org/10.1371/journal.pone.0044390.s001


  • Resource Title: Horn Fly Microbiome - Data Dictionary.

    File Name: HornFlyMicrobiome_DataDictionary.csv

    Resource Description: Machine-readable CSV format data dictionary defining data elements for Supplemental Table S1, Bacterial flora in adult female and male horn flies and horn fly eggs.


  • Resource Title: Supporting Information: Table S1.

    File Name: pone.0044390.s001.xls

    Resource Description: The file contains bacterial species load in the adult female horn flies, the adult male horn flies and horn fly eggs. Bacterial load was determined by dividing the 16S rDNA numbers representing the tally for each species by the total tally detected in each life stage. We report the average from the three replicates in the text.

    http://dx.doi.org/10.1371/journal.pone.0044390.s001

    (XLS)

    Direct data download: http://journals.plos.org/plosone/article/asset?unique&id=info:doi/10.1371/journal.pone.0044390.s001


  • Resource Title: Supporting Information: Figure S1.

    File Name: pone.0044390.s002.jpg

    Resource Description: Rarefaction curves plotted at 0.03 divergence level for all nine samples of horn fly. The rarefaction curves imply a depth of coverage of approximately 10000 sequences/sample for the female and egg samples. The male horn fly samples were only sampled to about one third of the depths of either the female or egg samples.

    http://dx.doi.org/10.1371/journal.pone.0044390.s002

    (JPG)

    Download image directly at http://journals.plos.org/plosone/article/asset?unique&id=info:doi/10.1371/journal.pone.0044390.s002

Funding

USDA-ARS: 58-6205-0-038

University of Maryland: 58-6205-0-038

USDA: 34-WT-10-5000-1992-IA

History

Data contact name

Palavesam, Azhahianambi

Data contact email

nambibio@gmail.com

Publisher

PLoS ONE

Temporal Extent Start Date

2012-02-01

Theme

  • Not specified

Geographic Coverage

{"type":"FeatureCollection","features":[{"geometry":{"type":"Point","coordinates":[-91.1048504,30.2668235]},"type":"Feature","properties":{}}]}

Geographic location - description

Louisiana State University Agricultural Center, St. Gabriel Research Station, 5755 LSU Ag Road St. Gabriel, LA 70776

ISO Topic Category

  • biota
  • farming

Ag Data Commons Group

  • Veterinary Pest Genomics Center

National Agricultural Library Thesaurus terms

microbiome; Haematobia irritans; ribosomal DNA; sequence analysis; classification; bacteria; females; males; eggs; nucleotide sequences; sampling; rumen; virulent strains; Wolbachia; Rikenella; hematophagous insects; cattle; Louisiana; oviposition; imagos; cages; water; paper; dry ice; freezing; extraction; insecticides; administrative management; insecticide resistance; insecticide residues; livestock and meat industry; control methods; livestock products; organic foods; organic production; livestock farming; blood; supply; nutrients; blood meal; Biological Sciences; Insects and Entomology

OMB Bureau Code

  • 005:18 - Agricultural Research Service

OMB Program Code

  • 005:040 - National Research

ARS National Program Number

  • 104

Pending citation

  • No

Public Access Level

  • Public

Preferred dataset citation

Palavesam, Azhahianambi; Guerrero, Felix D.; Heekin, Andrew M.; Wang, Ju; Dowd, Scot E.; Sun, Yan; Foil, Lane D.; Pérez de León, Adalberto A. (2015). Data from: Pyrosequencing-Based Analysis of the Microbiome Associated with the Horn Fly, Haematobia irritans. PLoS ONE. https://doi.org/10.15482/USDA.ADC/1529230