,Mediterranean fruit fly males reared for sterile insect technique (SIT) have been shown to have differences in their microbiomes relative to other population sources, which has been postulated to be a factor in the how well flies compete with wild conspecifics. To identify baseline performance metrics on the effects of irradiation on the gut microbiome of mass-reared flies in Hawai’i, a study was performed to assess performance metrics and microbiome (bacterial 16S-rRNA) variation across multiple timepoints. Mediterranean fruit fly pupae were selected from mass-reared trays intended for release, and paired samples were either irradiated or remained as controls and transported to the laboratory for evaluation.,

Fiberglass microscope slides were systematically distributed within a 5000 L larval rearing tank during a standard rearing attempt to assess biofilm development.Culture-based counts of bacterial densities were measured over 24 days for 2 media types, TCBS and Marine Agar (MA).Scanning electron microscopy (SEM) analyses of cell densities was carried out. SEM images revealed changes in dominant morphologies of bacterial populations. Succession of microbial species during biofilm development was tracked using denaturing gradient gel electrophoresis (DGGE) profiling of bacterial 16S rRNA genes in combination with statistical ordination analysis. Partial and complete 16S rDNA sequences of bacterial isolates were obtained using the primers 27f, 63f, 339f, 907r, 1387r, 1492r. Sequencing of re-amplified DGGE bands was performed with the 1392r primer (only sequences affiliated with a- and g-Proteobacteria were retrieved). Sequences were aligned to the closest relatives in the NCBI database using the BLAST algorithm.Fluorescence in situ hybridization (FISH) identified a-Proteobacteria as being numerically abundant in the biofilm. To evaluate if the biofilm is a reservoir for potentially pathogenic bacteria that cause mass larval mortalities. Trends between the DGGE profiles, SEM and plating on MA were analysed in the paper outlined in 'Lineage'.Identification of isolates derived from day 24 biofilm samples (and accession numbers): Vibrio alginolyticus (AY332566); V. campbellii (AY738126); V. harveyi (AY264924); V. harveyi (AY750578); Shewanella algae (AB205581).Phylogenetic identity of dominant bands from the DGGE bacterial profile of biofilm samples (and database accession number):Bacterium K2-11 (AY345438); Erythrobacter sp. (AB094461); Loktanella agnita (AY682198); Marine bacterium (AF388307); Marinobacterium jannaschii (AB006765); Oceanospirillum sp. (AJ302699); Rhodobacteraceae bacterium (AY962292); Uncultured agrobacterium (AF194391); Uncultured bacterium (AY845232); Uncultured gamma-proteobacterium (AJ318111); Vibrio harveyi (AY264924); Vibrio harveyi (AY264933); ¿-proteobacterium JC2042 (AY207502).

,Source Insects and Microbes,Sitophilus oryzae that were 4–8-week-old and collected from a food facility in eastern Kansas in 2012 were used. Adults were reared continuously in the laboratory at CGAHR and subcultured on tempered whole organic wheat, with 75 mixed-sex adults added to 300 g of wheat. Adults were sieved off after a 1-week oviposition period. Colonies were maintained in a chamber set at 27.5˚C, 65% RH, and 14:10 (L:D) h photoperiod. The fungus, Aspergillus flavus, was isolated from field-collected S. oryzae after allowing adults to forage on potato dextrose agar dishes (100 × 15 mm) for 5 d. From the initial dish, A. flavus was re-plated on a new dish in multiple successive rounds until it was a pure isolate. This was used to create A. flavus-inoculated grain, which was then used for the treatments below.,Treatments and Preparation of Grain Masses,At the outset of the experiment, the grain moisture of the wheat for the experiments was determined to be 10.8% using a moisture meter (DICKEY-john, GAC2100, Auburn, IL, USA). A total of 300 g of organic, whole untampered organic wheat (Heartland Mills, Marienthal, KS, USA) was added to each pint mason jar (950 mL; 8.5 D × 17 cm H). In order to assess the effect of singly or multiply infested grain by insects and microbes on the microclimate and fitness outcomes, each grain mass was assigned to one of our treatments: control (no insects or added microbes; Ctrl), the addition of 75 mixed-sex S. oryzae adults only (SO), inoculation with 11.6 g of A. flavus inoculated grain only (AF, details below), and finally the addition of both S. oryzae (75 adults) and A. flavus (11.6 g of inoculated grain; SO + AF). In each grain mass, a datalogger (Hobo® U10-003 Temp/RH Data Logger, Onset, Bourne, MA, USA) was attached below the lid and recorded temperature and relative humidity every 5 min. The experiment was allowed to run for 60 d in an environmental chamber set to 30°C, 60% RH, and 14:10 L:D photoperiod. At the end of the period, all jars were immediately frozen to stop halt reproduction and microbial activity. There were n = 5 replicate grain masses per treatment. For the purposes of looking at changes in abiotic variables, the 60 d period was divided into equal 20 d increments, and labeled early (first 20 d), mid (second 20 d), and late (final 20 d).,Preparation of A. flavus-inoculated grain,In order to inoculate treatments with A. flavus, an inoculum was prepared from wheat that had already undergone a complete colonization process. Briefly, 600 g of grain was added to a stainless-steel pot filled with water and placed on a hot plate at 500°C. It was allowed to boil for 15 min, then the water was drained, and the grain was evenly spread out on sterile wipes (38.1 × 42.5 cm, 3 ply, Tech wipes, Skilcraft, NIB, Alexandria, VA). The grain was allowed to dry inside a laminar fume hood (for ca. 3 h). Subsequently, the grain was divided (in 300 g lots) and placed in two separate sterile mason jars (950-mL capacity). A single hole was drilled through each lid and lined with a cotton ball. The jars were then sealed with aluminum foil and were autoclaved (533LS, Getinge, Rochester, NY, USA) for 30 min. To inoculate with A. flavus, a 3-inch strip of agar containing a pure culture of A. flavus grown on PDA for 7 days at 30°C, 60% RH, and 14:10 L:D photoperiod as above (Ponce et al., 2023; Ponce et al., 2024) was placed into each jar containing the grain. The jars were maintained at room temperature for roughly 10 days or until the A. flavus evenly covered as much of the grain as possible. Inoculated grain was used within 10–15 days of preparation. A total of 11.6 g of this inoculated grain was then added to 300 g for the A. flavus treatments above.,Grain Moisture & Progeny Production,At the end of 60 d, grain moisture readings were taken from 20 g of every replicate and each treatment after allowing grain masses to reach room temperature by using a moisture meter (DICKEY-john, GAC2100,

Data were generated as part of a NASA-funded study (Turek F (PI) et al. Effects of Spaceflight on Gastrointestinal Microbiota in Mice: Mechanisms and Impact on Multi-System. NASA NRA: NRA NNH14ZTT002N). As part of the study, we requested and received samples from RR1. We generated 16S rRNA gene amplicon sequence data from DNA extracted from fecal samples, and compared these data to similar data generated on shuttle mission STS-135 and from ground-based studies of radiation. We assessed effect of flight conditions and radiation.

,Study aim was to determine establishment success of antibiotic resistance bacteria in laboratory and mass-reared Mediterranean fruit fly (Ceratitis capitata). Experiments conducted in Hilo, HI, USA by the USDA ARS. Datasets include two different datatypes: 1) data from plate counts (CFUs) on nutrient-rich and antibiotic-amended data and 2) amplicon sequence variant (ASVs) count data from full-length 16S rRNA sequencing using PacBio Kinnex mas-Seq on a PacBio Revio sequencer and associated taxonomy and .fasta files. Experiments were performed with two different fly lines, one maintained by the USDA ARS and another by California Department of Agriculture (CDFA). Experiments were performed with a target Enterobacter strain that was transformed with mScarlet and antibiotic resistance selection markers. Experiments evaluated the impact of fly age and inoculation formulation (via diet) on the conditions in which bacteria would be established under different dietary conditions. Data show that liquid diets support establishment of bacteria, regardless of age class and source. Sequence data suggest additional Enterobacter strains are present in flies beyond the target strain and these can be distinguished via full-length 16S rRNA sequencing, but not by shorter amplicon fragments.,

To evaluate effects of human influence on the health of Puget Sound's pelagic ecosystems, we propose a sampling program across multiple oceanographic basins measuring key attributes of the pelagic foodweb. We will quantify seasonal abundance and composition of pelagic biota from lower trophic levels (e.g., bacteria and phytoplankton) to middle trophic levels (e.g., zooplankton, small pelagic fishes, and jellyfish), as well as assess the individual condition of forage fish and juvenile salmon. The goals of this program: 1) Determine how foodweb endpoints vary across natural and anthropogenic gradients. 2) Determine how these characteristics vary across Puget Sound. 3) Evaluate a number of biological metrics for monitoring ecosystem health. These outputs will improve our basic understanding of pelagic ecology in Puget Sound, better define what comprises a healthy pelagic ecosystem in Puget Sound, determine foodweb-relevant indicators that are sensitive to human influence, and help prioritize regional protection and restoration efforts. Work is conducted by NOAA personnel and contractors in collaboration with tribal partners (Squaxin, Port Gamble/S'Klallam), undergraduate research interns, and citizen volunteers. Filemaker Pro database containing microbial abundance, heterotrophic production, chlorophyll a, and inorganic nutrients.