Microbiome analyses of poultry feeds: Part II. Comparison of different poultry feeds.

Within the realm of poultry feed mill operations, the persistent concern over microbial feed quality necessitates the establishment of a robust baseline for enhancing and sustaining the standards of commercial feeds. This dual-phase investigation, comprising Parts I, was previously published, and the current study presented here as Part II aimed to illuminate this baseline using 16S rRNA gene sequencing. In Part II, nine distinct commercial poultry feeds formulated as starters, growers, starter/growers, or supplements, the selected feeds underwent genomic DNA extraction, amplification with custom dual-indexed primers, and subsequent Illumina MiSeq sequencing. Through data analysis in QIIME2-2021.4 and R Studio, the study unveils alpha (Kruskal-Wallis) and beta (ANOSIM) diversity, taxonomic differences (ANCOM), and core microbiomes (core_members), deeming main and pairwise effects statistically significant at p < 0.05 and Q < 0.05. Notably, the investigation identified 30% common core microbial members across the nine feed types, shedding light on potential foodborne poultry pathogens such as Helicobacter and Campylobacter. Probiotic-associated feeds exhibited distinct microbial communities, emphasizing the need to explore their impact on the early poultry gastrointestinal tract (GIT) further.

Microbiome analyses of poultry feeds: Part I. Comparison of five different DNA extraction methods.

Given extensive variability in feed composition, the absence of a dedicated DNA extraction kit for poultry feed underscores the need for an optimized extraction technique for reliable downstream sequencing analyses. This study investigates the impact of five DNA extraction techniques: Qiagen QIAamp DNA Stool Mini Kit (Qiagen), modified Qiagen with Lysing Matrix B (MQ), modified Qiagen with celite purification (MQC), polyethylene glycol (PEG), and 1-Day Direct. Genomic DNA amplification and Illumina MiSeq sequencing were conducted. QIIME2-2021.4 facilitated data analysis, revealing significant diversity and compositional differences influenced by extraction methods. Qiagen exhibited lower evenness and richness compared to other methods. 1-Day Direct and PEG enhanced bacterial diversities by employing bead beating and lysozyme. Despite similar taxonomic resolution, the Qiagen kit provides a rapid, consistent method for assessing poultry feed microbiomes. Modified techniques (MQ and MQC) improve DNA purification, reducing bias in commercial poultry feed samples. PEG and 1-Day Direct methods were effective but may require standardization. Overall, this study underscores the importance of optimized extraction techniques in poultry feed analysis, with potential implications for future standardization of effective methods.

In silico genome analysis of an acid mine drainage species, Acidiphilium multivorum, for potential commercial acetic acid production and biomining.

The genome of Acidiphilium multivorum strain AIU 301, acidophilic, aerobic Gram-negative bacteria, was investigated for potential metabolic pathways associated with organic acid production and metal uptake. The genome was compared to other acidic mine drainage isolates, Acidiphilium cryptum JF-5 and Acidithiobacillus ferrooxidans ATCC 23270, as well as Acetobacter pasteurianus 386B, which ferments cocoa beans. Plasmids between two Acidiphilium spp. were compared, and only two of the sixteen plasmids were identified as potentially similar. Comparisons of the genome size to the number of protein coding sequences indicated that A. multivorum and A. cryptum follow the line of best fit unlike A. pasteurianus 386B, which suggests that it was improperly annotated in the database. Pathways between these four species were analyzed bioinformatically and are discussed here. A. multivorum AIU 301, shares pathways with A. pasteurianus 386B including aldehyde and alcohol dehydrogenase pathways, which are used in the generation of vinegar. Mercury reductase, arsenate reductase and sulfur utilization proteins were identified and discussed at length. The absence of sulfur utilization proteins from A. multivorum AIU 301 suggests that this species uses previously undefined pathways for sulfur acquisition. Bioinformatic examination revealed novel pathways that may benefit commercial fields including acetic acid production and biomining.