Next-generation sequencing has been used to strengthen knowledge about taxonomic diversity and ecology of fungi within food ecosystems. However, primer amplification and identification bias could edge our understanding into the fungal ecology. The aim of this study is to compare the performance of two primer pairs over two nuclear ribosomal RNA (rRNA) regions of the fungal kingdom, namely the ITS2 and 26S regions. Fermented cocoa beans were employed as biological material and the fungal ecology during fermentation was studied using amplicon-based sequencing tools, making use of a manually curated 26S database constructed in this study, and validated with SILVA's database. To explore potential biases introduced by PCR amplification of fungal communities, a mock community of known composition was prepared and tested. The relative abundances observed for ITS2 suggest that species with longer amplification fragments are underestimated and concurrently species that render shorter amplification fragments are overestimated. However, this correlation between amplicon length and estimation is not valid for all the species analysed. Variability in the amplification lengths contributed to the preferential amplification phenomenon. DNA extracted from twenty fermented cocoa bean samples were used to assess the performance of the two target regions. Overall, the metataxonomic data set recovered similar taxonomic composition and provided consistent results in OTU richness among biological samples. However, 26S region provided higher alpha diversity index and greater fungal rRNA taxonomic depth and robustness results compared with ITS2. Based on the results of this study we suggest the use of the 26S region for targeting fungi. Furthermore, this study showed the efficacy of the manually curated reference database optimized for annotation of mycobiota by using the 26S as a gene target.

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Social marketing materials, including billboards and television and radio campaigns, are used by the LSU AgCenter SNAP-Ed program to encourage healthy eating behaviors and to supplement information provided in direct education lessons. An evaluation conducted in 2018 found campaign messages were previously found to be effective in encouraging healthful eating behaviors, but changes were since made to targeting and implementation. This study aimed to assess exposure, unaided recall, and behavior change outcomes of a social marketing campaign on a SNAP-Ed eligible audience. A survey was distributed to n = 78 SNAP-Ed eligible adults through channels used for recruitment of SNAP-Ed participants. The survey first assessed unaided recall of social marketing messages, then prompted participants with titles of campaign messages and assessed recall. Survey questions also asked about changes to attitudes and behaviors after exposure to campaign messages. Although few (n = 13, 17%) respondents recalled messages unaided, many remembered billboard (n = 55, 70%) and television (n = 35, 45%) messages. Only 5% (n = 6) of participants reported hearing a radio message. Many participants who specifically recalled messages about breakfast and fruits and vegetables said they changed their behavior in response to those messages (n = 12, 41% and n = 19, 68%, respectively). Though the generalizability of our findings are limited by the convenience sampling strategy used, our results demonstrated continued reach and effectiveness of the LSU AgCenter social marketing campaign among our sample of participants. Findings also suggested that radio messaging may not be effective in reaching the target audience. Supplemental Nutrition Assistance Program - Education

Widiarti R, Zamani NP, Bengen DG, Madduppa H. 2022. Molecular characterization of toxic benthic dinoflagellate, Prorocentrum lima in west Indonesian waters using LSU 28S rDNA gene. Biodiversitas 23: 3257-3263. Prorocentrum lima is one of the toxic benthic dinoflagellates known to cause Diarrhetic Shellfish Poisoning (DSP), which is also associated with ciguatoxin-producing species, Gambierdiscus toxicus that causes Ciguatera Fish Poisoning (CFP). P. lima has a wide range of morphological variability and genetic diversity, but such research has never been reported from Indonesian waters yet. This study aimed to determine the molecular characteristics of P. limain west Indonesian waters, namely Bintan Island, Belitung Island, Seribu Islands, and Karimunjawa Islands. Molecular characterization was conducted by amplification on large subunit (LSU)28S rDNA gene. Extraction was conducted using freeze-thaw which was continued with single cell PCR method. Genetic distance values and phylogenetic analysis were analyzed using MEGA software. Based on molecular analysis, P. lima from this research was divided into two subclades, namely subclade A from Seribu Islands and Belitung Island, and subclade B from Karimunjawa Islands and Bintan Island. P. lima from Bintan Island showed a closer relationship with the reference sequence from the Genbank. Observation of molecular characters of P. lima showed that the genetic diversity of P. lima depended on the variation of the island’s morphogenesis type.These findings could support a further study on the distribution of P. lima in Indonesian waters, related to the genetic variation and toxin production, since Indonesia consists of many small islands.