The first ITS2 sequence data set of eDNA from honey of Malaysian giant honeybees (Apis dorsata) and stingless bees (Heterotrigona itama) reveals plant species diversity.

Pollen is a useful tool for identifying the provenance and complex ecosystems surrounding honey production in Malaysian forests. As native key pollinators in Malaysia, Apis dorsata and Heterotrigona itama forage on various plant/pollen species to collect honey. This study aims to generate a dataset that uncovers the presence of these plant/pollen species and their relative abundance in the honey of A. dorsata and H. itama. The information gathered from this study can be used to determine the geographical and botanical origin and authenticity of the honey produced by these two species. Sequence data were obtained for both A. dorsata and H. itama. The raw sequence data for A. dorsata was 5 Mb, which was assembled into 5 contigs with a size of 6,098,728 bp, an N50 of 15,534, and a GC average of 57.42. Similarly, the raw sequence data for H. itama was 6.3 Mb, which was assembled into 11 contigs with a size of 7,642,048 bp, an N50 of 17,180, and a GC average of 55.38. In the honey sample of A. dorsata, we identified five different plant/pollen species, with only one of the five species exhibiting a relative abundance of less than 1%. For H. itama, we identified seven different plant/pollen species, with only three of the species exhibiting a relative abundance of less than 1%. All of the identified plant species were native to Peninsular Malaysia, especially the East Coast area of Terengganu. Our data offers valuable insights into honey's geographical and botanical origin and authenticity. Metagenomic studies could help identify the plant species that honeybees forage and provide preliminary data for researchers studying the biological development of A. dorsata and H. itama. The identification of various flowers from the eDNA of honey that are known for their medicinal properties could aid in regional honey with accurate product origin labeling, which is crucial for guaranteeing product authenticity to consumers.

Huda N ，Ullah S ，Wahab RA ，Lani MN ，Daud NHA ，Shariff AHM ，Ismail NI ，Hamid AAA ，Mohamad MAN ，Huyop F ... -  《-》

Using pollen DNA metabarcoding to trace the geographical and botanical origin of honey from Karangasem, Indonesia.

The unique floral fingerprint embedded within honey holds valuable clues to its geographical and botanical origin, playing a crucial role in ensuring authenticity and detecting adulteration. Honey from native Apis cerana and Heterotrigona itama bees in Karangasem, Indonesia, was examined utilizing pollen DNA metabarcoding for honey source identification. In this study, we used ITS2 amplicon sequencing to identify floral DNA in honey samples. The finding reveals distinct pollen signatures for each bee species. Results analysis showed A. cerana honey generated 179,267 sequence reads, assembled into Amplicon Sequence Variants (ASVs) with a total size of 485,932 bp and an average GC content of 59 %. H. itama honey generated 177,864 sequence reads, assembled into ASVs with a total size of 350,604 bp and an average GC content of 57 %. A. cerana honey exhibited a rich tapestry of pollen from eleven diverse genera, with Schleichera genus dominating at an impressive relative read abundance of 72.8 %. In contrast, H. itama honey displayed a remarkable mono-dominance of the Syzygium genus, accounting for a staggering 99.95 % of its pollen composition or relative read abundance, highlighting their distinct foraging preferences and floral resource utilization. Notably, all identified pollen taxa were indigenous to Karangasem, solidifying the geographical link between honey and its origin. This study demonstrates pollen DNA metabarcoding may identify honey floral sources. By using pollen profiles from different bee species and their foraging patterns, we may protect consumers against honey adulteration and promote sustainable beekeeping in Karangasem district. Future research could explore expanding the database of reference pollen sequences and investigating the influence of environmental factors on pollen composition in honey. Investigating this technology's economic and social effects on beekeepers and consumers may help promote fair trade and sustainable beekeeping worldwide.

Ullah S ，Huyop F ，Wahab RA ，Sujana IGA ，Antara NS ，Gunam IBW ... -  《Heliyon》

Physicochemical characteristics, antioxidant properties and bacterial profiling of three Malaysian honey varieties: a study using multivariate analysis.

The physicochemical characteristics, antioxidant properties, and bacterial profiles of Heterotrigona itama, Apis dorsata, and Apis mellifera honey of Malaysian origin were studied and the results were assessed using Pearson correlation analysis and canonical correspondence analysis (CCA). The analysis showed that the Mellifera honey was characterized by high total sugar content (717.5 g kg-1 ); the Itama honey was characterized by high free acidity (347.5 meq kg-1 ); and the Dorsata honey was characterized by high radical scavenging activity (RSA; 69.3%RSA), total phenolic content (TPC; 1284.5 mg GAE kg-1 ) and hydroxymethylfurfural (HMF; 51.5 mg kg-1 ). Pearson correlation analysis showed that the TPC was positively correlated with HMF as well as RSA. The bacterial profile of Mellifera honey was significantly less diverse than Itama and Dorsata honey. Mellifera and Itama honey seemed to harbour a significant abundance of lactic acid bacteria, with relative abundance of 0.339 and 0.195, which suggests that it possesses probiotic potential. The Dorsata honey, however, contained pathogenic Clostridium genus, constituting 5% of its bacterial profile. The characteristic properties of the three investigated honey, Itama, Dorsata, and Mellifera were identified. The distinctive characteristics of the honey varieties informed the development of appropriate processing strategies for retaining the beneficial qualities of raw honey. It is hoped that the results of this study will shine light on Malaysian honey in a highly competitive global market. © 2022 Society of Chemical Industry.

Yap SK ，Chin NL ，Abdul Karim Shah NN ，Than LTL ，Niranjan K ... -  《-》

Physicochemical homogeneity of stingless bee honey (Heterotrigona itama) produced in the west coast, east coast and inland area of Peninsular Malaysia.

Ramlan NAFM ，Mohamad Azman E ，Muhammad K ，Jusoh AZ ，Johari NA ，Yusof YA ，Zawawi N ... -  《-》

Exploring the genome of Lactobacillaceae spp. Sy-1 isolated from Heterotrigona itama honey.

Honey produced by Heterotrigona itama is highly preferred among consumers due to its high-value as a functional food and beneficial lactic acid bacteria (LAB) reservoir. Fructophilic lactic acid bacteria (FLAB) are a group of LAB with unique growth characteristics and are regarded as promising producers of bioactive compounds. Hence, it is not surprising that LAB, especially FLAB, may be involved with the excellent bioactivity of H. itama honey. With the trending consumer preference for H. itama honey coupled with increasing awareness for healthy food, the genomic background of FLAB isolated from this honey must, therefore, be clearly understood. In this study, one FLAB strain designated as Sy-1 was isolated from freshly collected H. itama honey. Its FLAB behavior and genomic features were investigated to uncover functional genes that could add value to functional food. The fructophilic characteristics of strain Sy-1 were determined, and the genome was sequenced using Illumina iSeq100 and Oxford Nanopore. The average nucleotide identity and phylogenetic analyses based on 16S rRNA, 92 core genes, and whole-genome sequence were performed to unravel the phylogenetic position of strain Sy-1. NCBI Prokaryotic Genome Annotation Pipeline annotated the genome, while the EggNOG-mapper, BLASTKoala, and GHOSTKoala were used to add functional genes and pathways information. Strain Sy-1 prefers D-fructose over D-glucose and actively metabolizes D-glucose in the presence of electron acceptors. Genomic annotation of strain Sy-1 revealed few genes involved in carbohydrate transport and metabolism, and partial deletion of adhE gene, in line with the characteristic of FLAB. The 16S rRNA gene sequence of strain Sy-1 showed the highest similarity to unknown LAB species isolated from the gut of honeybees. The phylogenetic analyses discovered that strain Sy-1 belonged to the Lactobacillaceae family and formed a separate branch closer to type strain from the genera of Acetilactobacillus and Apilactobacillus. The ANI analysis showed the similarity of the closest relative, Apilactobacillus micheneri Hlig3T. The assembled genome of Sy-1 contains 3 contigs with 2.03 Mbp and a 41% GC content. A total of 1,785 genes were identified, including 1,685 protein-coding genes, 68 tRNA, and 15 rRNA. Interestingly, strain Sy-1 encoded complete genes for the biosynthesis of folate and riboflavin. High-performance liquid chromatography analysis further confirmed the high production of folic acid (1.346 mg/L) by Sy-1. Based on phylogenetic and biochemical characteristics, strain Sy-1 should be classified as a novel genus in the family of Lactobacillaceae and a new member of FLAB. The genome information coupled with experimental studies supported the ability of strain Sy-1 to produce high folic acid. Our collective findings support the suitable application of FLAB strain Sy-1 in the functional food and pharmaceutical industries.

Syed Yaacob SN ，Huyop F ，Misson M ，Abdul Wahab R ，Huda N ... -  《-》