No abstract

This is the first assay that describes the isolation and identification of strains and species of Lactobacillus from the honey stomach of the Asiatic giant honeybee, Apis dorsata. Samples of honeybees were collected from A. dorsata colonies in different bee trees, and Lactobacillus was isolated from honey stomachs using selective media. The isolates were Gram-stained and tested for catalase reaction. The 16S rRNA genes from extracted DNA of bacterial colonies were amplified with polymerase chain reaction using lactobacilli genus primers (27F and 1492R). All bacterial 16S rRNA genes were sequenced and deposited in GenBank. The 34 isolated strains yielded three distinct rRNA sequences of 15 different strains. Lactobacillus sequences isolated from the bees' honey stomachs were comprised of Lactobacillus kunkeei related-sequences (56%) with other abundant sequences being related to other Lactobacillus sp. (38%) and Lactobacillus vermiform (6%). These strains can be good candidates for potential application as probiotics in honeybees and also as natural food preservatives, which, in turn, may be useful in the food industry.Apis dorsata / honey stomach / Lactobacillus bacteria / probiotics

Apis dorsata colonies often form dense aggregations, with over 100 colonies sometimes seen in the same tree. Reasons for these aggregations are unknown, but one reasonable hypothesis is that colonies form a related family group. Here we show that 7 adjacent colonies sampled from a single branch of a tree (near Alor Setar in northern peninsular Malaysia) containing over 120 colonies were not related as mother/daughter. Thus the notion that aggregations arise through splitting of the first-arriving colonies can be rejected.

This study aimed to isolate and identify Lactobacillus in the honey stomach of honeybee Apis dorsata. Samples of honeybee were collected from A. dorsata colonies in different bee trees and Lactobacillus bacteria isolated from honey stomachs. Ninety two isolates were Gram-stained and tested for catalase reaction. By using bacterial universal primers, the 16S rDNA gene from DNA of bacterial colonies amplified with polymerase chain reaction (PCR). Forty-nine bacterial 16S rDNA gene were sequenced and entrusted in GenBank. Phylogenetic analysis showed they were different phylotypes of Lactobacillus. Two of them were most closely relevant to the previously described species Lactobacillus plantarum. Other two phylotypes were identified to be closely related to Lactobacillus pentosus. However, only one phylotype was found to be distantly linked to the Lactobacillus fermentum. The outcomes of the present study indicated that L. plantarum, L. pentosus, and L. fermentum were the dominant lactobacilli in the honey stomach of honeybee A. dorsata collected during the dry season from Malaysia forest area - specifically “Melaleuca in Terengganu”.

-Capped brood ( capped within 36 h) and adult workers of Apis dorsata were removed from naturally occurring colonies and kept incubated in laboratory hoarding cages at constant temperatures ranging from 26 to 45 o C to study mortality, survivorship, and water and syrup consumption. Capped brood died at temperatures above 36 o C. Below 30 o C adults tended to emerge deformed. Low temperatures delayed development. The optimal temperature for complete emergence of healthy adult workers was 34 o C. Adult workers survived well from about 26 to 36 o C. At 38 o C they died within 5 days and at 45 o C they died within 48 hours. Although syrup (1 sugar: 1 water W/W) consumption did not change over the range of temperatures used, water consumption rose rapidly above 38 o C to over 3 ml/bee in 48 hours at 45 o C. Nest temperature control is critical for survival of brood of A. dorsata and adult worker bees have tight constraints on their abilities to endure high temperatures. Water availability is vital for cooling the colony under hot, tropical conditions, and rearing healthy brood.Apis dorsata/ optimal temperature/ thermoregulation/ tropical Asia/ brood/ workers