Identification of Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus fermentum from honey stomach of honeybee

Abstract

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”.

Microorganisms in Honey

“…This access has been carried out mainly by sequencing the coding region of the 16S subunit of the bacterial ribosome [53,121,130], both from genomic DNA from microorganisms growing on selective media as Man-Rogosa-Sharpe agar, Sabouraud dextrose agar, and Candida agar [117,120,131,132], such as process-independent culture as specific PCR [68], denaturing gradient gel electrophoresis [124,125], mixed and deep 16S sequencing [49,128], pyrosequencing [53,116,121], and clone library [115,118,120,122]. While culturedependent methods are ideal for quantification of microorganisms and phenotypic testing, culture-independent methods generally have greater coverage in relation to the amount of different species accessed and are ideal for fingerprinting studies, and the identification of these species may be performed by real-time PCR analysis [49,68,125,128].…”

“…Several microorganisms present in the honey and in the gut of honeybees have antagonistic effects on honeybees and human pathogens, especially of Bacillus genus [123,134], lactic acid bacteria as Lactobacillus [71,121,124,[130][131][132]135], Enterococcus [130], Bifidobacteria [116,132,[136][137][138], and Acetobacteraceae [117,121,133]. These same microorganisms can be accessed for other purposes, such as its potential as fermenters [116,130,133] or probiotics [116].…”

Alterations in honey bee gut microorganisms caused by Nosema spp. and pest control methods

“…Six of the 10 isolated Lactobacillus species identified in the study had been previously reported as honey bee endophytes including L. plantarum , L. buchneri , L. gasseri , L. fermentum , L. pentosaceus and L. sanfranciscensis . These Lactobacillus species are linked with bee health due to their antimicrobial properties, high lactic acid yield and biofilm formation, among others …”

“…Bacterial colonies per plate were then randomly picked according to morphological characteristics (white, round) and subcultured. As a first Lactobacillus selection filter, isolates were subjected to a catalase test with 3% H 2 O 2 , which should be negative because Lactobacillus isolated from bees do not produce catalase, as indicated by the lack of bubbles observed by microscopy . Isolated strains were grown under standard conditions for fatty acid identification in MRS (48 h at 35 °C) according to the Microbial Identification System Libraries (MIDI; Microbial ID, Inc., Newark, DE, USA).…”

Exact identification of six starter‐strain candidates of Streptococcus thermophilus by analysing genotypic and industrial properties

“…The primers used for the amplification of 16S rRNA gene were 27F (5′‐AGAGTTTGATCCTGGCTCA‐3′) and 1492R (5′‐GGTTACCTTGTTACGACTT‐3′) (Tajabadi et al . ) and for lacZ gene were F (5′‐CACTATGCTCAGAATACA‐3′) and R (5′‐CGAACAGCATTGATGTTA‐3′) (Giraffa et al . ).…”