Identification of Lactobacillus species from honey stomach of honeybee and their antagonistic properties against bacteria pathogens

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Introduction

The genus Lactobacillus is one of the most important genera of lactic acid bacteria (LAB) and includes over 174 species (Dsmz, 2008). Lactobacillus bacteria are catalase negative, Gram positive and usually rod shaped. They are found in a wide variety of habitats, including vegetable and the gastrointestinal tracts of animals, like honeybees (Tannock, 2004; Schrezenmeir and de Vrese, 2001; Fujisawa and Mitsuoka, 1996) and are employed in manufacturing of fermented foods, such as soybean products, fish products, vegetable products, bread, alcoholic beverages, dairy, and for cheese production (Kandler et al., 1986; Rhee et al., 2011; Caplice and Fitzgerald, 1999). The dominant LAB species in the honey stomach of honeybees’ A. melliferaA. dorsata, and bumblebees have been reported to belong to the Lactobacillus genus (Olofsson and Vasquez, 2008; Vásquez et al., 2009; Tajabadi et al., 2011b).

Microorganisms present in honey are those surviving the low pH and antimicrobial compounds found in honey. The microorganisms probably originate from pollen, the digestive tracts of honeybees, flowers, dirt, dust, and the air. Gilliam (1997) discovered that honeycomb was dominated mainly by bacteria and yeast, which all arrived from the bees or the raw materials (nectar) (Gilliam, 1997). Larvae could be sterile initially, but since they eat pollens and nectar supplied by the workers, hence they are inoculated prior to pupation (Lee and Kime, 1984).

The distinguishable phenotypic property for detection and identification of LAB microbiota has been leading to a trend towards using molecular-based identification methods. Different approaches of rDNA gene sequencing (Wallbanks et al., 1990; Rachman et al., 2003; Settanni et al., 2005; Yoshiyama and Kimura, 2009; Snell-Castro et al., 2005) have been employed to identify Lactobacillus species. Within the recent years, sequencing of 16S rDNA and its use has been considered as the “gold standard” for identification and phylogenetic analysis of LAB (Ludwig and Schleifer, 1999).

The use of Lactobacillus species as probiotics has been found to enhance the immunity of honeybees, helping them to survive against the effect of pathogens and bring advantageous properties for honeybee health (Evans and Lopez, 2004; Forsgren et al., 2010). Therefore, the present study aimed to isolate and identify lactobacilli strains from the honey stomach of honeybees A. dorsata collected during the dry season from Malaysia forest area specifically “Melaleuca in Terengganu”.

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