Abstract
Pathogen infection of honeybees can lead to economic losses in apiculture. The earlier the pathogen contamination can be found the better it will likely be for the treatment of the infected colony and prevention of the spread of the pathogen within and between colonies. A total of 50 colonies of Apis cerana were sampled in Samut-Songkhram (five colonies) and Chumphon (45 colonies) provinces in the central and the south of Thailand, respectively. Diagnostic multiplex polymerase chain reaction (PCR) revealed that 20, 6, 4, 20 and 0% of the samples were infected by Paenibacillus larvae, Nosema ceranae, Nosema apis, Ascosphaera apis and Sacbrood virus (Morator aetatulus), respectively. Positive amplified PCR products of target genes were sequenced. A phylogenetic tree was constructed by the neighbor-joining (NJ) distance based, using the Phylogenetic Analysis Using Parsimony (PAUP 4.0b10) program. The phylogenetic relationship of each pathogen, based on the partial sequence of the 16S rRNA of P. larvae, N. ceranae and N. apis, and of the 5.8S rRNA of A. apis, revealed a significant difference from the non-Thai isolates of these pathogens, but no significant geographical isolation between the different Thai apiaries, although it separated some into closely related clusters. In order to reduce the use of antibiotics in an apiary, bacteria in the gut of healthy bees were focused. Interestingly, Bifidobacterium species, Lactobacillus species, Bacillus species, Lactobacillus spp. and other lactic acid bacteria, were isolated from larvae and adult workers, but gave conflicting preliminary identities based on their biochemistry-morphology versus sequence analysis of a partial fragment (1.4 kb) of their 16S rRNA. Key words: Apis cerana indica, bee pathogens, gut bacteria, multiplex polymerase chain reaction (PCR), 16S rRNA.
Highlights
Honey bees (Apis species) are environmentally and economically important insects, with the latter being crop pollination and essential products used for food and traditional medicine (Moritz et al, 2010)
The deleterious bee diseases that are frequently found in apiaries are American foulbrood (AFB) caused by P. larvae, microsporidosis caused by N. ceranae and N. apis, chalkbrood disease caused by A. apis, and sacbrood disease caused by the Sacbrood virus (SBV)
The detection of SBV contamination in A. mellifera adults using a one-step RTPCR loop-mediated isothermal amplification primer set (RT-LAMP), where the SBV-specific primers were designed based on the SBV pol gene sequence
Summary
Honey bees (Apis species) are environmentally and economically important insects, with the latter being crop pollination and essential products used for food and traditional medicine (Moritz et al, 2010). Is a diverse array of pathogen-caused bee diseases, some of which can cause serious morbidity and mortality to bee colonies and lead to the economic loss to bee farmers. It would be much better if pathogen infections could be detected at an early stage. Contaminated microorganisms within the bees and in the nectar stored in forager’s honey crop still remain (Anderson et al, 2013)
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