Abstract
Apis mellifera colonies were sampled for over twelve months to study the Nosema infection in different apiaries of Himachal Pradesh, India. We found that the infection incidence was highest in the winter season (48.40 %) followed by autumn and rainy months. The infection was diagnosed conventionally through the microscopic analysis detection of Nosema species spores. However, with the recent findings that both N. ceranae and N. apis affect western honey bees, more sensitive and species specific molecular techniques are required that reliably differentiate between species of microsporidia. Therefore, molecular studies were conducted to precisely differentiate between Nosema species infecting honeybee colonies in India. Species specific multiplex PCR amplification using previously reported primers derived from 16S rRNA fragments were performed in a single reaction mixture. A single diagnostic band size of only 143 bp for N. ceranae was observed in the agarose gel. Furthermore, a diagnostic band size of 252 bp was observed with primer NoS, thus reconfirming the presence of N. ceranae. This study appears to be the first to report the detect of N. ceranae species from India in samples of A. mellifera.