Development of slide agglutination method for detection of Nosema mylitta in tasar silkworm using polyclonal antibody produced against proteins of pebrine spores.
Keywords:Antheraea mylitta, Nosema mylitta, Spore proteins, Polyclonal antibody, Agglutination
Production and marketing of quality tasar cocoons is the source of livelihood for the majority of poor tribal peoples of central and north eastern states of India. Due to outdoor rearing of the tasar silkworms, the worms are amenable to different diseases including transovarially transmitted protozoan disease called pebrine caused by Nosema mylitta. The present study was aimed to develop polyclonal antibody based detection method for pebrine spores during mother moth examination which will be sensitive in nature in comparison with microscopic detection. In the present, the pebrine spores were purified from the pebrinized larvae. The spore proteins were purified and used for the production of polyclonal antibody in rabbit. The antiserum was used for the detection of pebrine spores both on glass slides and microscope. The visual and microscopic observation revealed that, the antibody in the dilution of 1:2000 showed agglutination of pebrine spores. Further, the different concentration of pebrine spores was subjected to agglutination. The results revealed that, the concentration up to 10-4 showed the agglutination of pebrine spores (single spore in the microscopic field). The results also confirms that, due to its low range of spore detection, antibody based pebrine detection method can be easily employed in breeding stations of the tasar industry which helps to produce disease fee layings.
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