Isolation and Characterization of Phages Active against Paenibacillus larvae Causing American Foulbrood in Honeybees in Poland.

Ewa Jończyk‐Matysiak ,Paweł Migdał,Barbara Bubak,Barbara Owczarek,Paweł Chorbiński,Wojciech Fortuna,Mirosława Skupińska,Marta Kaszowska,Kinga Świtała-Jeleń,Anna Mucha,Norbert Łodej,Martyna Cieślik,Andrzej Górski,Filip Orwat,Dominika Kula,Adam Roman,Natalia Bagińska,Sławomir Letkiewicz,Agnieszka Belter,Katarzyna Hodyra-Stefaniak,J Neuberg ,Ewa Popiela-Pleban ,Beata Weber‐Dąbrowska

doi:10.3390/v13071217

Abstract

The aim of this study was the isolation and characterization, including the phage effect on honeybees in laboratory conditions, of phages active against Paenibacillus larvae, the causative agent of American Foulbrood—a highly infective and easily spreading disease occurring in honeybee larva, and subsequently the development of a preparation to prevent and treat this dangerous disease. From the tested material (over 2500 samples) 35 Paenibacillus spp. strains were obtained and used to search for phages. Five phages specific to Paenibacillus were isolated and characterized (ultrastructure, morphology, biological properties, storage stability, and genome sequence). The characteristics were performed to obtain knowledge of their lytic potential and compose the final phage cocktail with high antibacterial potential and intended use of future field application. Preliminary safety studies have also been carried out on healthy bees, which suggest that the phage preparation administered is harmless.

Highlights

Pollination is the most important process necessary for obtaining seeds for many plants
Paenibacillus larvae subsp. larvae (ATCC-9545), Paenibacillus larvae (NRRL B-3685ATCC-49843), Kurthia spp. (ATCC-39312), and Paenibacillus alvei (ATCC-6344), were used during the optimization of bacterial culture methods, as well as for research related to bacteriophage search and determination of their lytic spectrum
From the tested material 35 bacterial strains were identified as those belonging to genus Paenibacillus such as: P. larvae (n = 29), P. pabuli (n = 2), P. chinjensis (n = 1), P. alvei (n = 1), and P. thiaminolitycus (n = 1)

Summary

Introduction

Pollination is the most important process necessary for obtaining seeds for many plants. This means that lowering its efficiency would result in the depletion of the plant flora across the whole planet. In particular, play a dominant role in this process [2,3], in which bees are one of the most important groups, which is necessary for the development of 75% of all crops that are used directly to produce food for humans worldwide [4]. In 2005, the global annual economic value of insect pollination was estimated to be 153 billion euros [5]. In 2005, around 10% of the total economic value of European agricultural output for human food, amounting to €22 million, was dependent upon insect pollination [6]

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