Morphological, biological and genomic characterization of lytic phages against Streptococcus agalactiae causing streptococcosis in tilapia.

Streptococcus agalactiae, a highly invasive pathogen causing streptococcosis, is a major disease imparting devastating effect in the aquaculture, worldwide. As bacteriophage therapy is getting more attention recently, as an alternative viable biocontrol agent to antibiotic and vaccine, this study aimed to isolate and purify obligately lytic bacteriophages and study its morphological, genetic and biological characteristics. Host range analysis of the four bacteriophages isolated in this study, such as Phage- 12 P, 15 F, 16 E and 20D exhibited 100% infectivity to S. agalactiae serotype Ia, a predominant serotype infecting fish. Morphotype of the phages was revealed by HR-TEM and found that the phage 20D belong to the family Myoviridae and the phages 12 P, 15 F, 16 E belonged to the family Siphoviridae with typical head and tail structure. Lytic potential of the phages were ascertained by multiplicity of infection and one step lytic curve and it is found that the phages exhibit high burst size at an MOI of 0.01. Random amplified polymorphic DNA revealed the genetic diversity of these four phages with distinct banding pattern. The phages were found to be lytic with the absence of genes coding for integrase, transposase and recombinase on PCR based screening. Phages exhibited stability and viability at various physic-chemical parameters such as temperature ranging from 4 to 45 °C, pH of 4-12 and salinity ranging from 0 to 6%. Thus the present study revealed that S. agalactiae specific phages such as Phage- 12 P, 15 F, 16 E and 20D are highly stable and potential to eliminate the S. agalactiae serotype Ia infecting fish. After the complete characterization of the phages by whole genome sequencing and exploring the defense function against S. agalactiae infection in vivo, it may be applied as a therapeutic agent against S. agalactiae infection in aquaculture.