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Recovery of Pseudomonas koreensis from eye lesions in golden mahseer, Tor putitora (Hamilton, 1822) in Uttarakhand, India.

Freshwater aquaculture in India faces significant economic loss due to bacterial infections (Swain et al. 2002; Shahi et al. 2013). Among these, the bacterial pathogens belonging to the genus Aeromonas, Pseudomonas, Edwardsiella and Flavobacterium are mainly responsible for severe mortality in cultured fishes. Within Pseudomonas, Pseudomonas fluorescens and Pseudomonas aeruginosa are known to cause disease in freshwater fishes (Saharia & Prasad 2001; Swain et al. 2007). Infection of fish by P. fluorescens, P. aeruginosa and Pseudomonas putida leads to the development of septicaemia and ulcerative condition, especially in fishes under stress or injured by inappropriate handling (Ostland et al. 1999; Nabi, Jabeen & Hasnain 2000). Although there are sporadic reports of bacterial diseases of cultured carps in India (Mohan & Shankar 1994; Swain et al. 2003), no published information is available about the bacterial pathogens from coldwater sector of northern India, where rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), and golden mahseer, Tor putitora (Hamilton, 1822), are the fish of economic importance. However, there are incidences of mass mortality of rainbow trout, which resulted in severe economic loss to fish growers. Despite that, till date very limited research effort has been made to investigate the bacterial pathogens in cultured O. mykiss and T. putitora in India (Shahi et al. 2013). Between March and May 2012, opaqueness of eye occurred in adult T. putitora (Fig. 1), at rearing sites in Uttarakhand. At the advanced stage of disease, mortality was observed in fishes (weight 210.5 12.5 g). Clinical signs recorded were haemorrhages in the periphery of eye, corneal opacity and internally pale liver. Ten moribund T. putitora with eye lesions were killed by overdose of tricaine methane sulphonate (MS-222). The diseased eye of each killed fish was surface disinfected with 70% ethanol and then collected aseptically for bacterial isolation. Each eye was pulverized separately in sterile normal saline, and the bacterial suspension was serially diluted for spread plating on tryptic soy agar (TSA) and incubated at 28 °C for 48 h. After 48 h of incubation, dense growth of green-yellow colonies was observed on TSA plates. No other colonies were observed on any of the plates, even after 96 h of incubation. Five green-yellow pigmented strains of bacteria were recovered from the infected eyes on TSA, and their identity was established by biochemical (Austin & Austin 2012) and molecular methods. All the isolated bacteria were Gram-negative, motile, non-spore-forming rods, 1.6– 2.0 lm in size. Cells produced a green-yellow fluorescent pigment on Pseudomonas agar F (PAF). Strains were positive for ß-haemolysin on sheep as well as T. putitora blood (5% erythrocytes) agar plates. Positive result was noted for catalase, oxidase, citrate, triple sugar iron (TSI), O/F, caseinase, lecithinase, Tween 80 and urease. Growth Correspondence N Shahi, DCFR, Bhimtal 263 136, Nainital, Uttarakhand, India (e-mail: shahineetu@rediffmail.com)