Experimental evaluation of pathogenicity of Lactococcus garvieae in black rockfish (Sebastes schlegeli)

Efficacy of protein A-HRP in an immunological study of black rockfish (Sebastes schlegeli Higendorf) humoral immune responses

“…The reaction was stopped after 10 min by the addition of 50 ml well ÿ1 of 2 M H 2 SO 4 , and the results were read spectrophotometrically at 450 nm with an ELISA reader (Benchmark Ò , Bio-Rad, USA). Negative sera were collected from healthy yearling rockfish and the positive sera were those used in the 2-DE immunoblot assay of L. garvieae KG9408 [3]. The positivity and negativity of ELISA was decided by determining the upper limit for negativity by adding the mean value of the optical densities of negative sera (normal rockfish sera) to three times the standard deviation (3!SD).…”

“…The membrane was blocked with 5% skim milk in PBS-T for 1 h at RT, washed three times with PBS-T, and then incubated with L. garvieae KG9408 antisera (1:20) collected from rockfish that had been previously infected with L. garvieae KG9408 (10 8 cfu ml ÿ1 ). After 4 h at RT, the membrane was washed three times with PBS-T [3] and the L. garvieae antigenic spots were visualised by incubation with protein A-HRP or the mouse polyclonal antibody, followed by incubation with goat anti-mouse-HRP as described above for the SDS-PAGE immunoblot assays. The stained gel and immunoblotted membrane were digitalised with an Agfa Arcus 1200Ô image scanner (Agfa-Gevaert, Mortsel, Belgium) and the acquired images were analysed using Phoretix 2D software (Ver.…”

“…This disease is common during the warmer months, in association with other component causes such as poor water quality [4]. Lactococcus garvieae has been reported as a causative agent of fish streptococcosis in several species, including black rockfish (Sebastes schlegeli) [3], rainbow trout (Oncorhynchus mykiss) [5], grey mullet (Mugil cephalus L.) [6] and yellowtail (Seriola quinqueradiata) [7]. L. garvieae has also been identified in bovine [8] and human [9] clinical samples, indicating that this pathogen may be an important emerging zoonotic agent.…”

Molecular characterization and comparative expression analysis of two teleostean pro-inflammatory cytokines, IL-1β and IL-8, from Sebastes schlegeli

“…Black rockfish (Sebastes schlegeli) is the second most largely cultured fish species in South Korea. Its aquaculture has been mainly affected by a combination of bacterial species (Han et al, 2011;Kang et al, 2004).…”

Isolation and molecular identification of the etiological agents of streptococcosis in Nile tilapia (Oreochromis niloticus) cultured in net cages in Lake Sentani, Papua, Indonesia

“…The causative agents of the disease have been attributed to multiple bacterial agents but most commonly Streptococcus and/or Lactococcus (Kang et al 2004; Plumb and Hanson 2011). Streptococcosis is a common disease affecting various species of fish worldwide (Roberts 2012) including the Asian seabass (Suanyuk et al 2010), barramundi (Bromage et al 1999), grouper (Bowater et al 2012), Japanese flounder (Nguyen et al 2002), rabbit fish (Yuasa et al 1999), rainbow trout (Eldar and Ghittino 1999), red drum (Eldar et al 1999; Shen et al 2005), red tilapia (Hernandez et al 2009; Musa et al 2009), seabream (Evans et al 2002), silver pomfret (Duremdez et al 2004), and wild mullet (Evans et al 2002).…”