Tryptic soy broth medium tsb

E. coli strains R1–R4 and K-12 were obtained as a kind gift from Prof. Jolanta Łukasiewicz at the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy (Polish Academy of Sciences). The reference bacterial strains of E.coli (K12 ATCC 25404, R2 ATCC 39544, R3 ATCC 11775, R4 ATCC 39543 were provided from (LGC Standards U.K.) and were used according to the recommendation of ISO 11133: 2014. These strains were used to test antibacterial activity of the synthesized agents [10 (link),11 (link),12 (link),13 (link),14 (link),15 (link),16 (link),17 (link),18 (link),28 (link),29 (link),30 (link),31 (link)].
Bacteria were grown in liquid medium or agar plates containing tryptic soy broth medium (TSB; Sigma-Aldrich, Saint Louis, MI, USA), using a methodology described previously [12 (link),13 (link),14 (link),15 (link),16 (link),17 (link),18 (link)].

Overnight cultures of V. alginolyticus ATCC 17749 were diluted to 1: 100 in Tryptic Soy Broth Medium (TSB) (Sigma, Shanghai, China), grown at 28℃ with shaking, and harvested in the logarithmic phase of growth. The cells were washed, resuspended, and diluted to the appropriate concentration in sterile PBS. Sixteen fish were challenged by intraperitoneal injection with 5×10⁶ colony forming units (CFUs) of V. alginolyticus (in 100 μL PBS) per fish, and sixteen other fish were injected with 100 μL of PBS per fish as a negative control. The liver, spleen, head kidney, kidney, intestine, muscle, brain, and gill were collected from four fish at each time point at 4, 8, 12, and 24 h post-injection (hpi), as previously reported (Wu et al., 2015 (link)), then preserved at -70℃ until examination.

Escherichia coli strains R1–R4 and K-12 were obtained as a kind gift from Prof. Jolanta Łukasiewicz at the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy 86 (Polish Academy of Sciences), where the microorganisms were cultured. The reference bacterial strains of E. coli (K12 ATCC 25404, R2 ATCC 39544, R3 ATCC 11775, R4 ATCC 39543 were provided by LGC Standards (U.K.) and were used according to the recommendation of ISO 11133: 2014. These strains were used to test the antibacterial activity of the synthesized agents [20 (link),21 (link),22 (link),23 (link),35 (link),47 (link),48 (link)]. Bacteria were grown in liquid medium or agar plates containing tryptic soy broth medium (TSB, Sigma-Aldrich Saint Louis, MI, USA), using a methodology described previously [24 (link),25 (link),49 (link),50 (link)].