Cadmium (Cd) compounds are widely distributed toxic environmental and industrial pollutants, and they may bring danger to growth and development of aquatic organisms. In China, the Oujiang color common carp, Cyprinus carpio var. color, is a very important fish, from an economic point of view, and is well used for fish culture in paddy fields. The purpose of this study was to show the low concentrations of cadmium-induced oxidative stress response and DNA damage in the livers of Cyprinus carpio var. color. Superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in liver were measured after exposure to Cd levels (0.41, 0.52, 0.69, 1.03 and 2.06 mg/L, respectively) for 7 days and compared with the control groups. DNA damage, including indicators of damage percentage, DNA tail length (TL) and DNA tail moment (TM) were also analyzed by comet assays. Results showed that MDA and GSH levels in all treatment groups increased significantly relative to the controls (P < 0.01). Treatment with Cd at concentration of 0.41 mg/L increased SOD activity, while treatment with Cd at concentrations >0.41 mg/L inhibited SOD activities. DNA damage percentage, TL and TM also significantly increased when the Cd level was >0.41 mg/L. Positive correlations were also found between DNA damage levels and MDA levels (r = 0.74 for DNA damage percentage, r = 0.83 for TL, r = 0.84 for TM; P < 0.01 for all) as well as between GSH and MDA levels (r = 0.77, P < 0.01). These results strongly suggested that Cd-induced DNA damage in the livers of Cyprinus carpio var. color was due to lipid peroxidation and oxidative stress.
Bacterial endospores can serve as phage genome protection shells against various environmental stresses to enhance microbial control applications. The genomes of polyvalent lytic phages PBSC1 and PBSC2, which infect both subsp. and NRS 248, were incorporated into endospores (without integration into the host chromosome). When PBSC1 and PBSC2 were released from germinating endospores, they significantly inhibited the growth of the targeted opportunistic pathogen Optimal endospore entrapment was achieved when phages were introduced to the fast-sporulating prespores at a multiplicity of infection of 1. Longer endospore maturation (48 h versus 24 h) increased both spore yield and efficiency of entrapment. Compared with free phages, spore-protected phage genomes showed significantly higher resistance toward high temperatures (60 to 80°C), extreme pH (pH 2 or pH 12), and copper ions (0.1 to 10 mg/liter). Endospore germination is inducible by low concentrations of l-alanine or by a germinant mixture (l-asparagine, d-glucose, d-fructose, and K) to trigger the expression, assembly, and consequent release of phage particles within 60 to 90 min. Overall, the superior resiliency of polyvalent phages protected by endospores might enable nonrefrigerated phage storage and enhance phage applications after exposure to adverse environmental conditions. Bacteriophages are being considered for the control of multidrug-resistant and other problematic bacteria in environmental systems. However, the efficacy of phage-based microbial control is limited by infectivity loss during phage delivery and/or storage. Here, we exploit the pseudolysogenic state of phages, which involves incorporation of their genome into bacterial endospores (without integration into the host chromosome), to enhance survival in unfavorable environments. We isolated polyvalent (broad-host-range) phages that efficiently infect both benign and opportunistically pathogenic strains and encapsulated the phage genomes in endospores to significantly improve resistance to various environmental stressors. Encapsulation by spores also significantly enhanced phage genome viability during storage. We also show that endospore germination can be induced on demand with nutrient germinants that trigger the release of active phages. Overall, we demonstrate that encapsulation of polyvalent phage genomes into benign endospores holds great promise for broadening the scope and efficacy of phage biocontrol.
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