Chinese traditional medicines have been used for several thousands of years in Asian countries, not only in humans but also in many animal species. These compounds prevent and control different types of diseases including internal diseases as well as some infectious diseases, where the aetiological agent is viral, bacterial, parasitic or mycotic. Rhizoma coptidis is believed to inhibit Shigella dysenteriae and that Radix isatidis can prevent flu caused by the influenza virus. It is thus hypothesized that some of these traditional herbal compounds will have anti-fungal activity. Saprolegniosis is a disease common in fish and their eggs in both fresh and brackish water; a newer, safer medication against Saprolegnia is needed after the prohibition in many countries of the extremely effective fungicide, malachite green. In the present study an attempt is made to identify herbal compounds that have anti-Saprolegnia activity. A strain of Saprolegnia, CCF1301, was isolated from the skin of infected grass carp (Ctenopharyngodon idella) and identified as Saprolegnia ferax by the 26S rDNA D1/D2 region and ITS region. This strain was used to evaluate the antifungal activity of thirty Chinese traditional herbal medicine extracts, and a modified dish dilution method was developed for the evaluation. Saprolegnia-infected rapeseeds with visible hyphae were transplanted onto prepared PDA plates containing 2 g L À1 herbal plant extracts and incubated at 20°C for 48 h. Each herbal plant species was tested in triplicate. Those herbal plant extracts that showed negative mycelium presence at 2 g L À1 were further tested for minimum inhibitory concentration (MIC) evaluation. The results showed that Syzygium aromaticum, Magnolia officinalis, Melaphis chinensis, Euphorbia fischeriana Steud, and Sophora flavescentis exhibited enhanced growth inhibition at 2 g L À1 and MIC values of 500, 62.5, 250, 62.5, 250 mg L À1 concentrations, respectively. It was obvious that Magnolia officinalis and Euphorbia fischeriana Steud exhibited the best antifungal activity. Since there is a high natural toxicity in Euphorbia fischeriana Steud, its applicability as the main ingredient in an aquaculture therapeutic formulation requires further research. Thus, Magnolia officinalis would appear to be the more valuable antifungal herbal species with which to pursue further research.
Both catalase and peroxiredoxin show high activities of H2O2 decomposition and coexist in the same organism; however, their division of labor in defense against H2O2 is unclear. We focused on the major peroxiredoxin (PrxA) and catalase (CatB) in Aspergillus nidulans at different growth stages to discriminate their antioxidant roles. The dormant conidia lacking PrxA showed sensitivity to high concentrations of H2O2 (>100 mM), revealing that PrxA is one of the important antioxidants in dormant conidia. Once the conidia began to swell and germinate, or further develop to young hyphae (9 h to old age), PrxA-deficient cells (ΔprxA) did not survive on plates containing H2O2 concentrations higher than 1 mM, indicating that PrxA is an indispensable antioxidant in the early growth stage. During these early growth stages, absence of CatB did not affect fungal resistance to either high (>1 mM) or low (<1 mM) concentrations of H2O2. In the mature hyphae stage (24 h to old age), however, CatB fulfills the major antioxidant function, especially against high doses of H2O2. PrxA is constitutively expressed throughout the lifespan, whereas CatB levels are low in the early growth stage of the cells developing from swelling conidia to early growth hyphae, providing a molecular basis for their different contributions to H2O2 resistance in different growth stages. Further enzyme activity and cellular localization analysis indicated that CatB needs to be secreted to be functionalized, and this process is confined to the growth stage of mature hyphae. Our results revealed differences in effectiveness and timelines of two primary anti-H2O2 enzymes in fungus.
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