Effect of the coexistence of endosulfan on the lindane biodegradation by Novosphingobium barchaimii and microbial enrichment cultures.

Organochlorine pesticides, especially lindane and endosulfan, have been demonstrated to be both biodegradable and frequently coexistent, but their inhibitory effect has never been studied. In this study, we investigated the effect of endosulfan coexistence on lindane degradation to a lindane-degrading isolate, Novosphingobium barchaimii strain LL02, and mixed enrichment cultures from two different inocula. Our results of the lindane degradation batch experiments demonstrated that endosulfan concentration above 20 mg L-1 causes significant inhibition to the lindane degradation efficiency of the strain LL02. Besides, the acidic conditions at pH 5.0 to 6.0 further decreased its lindane degradation rate constants by 57% compared to the neutral and alkaline conditions. For the mixed microbial cultures, the lindane degradation efficiency in the lindane/endosulfan co-contamination conditions decreased by 35.7%-50.7% compared to the lindane alone conditions. From our 16S rRNA amplicon sequencing results through the PacBio platform, most of the predominant bacteria in the lindane-enriched cultures were depressed in the lindane/endosulfan-enriched cultures. Moreover, bacteria of Burkholderia australis, Chujaibacter soli, Flavitalea flava, and one Rhodanobacteraceae bacterium were relatively highly abundant in the co-contamination enrichment cultures, suggesting their potential for lindane degradation under the endosulfan stress. Our results demonstrated that endosulfan coexistence causes inhibitory impacts on lindane biodegradation toward both lindane-degrading bacteria and mixed microbial cultures. The coexistence of multiple organochlorine pesticides on the biodegradation efficiencies should be carefully considered when applying bioremediation to remove organochlorine pesticide contamination.

Isolation of lindane- and endosulfan-degrading bacteria and dominance analysis in the microbial communities by culture-dependent and independent methods.

　　Bioremediation for lindane and endosulfan removal is a cost-effective approach, but its effectiveness depends on the ability to isolate degrading functionalized microorganisms. Researchers have isolated many lindane and endosulfan degrading bacteria from enrichment cultures based on culture-dependent methods during the past decades. However, it is unknown whether the isolated bacteria can reflect the indigenous predominant degraders in enriching cultures. In this study, we compared the culture-dependent method with selective medium isolation with culture-independent method (PacBio SMRT sequencing of full-length 16S rRNA amplicon) to analyze the bacterial communities from four distinct lindane (LA1 and LC1) and endosulfan (EA1 and EC1) enrichment cultures. From all the isolates we harvested from lindane (63 isolates) and endosulfan (61 isolates) enrichment cultures, their BLAST alignment can only match 5.49 % and 4.32 % of the bacterial operational taxonomic units (OTUs), respectively. Rhodanbacter lindaniclasticus and Pandoraea thiooxydans were the rarely seen potential degrading representatives that were simultaneously enriched and isolated. This study is the first comparative analysis of microbial communities from lindane and endosulfan enrichment culture using culture-dependent and culture-independent methods. Our results suggested that developing a target-specific and efficient microbial isolation method is necessary to harvest and study representative degrading bacteria in the community.