Differential roles of the fish chitinous membrane in gut barrier immunity and digestive compartments.

The chitin-based peritrophic matrix (PM) is a structure critical for both gut immunity and digestion in invertebrates. PM was traditionally considered lost in all vertebrates, but a PM-like chitinous membrane (CM) has recently been discovered in fishes, which may increase the knowledge on vertebrate gut physiology and structural evolution. Here, we show that in zebrafish, the CM affects ingestion behavior, microbial homeostasis, epithelial renewal, digestion, growth, and longevity. Young mutant fish without CM appear healthy and are able to complete their life cycle normally, but with increasing age they develop gut inflammation, resulting in gut atrophy. Unlike mammals, zebrafish have no visible gel-forming mucin layers to protect their gut epithelia, but at least in young fish, the CM is not a prerequisite for the antibacterial gut immunity. These findings provide new insights into the role of the CM in fish prosperity and its eventual loss in tetrapods. These findings may also help to improve fish health and conservation, as well as to advance the understanding of vertebrate gut physiology and human intestinal diseases.

Two novel mollusk short-form ApeC-containing proteins act as pattern recognition proteins for peptidoglycan.

The Apextrin C-terminal (ApeC) domain is a new protein domain largely specific to aquatic invertebrates. In amphioxus, a short-form ApeC-containing protein (ACP) family is capable of binding peptidoglycan (PGN) and agglutinating bacteria via its ApeC domain. However, the functions of ApeC in other phyla remain unknown. Here we examined 130 ACPs from gastropods and bivalves, the first and second biggest mollusk classes. They were classified into nine groups based on their phylogenetics and architectures, including three groups of short-form ACPs, one group of apextrins and two groups of ACPs of complex architectures. No groups have orthologs in other phyla and only four groups have members in both gastropods and bivalves, suggesting that mollusk ACPs are highly diversified. We selected one bivalve ACP (CgACP1; from the oyster Crossostrea gigas) and one gastropod ACP (BgACP1; from the snail Biomphalaria glabrata) for functional experiments. Both are highly-expressed, secreted short-form ACPs and hence comparable to the amphioxus ACPs previously reported. We found that recombinant CgACP1 and BgACP1 bound with yeasts and several bacteria with different affinities. They also agglutinated these microbes, but showed no inhibiting or killing effects. Further analyses show that both ACPs had high affinities to the Lys-type PGN from S. aureus but weak or no affinities to the DAP-type PGN from Bacillus subtilis. Both recombinant ACPs displayed weak or no affinities to other microbial cell wall components, including lipopolysaccharide (LPS), lipoteichoic acid (LTA), zymosan A, chitin, chitosan and cellulose, as well as to several PGN moieties, including muramyl dipeptide (MDP), N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc). Besides, CgACP1 had the highest expression in the gill and could be greatly up-regulated quickly after bacterial challenge. This is reminiscent of the amphioxus ACP1/2 which serve as essential mucus lectins in the gill. Taken together, the current findings from mollusk and amphioxus ACPs suggest several basic common traits for the ApeC domains, including the high affinity to Lys-type PGN, the bacterial binding and agglutinating capacity, and the role as mucus proteins to protect the mucosal surface.

Water-Stable Silver-Based Metal-Organic Frameworks of Quaternized Carboxylates and Their Antimicrobial Activity.

Three-dimensional (3D) and two-dimensional (2D) Ag-based zwitterionic metal-organic frameworks (MOFs) [Ag2(Cedcp)]n (1, 3D, H3CedcpBr denotes N-(carboxyethyl)-(3,5-dicarboxyl)-pyridinium bromide) and {[Ag4(Cmdcp)2(H2O)4]·4H2O}n (2, 2D, H3CmdcpBr denotes N-(carboxymethyl)-(3,5-dicarboxyl)-pyridinium bromide) have been prepared and investigated for antimicrobial activity via minimal inhibition concentration (MIC) test and killing kinetic assay. Both MOFs 1 and 2 show good water stability and solubility ascribed to their characteristic aromatic rings and positively charged pyridinium of the ligands, as well as the presence of Ag+ on their surface, leading to strong antimicrobial activity and a wide antimicrobial spectrum toward Gram-negative and positive bacteria. The results indicated that MOF 2 possesses a faster antibacterial activity (60 min) than MOF 1 (120 min). Scanning electron microscopy analysis further suggests that the Ag-based MOFs are capable of rupturing the bacterial membrane, leading to cell death. Moreover, both MOFs exhibit little hemolytic activity against mouse erythrocytes and show good biocompatibility in vitro, rendering MOFs 1 and 2 potential therapeutic agents for diseases caused by bacteria.