Mucosal immunity and microbiota change in the rainbow trout (Oncorhynchus mykiss) gills after being challenged with infectious hematopoietic necrosis virus.

Respiratory structures are crucial for vertebrate survival, as they serve not only to perform gas-exchange processes but also as entry points for opportunistic pathogens. Previous studies have demonstrated that fish contain gill mucosal-associated lymphoid tissue, and harbor a large number of commensal bacteria on their surface and contribute to maintaining fish health. However, by far, very limited information is known regarding the effects of viral infection on gill mucosal immunity and microbiota homeostasis. In this study, we conducted an infection model by bath with infectious hematopoietic necrosis virus (IHNV) and revealed a 27 % mortality rate among rainbow trout in the first two weeks after infection. Moreover, we found that diseased fish with the highest IHNV loads in gills exhibiting severe damage, as well as increased goblet cell counts in both primary lamellae (PL) and secondary lamellae (SL). Additionally, RT-qPCR and RNA-seq analyses revealed that IHNV infection induced a strong innate and adaptive antiviral immune responses. Interestingly, an antibacterial immune response was also observed, suggesting that a secondary bacterial infection occurred in trout gills after viral infection. Furthermore, 16S rRNA analysis of trout gills revealed a profound dysbiosis marked by a loss of beneficial taxa and expansion of pathobionts following IHNV infection. Overall, our finding demonstrates that IHNV infection induces significant changes of the microbial community in the fish respiratory surface, thus triggering local antiviral and bacterial mucosal immunity.

Boosting CO2 Conversion by Synergy of Lead-Free Perovskite Cs2SnCl6 and Plasma with H2O.

Although dielectric barrier discharge (DBD) plasma is a promising technique for CO2 conversion, realizing CO2-to-alcohol is still challenging via the use of H2O. Herein, for the first time, efficient CO2 conversion was achieved via the synergism between the Cs2SnCl6 photocatalyst and DBD plasma assisted by H2O. The CO2 conversion ratio of plasma photocatalysis was 6.5% higher than that of only the plasma and photocatalysis, implying that the synergism of plasma catalysis and photocatalysis was achieved. Furthermore, the DBD plasma assisted by the Cs2SnCl6 photocatalyst could convert CO2 and H2O to CO and a small amount of methanol and ethanol. The CO2 conversion ratio was enhanced by 50.6% in the presence of H2O, which was attributed to the improvement of charge transfer due to the increased electrical conductivity of the photocatalyst surface during plasma discharge. This work provides a new idea for developing an efficient system for CO2 utilization.

Expression analysis of Igs and mucosal immune responses upon SVCV infection in common carp (Cyprinus carpio L.).

The immunoglobulin (Ig) is a crucial component of adaptive immune system in vertebrates including teleost fish. Here complete cDNA sequence of IgD heavy chain gene from common carp (Cyprinus carpio) was cloned and analyzed. The full-length cDNA of IgD heavy chain gene contained an open reading frame (ORF) of 2460 bp encoding 813 amino acids. According to amino acids sequence, multiple alignment and phylogenetic analysis showed that carp Igs are closely related to those of Cyprinidae fish. Transcriptional expression of IgD as well as IgM, IgZ1 and IgZ2 showed similar expression patterns in different organs, this is, high expression level in systemic immune tissues (ie, head kidney, heart and spleen) and low expression in mucosal tissues (ie, gill, skin and gut). Following viral infection with spring viraemia of carp virus (SVCV), obvious pathological changes in skin, gill and gut mucosa and up-regulated expression of antiviral related genes in skin, gill, gut and spleen were observed, indicating that SVCV successfully infected common carp and activated the systemic and mucosal immune system. Interestingly, IgM showed a significant up-regulation only in systemic tissue (spleen), but not in mucosal tissues (gut, gills and skin), while increased expression of IgZ1 and IgZ2 was found in gut. In contrast, the expression of IgD increased significantly in spleen, gills and skin. These strongly suggest that fish Ig isotypes play different roles in mucosal and systemic immunity during viral infection. Common carp (Cyprinus carpio); Igs; Spring viraemia of carp virus (SVCV).

Dynamic Interaction Between Mucosal Immunity and Microbiota Drives Nose and Pharynx Homeostasis of Common Carp (Cyprinus carpio) After SVCV Infection.

The crosstalk between the immune system and microbiota drives an amazingly complex mutualistic symbiosis. In mammals, the upper respiratory tract acts as a gateway for pathogen invasion, and the dynamic interaction between microbiota and mucosal immunity on its surface can effectively prevent disease development. However, the relationship between virus-mediated mucosal immune responses and microbes in lower vertebrates remains uncharacterized. In this study, we successfully constructed an infection model by intraperitoneally injecting common carp (Cyprinus carpio) with spring viremia of carp virus (SVCV). In addition to the detection of the SVCV in the nose and pharynx of common carp, we also identified obvious histopathological changes following viral infection. Moreover, numerous immune-related genes were significantly upregulated in the nose and pharynx at the peak of SVCV infection, after which the expression levels decreased to levels similar to those of the control group. Transcriptome sequencing results revealed that pathways associated with bacterial infection in the Toll-like receptor pathway and the Nod-like receptor pathway were activated in addition to the virus-related Rig-I-like receptor pathway after SVCV infection, suggesting that viral infection may be followed by opportunistic bacterial infection in these mucosal tissues. Using 16S rRNA gene sequencing, we further identified an upward trend in pathogenic bacteria on the mucosal surface of the nose and pharynx 4 days after SVCV infection, after which these tissues eventually reached new homeostasis. Taken together, our results suggest that the dynamic interaction between mucosal immunity and microbiota promotes the host to a new ecological state.

Interactions Between Commensal Microbiota and Mucosal Immunity in Teleost Fish During Viral Infection With SVCV.

The mucosa of vertebrates is a particularly complex but dynamic environment in which the host constantly interacts with trillions of commensal microorganisms and pathogens. Although the internal and external mucosal microbiomes with immune defense of mammals have been well investigated, the relationship between mucosal microbes and their host's immune responses has not been systematically understood in the early vertebrates. In this study, we compared the composition and distribution of mucosal microbiota in common carp (Cyprinus carpio), and found that there were significant differences of microbiota between in the internal (gut) and external mucosal (buccal mucosa, gills and skin) tissues. Next, we successfully constructed an infection model with spring viremia of carp virus (SVCV). Specifically, following viral infection, the immune and antiviral related genes showed different up-regulation in all selected mucosal tissues while significant morphological changes were only found in external tissues including buccal mucosa, gills and skin. Using 16S rRNA gene sequence, we revealed that the abundance of Proteobacteria in mucosal tissues including buccal mucosa, gills and gut showed increased trend after viral infection, whereas the abundance of Fusobacteria significantly decreased in gut. In addition, the loss of dominant commensal microorganisms and increased colonization of opportunistic bacteria were discovered in the mucosal surfaces indicating that a secondary bacterial infection might occur in these mucosal tissues after viral infection. Overall, our results firstly point out the distribution of internal and external mucosal microbiota and analyze the changes of mucosal microbiota in common carp after SVCV infection, which may indicated that the potential role of mucosal microbiota in the antiviral process in early vertebrates.

Prevailing Role of Mucosal Igs and B Cells in Teleost Skin Immune Responses to Bacterial Infection.

The skin of vertebrates is the outermost organ of the body and serves as the first line of defense against external aggressions. In contrast to mammalian skin, that of teleost fish lacks keratinization and has evolved to operate as a mucosal surface containing a skin-associated lymphoid tissue (SALT). Thus far, IgT representing the prevalent Ig in SALT have only been reported upon infection with a parasite. However, very little is known about the types of B cells and Igs responding to bacterial infection in the teleost skin mucosa, as well as the inductive or effector role of the SALT in such responses. To address these questions, in this study, we analyzed the immune response of trout skin upon infection with one of the most widespread fish skin bacterial pathogens, Flavobacterium columnare This pathogen induced strong skin innate immune and inflammatory responses at the initial phases of infection. More critically, we found that the skin mucus of fish having survived the infection contained significant IgT- but not IgM- or IgD-specific titers against the bacteria. Moreover, we demonstrate the local proliferation and production of IgT+ B cells and specific IgT titers, respectively, within the SALT upon bacterial infection. Thus, our findings represent the first demonstration that IgT is the main Ig isotype induced by the skin mucosa upon bacterial infection and that, because of the large surface of the skin, its SALT probably represents a prominent IgT-inductive site in fish.

Mediation of Mucosal Immunoglobulins in Buccal Cavity of Teleost in Antibacterial Immunity.

The buccal mucosa (BM) of vertebrates is a critical mucosal barrier constantly exposed to rich and diverse pathogens from air, water, and food. While mammals are known to contain a mucosal associated lymphoid tissue (MALT) in the buccal cavity which induces B-cells and immunoglobulins (Igs) responses against bacterial pathogens, however, very little is known about the evolutionary roles of buccal MALT in immune defense. Here we developed a bath infection model that rainbow trout experimentally exposed to Flavobacterium columnare (F. columnare), which is well known as a mucosal pathogen. Using this model, we provided the first evidence for the process of bacterial invasion in the fish BM. Moreover, strong pathogen-specific IgT responses and accumulation of IgT+ B-cells were induced in the buccal mucus and BM of infected trout with F. columnare. In contrast, specific IgM responses were for the most part detected in the fish serum. More specifically, we showed that the local proliferation of IgT+ B-cells and production of pathogen-specific IgT within the BM upon bacterial infection. Overall, our findings represent the first demonstration that IgT is the main Ig isotype specialized for buccal immune responses against bacterial infection in a non-tetrapod species.

The predominant role of mucosal immunoglobulin IgT in the gills of rainbow trout (Oncorhynchus mykiss) after infection with Flavobacterium columnare.

Columnaris disease, induced by Flavobacterium columnare, seriously affects the health of freshwater fish species and damages the mucosal tissues, such as the fins, skin, and gills. Teleosts represent the first bony vertebrate to contain both innate and adaptive immune responses against pathogens. So far, three immunoglobulin isotypes (IgM, IgD, and IgT/IgZ) have been identified in teleost fish, and IgT in mucosal tissues of teleost fish was reported to perform a similar function to IgA in mammals during parasitic infection. However, very limited information is known about the function of IgT in gill mucosal tissues during bacterial infection. In the present study, rainbow trout (Oncorhynchus mykiss) was infected with F. columnare (Fc) via immersion. After Fc infection, the gill structure of rainbow trout showed serious hyperplasia symptoms on the secondary lamellae at 12 h post infection (hpi). Moreover, the mRNA expression levels of NOS2 and cathelicidin-1 were significantly upregulated immediately at 12 hpi and showed high expression throughout the experiment. IgT and IgM showed much higher mRNA expression levels at 28 days post infection (dpi) and 75 dpi, while IgD only showed high mRNA expression levels at 28 dpi. Importantly, the accumulation of IgT+ B cells and strong bacteria-specific IgT responses were detected in the gill lamellae of both infected fish (28 dpi) and survivor fish (75 dpi). Overall, our results suggest that IgT and IgT+ B cells play a central role in the adaptive immune responses of fish gill mucosa against bacterial infection.