Vibrio diabolicus challenge in Bathymodiolus azoricus populations from Menez Gwen and Lucky Strike hydrothermal vent sites.

Menez Gwen (MG) and Lucky Strike (LS) deep-sea hydrothermal vents are located at 850 m and 1730 m depths respectively and support chemosynthesis-based ecosystems partially differing in heavy metal concentration, temperature range, and faunistic composition. The successfully adapted deep-sea vent mussel Bathymodiolus azoricus is found at both vent locations. In such inhospitable environments survival strategies rely on the establishment of bacteria-vent animal symbiosis In spite of the toxic nature of deep-sea vents, the problem of microbial threat and the need for immunity exist in B. azoricus. This study aims at investigating the immune system of B. azoricus from MG and LS populations by comparing immune gene expressions profiles using the deep-sea vent-related Vibrio diabolicus. Expression of nineteen immune genes was analyzed from gill, digestive gland and mantle tissues upon 3 h, 12 h and 24 h V. diabolicus challenges. Based on quantitative-Polymerase Chain Reaction (qPCR) significant gene expression differences were found among MG and LS populations and challenge times MG mussels revealed that gill and digestive gland gene expression levels were remarkably higher than those from LS mussels. Expression of Carcinolectin, Serpin-2, SRCR, IRGs, RTK, TLR2, NF-κB, HSP70 and Ferritin genes was greater in MG than LS mussels. In contrast, mantle tissue from LS mussels revealed the highest peak of expression at 24 h for most genes analyzed. The activation of immune signaling pathways demonstrated that gene expression profiles are distinct between the two mussel populations. These differences may possibly ensue from intrinsic immune transcriptional activities upon which host responses are modulated in presence of V. diabolicus. mRNA transcript variations were assessed during 24 h acclimatization taking into account the partial depuration to which mussels were subjected to. Additionally, gene expression differences may reflect still accountable effects from the presence of vent remaining microfluidic environments within the tissues analyzed.

Site-related differences in gene expression and bacterial densities in the mussel Bathymodiolus azoricus from the Menez Gwen and Lucky Strike deep-sea hydrothermal vent sites.

The deep-sea hydrothermal vent mussel Bathymodiolus azoricus is a symbiont bearing bivalve that is found in great abundance at the Menez Gwen and Lucky Strike hydrothermal vent sites and in close vicinity of the Azores region near the Mid-Atlantic Ridge (MAR). The physiological relationships that vent mussels have developed with their physical and chemical environments are likely to influence global gene expression profiles providing thus the means to investigate distinct biological markers predicting the origin of Bathymodiolus sp. irrespectively of their geographical localization. Differences found at gene expression levels, and between fluorescence in situ hybridization (FISH) and 16S rRNA amplicon sequencing results provided experimental evidence for the distinction of both Menez Gwen and Lucky Strike vent mussel individuals based on bacterial and vent mussel gene expression signatures and on the constitutive distribution and relative abundance of endosymbiotic bacteria within gill tissues. Our results confirmed the presence of methanotroph endosymbionts in Menez Gwen vent mussels whereas Lucky Strike specimens seem to harbor a different bacterial morphotype when a methane monooxygenase gene specific probe was used. No qualitative differences could be visualized between Menez Gwen and Lucky Strike individuals when tested with a sulfur-oxidizing-related probe. Quantitative PCR (qPCR) studies revealed different gene expression profiles in both Menez Gwen and Lucky Strike mussel gill tissues for the immune genes selected. Genes encoding transcription factors presented noticeably low levels of fold expression whether in Menez Gwen or Lucky Strike animals whereas the genes encoding effector molecules appeared to have higher levels expression in gill tissues from Menez Gwen animals. The peptidoglycan recognition molecule encoding gene, PGRP, presented the highest level of transcriptional activity among the genes analyzed in Menez Gwen mussel gill tissues, seconded by carcinolectin and thus denoting the relevance of immune recognition molecules in early stage of the immune responses onset. Genes regarded as encoding molecules involved in signaling pathways were consistently expressed in both Menez Gwen and Lucky Strike mussel gill tissues. Remarkably, the immunity-related GTPase encoding gene demonstrated, in Lucky Strike samples, the highest level of expression among the signaling molecule encoding genes tested when expressions levels were compared between Menez Gwen and Lucky Strike animals. A differential expression analysis of bacterial genes between Menez Gwen and Lucky Strike mussels indicated a clear expression signature in the latter animal gill tissues. The bacterial community structure ensued from the 16S rRNA sequencing analyses pointed at an unpredicted conservation of endosymbiont bacterial loads between Menez Gwen and Lucky Strike samples. Taken together, our results support the hypothesis that B. azoricus exhibits different transcriptional statuses while living in distinct hydrothermal vent sites may result in distinct gene expressions because of physico-chemical and/or symbiont densities differences.

Regulation of immunocompetence by different androgen metabolites in a blenny with alternative reproductive tactics.

In Parablennius parvicornis, small reproductive males with relatively low expression of secondary sexual characters (M- morphotype) parasite on the parental investment of the larger nest-holder males which have fully developed secondary sexual characters (M+ morphotype). In comparison with M+ males, M- males have relatively low levels of androgens while having high blood cell percentages of lymphocytes and antigen responsiveness. Here we test the hypothesis that androgens are a causal factor for these differences in immunocompetence between morphotypes. After drawing an initial blood sample, males received a silastic implant containing either oil only (C), or oil with testosterone (T) or 11-ketotestosterone (KT). Males were re-caught 2 weeks later for drawing of the final blood sample. KT but not T induced the development of secondary sexual characters in M- males. M- males treated with KT showed lower swimming activity than the males treated with T or C implants, suggesting that KT also mediates behavioral changes in M- males. As expected, blood cell percentages of lymphocytes, but not of granulocytes, were higher in M- males than in M+ males. Overall, lymphocyte percentages increased in the C group which might have been a response to the surgery/treatment. In concordance with the hypothesis, lymphocyte percentages were suppressed in males treated with T in comparison with controls. However, no significant change was found in KT-treated males. This suggests that androgens modulate central, morphological and immunological traits by partly independent androgen mechanisms in P. parvicornis.

Diversity and seasonal changes in the ichthyofauna of rocky tidal pools from Praia Vermelha and São Roque, Santa Catarina

Este trabalho realizou-se entre Abril de 1998 e Maio de 1999 e visou o estudo das comunidades de peixes em duas poças de maré situadas nas praias de Praia Vermelha (PV) e São Roque (SR), Santa Catarina, Brasil. A poça de PV é maior e mais próxima do sub-mareal. Os exemplares foram coletados mensalmente, durante a baixa-mar, com o anestésico quinaldina. Em PV recolheram-se 1287 peixes pertencentes a 19 espécies enquanto que em SR foram capturados 553 peixes de 11 species. Em ambas as poças as espécies dominantes foram Bathygobius soporator (Gobiidae), Abudefduf saxatilis (Pomacentridae), Parablennius pilicornis (Blenniidae), Labrisomus nuchipinnis (Labrisomidae) e Epinephelus marginatus (Serranidae). A diversidade e o número de peixes foram maiores nos meses de Verão e correspondem a uma época de recrutamento para as espécies A. saxatilis e E. marginatus.