Fibronectin 1B Gene Plays an Important Role in Loach Barbel Air-Breathing

Loach (Misgurnus anguillicaudatus) is well known to perform air-breathing through the posterior intestine and skin. However, we find here for the first time a unique central vascular structure in the loach barbel, with a blood–gas diffusion distance as short as that of the posterior intestine. Under acute hypoxia, the distance of loach barbels became significantly shorter. Moreover, barbel removal significantly decreased air-breathing frequency of the loach. These findings imply that the barbel is another air-breathing organ of the loach. For further investigation of loach barbel air-breathing, a transcriptome analysis of barbels with air exposure treatment was performed. A total of 2546 differentially expressed genes (DEGs) between the T-XU (air exposure) and C-XU (control) group were identified, and 13 key DEGs related to barbel air-breathing were screened out. On this foundation, sequence, expression, and location analysis results indicated an important positive role of fibronectin 1b (fn1b) in loach barbel air-breathing. We further generated an fn1b-depletion loach (MT for short) using the CRISPR/Cas9 technique. It was indicated that depletion of fn1b could weaker barbel air-breathing ability. In conclusion, due to nonlethal and regenerative characteristics, the loach barbel, a newly discovered and fn1b-related fish air-breathing organ, can be a good model for fish air-breathing research.

Osmoregulatory Plasticity of Juvenile Greater Amberjack (Seriola dumerili) to Environmental Salinity

Osmotic costs in teleosts are highly variable, reaching up to 50% of energy expenditure in some. In several species, environmental salinities close to the isosmotic point (~15 psu) minimize energy demand for osmoregulation while enhancing growth. The present study aimed to characterize the physiological status related to osmoregulation in early juveniles of the greater amberjack, Seriola dumerili, acclimated to three salinities (15, 22, and 36 psu). Our results indicate that plasma metabolic substrates were enhanced at the lower salinities, whereas hepatic carbohydrate and energetic lipid substrates decreased. Moreover, osmoregulatory parameters, such as osmolality, muscle water content, gill and intestine Na+-K+-ATPase activities, suggested a great osmoregulatory capacity in this species. Remarkably, electrophysiological parameters, such as short-circuit current (Isc) and transepithelial electric resistance (TER), were enhanced significantly at the posterior intestine. Concomitantly, Isc and TER anterior-to-posterior intestine differences were intensified with increasing environmental salinity. Furthermore, the expression of several adeno-hypophyseal genes was assessed. Expression of prl showed an inverse linear relationship with increasing environmental salinity, while gh mRNA enhanced significantly in the 22 psu-acclimated groups. Overall, these results could explain the better growth observed in S. dumerili juveniles kept at salinities close to isosmotic rather than in seawater.

Comparative Histological Analysis of Intestines of Loach, Grass Carp and Catfish Provide Insights into Adaptive Characteristics in Air-Breathing Fish

Accessory respiratory is generally accepted to have evolved independently on numerous occasions in adaption to aquatic hypoxia in freshwater habitats. In general, the air-breathing organ in fish is believed to be structurally modified to supplement respiration. In this study, intuitive evidence for elaborate structural modifications of the intestine, an air-breathing organ in mud loach (Misgurnus anguillicaudatus), compared with two other obligate aquatic breathers, grass carp (Ctenopharyngodon idellus) and yellow catfish (Pelteobagrus fulvidraco), were directly provided by histological and morphometric methods. As a result, a sharply decreasing height of mucosal folds and thickness of muscularis were manifested in loach intestine from its anterior to posterior region. Compared with grass carp and yellow catfish, loach had the smallest ratios of mucosal fold height/muscularis thickness to intestinal lumen radius in the posterior intestine. These suggested that the posterior intestine is the air-breathing location for the loach. Furthermore, length density of capillary (0.46±0.05 μm−2) in the posterior intestine of the loach was significantly higher than those of grass carp and yellow catfish. Meanwhile, diffusion distance of air-blood barrier (1.34±0.04 μm) in the posterior intestine of the loach was significantly smaller than those of the other two fish species. In summary, the characteristics of highly vascularized, short diffusion distance of air-blood barrier, thinned and flattened made the posterior intestine a perfect air-breathing location for the loach.

Protease, Lipase and Amylase Activities in Barred Loach, Nemacheilus Fasciatus C.V.

Barred loach is wild fish in which there is no information related to its enzymatic digestive capacity yet. Therefore research on this subject needs to be conducted. The spectrophotometry method was used to study protease, lipase, and amylase activities in barred loach. The study used 89 fish with an average weight of 3.61 ± 0.26g, and 1.68 ± 0.21g. The results showed that pH 7.0 was the highest protease activity. Similarly, lipase activity found between pH 7.0 and pH 8.1. Still, the amylase activity found to be high at pH 8.1. Protease activity also found to be high in the posterior intestine than in the hepato-pancreas and anterior intestine. Lipase and amylase activities did not found differences between hepato-pancreas, anterior intestine, and posterior intestine. In conclusion, the protease, lipase, and amylase activities of barred loach found along the gastrointestinal tract in both large and small fish, which shows that barred loach can digest protein, fat, and carbohydrates in diets better.

Osmoregulatory role of the intestine in the sea lamprey (Petromyzon marinus)

Lampreys are the most basal vertebrates with an osmoregulatory strategy. Previous research has established that the salinity tolerance of sea lamprey increases dramatically during metamorphosis, but underlying changes in the gut have not been examined. In the present work, we examined changes in intestinal function during metamorphosis and seawater exposure of sea lamprey ( Petromyzon marinus). Fully metamorphosed juvenile sea lamprey had 100% survival after direct exposure to 35 parts per thousand seawater (SW) and only slight elevations in plasma chloride (Cl−) levels. Drinking rates of sea lamprey juveniles in seawater were 26-fold higher than juveniles in freshwater (FW). Na+-K+-ATPase (NKA) activity in the anterior and posterior intestine increased 12- and 3-fold, respectively, during metamorphosis, whereas esophageal NKA activity was lower than in the intestine and did not change with development. Acclimation to SW significantly enhanced NKA activity in the posterior intestine but did not significantly change NKA activity in the anterior intestine, which remained higher than that in the posterior intestine. Intestinal Cl− and water uptake, which were observed in ex vivo preparations of anterior and posterior intestine under both symmetric and asymmetric conditions, were higher in juveniles than in larvae and were similar in magnitude of those of teleost fish. Inhibition of NKA by ouabain in ex vivo preparations inhibited intestinal water absorption by 64%. Our results indicate drinking and intestinal ion and water absorption are important to osmoregulation in SW and that preparatory increases in intestinal NKA activity are important to the development of salinity tolerance that occurs during sea lamprey metamorphosis.

The effect of diet on the structure of gut bacterial community of sympatric pair of whitefishes (Coregonus lavaretus): one story more

In the Coregonus lavaretus complex may be found lacustrine sympatric pairs, which serves as an intriguing model for studying different aspects of fish evolutionary biology. One such sympatric whitefish pair inhabits Teletskoye Lake (West Siberia, Russia) and includes a “large” form (Coregonus lavaretus pidschian (Gmelin, 1789)) and a “small” form (C. l. pravdinellus (Dulkeit, 1949)). C. l. pravdinellus has a narrow trophic specialization and feeds on zooplankton, whereas the diet of C. l. pidschian is based on benthic prey. In the present study we aimed to address the question of how the gut microbial community reflects the divergence in diet of a sympatric pair of whitefish. Studied samples included the mucosa and content were collected for cardiac and pyloric stomach, anterior, middle, and posterior intestine, but only mucosa was collected for the pyloric caeca. In addition, water, sediment, macrophyte (environmental microbiota) and invertebrate (microbiota of prey) samples were collected in the same location. The V3–V4 region of the 16S rRNA genes was chosen for microbiome analysis and the software PICRUSt used to estimate the difference functional roles of the microbiota. The number of OTUs and Chao1 index in mucosa and content of cardiac and pyloric stomach were significantly different between whitefish. Significant differences were observed between whitefish for content from different parts of the intestine in terms of OTU number and Chao1 indices, whereas for mucosa from the same parts of intestine these differences were absent. No significant differences were found for diversity estimates of mucosa and content of different parts of the gut (there were a few exceptions) between whitefish. The form of whitefish and the segment of the digestive system were factors with a significant determinative effect on the structure of the microbiota from gut mucosa and content. The most dominant phyla in mucosa and content of cardiac and pyloric stomach was Proteobacteria (57.0–84.0%) for both whitefish. Throughout the intestine of C. l. pidschian the dominant phyla in mucosa were Proteobacteria (38.8%) and Firmicutes (15.6%), whereas for C. l. pravdinellus–Tenericutes (49.6%) and Proteobacteria (28.1%). For both forms, the phylum Spirochaetes was found in a significant amount (20.0–25.0%) in the mucosa of the posterior intestine. While for the content obtained from anterior, middle and posterior intestines, the dominant bacterial phyla were the same as those described for mucosa from the same parts of the intestine for both whitefish. The bacterial community of the prey and environment was significantly different from bacterial communities found for all parts of the gut mucosa for both whitefish, with the exception of the mucosa of the cardiac stomach. According to PICRUSt the highest level of differences between whitefish at the L3 level were found for the intestinal mucosa (75.3%), whereas the lowest one was registered for stomach content (38.8%).

Histological and Ultrastructural Studies on the Intestine of Guntea Loach, Lepidocephalichthys Guntea (Cypriniformes, Cobitidae)

The cellular organizations of intestine in Lepidocephalichthys guntea (Hamilton, 1822) have been described by light as well as scanning and transmission electron microscopy. The intestine is short and straight like, marked into anterior, middle and posterior region based on mucosal folds, number and size of columnar epithelial cells and mucous cells, thickness of submucosa and muscularis layer. The mucosa of anterior intestine forms high folds, which are lined with compactly arranged columnar epithelial cells and mucous cells. In the middle intestine, folds are pointless whereas the posterior intestine is without folds. The submucosa is formed of thin layer of connective tissue, contained collagen bundles and blood capillaries, comparatively well developed in the posterior intestine. By scanning electron microscopy, outlines of the luminal surface of anterior and middle intestine is embossed with oval or rounded columnar epithelial cells contained densely packed stubby microridges. The posterior intestine has closely set longitudinal folds characterized with minute blood capillaries and columnar epithelial cells having inconspicuous microridges. Ultrastructurally, the mucosal surface of the intestine consists of mucous cells with electron dense granules and columnar epithelial cells having numerous microvilli, mitochondria, endoplasmic reticulum, lysosomes and Golgi body. Cellular components of the anterior and middle intestine participate in the absorption whereas the presence of enormous blood vessels and capillary net work of posterior intestine probably responsible for air breathing.

Comparative histological description of the digestive and visual system development of larval chame Dormitator latifrons (Pisces: Eleotridae)

This study was focused on the morphological description of chame larvae from one to six days post-hatching (dph), in order to generate information that helps understanding their feeding ability in early stages. The larvae were obtained by hormonal induction of a broodstock, using implants of GnRH synthetic analogues, during the spawning season from September to November 2010. The samples were included in historesin, and stained with hematoxylin-eosin. It was described that, once the larva hatched has a large amount of yolk it is gradually absorbed (from 1 to 3 dph). At day four, there is an open oral cavity with dental structures, the eye development is complete and there was pigmentation on it, the gills are observable and the foregut was differentiated in anterior and posterior intestine, showing the presence of digestive vacuoles in the anterior intestine. By five dph the larvae has developed muscle fibers and a structure identified as the stomach. At sixth dph, it is possible to identify the pancreas, the intestinal folds, and the brush border membrane, and there were food particles and bacteria rests in the gut. Based on the above, it is concluded that the larvae of D. latifrons from the fourth dph fully developed organs and systems that help the search of food, so they are capable of starting with exogenous feeding.

The role of the rectum in osmoregulation and the potential effect of renoguanylin on SLC26a6 transport activity in the Gulf toadfish (Opsanus beta)

Teleosts living in seawater continually absorb water across the intestine to compensate for branchial water loss to the environment. The present study reveals that the Gulf toadfish ( Opsanus beta) rectum plays a comparable role to the posterior intestine in ion and water absorption. However, the posterior intestine appears to rely more on SLC26a6 (a HCO3−/Cl− antiporter) and the rectum appears to rely on NKCC2 (SLC12a1) for the purposes of solute-coupled water absorption. The present study also demonstrates that the rectum responds to renoguanylin (RGN), a member of the guanylin family of peptides that alters the normal osmoregulatory processes of the distal intestine, by inhibited water absorption. RGN decreases rectal water absorption more greatly than in the posterior intestine and leads to net Na+ and Cl− secretion, and a reversal of the absorptive short-circuit current ( ISC). It is hypothesized that maintaining a larger fluid volume within the distal segments of intestinal tract facilitates the removal of CaCO3 precipitates and other solids from the intestine. Indeed, the expression of the components of the Cl−-secretory response, apical CFTR, and basolateral NKCC1 (SLC12a2), are upregulated in the rectum of the Gulf toadfish after 96 h in 60 ppt, an exposure that increases CaCO3 precipitate formation relative to 35 ppt. Moreover, the downstream intracellular effects of RGN appear to directly inhibit ion absorption by NKCC2 and anion exchange by SLC26a6. Overall, the present findings elucidate key electrophysiological differences between the posterior intestine and rectum of Gulf toadfish and the potent regulatory role renoguanylin plays in osmoregulation.