Swim Bladder Research Articles

자동차 시트에서 자동으로 좌우 엉덩이 부분 독립제어가 가능한 Air Bladder를 이용한 척추 골반 균형 개선 효과에 대한 연구

자동차 시트에서 자동으로 좌우 엉덩이 부분 독립제어가 가능한 Air Bladder를 이용한 척추 골반 균형 개선 효과에 대한 연구

Zooplankton functional traits in a tropical estuarine system: Are lower and upper estuaries functionally different?

Functional diversity measures help to understand the underlying mechanisms explaining the relationship between organisms and environment. This work examined the mesozooplankton community of an estuary under the hypothesis that the upper (< 25 psu) and lower (> 25 psu) zones are functionally different in terms of feeding strategies, trophic groups, camouflage, buoyancy, and defense against predators, due to the salinity gradient. We used the ‘community-weighted mean trait values’ (CWM) and a fuzzy correspondence analysis (FCA) in combination with three functional indices (FRic: functional richness, FEve: functional evenness, FDiv: functional divergence) to test our hypothesis. Zooplankton samples were taken in the tropical Sontecomapan estuary, southern Gulf of Mexico, in June 2018 and October 2019. A total of 21 zooplankton groups were recognized, all of them present in the lower estuary. Results showed significant differences (p< 0.05) in the FRic and FDiv values between the two zones, as well in the CWM metric. Results of the FCA showed that the lower estuary was characterized by herbivorous filter feeders that use watery bodies, flattened body forms, mucus houses, or ‘wings’ for buoyancy; highly transparent bodies (associated with gelatinous body structure) for camouflage and predators’ avoidance and, shells or carapaces as antipredator tactics. The upper estuary was characterized by omnivorous cruising predators that use their appendages or swim bladders for buoyancy, with ‘medium’ and ‘low’ levels of transparency, associated with chitinous and skin-protected body structures, which provide alternative ways for camouflage and spines or fast swimming behavior to protect from predators. These findings evidenced that the upper and lower zones of an estuary exhibit differences in the function of the zooplankton community and shed new light on the comprehension of estuaries’ ecological function.

Infestation with metacercarial stage of Isoparorchis hypselobagri (Billet, 1898) in cage cultured Ompok bimaculatus vis-a-vis host and environmental interaction in a large tropical reservoir

The present study reports the occurrence of metacercariae of Isoparorchis hypselobagri in a food fish Ompok bimaculatus cultured in cages at Maithon reservoir, India. The metacercarial parasite was initially identified morphologically. Molecular identification through partial amplification of 18S rRNA and cytochrome oxidase subunit 1 (COI) gene and sequencing further confirmed the parasite species. The patterns of infection in terms of intensity, mean intensity, and prevalence of metacercariae in O. bimaculatus was recorded. Influence of host size, season and water quality on infection rate was tested through Generalized linear model (GLM). The prevalence varied from 42.2% to 88.8% and intensity of infection ranged from 1 to 10. PCA analysis showed that the metacercarial parasite was mostly located near the air bladder, around kidney and near kidney of the host. GLM results showed that the size of the fish influenced the intensity of infection. Higher water temperature reduced the parasitic growth whereas dissolved oxygen was positively related with intensity of the parasite infection. The occurrence of the first intermediate host snails indicated that they might be the possible source of cercaria in the cages. This study is the first report of I. hypselobagri infections in inland cage culture from India and has implications for understanding the ecological and taxonomic patterns and pathology in caged fish in tropical reservoirs. The high infestation rate and large size of parasite presents serious threat for production of this high value fish species in cages. The presence of parasites not only affects growth and survival but also leads to reduced consumer acceptance. The control of snail vectors might play a significant role in prevention of parasitic diseases in cage culture system. Hence, before selection of candidate fish species and site for cage culture, management of such parasitic disease should be duly considered. This will not only prevent economic losses due to parasitic diseases but also ensure consumer safety.

Combined Reproductive Effects of Imidacloprid, Acetochlor and Tebuconazole on Zebrafish (Danio rerio)

Pesticides usually occur as mixtures of multiple chemicals in the natural aquatic ecosystem, so research based on the toxicity data of a single compound on aquatic organisms is not enough to accurately assess the actual toxicity risk of pesticides. There is still a gap in the research on the reproductive toxicity of combined insecticides, herbicides and fungicides on zebrafish (Danio rerio). In this study, zebrafish were used to systematically investigate the separate and combined reproductive toxicity of imidacloprid (IMI), acetochlor (ACT) and tebuconazole (TBZ), which are commonly used in rice fields. Adult zebrafish were exposed to the three pesticides individually and in combination for 28 days, and the number, heartbeat, deformation rate, body length, and swim bladder development of F1 offspring embryos were observed and the reproductive hormones testosterone (T), estradiol (E2), and vitellogenin (VTG) contents and the expressions of nine reproductive genes (ar, esr2a, vtg1, gr, star, fshr, hmgcrb, 3βhsd and vasa) in the testes of the male and the ovaries of the female F0 zebrafish adults were measured to evaluate the individual and combined effects. The results showed that exposure to the mixtures of IMI, ACT and TBZ resulted in a decrease in heartbeat, body length and swim bladder development and an increase in the deformity rate of F1 offspring embryos compared to the individual exposure groups. In the combined exposure group, the content of T decreased significantly and the content of VTG increased significantly in the testes of the males; the content of T significantly increased, while the content of E2 and VTG significantly decreased in the ovaries of the females, indicating that combined exposure showed a more obvious endocrine-disrupting effect compared to the individual exposures. In addition, the expression of nine reproductive genes was significantly altered compared to the individual exposure groups. Therefore, our results indicated that the mixture of IMI, ACT and TBZ caused fewer number of F1 embryos, higher developmental defects of F1, greater disruption in the content of reproductive hormones and the expression of reproductive genes compared to the individual pesticides at the corresponding doses. Therefore, the presence of pesticides in mixtures in the real water environment is likely to increase the toxic reproductive effects on zebrafish and cause more serious impacts on aquatic ecosystems.

Developmental safety of nirmatrelvir in zebrafish (Danio rerio) embryos.

Nirmatrelvir, in combination with ritonavir, is one of the first orally available antiviral treatment for coronavirus disease 2019 (COVID-19). Symptomatic pregnant women are at increased risk for severe illness and complications that can affect the developing baby. No malformations or lower embryo-fetal survival have been observed when nirmatrelvir were administered to pregnant rats and rabbits. Safety evaluation of drugs used for treating COVID-19 also in pregnancy is urgent for public health, then in this study we further investigated nirmatrelvir developmental toxicity using zebrafish as in vivo model. Using the standardized Fish Embryo Toxicity (FET) test, we first determined the lethal concentration 50 (LC50), exposing embryos from gastrula stage up to 120 hr post fertilization (hpf) and daily recording lethality. Then, we exposed embryos to five doses comprising the human therapeutic one and up to the LC50 (25 μM). Morphology was evaluated at 72 and 120 hpf. Nirmatrelvir did not affect survival rate and did not induce morphological defects up to the human therapeutic dose. Exposure at higher doses (2.4× and 3× the human Cmax ) however resulted in decreased hatching rate, reduced growth, slower heartbeat with pericardial edema, reduction of eye dimension, absence of the swim bladder and disruption of the anterior-posterior axis, with lack of tail detachment, spinal curvature and straight and smaller head. Our findings in zebrafish embryos add further information about developmental nirmatrelvir safety. Further studies are needed for pharmacological safety assessment of nirmatrelvir exposure during pregnancy.

Adverse effects of thyroid-hormone-disrupting chemicals 6-propyl-2-thiouracil and tetrabromobisphenol A on Japanese medaka (Oryzias latipes)

Thyroid-hormone-disrupting chemicals are increasingly attracting attention because of their potential harmful effects on animal health, including on fishes. Here, we investigated the effects of exposure to the thyroid-hormone-disrupting chemicals 6-propyl-2-thiouracil (PTU) and tetrabromobisphenol A (TBBPA) on swim bladder inflation, eye development, growth, swimming performance, and the expression of thyroid-related genes in Japanese medaka (Oryzias latipes). PTU exposure resulted in reductions in eye size, growth, and swim bladder inflation, and these effects led to poorer swimming performance. These phenotypic effects were accompanied by increased expression of the thyroid-stimulating hormone subunit beta (tshβ) paralog tshβ-like, but there were no significant changes in expression for tshβ, deiodinase 1 (dio1), deiodinase 2 (dio2), and thyroid hormone receptor alpha (trα) and beta (trβ). For PTU exposure, we identified the key event (swim bladder inflation reduction) and an adverse outcome (swimming performance reduction). No significant effects from TBBPA exposure were seen on swim bladder inflation, eye development, growth, or swimming performance. However, expression of tshβ-like and tshβ (significantly enhanced) and trα and trβ (significantly reduced) were affected by TBBPA exposure albeit not in dose-dependent manners. There were no effects of TBBPA on the expression of dio1 and dio2. We thus show that the two thyroid-hormone-disrupting chemicals PTU and TBBPA differ in their effect profiles with comparable effects on the studied phenotypes and thyroid-related gene expression to those reported in zebrafish.

Early ontogeny of the commercially valuable fish red-bellied pacu Piaractus brachypomus (Characiformes, Serrasalmidae) from the Amazon, Brazil.

The initial development of the red-bellied pacu Piaractus brachypomus is described using morphological, meristic and morphometric characteristics. A total of 127 individuals were analysed (47 in the yolk-sac, 35 in pre-flexion, five in flexion, 20 in post-flexion and 20 in juvenile) with standard length varying between 2.92 and 48.61 mm. The larvae are born poorly developed and have a discoidal yolk at ~6.33 mm standard length. During early ontogeny, the mouth passes from terminal to subterminal and the anal opening reaches the vertical line over the midline region of the body. There are changes in body shape from long and moderate to deep, head length from small to large, and eye diameter from moderate to large. Dendritic chromatophores were present in the ventral, dorsal and upper part of the swim bladder in the early larval stages. Rounded spots are evident all over the body in juveniles. The total number of myomeres ranges from 39 to 41 (20-23 pre-anal, 17-20 post-anal). Through the morphometric relationships, it was evidenced that the greatest changes during the initial ontogeny of P. brachypomus occur in the transition from the post-flexion stage to the juvenile period, indicating changes in behaviour, foraging and physiology.

Developmental toxicity and thyroid hormone-disrupting effects of acetyl tributyl citrate in zebrafish and Japanese medaka

Chemical pollution from plasticizers in water environments is a serious environmental problem worldwide. Phthalate plasticizers have potential endocrine-disrupting effects in vertebrates. In this study, the effects of a non-phthalate acetyl tributyl citrate (ATBC) plasticizer on endocrine hormone activity (according to thyroid-related gene expression) and the developmental toxicity of ATBC were determined in zebrafish (Danio rerio) and Japanese medaka (Oryzias latipes) embryos. ATBC exposure increased the mortality of zebrafish and Japanese medaka at concentrations of 443.05 and 1,937.41 μg/L, respectively. Body curvature, edema, and growth inhibition were also observed after ATBC exposure in both species. Importantly, both species exhibited suppressed thyroid-related gene mRNA expression after ATBC exposure. ATBC exposure significantly suppressed the expression of thyroid-stimulating hormone beta subunit (tshβ), iodothyronine deiodinase 1 (dio1), dio2, and thyroid hormone receptor alpha (trα) in zebrafish. In Japanese medaka, ATBC exposure suppressed the mRNA expression of dio2, trα, and trβ but not tshβ and dio1. In summary, ATBC exposure inhibited thyroid-related gene expression and led to improper swim bladder inflation in the test species. This is the first report of a non-phthalate ATBC plasticizer inducing abnormal embryo development and disrupting thyroid hormone activity in fish.

Properties of Grass Carp (Ctenopharyngodon idella) Collagen and Gel for Application in Biomaterials.

The biochemical properties of collagens and gels from grass carp (Ctenopharyngodon idella) were studied to explore the feasibility of their application in biomaterials. The yields of skin collagen (SC) and swim bladder collagen (SBC) extracted from grass carp were 10.41 ± 0.67% and 6.11 ± 0.12% on a wet basis, respectively. Both collagens were characterized as type I collagen. Denaturation temperatures of SC and SBC were 37.41 ± 0.02 °C and 39.82 ± 0.06 °C, respectively. SC and SBC had high fibril formation ability in vitro, and higher values of salinity (NaCl, 0-280 mM) and pH (6-8) in formation solution were found to result in faster self-assembly of SC and SBC fibrils as well as thicker fibrils. Further tests of SC gels with regular morphology revealed that their texture properties and water content were affected by pH and NaCl concentration. The hardness, springiness, and cohesiveness of SC gels increased and the chewiness and water content decreased as pH increased from 7 to 8 and NaCl concentration increased from 140 to 280 mM. These properties suggest that collagens from grass carp may be useful in biomaterial applications in the future.

Non-additive mixture effects of benzo[a]pyrene and pesticides in vitro and in vivo: Role of AhR signaling

Polycyclic aromatic hydrocarbons (PAHs) and pesticides are two major groups of environmental contaminants which humans are simultaneously exposed to. However, potential mixture interactions of these groups of chemicals are not well-studied. In this study, the effects of binary mixtures of the PAH benzo[a]pyrene (B[a]P) and the commonly used pesticides chlorpyrifos, paraquat and tebuconazole on human liver HepG2 cells were investigated. The results showed that binary mixtures of B[a]P and paraquat or tebuconazole mainly caused additive effects on cell viability and cytochrome P4501a1 (CYP1A1) expression compared to single compound exposures. In contrast, the binary mixture with chlorpyrifos interacted antagonistically on cell viability and ROS production, whereas synergistic effects were observed for induction of CYP1A1 expression. B[a]P and chlorpyrifos also inhibited the activity of recombinant human CYP1A1 enzyme. To verify the synergistic in vitro results, zebrafish (Danio rerio) embryos were exposed to binary mixtures of B[a]P and chlorpyrifos. The mixtures caused synergistic induction of CYP1A expression, as well as synergistic developmental toxicity on multiple endpoints including non-inflated swim bladder, yolk-sac and pericardial edema, and spinal deformation. The effects were reduced upon morpholino-mediated knockdown of the aryl hydrocarbon receptor (AhR), indicating an AhR-dependence of the synergistic toxicity. Altogether, these data suggest that the combination of AhR activation and CYP1A1 inhibition is responsible for the underlying non-additive interaction between B[a]P and chlorpyrifos in vitro and in vivo.

A novel approach reveals underestimation of productivity in the globally important macroalga, Ascophyllum nodosum

Ascophyllum nodosum (L.) Le Jolis (Fucales, Fucaceae) is a modular intertidal brown alga that has the particularity of forming an air bladder once a year at the apical tip of the growing shoots. This characteristic provides a means for aging and estimating the growth of individuals. While it has long been recognized that growth can occur in older parts of the frond, this has not been properly assessed until now and has largely been overlooked when calculating the productivity of the species. Recent studies have suggested that the growth and elongation of older segments is minimal and thus has been used to infer past environmental conditions. Here we assessed the length and mass of successive internodal segments from 25 sites spread over both sides of the North Atlantic, covering a wide portion of the distribution of the species. By calculating the ratio of the mass and length of a segment divided by the segment produced the following year, we established that internodal segments continue accumulating mass for 1–5 years and increase in length for 1–3 years at most sites. Segments can almost triple their mass during their second year and more than double their length. These results indicate that previous productivity and growth estimates for A. nodosum based on apical growth alone greatly underestimate the true productivity of the species and its role in coastal carbon cycling. Furthermore, because they grow over several years, internodal segments should not be used to infer past environmental conditions or to reconstruct growth patterns over time.

Generation and characterization of a novel gne Knockout Model in Zebrafish.

GNE Myopathy is a rare, recessively inherited neuromuscular worldwide disorder, caused by a spectrum of bi-allelic mutations in the human GNE gene. GNE encodes a bi-functional enzyme responsible for the rate-limiting step of sialic acid biosynthesis pathway. However, the process in which GNE mutations lead to the development of a muscle pathology is not clear yet. Cellular and mouse models for GNE Myopathy established to date have not been informative. Further, additional GNE functions in muscle have been hypothesized. In these studies, we aimed to investigate gne functions using zebrafish genetic and transgenic models, and characterized them using macroscopic, microscopic, and molecular approaches. We first established transgenic zebrafish lineages expressing the human GNE cDNA carrying the M743T mutation, driven by the zebrafish gne promoter. These fish developed entirely normally. Then, we generated a gne knocked-out (KO) fish using the CRISPR/Cas9 methodology. These fish died 8-10days post-fertilization (dpf), but a phenotype appeared less than 24h before death and included progressive body axis curving, deflation of the swim bladder and decreasing movement and heart rate. However, muscle histology uncovered severe defects, already at 5 dpf, with compromised fiber organization. Sialic acid supplementation did not rescue the larvae from this phenotype nor prolonged their lifespan. To have deeper insights into the potential functions of gne in zebrafish, RNA sequencing was performed at 3 time points (3, 5, and 7 dpf). Genotype clustering was progressive, with only 5 genes differentially expressed in gne KO compared to gne WT siblings at 3 dpf. Enrichment analyses of the primary processes affected by the lack of gne also at 5 and 7 dpf point to the involvement of cell cycle and DNA damage/repair processes in the gne KO zebrafish. Thus, we have established a gne KO zebrafish lineage and obtained new insights into gne functions. This is the only model where GNE can be related to clear muscle defects, thus the only animal model relevant to GNE Myopathy to date. Further elucidation of gne precise mechanism-of-action in these processes could be relevant to GNE Myopathy and allow the identification of novel therapeutic targets.

Triplophysadaryoae, a new nemacheilid loach species (Teleostei, Nemacheilidae) from the Syr Darya River basin, Central Asia.

Triplophysadaryoae, new species, is described from the Sokh River, a former tributary of Syr Darya that today fails to reach the river, in the Sokh District, an exclave of Uzbekistan, surrounded by Kyrgyzstan. Triplophysadaryoae is distinguished from other species of Triplophysa in Central Asia by a truncate caudal fin with 13 or 14 branched rays, body without obvious mottling, dorsal-fin origin opposite to pelvic-fin insertion, and absence of the posterior chamber of the air bladder. Molecular data suggest that Triplophysadaryoae is closely related to T.ferganaensis from the Shakhimardan stream, a small tributary of Syr Darya in the Yordon village, another exclave of Uzbekistan in Kyrgyzstan. The two species were separated by a Kimura 2-parameter genetic distance of 2.8% in the mitochondrial DNA cytochrome c oxidase subunit I barcode region; they are also distinguished morphologically. A key to the species of Triplophysa in the Syr Darya basin and adjacent regions is provided.

Gsx2, but not Gsx1, is necessary for early forebrain patterning and long-term survival in zebrafish.

Homeobox transcription factor encoding genes, genomic screen homeobox 1 and 2 (gsx1 and gsx2), are expressed during neurodevelopment in multiple vertebrates. However, we have limited knowledge of the dynamic expression of these genes through developmental time and the gene networks that they regulate in zebrafish. We confirmed that gsx1 is expressed initially in the hindbrain and diencephalon and later in the optic tectum, pretectum, and cerebellar plate. gsx2 is expressed in the early telencephalon and later in the pallium and olfactory bulb. gsx1 and gsx2 are co-expressed in the hypothalamus, preoptic area, and hindbrain, however, rarely co-localize in the same cells. gsx1 and gsx2 mutant zebrafish were made with TALENs. gsx1 mutants exhibit stunted growth, however, they survive to adulthood and are fertile. gsx2 mutants experience swim bladder inflation failure that prevents survival. We also observed significantly reduced expression of multiple forebrain patterning distal-less homeobox genes in mutants, and expression of foxp2 was not significantly affected. This work provides novel tools with which other target genes and functions of Gsx1 and Gsx2 can be characterized across the central nervous system to better understand the unique and overlapping roles of these highly conserved transcription factors.

Comparing Body Density of Lumpfish (Cyclopterus lumpus) to Different Operational Welfare Indicators

Farmed lumpfish (Cyclopterus lumpus) are commonly used as cleaner fish in the salmonid aquaculture industry, but a knowledge gap exists with regards to their body density. Filling this knowledge gap is of importance, as the lumpfish has no swim bladder and thus relies on alternative methods for buoyancy, i.e., the body density difference between the fish and its surroundings. The aims of this study were to measure the body density of lumpfish and investigate the correlation between body density and different operational welfare indicators. A total of 138 lumpfish were sampled at five different aquaculture sites situated in the Faroe Islands. Weight in water and air was measured, body density was calculated, and operational welfare was assessed. The average body density of the juvenile lumpfish was 1.030 g mL−1. Fulton’s K, stomach score, and length were negatively correlated to body density, while the hepatosomatic index was positively correlated to body density. Liver colour was correlated to body density, but the groupings were too broad for a final definitive conclusion. The knowledge gained from this study might help the industry improve their understanding of the operational welfare indicators used for lumpfish. Additionally, the knowledge might also help the aquaculture industry improve their husbandry and feeding practices.

Effects of dietary collagen cofactors and hydroxyproline on the growth performance, textural properties and collagen deposition in swim bladder of Nibea coibor based on orthogonal array analysis

The global demand for collagen is continuously growing over years. Nutritional regulation of quality and collagen deposition in the swim bladder, an ideal collagen source of marine product, is a promising method for improving the economic value of fish. In this study, to enhance growth efficiency and collagen deposition in swim bladder of an important economic marine fish, the chu’s croaker (Nibea coibor), a L9(34) orthogonal array was designed with three levels for each factor, which are hydroxyproline (Hyp) and the key cofactors for proline hydroxylation during collagen post-translation modification, such as A1-A3 (Hyp): 0, 10 and 20 g/kg; B1-B3 (vitamin C, VC): 0, 0.20 and 0.40 g/kg; C1-C3 (Fe2+): 0, 0.20 and 0.40 g/kg; D1-D3 (α-ketoglutarate, AKG): 0, 10 and 20 g/kg, to formulate the trial feeds. An 8-week feed trial was carried out on 405 fish (initial weight 269.04 ± 3.18 g) which were randomly distributed into the nine treatment groups with three replicates of 15 fish per cage. Results indicated that appropriate increase of dietary Hyp and VC level is more beneficial to collagen deposition and textural properties in swim bladder, while Hyp and AKG exerted more significant effects on the growth performance. Gene expression analysis of collagen metabolism indicated that Hyp mainly improved the synthesis of type I collagen and inhibited collagen degradation, while VC and AKG mainly play an important role in hydroxylation of proline residues on collagen. The best combination for growth performance and collagen deposition in swim bladder were suggested to be A2B2C1D3 and A2B2C1D1, respectively. The results demonstrated that appropriate dietary supplementation of Hyp and cofactors can improve growth and collagen production of swim bladder. Our result would provide reference data for nutritional regulation of the species and optimization of the commercial feed to provide fish products with high quality.

The Zebrafish (Danio rerio) Embryo Model as a Tool to Assess Drinking Water Treatment Efficacy for Freshwater Impacted by Crude Oil Spill.

Traditional approaches toward evaluating oil spill mitigation effectiveness in drinking water supplies using analytical chemistry can overlook residual hydrocarbons and treatment byproducts of unknown toxicity. Zebrafish (Danio rerio) were used to address this limitation by evaluating the reduction in toxicity to fish exposed to laboratory solutions of dissolved crude oil constituents treated with 3 mg/L ozone (O3 ) with or without a peroxone-based advanced oxidation process using 0.5 M H2 O2 /M O3 or 1 M H2 O2 /M O3 . Crude oil water mixtures (OWMs) were generated using three mixing protocols-orbital (OWM-Orb), rapid (OWM-Rap), and impeller (OWM-Imp) and contained dissolved total aromatic concentrations of 106-1019 µg/L. In a first experiment, embryos were exposed at 24 h post fertilization (hpf) to OWM-Orb or OWM-Rap diluted to 25%-50% of full-strength samples and in a second experiment, to untreated or treated OWM-Imp mixtures at 50% dilutions. Toxicity profiles included body length, pericardial area, and swim bladder inflation, and these varied depending on the OWM preparation, with OWM-Rap resulting in the most toxicity, followed by OWM-Imp and then OWM-Orb. Zebrafish exposed to a 50% dilution of OWM-Imp resulted in 6% shorter body length, 83% increased pericardial area, and no swim bladder inflation, but exposure to a 50% dilution of OWM-Imp treated with O3 alone or with 0.5 M H2 O2 /M O3 resulted in normal zebrafish development and average total aromatic destruction of 54%-57%. Additional aromatic removal occurred with O3 + 1 M H2 O2 /M O3 but without further attenuation of toxicity to zebrafish. This study demonstrates using zebrafish as an additional evaluation component for modeling the effectiveness of freshwater oil spill treatment methods. Environ Toxicol Chem 2022;41:2822-2834. © 2022 SETAC.

Decellularized fish swim bladder patch loaded with mesenchymal stem cells inhibits neointimal hyperplasia.

We previously showed decellularized fish swim bladder can be used as vascular patch and tube graft in rats, mesenchymal stem cells (MSCs) have showed the capability to inhibit neointimal hyperplasia in different animal models. We hypothesized that decellularized fish swim bladder patch loaded with MSCs (bioinspired patch) can inhibit neointimal hyperplasia in a rat aortic patch angioplasty model. Rat MSCs were grown in vitro and flow cytometry was used to confirm their quality. 3.6 × 105 MSCs were mixed into 100 μl of sodium alginate (SA)/hyaluronic acid (HA) hydrogel, two layers of fish swim bladders (5 mm × 5 mm) were sutured together, bioinspired patch was created by injection of hydrogel with MSCs into the space between two layers of fish swim bladder patches. Decellularized rat thoracic aorta patch was used as control. Patches were harvested at days 1 and 14 after implantation. Samples were examined by histology, immunohistochemistry, and immunofluorescence. The decellularized rat thoracic aorta patch and the fish swim bladder patch had a similar healing process after implantation. The bioinspired patch had a similar structure like native aorta. Bioinspired patch showed a decreased neointimal thickness (p=.0053), fewer macrophages infiltration (p=.0090), and lower proliferation rate (p=.0291) compared to the double layers fish swim bladder patch group. Decellularized fish swim bladder patch loaded with MSCs can inhibit neointimal hyperplasia effectively. Although this is a preliminary animal study, it may have a potential application in large animals or clinical research.

Molecular Identification of Commercial Fish Maws by DNA Sequencing of 16S rRNA and Cytochrome c Oxidase I Genes

Fish maws (dried swim bladders) have long been used for medicinal tonics and as a valuable food resource in Southeast Asia. However, it is difficult to identify the original species of fish maws sold in markets due to a lack of taxonomic characteristics. In the present study, 37 kinds of commercial fish maws from various medicinal material markets were examined, and gene sequences were successfully obtained from ca. 95% of the samples. Partial sequences of the 16S rRNA gene and cytochrome c oxidase I (COI) gene were obtained and used to investigate the origin of these commercial fish maws. Thirty-five specimens belonged to nine species: five croakers and four noncroakers. All species identification was supported by both high homogeneity (98 to 100%) and clear clustering with low within-group Kimura two-parameter divergence scores (0 to 0.04 for 16S rRNA and 0 to 0.07 for COI) and high between-group divergence scores (0.07 to 0.15 for 16S rRNA and 0.11 to 0.24 for COI). Croakers were the predominant species, accounting for 74% of the total fish maw specimens. The large demand for croakers has put some species at the risk of extinction due to overfishing. As a valuable food, fish maw has progressively become more popular and has been used as a substitute for shark fin. The identification results allowed us to learn more about the fish species available on the fish maw market and provided an indicator for possible control of threatened or endangered fish species. A probable correlation between the molecular characteristics and morphological features of fish maws was also found and could provide both consumers and merchants with an important reference for identifying the origin of fish maws.

Bioaccumulation of Hexavalent Chromium in Commercially Edible Fish Grass Carp, Ctenopharyngodon idella

The current research was conducted with the aim to investigate the rate of chromium accumulation in various tissues of commercially valuable freshwater fish, Grass Carp, Ctenopharyngodon idella. Investigation included various tissues of skin, muscles, liver, swim bladder, intestine, and gills in fish. For this purpose, the fish were exposed to sub lethal concentration of hexavalent chromium in the form of potassium dichromate with a dose of 120 mg/L for 40, 20 days and 10 mg/L for 7, 25, 40 and 60 days. Forty fish of the same weight and length (70.45±2.91 g and 7.32±0.16 inches) were used in the present research. Chromium was estimated after acid digestion of the sample tissues and further analyzed through atomic absorption spectrophotometer (Spectra AA6300 China). The results showed that in all the experiments intestine remained the highly accumulative tissue and the accumulation of chromium in the intestine increased with the increase in exposure time i.e., 0.63±0.21 after 7 days, 0.83±0.35 after 25 and 1.63±0.44 after 60 days. Finding of this research work supports that the route of metal uptake was mainly oral instead of absorption by gills or skin.