Cells In Fish Research Articles

Xylanase enhances gut microbiota-derived butyrate to exert immune-protective effects in a histone deacetylase-dependent manner

BackgroundCommensal bacteria in the intestine release enzymes to degrade and ferment dietary components, producing beneficial metabolites. However, the regulatory effects of microbial-derived enzymes on the intestinal microbiota composition and the influence on host health remain elusive. Xylanase can degrade xylan into oligosaccharides, showing wide application in feed industry.ResultsTo validate the immune-protective effects of xylanase, Nile tilapia was used as the model and fed with xylanase. The results showed that dietary xylanase improved the survival rate of Nile tilapia when they were challenged with Aeromonas hydrophila. The transcriptome analysis showed significant enrichment of genes related to interleukin-17d (il-17d) signaling pathway in the xylanase treatment group. High-throughput sequencing revealed that dietary xylanase altered the composition of the intestinal microbiota and directly promoted the proliferation of Allobaculum stercoricanis which could produce butyrate in vitro. Consequently, dietary xylanase supplementation increased the butyrate level in fish gut. Further experiment verified that butyrate supplementation enhanced the expression of il-17d and regenerating islet-derived 3 gamma (reg3g) in the gut. The knockdown experiment of il-17d confirmed that il-17d is necessary for butyrate to protect Nile tilapia from pathogen resistance. Flow cytometry analysis indicated that butyrate increased the abundance of IL-17D+ intestinal epithelial cells in fish. Mechanistically, butyrate functions as an HDAC3 inhibitor, enhancing il-17d expression and playing a crucial role in pathogen resistance.ConclusionDietary xylanase significantly altered the composition of intestinal microbiota and increased the content of butyrate in the intestine. Butyrate activated the transcription of il-17d in intestinal epithelial cells by inhibiting histone deacetylase 3, thereby protecting the Nile tilapia from pathogen infection. This study elucidated how microbial-derived xylanase regulates host immune function, providing a theoretical basis for the development and application of functional enzymes.7BVRPuQYn3eQdwFeU5j8vPVideo

Immunity Induction in Catfish (Clarias gariepinus) with Feed Containing Red Ginger Extract after High Temperature Treatment

This study aims to evaluate the effect of feeding feed containing red ginger extract on the immunity and survival of catfish (Clarias gariepinus) fingerlings exposed to high temperature conditions. The research was carried out experimentally with a Completely Randomized Design (CRD) consisting of four treatment doses of red ginger extract (A: 0 ml/kg, B: 30 ml/kg, C: 40 ml/kg, D: 50 ml/kg) and three replications. Parameters observed included survival rate, white blood cell count, red blood cell count. The results showed that the addition of red ginger extract to feed was able to increase catfish survival by up to 90% at a dose of 50 ml/kg. A significant increase in the number of white blood cells and red blood cells in fish given red ginger extract indicates a better immune response. Red ginger extract has proven effective in helping fish adapt to thermal stress by increasing immunity and resistance to high water temperatures.

Immune Responses of Club Cells in Fish: A Review

The primary line of defense against pathogens from the environment is often fish epidermis tissue. Nevertheless, little is understood about the physiological mechanisms that underlie the non-specific and/or specific protection that these cells can offer. The exact nature of the relationship between the evolution of ostariophysan fish club cells and chemical warning signals is still unknown and controversial. Fish epidermis layer cells comprise mucus cells, lymphocytes, macrophage cells, cuboidal and squamous epithelial cells, and cells specific to certain fish species. Club cells, also called "alarm cells," are chemical alarms that sound in the event of a potentially hazardous scenario. These cells will burst in the presence of a predator, releasing pheromones that, if the skin is physically damaged, trigger an avoidance and terror reaction. In sturgeon larvae, mucus cells were visible in week 1, but club cells did not appear until week 4. Due to their later development during ontogenesis and after wounding, club cells may not have as much of a protective effect during wound healing as filament or mucus cells. Club cells are mostly found in the epidermis of the skin, and it is thought that when they work in tandem with mucus and goblet cells to fight infections, they serve as the body's first line of defense.

Soluble adenylyl cyclase is an acid-base sensor in rainbow trout red blood cells that regulates intracellular pH and haemoglobin-oxygen binding.

To identify the physiological role of the acid-base sensing enzyme, soluble adenylyl cyclase (sAC), in red blood cells (RBC) of the model teleost fish, rainbow trout. We used: (i) super-resolution microscopy to determine the subcellular location of sAC protein; (ii) live-cell imaging of RBC intracellular pH (pHi) with specific sAC inhibition (KH7 or LRE1) to determine its role in cellular acid-base regulation; (iii) spectrophotometric measurements of haemoglobin-oxygen (Hb-O2) binding in steady-state conditions; and (iv) during simulated arterial-venous transit, to determine the role of sAC in systemic O2 transport. Distinct pools of sAC protein were detected in the RBC cytoplasm, at the plasma membrane and within the nucleus. Inhibition of sAC decreased the setpoint for RBC pHi regulation by ~0.25 pH units compared to controls, and slowed the rates of RBC pHi recovery after an acid-base disturbance. RBC pHi recovery was entirely through the anion exchanger (AE) that was in part regulated by HCO3 --dependent sAC signaling. Inhibition of sAC decreased Hb-O2 affinity during a respiratory acidosis compared to controls and reduced the cooperativity of O2 binding. During invitro simulations of arterial-venous transit, sAC inhibition decreased the amount of O2 that is unloaded by ~11%. sAC represents a novel acid-base sensor in the RBCs of rainbow trout, where it participates in the modulation of RBC pHi and blood O2 transport though the regulation of AE activity. If substantiated in other species, these findings may have broad implications for our understanding of cardiovascular physiology in vertebrates.

Herbicide paraquat dichloride augments the expression of the c-abl-noxa-apoptotic pathway, instigating ovarian atresia in edible fish, Channa punctatus.

Ovarian cells of animals undergo atresia to regulate egg numbers and their release. However, unrestrained atretic activity decreases the chances of oocyte maturity and further process, ultimately resulting in a reduced population. Edible fishes are a critical source of nutrition and it is required to regulate their population. The application of herbicides at fish production sites is a threat to their population by altering female reproductive activity. In concern to this, here, this 60-day study investigates herbicide paraquat dichloride (PD)-induced atresia in ovarian cells of Channa punctatus (C. punctatus). Further, 96-h LC50 of PD for C. punctatus was calculated as 58.40mg/L. At the end of each exposure period, an increase in intracellular ROS in blood cells and activities of SOD and CAT in ovary tissue were observed in PD-treated fish with increasing concentration. The frequency of micronuclei in blood cells subsequently increased with the highest PD concentration. A significant (p < 0.05) increase in the relative expression of target genes was observed at the 30th and 60th day of the sampling periods in the fish treated with the highest concentration of PD. The histological study confirmed the occurrence of atretic ovary cells in PD-exposed fish. From the results it can be concluded that PD enhances the uncontrolled death of ovarian cells and has consequences of impairing the ovary health and reproductive potential of fish, hence reducing the fish population.

Anti-Müllerian hormone deficiency leads to two distinct ovarian phenotypes with different alterations of sex hormones in gynogenetic carp

Anti-Müllerian hormone (Amh) plays conserved roles in gonadal development and function in vertebrates. Previous studies have demonstrated that amh deficiency results in severe gonadal hypertrophy in teleost. Here, we report a novel phenotype of gonadal atrophy caused by amh knockout in fish. Firstly, it was found that the amh gene had three alleles and was expressed in the cytoplasm of follicle cells in gynogenetic amphiploid gibel carp (Carassius gibelio). Subsequently, we constructed a mutant line of amh and identified two distinct ovarian abnormalities in gibel carp. Most mutants exhibited hypertrophic ovaries with increased proliferation and decreased differentiation of early germ cells, which is consistent with previously reported findings. Intriguingly, a small number of mutants displayed atrophic ovaries with decreased proliferation and premature differentiation of germ cells, which represents a novel phenotype in fish species. Transcriptome analysis revealed that these two distinct ovaries of amh mutants showed different alterations in the sex hormone biosynthesis pathway, when compared with wild-type ovaries. Furthermore, diverse gonadal sex hormone levels were also detected in these two abnormal ovaries, which were consistent with the transcriptional expressions of corresponding genes in the sex hormone synthesis pathway. These findings identify a novel atrophic ovarian phenotype of amh mutants and confirm the amh function in controlling the balance between proliferation and differentiation of germ cells in fish.

Both rapid hyperplasia and hypertrophy promoted skeletal muscle growth in mandarin fish Siniperca chuatsi, compared to big-eye mandarin fish Siniperca kneri

As important cultured fishes in China, mandarin fish, Siniperca chuatsi and big-eye mandarin fish, Siniperca kneri share similar external morphology and genomes, but have significant differences in growth rates. In this study, we used 6 mph (month post hatching) mandarin fish and big-eye mandarin fish to compare differences in skeletal muscle growth over a one-month period, and use transcriptomes to explore the reasons for the growth differences. The results showed that the different value in number of muscle fibers (2.72 times), density (2.56 times), area (1.95 times) and diameter (2.73 times) of mandarin fish are significantly higher than that of big-eye mandarin fish (P<0.05). Hypertrophy (Contribution rate: 97.9%) is the dominant factor in big-eye mandarin fish, and both hypertrophy and hyperplasia (Contribution rate: 2.1%) were significantly different from mandarin fish (P<0.05). Transcriptome results showed that differential genes were enriched in proliferation of satellite cells, protein synthesis and degradation. Further analysis of the transcriptome results showed that the number of Pax7-positive cells per unit area of mandarin fish was 2.3 fold higher that of big-eye mandarin fish (P<0.05). Myod, pax3, pax7, and myf5 mRNA expression of mandarin fish were significantly higher than those of big-eye mandarin fish(P<0.05). In muscle fiber fusion, mef2, myog, and myomaker mRNA expression of mandarin fish were significantly higher than that of big-eye mandarin fish (P < 0.05). In protein deposits, the results showed that the muscle fiber area of mandarin fish was 2.85 times larger than that of big-eye mandarin fish by comparing the muscle fiber area with the same number of nuclei. Polr2a, mtorc1, 4e-bp1, s6, e1, murf, foxo3, atg5, beclin, and atg14 mRNA expression of mandarin fish were significantly higher than that of big-eye mandarin fish (P < 0.05), while eef2, eif2α, gcn2, eif2b, capn and cast of mandarin fish was significantly lower than that of big-eye mandarin fish (P < 0.05). This work verified that the skeletal muscle growth rate of mandarin fish is faster than that of big-eye mandarin fish. The greater number and higher activity of satellite cells of mandarin fish increased hyperplasia efficiency, and the faster fusion rate and protein precipitation rate increased hypertrophy efficiency. Both hyperplasia and hypertrophy contribute to faster skeletal muscle growth of mandarin fish than that of big-eye mandarin fish.

Changes in gill neuroepithelial cells and morphology of threespine stickleback (Gasterosteus aculeatus) to hypoxia and simulated ocean acidification.

Coastal marine environments are characterized by daily, seasonal and long-term changes in both O2 and CO2, driven by local biotic and abiotic factors. The neuroepithelial cells (NECs) of fish are thought to be the putative chemoreceptors for sensing oxygen and CO2, and, thus, NECs play a key role in detecting these environmental changes. However, the role of NECs as chemosensors in marine fish remains largely understudied. In this study, the NECs of marine threespine sticklebacks (Gasterosteus aculeatus) were characterized using immunohistochemistry. We then determined if there were changes in NEC size and density, and in gill morphology in response to either mild (10kPa) or moderate (6.8kPa) hypoxia and two levels of elevated CO2 (1,500 and 3,000 µatm). We found that the NECs of stickleback contained synaptic vesicles and were innervated, and were 50-300% larger and 2 to 4 times more abundant than in other similar sized freshwater fishes. NEC size and density were largely unaffected by exposure to hypoxia, but there was a 50% decrease in interlamellar cell mass (ILCM) in response to mild and moderate hypoxia. NECs increased in size, but not abundance in response to elevated CO2. Moreover, fish exposed to moderate or elevated CO2 had 53-78% larger ILCMs compared to control fish. Our results demonstrated that adult marine sticklebacks have NECs that can respond to environmentally relevant pCO2 and likely hypoxia, which highlights the importance of NECs in marine fishes under the heterogeneity of environmental conditions in coastal areas.

A consistency comparison between next-generation sequencing and the FISH method for gene rearrangement detection in B-cell lymphomas

Objective: To compare the consistency of lymphoma multigene detection panels based on next-generation sequencing (NGS) with FISH detection of B-cell lymphoma gene rearrangement. Methods: From January 2019 to May 2023, fusion genes detected by lymphoma-related 413 genes that targeted capture sequencing of 489 B-cell lymphoma tissues embedded in paraffin were collected from Henan Cancer Hospital, and the results were compared with simultaneous FISH detection of four break/fusion genes: BCL2, BCL6, MYC, and CCND1. Consistency was defined as both methods yielding positive or negative results for the same sample. The relationship between fusion mutation abundance in NGS and the positivity rate of cells in FISH was also analyzed. Results: Kappa consistency analysis revealed high consistency between NGS and FISH in detecting the four B-cell lymphoma-related gene rearrangement (P<0.001 for all) ; however, the detection rates of positive individuals differed for the four genes. Compared with FISH, NGS demonstrated a higher detection rate for BCL2 rearrangement, a lower detection rate for BCL6 and MYC rearrangement, and a similar detection rate for CCND1 rearrangement. No correlation was found between fusion mutation abundance in NGS and the positivity rate of cells in FISH. Conclusions: NGS and FISH detection of B-cell lymphoma gene rearrangement demonstrate overall good consistency. NGS is superior to FISH in detecting BCL2 rearrangement, inferior in detecting MYC rearrangement, and comparable in detecting CCND1 rearrangement.

Genotoxicity and metabolic changes induced via ingestion of virgin and UV-aged polyethylene microplastics by the freshwater fish Perca fluviatilis

The present study aims to compare and assess the toxicity induced by aged (irradiated with ultraviolet radiation for 120 days) polyethylene microplastics (PE-MPs) in comparison to virgin (non-irradiated) ones, after feeding the freshwater fish Perca fluviatilis. To this end, MPs mediated genotoxicity was assessed by the investigation of micronucleus nuclear abnormalities frequency in fish blood, and the degree of DNA damage in the liver and muscle tissues, while metabolic alterations were also recorded in both tissues. Results showed that both virgin and aged PE-MPs induced signaling pathways leading to DNA damage and nuclear abnormalities, as well as metabolites changes in all tissues studied. Metabolic changes revealed that the metabolism of nucleic acids, energy, amino acids, and neurotransmitters was more disrupted in the liver and by aged PE-MPs compared to muscles. Fish fed with aged PE-MPs exhibited greater DNA damage, while blood cells of fish fed with virgin PE-MPs seemed to be more vulnerable to nuclear abnormalities in relation to those fed with aged PE-MPs. Moreover, aged PE-MPs induced more acute overall effects on the metabolic profiles of fish tissues, and initiated stronger stress responses, inflammation, and cellular damages in fish tissues in relation to virgin ones. Characterization of both virgin and aged MPs revealed that the latter exhibited lower crystallinity and melting point, more irregular shapes and higher moiety of oxygen and carbonyl groups, which could be attributed for their observed higher toxicity. The research outcomes provide significant insights for advancing toxicological investigations in this field.

Identification and functional analyses of CD4-1+ cells in grass carp (Ctenopharyngodon idella)

In mammals, CD4 is found to be expressed on T cells and innate immune cells, however, teleost cells bearing CD4 have not been well identified and characterized. In this study, we identified two different CD4-1+ cell subsets in grass carp (Ctenopharyngodon idella): CD4-1+ lymphocytes (Lym) and CD4-1+ myeloid cells (Mye), both of which had the highest proportions in the head kidney. The mRNA expression analysis showed that CD4-1, CD4-2, TCRβ, CD3γ/δ, and LCK1 are highly expressed in CD4-1+ Lym and also expressed in CD4-1+ Mye. Furthermore, we found that CD4-1+ Lym have a Lym morphology and highly express T-cell cytokines, suggesting that they are CD4+ T cells equivalent to mammalian Th cells. On the other hand, CD4-1+ Mye were found to have a morphology of macrophage and highly express macrophage marker gene MCSFR, indicating that they are macrophages. In addition, functional analysis revealed that CD4-1+ Mye possess phagocytic ability and great antigen-processing ability. Taken together, our study sheds further light on the composition and function of CD4+ cells in teleost fish.

Histology and transcriptomic analysis reveal effects of dietary fish oil and soybean oil on lipid absorption and immune inflammation in the foregut of adult female spotted scat (Scatophagus argus)

The foregut is the main site of lipid absorption in fish, but the effect of different lipid sources on the foregut lipid absorption and metabolism in adult female spotted scat is still unclear. In this experiment, adult female spotted scat (length = 19.48 ± 1.13 cm, weight = 242.83 ± 50.90 g) were fed two iso-nitrogenous (45 % crude protein) and iso-lipidic (12.5 % crude lipid) diets configured with either 8 % fish oil (FO) or 8 % soybean oil (SO) for 60 days. Histological observations, serum measurements and transcriptome sequencing were used to investigate the effects of the oil sources on the foregut. Histological observation showed that the foregut folds of the SO group were shorter and narrower than those of the FO group, and nuclear vacuolation and villous atrophy were observed in the intestinal cells of fish in the SO group, indicating an inflammatory reaction. Both serum triglyceride and total cholesterol levels were significantly higher in the FO group than in the SO group. Transcriptome analysis revealed 1355 differentially expressed genes between the FO and SO groups. Genes related to lipid metabolism (adipor2 and creb3l3) and cholesterol absorption and efflux (npc1l1 and sphk2) were up-regulated, whereas genes related to fatty acid synthesis and transport (fad1 and mfsd2ab) were down-regulated in the FO group. Circadian rhythm related genes ciart and nfil3, which are also involved in lipid metabolism, were up- and down-regulated, respectively in the FO group. The immune-related genes steap4, ddit4 and cldn6 which can inhibit inflammation, were up-regulated in the FO group, whereas pro-inflammation gene rnf225 was up-regulated in the SO group. In conclusion, compared with SO, FO may enhance the lipid absorption and reduce the inflammatory response in the foregut of the spotted scat. The results of this study provide a reference for the selection of dietary oil sources for adult female spotted scat.

Effects of saline and alkaline stresses on the survival, growth, and physiological responses in juvenile mandarin fish (Siniperca chuatsi)

Saline-alkaline water is widely distributed and abundant worldwide. It plays a critical role in the advancement of the aquaculture and the comprehensive utilization of saline-alkaline resources. The mandarin fish (Siniperca chuatsi), a freshwater species with high economic value, and has potential for cultivation in such environments. We evaluated its tolerance to acute salinity and alkalinity, and investigated the growth and physiological changes under chronic saline and alkaline stress. Initially, the effects of acute salinity and alkalinity were estimated on mandarin fish (4.33 ± 0.31 g), and found that the LC50 for 96 h of salinity and alkalinity was 13.91 ppt and 13.45 mmol/L, respectively. Subsequently, over a 60-day period, we measured the survival, growth performance, and physiological responses in mandarin fish (22.53 ± 0.60 g) reared in freshwater (FW, the control group), saline water (SW; fixed salinity of 5 ppt) and alkaline water (AW; carbonate alkalinity of 8 mmol/L). The results indicated that neither SW nor AW group adversely affected survival rates, and growth performance in SW group was significantly superior to the FW group. Additionally, compared to the FW group， the AW group had a lower hepatosomatic index, while the SW group exhibited significantly higher meat content. In terms of physiological responses, the AW group showed elevated V-H+-ATPase (VHA) activity, whereas the SW group had decreased prolactin (PRL) levels. Furthermore, the mitochondria-rich cells (MRCs) in mandarin fish underwent morphological adaptations to saline and alkaline stress, indicating their role in osmoregulation and acid-base balance. Our results suggested that the mandarin fish demonstrated good tolerance to both salinity and alkalinity, indicating suitability for breeding in low saline and alkaline water. This research contributes to a broader understanding of saline and alkaline tolerance in aquaculture species and provides theoretical support for the cultivation of mandarin fish in saline-alkaline water. Moreover, the results of this study suggested that saline-alkaline areas and agricultural irrigation water in the world could be successfully utilized for sustainable aquaculture.

Comparative evaluation of neonicotinoids and their metabolites-induced oxidative stress in carp primary leukocytes and CLC cells

Neonicotinoids (NEOs) have been designed to act selectively on insect nicotinic acetylcholine receptors (nAChRs). However, nAChRs are also expressed in vertebrate immune cells, so NEOs may interfere with the immune system in exposed non-target animals. The present study shows that NEOs: imidacloprid and thiacloprid, and their main metabolites: desnitro-imidacloprid and thiacloprid amide, at sub-micromolar concentrations ranging from 2.25 to 20 μM, affect the immune cells of fish. This was found both in primary cultures of leukocytes isolated from the carp head kidney and in the continuous adherent carp monocyte/macrophage cell line. Moreover, the results revealed that the studied pesticides and metabolites generate oxidative stress in carp immune cells and that this is one of the most important mechanisms of neonicotinoid immunotoxicity. Significant increases were observed in the formation of ROS and malondialdehyde (MDA). The antioxidant status alteration was linked with decrease in antioxidant enzyme activity: superoxide dismutase (SOD), catalase (CAT), and non-enzymatic antioxidant glutathione (GSH). Importantly, the metabolites: desnitro-imidacloprid and thiacloprid amide showed significantly higher cytotoxicity towards fish leukocytes than their parent compounds, imidacloprid and thiacloprid, which emphasizes the importance of including intermediate metabolites in toxicology studies.

Finite Element Modeling of Crystallization with Temperature Jump to Improve Cryopreservation of Fish Germ Cells

This article is devoted to the further development of a viable technology for low-temperature cryopreservation of reproductive cells of sturgeon fish using acoustic–mechanical fields and intelligent control of the freezing process. Before vitrification begins, the piezoactuator acts on a mixture of cryoprotectant and reproductive cells. This promotes intensive mixing of the cryoprotector and its diffusion through the cell membrane. When vitrification is carried out directly, a phase transition phenomenon is observed, accompanied by crystal formation. This article presents a new mathematical model describing this process as developed by the authors. The corresponding boundary conditions are formulated. Numerical experiments were carried out using the finite element method. It has been established that during vitrification without the use of a cryoprotectant, a sharp temperature jump is observed at the front of the crystalline formation boundary. The use of a cryoprotectant leads to a slowdown in the process of crystal formation, that is, to a weakening of the effect of one of the most important cryoprotective factors. The comparison with full-scale experiments showed qualitative agreement with the experimental results, which indicates the adequacy of the proposed model. The results obtained can be used in the future during the vitrification process and the evaluation of the quality of cryofreezing. The application of a new methodological approach to methods of long-term preservation at low temperatures of the genetic and reproductive material of hydrobionts using acoustic and mechanical effects and an intelligent control module opens up great opportunities for the creation of new cost-effective biotechnologies that make it possible to make the transition to a new type of aquatic farms, increase the stability of aquaculture, in general, to make environmental protection measures to save rare and endangered species more effective.

Preliminary Trial of Male to Female Sex Reversal by 17β-Estradiol in Combination with Trilostane in Spotted Scat (Scatophagus argus)

The spotted scat (Scatophagus argus) is an important ornamental species with sexually biased ornamental values that favor males. Therefore, it makes sense to breed mono-male fingerlings as ornamental fish. The spotted scat has an XX/XY sex determination system; therefore, the first step in producing genetically all-male offspring should be the induction of a fertile sex-reversed XY pseudo-female, which would then be mated with a normal XY male to produce a YY super-male. However, the XY pseudo-female produced by estradiol (E2) treatment failed due to ovarian malformations. Here, male to female sex reversal was induced in spotted scat through a combination of E2 and an androgen inhibitor (trilostane, TR). Spotted scat fingerlings of approximately 2.8–3.5 cm were fed a diet containing both E2 (300 µg/g) and TR (300 µg/g). The fish were treated for 90 days and then fed a normal diet until they reached one year of age, when they were sampled. Twenty-eight treated XY individuals were identified using sex-linked markers. According to their gonadal histological characteristics, these treated XY fish could be divided into three groups: males with testes (n = 21), intersex individuals with ovaries–testes (n = 3), and fully sex-reversed individuals with ovaries (n = 4). All treated XX fish (n = 8) developed into normal females. There were no obvious abnormalities in the ovaries of the XY sex-reversed fish compared to the treated XX fish. Serum 11-ketotestosterone (11-KT) levels were significantly lower in XY-reversed females and treated XX females than in XY males. Compared with XY male fish, the expression of female (42sp50, foxl2, figla, zar1, and zp2) and male (dmrt1, gsdf, amh and cyp11b2) biased genes was up- and down-regulated, respectively, in the gonads of XY-completely sex-reversed fish. Immunohistochemical results indicate that 42Sp50 was expressed in oocytes of XY-partially and completely sex-reversed fish, while strong Gsdf signals were mainly detectable in testicular somatic cells of XY-partially sex-reversed fish and XY male fish. XY females were successfully produced by the combined treatment of E2 and TR in the spotted scat. Their fertility will be tested in the future.

Features of SARS-CoV-2 Replication in Various Types of Reptilian and Fish Cell Cultures.

SARS-CoV-2 can enter the environment from the feces of COVID-19 patients and virus carriers through untreated sewage. The virus has shown the ability to adapt to a wide range of hosts, so the question of the possible involvement of aquafauna and animals of coastal ecosystems in maintaining its circulation remains open. the aim of this work was to study the tropism of SARS-CoV-2 for cells of freshwater fish and reptiles, including those associated with aquatic and coastal ecosystems, and the effect of ambient temperature on this process. In a continuous cell culture FHM (fathead minnow) and diploid fibroblasts CGIB (silver carp), SARS-CoV-2 replication was not maintained at either 25 °C or 29 °C. At 29 °C, the continuous cell culture TH-1 (eastern box turtle) showed high susceptibility to SARS-CoV-2, comparable to Vero E6 (development of virus-induced cytopathic effect (CPE) and an infectious titer of 7.5 ± 0.17 log10 TCID50/mL on day 3 after infection), and primary fibroblasts CNI (Nile crocodile embryo) showed moderate susceptibility (no CPE, infectious titer 4.52 ± 0.14 log10 TCID50/mL on day 5 after infection). At 25 °C, SARS-CoV-2 infection did not develop in TH-1 and CNI. our results show the ability of SARS-CoV-2 to effectively replicate without adaptation in the cells of certain reptile species when the ambient temperature rises.

Identification and analysis of senescence-related genes in caudal fin cells of triploid crucian carp

This research aims to identify the hub genes associated with the senescence of triploid caudal fin cells. Transcriptomic data are obtained from the high and low generation (P6, P60) of triploid crucian carp caudal fin cells by high-throughput sequencing technology. Initially, all differential genes between the high and low generations are screened, yielding 4140 significantly upregulated genes and 3724 significantly downregulated genes. Subsequently, an aging gene set containing 950 genes is downloaded from the CellAge database to extract the differentially expressed genes associated with caudal fin cell aging, totaling 29 genes. GO and KEGG enrichment analyses are performed on these 29 aging differential genes. The GO analysis shows enrichment mainly in cellular processes related to aging, such as regulation of cell division, chromatin organization, cell cycle regulation. KEGG analysis reveals that the 29 aging-related genes are primarily involved in cell cycle and cellular senescence pathways. A PPI network of aging-related genes is constructed using the STRING database and Cytoscape software. Top-ranked genes were identified by using Degree, MCC, MNC, and Closeness algorithms in the Cytohubba plugin in Cytoscape, resulting in hub genes EZH2, JUN, MYD88, RBL2, BMP4, CCND1, NFKB2, MMP9. Lastly, qRT-PCR validation of these eight hub genes further confirmed the involvement of four genes: EZH2, RBL2, BMP4, and CCND1. The hub gene screened in this study may become a potential biomarker of fish caudal fin cell senescence, which provides a valuable experimental basis for the senescence of fish caudal fin cells, especially the senescence of caudal fin cells in polyploid fish, and the reproduction and breeding improvement of polyploid fish. It also provides meaningful data for elucidating the molecular mechanism of polyploid formation in animals, as well as the formation of aging and tumour in human beings.

Thymic Nurse Cells in Fish

Under the transmission electron microscope, the thymic nurse cells (TNC) are evident in adult Pseudopocryptes lanceolatus (Bloch and Schneider), an Indian teleost gobiid fish. The thymocytes are the chief constituents of the the TNCs. The microscopic features of the TNC in this fish are described in this communication.

Phagocytic plasma cells in teleost fish provide insights into the idea that B cells evolved from ancient phagocytic cells

Phagocytic plasma cells in teleost fish provide insights into the idea that B cells evolved from ancient phagocytic cells