Largemouth Research Articles

Dietary sodium acetate and sodium butyrate improve high-carbohydrate diet utilization by regulating gut microbiota, liver lipid metabolism, oxidative stress, and inflammation in largemouth bass (Micropterus salmoides)

BackgroundAdequate level of carbohydrates in aquafeeds help to conserve protein and reduce cost. However, studies have indicated that high-carbohydrate (HC) diet disrupt the homeostasis of the gut–liver axis in largemouth bass, resulting in decreased intestinal acetate and butyrate level.MethodHerein, we had concepted a set of feeding experiment to assess the effects of dietary sodium acetate (SA) and sodium butyrate (SB) on liver health and the intestinal microbiota in largemouth bass fed an HC diet. The experimental design comprised 5 isonitrogenous and isolipidic diets, including LC (9% starch), HC (18% starch), HCSA (18% starch; 2 g/kg SA), HCSB (18% starch; 2 g/kg SB), and HCSASB (18% starch; 1 g/kg SA + 1 g/kg SB). Juvenile largemouth bass with an initial body weight of 7.00 ± 0.20 g were fed on these diets for 56 d.ResultsWe found that dietary SA and SB reduced hepatic triglyceride accumulation by activating autophagy (ATG101, LC3B and TFEB), promoting lipolysis (CPT1α, HSL and AMPKα), and inhibiting adipogenesis (FAS, ACCA, SCD1 and PPARγ). In addition, SA and SB decreased oxidative stress in the liver (CAT, GPX1α and SOD1) by activating the Keap1-Nrf2 pathway. Meanwhile, SA and SB alleviated HC-induced inflammation by downregulating the expression of pro-inflammatory factors (IL-1β, COX2 and Hepcidin1) through the NF-κB pathway. Importantly, SA and SB increased the abundance of bacteria that produced acetic acid and butyrate (Clostridium_sensu_stricto_1). Combined with the KEGG analysis, the results showed that SA and SB enriched carbohydrate metabolism and amino acid metabolism pathways, thereby improving the utilization of carbohydrates. Pearson correlation analysis indicated that growth performance was closely related to hepatic lipid deposition, autophagy, antioxidant capacity, inflammation, and intestinal microbial composition.ConclusionsIn conclusion, dietary SA and SB can reduce hepatic lipid deposition; and alleviate oxidative stress and inflammation in largemouth bass fed on HC diet. These beneficial effects may be due to the altered composition of the gut microbiota caused by SA and SB. The improvement effects of SB were stronger than those associated with SA.

Hepatic transcriptome profiling of largemouth bass (Micropterus salmoides Lacépède) injected with Flavobacterium covae or lipopolysaccharide.

Flavobacterium covae (columnaris) is the most detrimental bacterial disease affecting the largemouth bass (Micropterus salmoides Lacépède) aquaculture industry. In the current study, fish received an intraperitoneal injection of either 1× PBS (100 μL), LPS in PBS (100 μL, 10 μg/mL), or F. covae (100 μL, 2.85 × 1011 CFU/mL) to simulate immunological challenges. After 24 h post-injection, liver tissue from the control and treated groups were then collected for transcriptome analysis. Results of the Gene Ontology (GO) and KEGG pathway analyses for the F. covae and LPS-injected groups found differentially expressed genes (DEGs) enriched primarily in toll-like receptors (TLRs), cytokine-cytokine receptors, complement and coagulation cascades, and the PPAR signalling pathways. This suggests that the liver immune system is enhanced by these five combined pathways. Additionally, the DEGs TLR5, MYD88, and IL-1 were significantly upregulated in F. covae and LPS-injected fish compared to the controls, whereas IL-8 was downregulated. The upregulation of TLR5 was unexpected as F. covae lacks flagellin, the protein that binds to TLR5. Additionally, it is unknown whether the TLR5 is upregulated by LPS. Further research into the upregulation of TLR5 is warranted. These results provide insight into immune responses and associated pathways contributing to the immune system in the liver during columnaris infection and induced response to LPS in largemouth bass.

A nanobody-mediated drug system against largemouth bass virus delivered by bacterial nanocellulose in Micropterus salmoides

Diseases caused by pathogens severely hampered the development of aquaculture, especially largemouth bass virus (LMBV) has caused massive mortality and severe economic losses to the culture of largemouth bass (Micropterus salmoides). Considering the environmental hazards and human health, effective and environmentally friendly therapy strategy against LMBV is of vital importance and in pressing need. In the present study, a novel nanobody (NbE4) specific for LMBV was selected from a phage display nanobody library. Immunofluorescence and indirect ELISA showed that NbE4 could recognize LMBV virions and had strong binding capacity, but RT-qPCR evidenced that NBE4 did not render the virus uninfectious. Besides, antiviral drug ribavirin was used to construct a targeted drug system delivered by bacterial nanocellulose (BNC). RT-qPCR revealed that NbE4 could significantly enhance the antiviral activity of ribavirin in vitro and in vivo. The targeted drug delivery system (BNC-Ribavirin-NbE4, BRN) reduced the inflammatory response caused by LMBV infection and improved survival rate (BRN-L, 33.3 %; BRN-M, 46.7 %; BRN-H, 56.7 %)compared with control group (13.3 %), ribavirin group (RBV, 26.7 %) and BNC-ribavirin (BNC-R, 40.0 %), respectively. This research provided an effective antiviral strategy that improved the drug therapeutic effect and thus reduced the dosage.

Development and characterization of 40 InDel markers from the major histocompatibility complex genes of largemouth bass (Micropterus salmoides).

The genetic improvement in growth and food habit domestication of largemouth bass (Micropterus salmoides) have made breakthroughs in past decades, while the relevant work on disease resistance were rarely carried out. Major histocompatibility complex (MHC) genes, which are well known as their numbers and high polymorphisms, have been used as candidate genes to mine disease-resistant-related molecular markers in many species. In present study, we developed and characterized 40 polymorphic and biallelic InDel markers from the major histocompatibility complex genes of largemouth bass. The minor allele frequency, observed heterozygosity, expected heterozygosity and polymorphic information content of these markers ranged from 0.0556 to 0.5000, 0.1111 to 0.6389, 0.1064 to 0.5070, and 0.0994 to 0.3750, respectively. Three loci deviated significantly from Hardy-Weinberg equilibrium, while linkage disequilibrium existed at none of these loci. These InDel markers might provide references for the further correlation analysis and molecular assisted selection of disease resistance in largemouth bass.

Population demographics and angler exploitation of Suwannee Bass in two Georgia rivers

AbstractObjectiveSuwannee Bass Micropterus notius are a unique black bass that are endemic to riverine systems in Georgia and Florida, USA. Despite their “near threatened” status by the International Union for Conservation of Nature's Red List, Suwannee Bass are a popular sport fish that can be harvested throughout their range under a single harvest regulation with all black basses (genus Micropterus). However, a single harvest regulation may have disproportion effects on Suwannee Bass because of their rarity, limited distribution, and sex‐specific growth patterns.MethodsBoat electrofishing was used to collect Suwannee Bass and Largemouth Bass M. nigricans from both the Withlacoochee and Ochlockonee rivers to assess population demographics and angler exploitation. Common fisheries assessment metrics were calculated and annual exploitation rate was estimated from high reward (US$100) tags under simulated variability in tag loss and angler reporting rates.ResultWe observed different relative abundance and size structure among river systems. Electrofishing catch per unit effort of Suwannee Bass was higher in the Withlacoochee River (6.89 fish/h) than the Ochlockonee River (2.01 fish/h). Length‐frequency histograms suggested a wide breadth of lengths and presumed ages except for a lack of stock size Suwannee Bass in the Ochlockonee River. Eight percent of monetary reward tags were returned from anglers, yielding an overall exploitation estimate of 3.2%.ConclusionAnnual angler exploitation rates of Suwannee Bass and Largemouth Bass were low (<9.1%) in the Withlacoochee and Ochlockonee rivers of Georgia. Low exploitation coupled with a wide breadth of lengths and ages suggest these populations are currently not at risk of overexploitation.

Differing physiological performance of coexisting cool- and warmwater fish species under heatwaves in the Midwestern United States.

Heatwaves are becoming more frequent and intensified with climate change. Freshwater ecosystems are among the most threatened, within which, differing responses between cool- and warmwater species to heatwaves can lead to fundamental changes in communities. Physiological experiments can identify potential mechanisms underlying the impacts of such heatwaves on fish communities. In the current study, we quantified the oxygen consumption rate, aerobic scope and swimming performance of cool- and warmwater fish species following the simulation of short-term heatwaves currently occurring in streams in the Midwestern United States. The coolwater predator walleye (Sander vitreus) showed clear thermal disadvantages relative to the warmwater predator largemouth bass (Micropterus salmoides), based on a high metabolic cost during the heatwave, low metabolic activity when encountering prey, and reduced swimming performance following the heatwave. Largemouth bass also showed a thermal advantage relative to the warmwater prey fathead minnow (Pimephales promelas) related to swimming performance and energetic costs, highlighting differing thermal responses between predators and prey. This study demonstrates the importance of considering short-term extreme thermal events in the response of aquatic communities to climate stressors.

Effects of dietary D-mannose supplementation on growth performance, intestinal digestive capacity, gut microbiota, and ammonia tolerance of largemouth bass Micropterus salmoides

To investigate the effects of D-mannose on the growth performance, intestinal digestion capacity, intestinal microbiota composition, and ammonia tolerance of largemouth bass, experiments were conducted using largemouth bass fry with an average body weight of 7.41±0.05 g. Three experimental groups were designed, with 3 replicates per group and 30 fish per replicate. The fish were fed with feed containing different concentrations of D-mannose (0.00%, 1.00%, and 2.00% - Control, 1% mannose, 2% mannose) and were given full satiation twice a day for 56 days. The results indicated that the addition of 2% D-mannose in the feed enhanced the growth performance of the largemouth bass while reducing the feed conversion ratio. The addition of D-mannose in the feed increased the activity of digestive enzymes in the intestines (lipase, α-amylase, pepsin, alkaline phosphatase, Na+/K+ATPase, gamma-glutamyl transpeptidase, and creatine kinase). It also led to an increase in intestinal villi height and intestinal wall thickness; Adding D-mannose reduced the abundance of Staphylococcus and Achromobacter in the gut microbiota, while increasing the abundance of Aurantimicrobium. After being subjected to ammonia stress for 96 hours, the addition of D-mannose in the feed resulted in a decrease in mortality rate and neuronal nitric oxide synthase activity, while simultaneously enhancing the activity of liver argininosuccinate synthetase, argininosuccinate lyase, arginase, ornithine transcarboxylase, and inducible nitric oxide synthase. Adding D-mannose to the feed promotes the growth of largemouth bass, enhances their digestive capacity, and has a positive impact on their intestinal microbiota composition. Furthermore, it has been demonstrated that it can enhance the tolerance of largemouth bass to ammonia.

Improved traits of proximate composition, liver antioxidant capacity and feeding habits in diploid hybrids from female Micropterus salmoides × male Lepomis cyanellus

Distant hybridization has been widely used as an effective strategy for improving fish germplasm. The largemouth bass (Micropterus salmoides, LB) is a popular fish worldwide due to its excellent performance in aquaculture, edibility, and sport fishing. However, the carnivorous nature of LB increases the cost and difficulty of culture and poses a threat to wild fishery resources. At present, there is a lack of research on improving the germplasm of LB. In this study, hybrid offspring (LG) were produced by crossing female LB and male green sunfish (Lepomis cyanellus, GS). The biological characteristics of the LG, such as muscle proximate composition, liver antioxidant enzyme activity, and pharyngeal tooth structure, were analyzed to evaluate their potential for aquaculture. The proximate composition analysis revealed that LG has a higher crude protein content of 20.07% compared to LB (18.1%) and GS (18.7%) (P < 0.05), and a lower crude fat content of 0.57% than LB (1.17%) (P < 0.05). Furthermore, LG had a significantly higher essential amino acid content (7.20%) than LB (6.43%) (P < 0.05), but not significantly different from GS (6.82%) (P > 0.05). In terms of fatty acid content, LG had significantly lower levels of saturated fatty acids (0.083%) compared to LB (0.153%) and GS (0.169%) (P < 0.05). Additionally, the amount of arachidonic acid (C20:4n6, ARA) in LG (0.058%) was significantly higher than LB (0.019%) and GS (0.043%) (P < 0.05). Regarding antioxidant enzyme activities in the liver, LG exhibited significantly higher superoxide dismutase (SOD) activity than LB and GS (P < 0.05). Electron microscopic observation of the pharyngeal tooth showed that the tooth number of LG (160.67 ± 3.93) was significantly lower than that of LB (181.67 ± 4.97) (P < 0.05) and significantly higher than that of GS (108.00 ± 3.58) (P < 0.05), while the tooth diameter of LG (0.076 ± 0.023) was significantly higher than that of LB (0.057 ± 0.016) (P < 0.05) and significantly lower than that of GS (0.108 ± 0.044) (P < 0.05). In conclusion, this study found that LG exhibited improved traits in proximate composition and hepatic antioxidant enzyme activity. The pharyngeal tooth characteristics indicated that LG may have different food selection and processing capabilities. This is the first report on biological characteristics in offspring produced from a distant hybridization between LB and GS, providing new insights into the genetic improvement and germplasm innovation of LB.

Comparative transcriptomics and metabolomics reveal specific biochemical responses in cultured largemouth bass (Micropterus salmoides) juveniles with sexual dimorphism

Currently, it has been well established that growth, bodily form, physiological responses, and metabolic features of teleost fish may display sexual difference during intensive farming. Herein, the study aims to mainly examine the disparities of histomorphological and molecular responses in representative tissues liver and caudal vein blood from male and female cultured largemouth bass (Micropterus salmoides) juveniles by tracking a growth period of 135 days for reaching sexual maturity. The liver indices of a majority of the females, such as mass ratio of hepatic and visceral tissues (MRHV), hepatosomatic index (HSI), and hepatic lipid droplet abundance, were obviously greater than the males. Observational outcomes on size and distribution of accumulated lipid droplets in the liver of the females were more obvious and distinct from the males. Hepatic transcriptome sequencing results also demonstrated that glycerophospholipid metabolism-related/lipid transport-related genes (e.g. chkb, vtg3), PPAR signaling pathway, and aromatic amino acids metabolism (e.g. tyrosine, phenylalanine) were more active in the females; whereas, genes or pathways related to long-chain lipid transport (e.g. slc27a6), fatty acid biosynthesis and decomposition (e.g. ptgdsa), tryptophan metabolism, and immunological/inflammatory responses (e.g. mst1), as well as mTOR/PI3K-Akt/FoxO/AMPK signaling pathways in the males, were more evidently up-regulated than the females. Serum non-targeted metabolomics results exhibited that 5-hydroxyindoleacetic acid (growth factor derivative) and adipoyl-l-carnitine (lipid transporter) in the males were higher in quantity while L-kynurenine (tryptophan metabolism) and l-carnitine (lipid transporter) in the females higher than the males. The findings preliminarily showed that activity of PPAR signaling pathway and high expression of chkb gene might be crucial mechanisms in gained hepatic lipids in the females, while the actions of metabolites l-carnitine/adipoyl-l-carnitine might be more conducive to lipid transport in the liver of the males.

Rapid and facile detection of largemouth bass ranavirus with CRISPR/Cas13a

Largemouth bass ranavirus (LMBV) is an epidemic disease that seriously jeopardizes the culture of largemouth bass（Micropterus salmoides）, and it has a very high incidence in largemouth bass. Once an outbreak occurs, it may directly lead to the failure of the culture, resulting in substantial economic losses, but there is no effective vaccine or special effective drug yet. Consequently, it is important to establish an accurate, sensitive, convenient and specific detection approach for preventing LMBV infection. The recombinant enzyme-assisted amplification (RAA) technology was used in combination with clustered regularly interspaced short palindromic repeats (CRISPR), and associated protein 13a (CRISPR/Cas13a) to detect LMBV. We designed RAA primers and CRISPR RNA (crRNA) that targeted the conserved region in the LMBV main capsid protein (MCP) gene, amplified sample nucleic acids using the RAA technology, performed CRISPR/Cas13a fluorescence detection and evaluated the sensitivity and specificity of the established method with qPCR as a control method. This technique was able to determine the results by collecting fluorescence signals, visualizing fluorescence by UV excitation and combining with lateral flow strips (LFS). The sensitivity and specificity of the established method were consistent with the qPCR method. Besides, it was performed at a constant temperature of 37 °C and the sensitivity of the reaction system was 3.1 × 101 copies/μL, with no cross-reactivity with other common aquatic pathogens. Further, the positive detection rate of the proposed method in 32 clinical samples was consistent with that of qPCR. In conclusion, our established RAA-CRISPR/Cas13 method for detecting LMBV is sensitive, simple and specific, which is applicable in the rapid on-site detection and epidemiological monitoring of LMBV.

DL-Methionyl-DL-Methionine/DL-Methionine Supplementation Alleviated the Adverse Effects of Dietary Low Fishmeal Levels on Growth and Intestinal Health of Micropterus salmoides.

DL-methionyl-DL-methionine (AQUAVI® Met-Met) (Met-Met) (0.10%, 0.20%, 0.30%, and 0.40%) or DL-methionine (DL-Met) (0.10%, 0.20%, 0.30%, and 0.40%) were added to a low-fishmeal diet in an attempt to reduce fishmeal in the diet of Micropterus salmoides (M. salmoides). The fish were randomly allocated into ten experimental groups (n = 100), each with 4 replicates of 25 fish (16.39 ± 0.01 g) each. Compared to 25% FM, 0.40% of DL-Met and 0.10% of Met-Met promoted growth, and 0.10% of Met-Met decreased FCR. Compared to 25% FM, the supplementation of Met-Met or DL-Met improved the intestinal antioxidant capacity by upregulating the NF-E2-related factor 2-mediated antioxidant factors and enzyme activities and nuclear factor kappa-B-mediated anti-inflammatory factors while downregulating the pro-inflammatory factors, thereby exerting anti-inflammatory effects. Moreover, 0.10% of the Met-Met diet affected the Firmicutes-to-Bacteroidota ratio, increased the levels of Proteobacteria, changed the composition of intestinal flora (Roseburia, Lachnospiraceae_NK4A136_group, and unclassified_Oscillospiraceae), and enhanced intestinal dominant bacteria (Caldicoprobacter, Pseudogracilibacillus, and Parasutterella), leading to improved gut health. In summary, the supplementation of DL-Met or Met-Met alleviated the adverse effect of fishmeal reduction (from 40 to 25%) on the growth performance and intestinal health of M. salmoides.

IFN-γ enhances protective efficacy against Nocardia seriolae infection in largemouth bass (Micropterus salmoides).

Nocardia seriolae adversely impacts a diverse range of fish species, exhibiting significant pathogenic characteristics that substantially impede the progress of aquaculture. N. seriolae infects in fish has a long incubation period, and clinical symptoms are not obvious in the early stages. There is presently no viable and eco-friendly approach to combat the spread of the disease. According to reports, N. seriolae primarily targets macrophages in tissues after infecting fish and can proliferate massively, leading to the death of fish. Interferon-gamma (IFN-γ) is a crucial molecule that regulates macrophage activation, but little is known about its role in the N. seriolae prevention. IFN-γ was first defined as largemouth bass (Micropterus salmoides, MsIFN-γ), which has a highly conserved IFN-γ characteristic sequence through homology analysis. The recombinant proteins (rMsIFN-γ) were obtained in Escherichia coli (E. coli) strain BL21 (DE3). The inflammatory response-inducing ability of rMsIFN-γ was assessed in vitro using monocytes/macrophages. Meanwhile, the protective effect of MsIFN-γ in vivo was evaluated by N. seriolae infection largemouth bass model. In the inflammatory response of the monocytes/macrophages activated by rMsIFN-γ, various cytokines were significantly increased. Interestingly, interleukin 1β (IL-1β) and tumor necrosis factor alpha (TNF-a) increased by 183- and 12-fold, respectively, after rMsIFN-γ stimulation. rMsIFN-γ improved survival by 42.1% compared with the control. The bacterial load in the liver, spleen and head kidney significantly decreased. rMsIFN-γ was also shown to better induce increased expression of IL-1β, TNF-α, hepcidin-1(Hep-1), major histocompatibility complex I (MHCI), and MHC II in head kidney, spleen and liver. The histopathological examination demonstrated the transformation of granuloma status from an early necrotic foci to fibrosis in the infection period. Unexpectedly, the development of granulomas was successfully slowed in the rMsIFN-γ group. This work paves the way for further research into IFN-γ of largemouth bass and identifies a potential therapeutic target for the prevention of N. seriolae.

Psychological commitment of freshwater anglers and its relation to their preferences for stocking and other management actions

Understanding stakeholder diversity can help natural resource managers tailor activities to achieve greater stakeholder satisfaction. Stakeholder diversity can be described by the concept of recreational specialization. Centrality-to-lifestyle, one subdimension of specialization that measures the psychological importance of a recreational activity to an angler, has been shown to explain many human dimensions and behaviors of recreational fishers and to correlate with preferences for management actions. We surveyed 9911 anglers in Florida, USA to examine how centrality-to-lifestyle relates to preferences for stocking and other management tools. We found that most anglers support stocking and that anglers of greater centrality-to-lifestyle had more positive views toward stocking than less central anglers. Participants, regardless of level of centrality-to-lifestyle, generally preferred stocking of one species, Florida largemouth bass ( Micropterus salmoides), and they preferred habitat management above stocking, with no relation to centrality. The results suggest stocking can improve the satisfaction of anglers of all commitment levels, but habitat improvement could do so even more. Managers might consider prioritizing habitat management over stocking in systems where natural recruitment is ample to increase overall angler satisfaction, and where the anglers will, on average, support such actions independent of the degree of centrality.

Effects of partial replacement of fishmeal with Spirulina platensis powder and addition of Spirulina platensis polysaccharide on growth, nutrition, antioxidant capacity and gut microbiota of Micropterus salmoides

Fish is currently considered a primary protein in the human diet, and the high demand for fish will stimulate the growth of aquaculture. However, a lack of high-quality fish feed (fishmeal) has led to a decline in the quality and yield of aquatic products. Therefore, an alternative high-quality protein source for fish meal is required to sustain the industry. This study assessed the effects of incorporating Spirulina platensis (SP) dried powder as a partial replacement for fishmeal and supplementing with S. platensis purified polysaccharides (PSP) on the growth, nutritional value, and health status of the largemouth bass, Micropterus salmoides, to address the challenges of fishmeal scarcity and promote sustainable development in fish aquaculture. Juveniles were fed four isonitrogenous and isolipidic diets containing SP at 0% (SP0), 3% (SP3) and 6% (SP6) and PSP at 0.3% for 56 days. The results showed that both the SP3 and SP6 groups displayed significantly (P < 0.05) improved growth performance and lower hepatosomatic indices than the control diet (SP0). Whole-body crude protein and protein efficiency significantly increased in both the SP3 and SP6 groups compared with SP0 (P < 0.05). Conversely, crude lipid content notably (P < 0.05) decreased in SP3 (5.96%) and SP6 (5.41%) in contrast to SP0 (6.59%). The muscle amino acid composition of the PSP, SP3, and SP6 groups also improved significantly (P < 0.05) compared to that of the SP0. Liver SOD activity was significantly increased from 1.11 U/mg in SP0 to 1.22–1.24 U/mg in SP3 and SP6 (P < 0.05), while MDA levels declined accordingly. Serum triglyceride levels were significantly lower in SP3 than in SP0 (P < 0.05) but remained stable in the SP6 and PSP groups. Low-density lipoprotein cholesterol decreased significantly in the SP3 and SP6 groups, but no significant effect was observed in the PSP group. Alanine aminotransferase activity was significantly (P < 0.05) lower in the PSP, SP3, and SP6 groups than in SP0. Aspartate aminotransferase activity in PSP and SP3 was significantly lower than in SP0 (P < 0.05), but showed no significant difference with SP6 (P > 0.05). Intestinal trypsin and amylase activities significantly improved in the PSP, SP3, and SP6 groups (P < 0.05), and lipase activities in PSP and SP3 increased significantly compared to those in SP0 (P < 0.05). The intestinal microbiome displayed significantly higher Shannon index scores for SP3 (0.83) and SP6 (0.93) than for SP0 (0.61). The relative abundances of the phylum Fusobacteria and genus Cetobacterium were significantly increased, whereas Proteobacteria and Plesiomonas were significantly decreased in the PSP and SP3 groups compared with that in SP0 (P < 0.05). The present study indicated that the replacement of fishmeal with 3–6% SP and 0.3% PSP addition could improve the growth performance, protein efficiency, muscle nutritional value, liver health, antioxidant capacity, intestinal digestive enzyme activity, and intestinal microbiota diversity of the largemouth bass. Therefore, the partial replacement of SP or the addition of PSP has great potential in promoting healthy aquaculture of largemouth bass.

Morphogenesis of largemouth bass ranavirus (LMBRaV) in the epithelioma papulosum cyprinid cell line

Largemouth bass ranavirus (LMBRaV) belongs to the Ranavirus genus of the Iridoviridae family. It is a highly pathogenic virus that causes mass mortality in largemouth bass. In recent years, outbreaks of LMBRaV have been found in various provinces throughout China. Previous research mainly focused on virus isolation, identification, and detection, while the morphological change of the virus was still unknown. In this study, the ultrastructural morphogenesis of LMBRaV in epithelioma papulosum cyprinid (EPC) cells was observed and studied by using transmission electron microscopy. EPC cells were infected with LMBRaV (MOI=0.1) and then examined at 2 h, 4 h, 6 h, 12 h, 24 h, 48 h, 72 h and 96 h post infection. LMBRaV entered cells through endocytosis or direct penetration of cell membrane. After entering, the virus was observed in vesicles or lysosomes. After capsid uncoating, the virus genomes passed through the nuclear membrane and entered the cell nucleus. Virus genomes completed replication in the nucleus then transferred into the cytoplasm. In the cytoplasm, the progeny virus was assembled in the viromatrix and then aggregated in pseudocrystalline array. Finally, mature virus particles released through budding release from the cell membrane. Mature virus particles had a hexagonal shape and a diameter of approximately 150 nm. This study revealed the process of morphogenesis of LMBRaV in EPC cell line, providing essential information for further research on pathogenic mechanisms and immunological prevention of LMBRaV.

Isolation, Identification, and Pathogenicity Characterization of a Largemouth Bass Ranavirus from Fujian, China

This study aimed to isolate and characterize the pathogenic agent of a suspected viral disease of largemouth bass (Micropeterus salmoides), which broke out in an aquatic farm in Fujian, China. The diseased bass had ulcer skin and muscles, bleeding fin, lesion liver, and enlarged spleen. Histopathological analysis showed multifocal necrosis on the gill, hepatopancreas spleen, and kidney. In the epithelioma papulosum cyprinid (EPC) cell line, stable cytopathic effects (CPE) were observed. Electron microscopy analysis showed icosahedral virus particles in the cytoplasm with a 150-160 nm diameter. PCR amplification revealed that the isolated virus belongs to largemouth bass ranavirus (LMBV) and was designated as LMBV FJ_22109. Further phylogenetic analysis determined that LMBV FJ_22109 is a variant of European and American strains. The intraperitoneal injection of LMBV FJ_22109 resulted in a regress infection, demonstrating its ability to induce pathogenicity in largemouth bass. The cumulative mortality rate was 100% at a culture temperature of 29°C, and 47% at a culture temperature of 17°C. Thus, it confirmed that LMBV FJ_22109 is indeed the causative agent of the suspected viral disease, and its virulence could be influenced by temperature, which will be beneficial for preventing and managing the disease.

Counteracting effects of “hook avoidance” and “hook habituation” on angler catch rates in a catch‐and‐release fishery

AbstractCatch‐and‐release (C&amp;R) angling is often used to maintain high catch rates but fish vulnerability to capture may decrease following hooking, thereby decreasing angler catch per unit effort (CPUE) (hyperdepletion). To determine if fish post‐capture response affected recapture probability and population‐level CPUE, individual capture histories of Largemouth Bass in two lakes were compared before and after doubling angling effort in a Before‐After Control‐Impact (BACI) analysis. Previous capture and day‐of‐season both affected recapture probability. Counteracting effects of previous capture and reduced late‐season catch rates caused no hyperdepletion of angler CPUE. Our results highlight the complexity of fish behavioral responses to angling and suggest that hyperdepletion of angling catch rates may not be an issue in C&amp;R fisheries.

The antiviral role of largemouth bass STING against iridovirus infection

Stimulator of interferon gene (STING) plays a crucial role in the innate immune response against viral and bacterial pathogens. However, its function in largemouth bass iridovirus (LMBV) infection remains uncertain. Here, a STING homolog (MsSTING) from largemouth bass (Micropterus salmoides) was cloned and characterized. MsSTING encoded a 407-amino-acid polypeptide, which shared 84.08% and 41.45% identity with golden perch (Perca flavescens) and human (Homo sapiens) homologs, respectively. MsSTING contained four transmembrane domains and a conserved C-terminal domain. The mRNA level of MsSTING was significantly increased in response to LMBV infection in vitro. Subcellular localization observation indicated that MsSTING encoded a cytoplasmic protein, which co-localized predominantly with endoplasmic reticulum (ER) and partially with mitochondria. Moreover, its accurate localization was dependent on the N-terminal transmembrane motif (TM) domains. MsSTING was able to activate interferon (IFN) response, evidenced by the activation of IFN1, IFN3 and ISRE promoters by its overexpression in vitro. Mutant analysis showed that both the N-terminal and C-terminal domain of MsSTING were essential for its activation on IFN response. In addition, overexpression of MsSTING inhibited the transcription and protein levels of viral core genes, indicating that MsSTING exerted antiviral action against LMBV. Consistently, the inhibitory effects were significantly attenuated when the N-terminal or C-terminal domains of MsSTING was deleted. Furthermore, MsSTING overexpression upregulated the transcriptions of interferon-related genes and pro-inflammatory factors, including TANK-binding kinase 1(TBK1), interferon regulatory factor 3 (IRF3), interferon regulatory factor 7 (IRF7), interferon stimulated exonuclease gene 20 (ISG20), interferon-induced transmembrane protein 1(IFITM1), interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and interleukin 6 (IL-6). Together, MsSTING exerted antiviral action upon LMBV infection through positive regulation the innate immune response.

Effects of dietary binder sources on growth, digestive enzymes activity, and gut barrier function and microbial composition of larval largemouth bass (Micropterus salmoides)

A 4-week feeding trail was conducted to explore the impacts of dietary binder sources on growth performance, digestive enzymes activities, intestinal microbiota, and expression of genes involved in tight junctions and nutrient sensing pathway of larval largemouth bass with initial body weight of 8.43 ± 0.01 mg (13 day after hatching). Five isolipidic micro-diets were formulated with different binder sources including sodium alginate (SA), carrageenan (CG), hydroxypropyl methyl cellulose (HPMC), gelatin (GT), and starch (SC). The results revealed that the highest growth performance was observed in the CG group, and meanwhile, the promotion effects of SA and GT was statistically higher than that of HPMC and SC groups. SA, CG and GT had positive effects on digestive enzymes activity compared to other groups. Gene expression results showed that SA, CG and GT groups up-regulated the expression of tight junction genes, zo-1, claudin-4 and claudin-7, and activated the genes involved in the TOR signaling pathway, including tor, akt1, s6kβ1 and s6. Among all the groups, the CG had the most significant activation effect on TOR signaling pathway. Bacterial 16S rRNA V3–4 region analysis showed that the addition of different dietary binders influenced the diversity of intestinal microbial communities. The inclusion of SA and CG groups reduced the relative abundance of Proteobacteria and Actinobacteria at the phylum level, while at the genus level, the relative abundance of Rhodococcus and Achromobacter increased and the abundance of Plesiomonas was inhibited. In summary, dietary binder sources affected the growth, digestive function, gut barrier function and microbial composition, and nutrient sensing, and SA and CG could be suggested as the optimal dietary binders for formulation of micro-diets for largemouth bass larvae.

The difference of the composition and digestive enzymes of gut microbiome in herbivorous blunt snout bream (Megalobrama amblycephala) and carnivorous largemouth bass (Micropterus salmoides)

Blunt snout bream (Megalobrama amblycephala) and largemouth bass (Micropterus salmoides) are a herbivorous and a carnivorous freshwater fish species respectively. These fish possess distinct intestinal microorganisms and digestive enzymes that play a vital role in nutrient digestion. As a result, herbivorous and carnivorous fish exhibit significant differences in their digestive strategies. In this study, we investigated the compositions and functions of the gut flora of herbivorous M. amblycephala and carnivorous M. salmoides by 16S rRNA sequencing. PCoA analysis revealed that the gut flora of M. amblycephala and M. salmoides formed two distinct clusters. OTU analysis found that M. amblycephala and M. salmoides shared a small number, but high abundance, of core microorganisms, and that a large number of microorganisms differed at the phylum and genus levels between the two species. The cellulose-degrading bacteria Longivirga, Flavobacterium, Clostridium_sensu_stricto_12, and Bacillus were dominant in M. amblycephala, while protein-degrading bacteria Clostridium_sensu_stricto_1, Lactococcus, Streptococcus and Proteiniclasticum were dominant in M. salmoides. PICRUSt analysis showed significant differences in carbohydrate and protein metabolism functions as well. In addition, cellulase and amylase activities were significantly higher in M. amblycephala than in M. salmoides, while trypsin activity was significantly higher in M. salmoides than in M. amblycephala. These results indicated that dietary patterns influenced the composition and function of the bacterial community and the activity of digestive enzymes in the host’s gut.