The impacts of thermal heterogeneity across microhabitats on post-settlement selection of intertidal mussels.

Rapid genetic selection is critical for allowing natural populations to adapt to different thermal environments such as those that occur across intertidal microhabitats with high degrees of thermal heterogeneity. To address the question of how thermal regimes influence selection and adaptation in the intertidal black mussel Mytilisepta virgata, we continuously recorded environmental temperatures in both tidal pools and emergent rock microhabitats and then assessed genetic differentiation, gene expression patterns, RNA editing level, and cardiac performance. Our results showed that the subpopulations in the tidal pool and on emergent rocks had different genetic structures and exhibited different physiological and molecular responses to high-temperature stress. These results indicate that environmental heterogeneity across microhabitats is important for driving genetic differentiation and shed light on the importance of post-settlement selection for adaptively modifying the genetic composition and thermal responses of these intertidal mussels.

Surviving hot summer: Roles of phenotypic plasticity of intertidal mobile species considering microhabitat environmental heterogeneity.

For species inhabiting warming and variable thermal environment, coordinated changes in heat tolerance to temperature fluctuations, which largely depend on phenotypic plasticity, are pivotal in buffering high temperatures. Determining the roles of phenotypic plasticity in wild populations and common garden experiments help us understand how organisms survive hot summer and the warming world. We thus monitored the operative temperature of the intertidal limpets Cellana toreuma in both emergent rock and tidal pool microhabitats from June to October 2021, determined the variations of upper thermal limits of short-term acclimated and long-term acclimated limpets from different microhabitats (emergent rock and tidal pool), and further calculated the relationship between the upper thermal limits and acclimation capacity. Our results indicated that living on the emergent rock, limpets encountered more extreme events in summer. For the short-term acclimated samples, limpets on the emergent rock exhibited obvious variations of sublethal thermal limit (i.e., Arrhenius Break Point of cardiac performance, ABT) during summer months, however, this variation of ABT was absent in the limpets in the tidal pool. After the laboratory long-term acclimation, the ABTs and FLTs (Flat Line Temperature of cardiac performance, as an indicator of lethal temperature) of limpets both on the rock and in the tidal pool increased significantly in October, implying the potential existence of selection during the hot summer. Our results further showed that environmental temperature was an important driver of phenotypic plasticity. This study highlighted the changes in the thermal tolerance of intertidal limpets during summer in different microhabitats.

Microhabitat-specific diurnal metabolomic responses of the intertidal limpet Cellana toreuma to winter low temperature.

High-throughput determination of circadian rhythms in metabolic response and their divergent patterns in various microhabitats are crucial for understanding how organisms respond to environmental stresses. A mid-intertidal limpet Cellana toreuma was collected at various time points across both daytime and nighttime in winter during low tide for investigating the diurnal metabolomic responses to cold stress and elucidating the divergent metabolic responses to temperature variations across microhabitats. Temperatures of emergent rock microhabitats were lower than the tidal pool and even aggravated at night. A series of metabolomic responses exhibited coordinated diurnal changes in winter. Metabolic responses which were associated with cellular stress responses and energy metabolism of emergent rock microhabitat individuals were highly induced compared to the tidal pool ones. This study shed light on the diurnal patterns of metabolomic responses of intertidal molluscs in the field and emphasized the variations in metabolic responses between microhabitats.

Correction to: The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

In the end of the Discussion section, following information should be included.

The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

Hypoxia stress may affect the fish intestine and thereby threaten the growth and survival of the fish. Teprenone is a clinically effective agent in protecting gastrointestinal mucosa. This study aims to assess the effect of teprenone in the intestine of Chinese sea bass Lateolabrax maculatus under intermittent hypoxic stress. L. maculatus juveniles were either raised under intermittent hypoxic condition or normal condition (NC). Part of the hypoxic-intervened fish were treated with teprenone at different concentrations (HTs), and the rest were regarded as hypoxic control (HC). Histological analysis was performed on the epithelial tissue of the fish intestine. High-throughput sequencing technology was used to analyze the diversity and composition of the microbial community in L. maculatus intestine. Reduced villi length and goblet cell, exfoliated enterocyte, and improper arrangement of villi were observed in HC compared with NC and HTs. Proteobacteria, Firmicutes, and Bacteroidetes represented the most abundant phyla in each sample. Significantly higher microbial diversity was detected in HC compared with NC (P < 0.05). At the phylum level, HC presented significantly decreased relative abundance of Proteobacteria, and significantly increased relative abundance of Bacteroidetes, Chloroflex, and Cyanobacteria compared with NC (P < 0.05). At the class level, HC showed significantly reduced relative abundance of Alphaproteobacteria and Bacilli, and significantly increased relative abundance of Clostridia, Gammaproteobacteria, and Bacteroides (P < 0.05). Teprenone protects the intestine from epithelial damages and maintains the microbial harmony in L. maculatus under intermittent hypoxic stress.

Transcriptomic analysis of juvenile Chinese sea bass (Lateolabrax maculatus) anesthetized by MS-222 (tricaine methanesulfonate) and eugenol.

MS-222 (tricaine methanesulfonate) and eugenol are the two frequently used fish anesthetics. This study intends to analyze the regulation of these anesthetics in Chinese sea bass, Lateolabrax maculatus, through transcriptomic analysis. L. maculatus were exposed to MS-222 or eugenol, and those without any treatments were regarded as controls. Gills and livers were extracted for transcriptomic analysis after recovery in fresh water for 6 h. Identified genes were assigned to NR, COG, SWISS, GO, and KEGG database for predicting gene functions. A FDR ≤ 0.05 and |log2(FC)| ≥ 1 were applied to determined differentially expressed gene (DEG). A total of 45,626 unigenes were annotated using at least one database. The eugenol-treated liver group presented less DEGs compared with that treated by MS-222. Both the MS-222- and eugenol-treated liver groups presented notable DEGs that participated in human disease and metabolism pathways. The eugenol group showed more pathways related to detoxification activity and xenobiotics biodegradation, and those from the MS-222 group were related to organismal system such as reproduction. By comparing gill and liver samples using the same drug, the enriched pathways were generally consistent among the three comparisons. In conclusion, eugenol and MS-222 could change the pathways related to metabolism and immunity in L. maculatus. MS-222 may trigger more damages on the fish liver and reproduction.

Protection of teprenone against hypoxia and reoxygenation stress in stomach and intestine of Lateolabrax maculatus.

Teprenone (geranylgeranylacetone) is one kind of safe and effective agent in gastrointestinal mucosa, which have been widely used in human and veterinary, but rarely used in aquaculture animals. In this study, Lateolabrax maculatus, an important economic fish species in southern China, was taken as the object of study to investigate the protective effect of teprenone on intestinal stress. The present study was designed to investigate the potential mechanism underlying the protection offered by teprenone to protect the gastrointestinal tract against hypoxia and reoxygenation injury of L. maculatus. (a) For oxidative stress parameters, SOD, CAT, and T-AOC in control group were higher than those in teprenone group. MDA content was significantly higher than that in teprenone group at N and 12h time points in intestine (P < 0.05), and at 12, 24, and 48 h time points in stomach. (b) For immune-associated proteins, LZM activity in the control group was lower than that in the teprenone group, and the difference between the two groups in stomach and intestine was significant at 12.48 h and 6.48 h time points, respectively (P < 0.05). Compared with time point N, the content of HSP70 in the control group increased at 0 h in intestine. At 0-48 h, intestine HSP70 content in the control group showed a gradually decreasing trend, which was higher than that in the teprenone group. (c) For apoptosis-related factors, the activity of Cyt-C, caspase9, and caspase3 increased first and then decreased in both groups. The content of Cyt-C in the control group was significantly higher than that in the teprenone group at N-3.6 h, and 3.48 h time points in stomach and intestine, respectively (P<0.05). The activity of caspase9 and caspase3 was higher than that in the teprenone group at N-48 h. Results indicated that acute hypoxia and reoxygenation cause the expression levels of oxidative stress and apoptosis-related factors in the stomach and intestine increased first and then decreased within 0-48 h. Acute hypoxia and reoxygenation also that causes the level of nonspecific immunity decreased first and then increased. A total of 400-mg/kg treatment of teprenone can protect stomach and intestinal tissues to a certain extent. It can effectively protect oxidative stress and apoptosis within 0-48 h after acute hypoxia and reoxygenation and enhance non-specific immunity.

Dietary effects of succinic acid on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei.

Organic acids acts as an growth promoter and antimicrobial agent in aquaculture. The present study investigated the effects of a natural organic acid - succinic acid (SA) on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei. The shrimps were firstly fed with diets containing different levels of SA: 0% (Control), 0.25% (SA1), 0.50% (SA2), and 1.0% (SA3) (w/w) for 56 days, followed by an acute ammonia stress for 48 h. The results indicated that dietary of SA improved the growth of shrimp, and increased the survival rate of shrimp after ammonia stress for 48 h. The amylase, lipase and pepsin activity increased in hepatopancreas in three SA group, while trypsin activity was only increased in the SA1 and SA2 groups. At 56 d, T-NOS activity, proPO and HSP70 gene expression level increased in the three SA group, PO activity increased in the SA1 and SA2 groups, T-AOC content and Toll gene expression level increased in the SA2 and SA3 groups, Trx and SOD gene expression level increased in the SA2 group, while Imd, GS and GDH gene expression level was no changes. After exposure to ammonia stress for 48 h, immune biochemical parameters (T-AOC and PO) and genes (proPO, HSP70, Trx and GDH) expression level increased in the three SA group, T-NOS activity, Toll, Imd and GS gene expression level increased in the SA2 and SA3 groups, while SOD gene expression level increased in the SA1 and SA2 groups. These results indicated that SA improved growth, enhanced digestive and immune capacities of L. vannamei against ammonia stress, and may be a potential feed additive for shrimp. The optimal dietary supplementation dosage is 0.50% (w/w) in diet.

A polysaccharide isolated from Agaricus blazei Murill (ABP-AW1) as a potential Th1 immunity-stimulating adjuvant.

In the present study, a low molecular weight polysaccharide, ABP-AW1, isolated from Agaricus blazei Murill was assessed for its potential adjuvant activity. ABP-AW1 is considered to create a 'depot' of antigen at a subcutaneous injection site. ICR mice were immunized with 100 µg ovalbumin (OVA) alone or with 100 µg OVA formulated in 0.9% saline containing 200 µg aluminum (alum) or ABP-AW1 (50, 100 and 200 µg) on days 1 and 15. Two weeks after the secondary immunization, splenocyte proliferation, the expression of surface markers, cytokine production and the OVA-specific antibody levels in the serum were determined. The OVA/ABP-AW1 vaccine, in comparison with OVA alone, markedly increased the proliferation of splenic lymphocytes and elicited greater antigen-specific CD4+ T cell activation, as determined by splenic CD4+CD69+ T cells and Th1 cytokine interferon (IFN)-γ release. The combination of ABP-AW1 and OVA also enhanced IgG2b antibody responses to OVA. In conclusion, these data indicated that ABP-AW1 significantly enhanced the humoral and cellular immune responses against OVA in the mice, suggesting that ABP-AW1 stimulated Th1-type immunity. We suggest that ABP-AW1 may serve as a new adjuvant.

Purification and identification of one glucan from golden oyster mushroom (Pleurotus citrinopileatus (Fr.) Singer).

One water-soluble glucan (PCP-W1) was purified from the crude polysaccharide of Pleurotus citrinopileatus by chromatography on DEAE Sepharose Fast Flow and Sephadex G-200 column, and PCP-W1 (Mw=45kDa), was predominantly composed of Glc. Partial acid hydrolysis, Smith degradation-periodate oxidation, methylation analysis, Fourier transform infrared spectroscopy (FTIR), gas chromatography-mass spectrometry (GC-MS) analysis and nuclear magnetic resonance spectroscopy (NMR) experiments were conducted to elucidate its structure. The results indicated that PCP-W1 had a glucan backbone consisting of (1→6)-linked-ß-d-glucopyranosyl residues, which were branched at O-3 position of the backbone with (1→3)-linked-ß-d-glucopyranosyl and non-reducing end 1-ß-d-glucopyranosyl residues. The repeating unit of the polysaccharide was established as.

Phytochemicals and bioactivities of Anemone raddeana Regel: a review.

Anemone raddeana, usually called as'"Toujian Liang" in China, is an Anemone herb belonging to the Ranunculaceae family. Until now there are in total 67 of chemical components identified including triterpenoids, steroids, lactones, fats and oils, saccharide and alkaloids. A broad spectrum of pharmacological activity of A. raddeana compounds have been reported, such as antitumor, antimicrobial, anti-inflammatory, sedative and analgesic activites, as well as anti-convulsant and anti-histamine effects. In view of this, we initiated this short review to present the phytochemical and pharmacological profile of A. raddeana to support future studies in this discipline.

Structural elucidation of a heteroglycan from the fruiting bodies of Agaricus blazei Murill.

One water-soluble polysaccharide (ABP-W1) was purified from the fruiting bodies of Agaricus blazei by DEAE Sepharose Fast Flow and Sepharose 6 Fast Flow column chromatography. Its molecular weight was about 3.9×10(2) kDa as determined by high-performance size-exclusion chromatography (HPSEC). The structural feature of ABP-W1 was investigated by a combination of chemical and instrumental analysis, including partial hydrolysis with acid, periodate oxidation-Smith degradation, acetylation, methylation analysis and nuclear magnetic resonance spectroscopy (NMR (1)H, (13)C). The results revealed that ABP-W1 had a backbone consisting of (1→6)-linked-α-D-galactopyranosyl and (1→2,6)-linked-α-D-glucopyranosyl, which was branched with one single terminal (1→)-α-D-glucopyranosyl at the O-2 position of (1→2,6)-linked-α-D-glucopyranosyl along the main chain in the ratio of 1:1:1. The observation of the complex-formation between ABP-W1 and Congo Red indicated that ABP-W1 probably existed in a triple-strand helical conformation in water. Based on the data obtained, ABP-W1 was composed of a repeating unit with a structure as below: [structure: see text].

The extraction process optimization and physicochemical properties of polysaccharides from the roots of Euphorbia fischeriana.

Response surface methodology (RSM) was used to determine the optimum extraction conditions for polysaccharides (EFP) from the roots of Euphorbia fischeriana. A Box-Behnken design (BBD) with four independent variables was investigated, such as extraction temperature (°C), water/solid ratio, extraction number (n), and extraction time (h). The results indicated optimum extraction conditions were extraction temperature of 97°C, water/solid ratio of 9:1, extraction number of 2 and extraction time of 2.4h, respectively. Under these conditions, the experimental value was 24.6±0.62, which was well in close agreement with value predicted by the model. The preliminary chemical analysis of EFP revealed the EFP contained 25.43% polysaccharides, 20.42% uronic acids, 2.54% sulfate radical and 23.41% proteins. And the neutral polysaccharides were mainly composed of glucose, arabinose, rhamnose, galactose, xylose, mannose in the ratio of 21:8:5:3:1:1.

Prevalence and dissemination of oqxAB in Escherichia coli isolates from animals, farmworkers, and the environment.

OqxAB has recently been identified as one of the mechanisms of plasmid-mediated quinolone resistance (PMQR). Compared to what is observed for other PMQR determinants, there is a paucity of data with regard to the prevalence and epidemiology of OqxAB and its contribution to resistance to different antimicrobials. In this study, the prevalence and dissemination of oqxAB and other PMQR genes in Escherichia coli isolates from animals, farmworkers, and the environment in 2002 in China were investigated. Of the 172 E. coli isolates, 39.0% carried oqxA, while only 4.1%, 2.9%, and 0.6% carried qnr (1 qnrB6 isolate, 5 qnrS1 isolates, and 1 qnrD isolate), qepA, and aac(6')-Ib-cr, respectively. Among the 33 isolates from farmworkers, 10 (30.3%) were positive for oqxA. oqxAB was associated with IS26 and was carried on the 43- to 115-kb IncF transferable plasmid. Transconjugants carrying oqxAB showed 4- to 16-fold increases in the MICs of quinolones, 16- to 64-fold increases in the MICs of quinoxalines, 8- to 32-fold increases in the MICs of chloramphenicol and trimethoprim-sulfamethoxazole, and 4- to 8-fold increases in the MICs of florfenicol compared to the levels for the recipient. The pulsed-field gel electrophoresis (PFGE) analysis showed that the high levels of prevalence and dissemination of oqxAB in E. coli in animal farms were primarily due to the transmission of plasmids carrying oqxAB, although clonal transmission between human and swine E. coli isolates was observed. It is concluded that oqxAB was widespread in animal farms in China, which may be due to the overuse of quinoxalines in animals. This study warrants the prudent use of quinoxalines in food animals.

Isolation and evaluation of immunological adjuvant activities of saponins from the roots of Pulsatilla chinensis with less adverse reactions.

In this study, the saponins (PCS) from the roots of Pulsatilla chinensis were evaluated for its haemolytic activity, acute toxicity and tested for potential adjuvant activity in mice immunized with ovalbumin (OVA) compared with that of Quil A saponin. The haemolytic activity of PCS was determined using 0.5% rabbit red blood cell with values of 15.41 and 7.42% at concentrations of 500 and 250microg/mL, respectively. The saponins were tested for their toxicity by lethality in mice and were found to be less toxic at the same dose than their counterpart Quil A. ICR mice were immunized subcutaneously with OVA 100microg in phosphate-buffered saline (PBS) alone or with OVA 100microg in the presence of Quil A (10microg) or PCS (50, 100 or 200microg) twice at a 2-week interval. Four weeks later, the ConA-, LPS-, and OVA-induced splenocyte proliferation, OVA-specific antibodies levels (IgG, IgG1, and IgG2a) in serum, IL-2, and TNF-alpha were significantly enhanced by PCS at a high dose compared to that induced by Quil A. The P values of various testing activities in saponin-treated groups were obviously differential to that in the OVA-immunized mice (p<0.05 or p<0.01). This finding suggested that PCS might have an effect on Th1 and Th2 helper T cells. In conclusion, the results indicated that PCS showed slight side effects and at an appropriate dose could be used as a vaccine adjuvant to increase immune responses.

Immunological adjuvant effect of a water-soluble polysaccharide, CPP, from the roots of Codonopsis pilosula on the immune responses to ovalbumin in mice.

In this study, one water-soluble polysaccharide, CPP, was purified from the root of Codonopsis pilosula. The immunomodulatory effect and the adjuvant potential of CPP on the cellular and humoral immune response of ICR mice against ovalbumin (OVA) were investigated. CPP was shown not to be lethal in vivo for mice in doses ranging from 0.5 to 4 mg. ICR Mice were immunized subcutaneously with 0.1 mg of OVA alone or with 0.1 mg of OVA dissolved in saline-containing aluminum hydroxide gel (Alum) (0.2 mg), QuilA (0.01 and 0.02 mg) or CPP (0.5, 1 or 2 mg) on days 1 and 15. Two weeks later (day 28), concanavalin A (ConA)-, lipopolysaccharide (LPS)-, and OVA-stimulated splenocyte proliferation, and OVA-specific serum antibodies were measured. CPP significantly enhanced the ConA-, LPS-, or OVA-induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 1 mg (P<0.05 or P<0.01). The OVA-specific IgG, IgG1, and IgG2b antibody levels in serum were also significantly enhanced by CPP compared with OVA control group (P<0.05 or P<0.01). The results suggest that CPP could be a safe efficacious adjuvant for use in vaccines against both pathogens and cancer.

Structural elucidation and immunological activity of a polysaccharide from the fruiting body of Armillaria mellea.

The water-soluble polysaccharide (AMP), with a molecular mass of 7.8x10(3)Da as determined by high-performance size-exclusion chromatography (HPSEC), was obtained from the fruiting body of Armillaria mellea. Methylation, Smith degradation, acetolysis, (1)H and (13)C NMR spectroscopy and acid hydrolysis studies were conducted to elucidate its structure. The results indicated that AMP consisted of a backbone composed of (1-->6)-linked-alpha-D-glucopyranosyl, (1-->2,6)-linked-alpha-D-glucopyranosyl and (1-->6)-linked-alpha-D-galactopyranosyl residues in the ratio of 3:1:1, and terminated with one single terminal (1-->)-beta-D-glucopyranosyl at the O-2 position of (1-->2,6)-linked-alpha-D-glucopyranosyl, on average, along the main chain. Preliminary tests in vitro showed that AMP has stimulating effects on murine lymphocyte proliferation induced by concanavalin A or lipopolysaccharide in a dose-dependent manner. It is a possible potential immunopotentiating agent for use in health-care food or medicine.

Sulfated modification of the water-soluble polysaccharides from Polyporus albicans mycelia and its potential biological activities.

In this study, three chemically sulfated polysaccharides (SPAPs) were derived from one water-soluble polysaccharide (PAP) of Polyporus albicans mycelia by chlorosulfonic acid-pyridine method. The effects of polysaccharides on the immune function were examined after the mice were intragastrical administrated with polysaccharides at three doses of 100, 200, and 300 mg/kg body weight for 7 days. The results showed that both the lymphocytes proliferation and macrophage function were significantly enhanced by SPAP in all groups along with the increase of the substitution degree and dose (P<0.01). It indicated that SPAP could be a potential immunostimulants used in the food and pharmaceutical industry.

Purification, structure and immunobiological activity of a water-soluble polysaccharide from the fruiting body of Pleurotus ostreatus.

The water-soluble polysaccharide (POP), with a molecular mass of 2.4x10(4)Da, was obtained from the fruiting body of Pleurotus ostreatus. Structure features of the purified polysaccharide were investigated by a combination of chemical and instrumental analysis, such as methylation analysis, Smith degradation, GC-MS, (13)C and (1)H NMR and FTIR. The results indicated that the backbone of POP was composed of (1-->6)-linked-alpha-D-galactopyranosyl and (1-->2,6)-linked-alpha-D-galactopyranosyl residues, which were terminated with a single terminal (1-->)-beta-D-glucose residue at the O-2 position of galactosyl along the main chain in the ratio of 1:1:1. Preliminary tests in vitro showed POP is capable of enhancing concanavalin A (ConA)- or lipopolysaccharide (LPS)-induced lymphocyte proliferation, which suggested that POP could be a potential immunostimulating agent for use in functional foods or medicine against both pathogens and cancer.