ß-glucan improves intestinal health of pearl gentian grouper via activation of the p38 mitogen-activated protein kinase signaling pathway.

Our previous study has demonstrated that supplementation of yeast ß-glucan improves intestinal health in pearl gentian grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀), accompanied by the activation of the mitogen-activated protein kinase (MAPK) signaling pathway. In this study, we investigated the effects of perturbing p38 MAPK activity using an inhibitor on the intestinal health of ß-glucan-injected pearl gentian grouper to elucidate the potential molecular mechanism underlying the protective effects of ß-glucan on the fish gut. The pearl gentian grouper was categorized into four groups: PBS injected (CD group), ß-glucan injected at a dose of 80 mg/kg (ßG group), p38 MAPK inhibitor SB203580 injected at a dose of 1 mg/kg (SB203580 group), and a combination of ß-glucan (80 mg/kg) and SB203580 (1 mg/kg) injected together (ßG + SB203580 group). The results revealed that the introduction of SB203580 significantly suppressed the ß-glucan-induced increase in p38α and p38ß mRNA expression, as well as the phosphorylation of p38 MAPK. Both the ßG group and SB203580 group exhibited reduced plica height and muscularis thickness. The ßG + SB203580 group displayed a significant reduction in mucin cell level; interleukin 1ß (il1ß) mRNA expression; induced nitric oxide synthase, tumor necrosis factor α, and IL1ß concentration; catalase and total antioxidant capacity activities. Additionally, there was a significant increase in the levels of intestinal malondialdehyde in the ßG + SB203580 group compared to the ßG group. The inhibition of the p38 MAPK signaling halted the trend of apoptosis-related caspase molecular expression induced by ß-glucan. In conclusion, ß-glucan injection resulted in elevated levels of mucous cells, nonspecific immunity, antioxidant capacity, and anti-apoptosis in grouper by modulating the p38 MAPK pathway. This study offers insights into the potential molecular mechanism underlying the protective effects of ß-glucan on intestinal health in pearl gentian grouper.

ß-glucan regulates the intestinal immunity of pearl gentian grouper via the nuclear factor kappa B signaling pathway.

The preceding study observed that yeast ß-glucan supplementation enhanced intestinal health and augmented disease resistance in pearl gentian grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀), which occurred concurrently with the activation of the nuclear factor kappa B (NFκB) signaling pathway. Thus, we hypothesized that ß-glucan improves intestinal health in grouper by modulating the NFκB pathway. Accordingly, the present study examined the effects of NFκB pathway disruption using a specific inhibitor on the intestinal health of pearl gentian grouper that had been injected with ß-glucan. The experimental groups were as follows, (1) CD group: PBS injected; (2) ßG group: ß-glucan injected at a dose of 80 mg/kg; (3) PDTC group: NFκB inhibitor PDTC injected at a dose of 30 mg/kg; (4) ßG + PDTC group: a combination of ß-glucan (80 mg/kg) and PDTC (30 mg/kg) injected together. The results demonstrated that ß-glucan-induced increases in mRNA expression levels of NFκB inhibitor α (iκbα) and p65, the degradation and phosphorylation of IκBα, and the phosphorylation of NFκB p65 were significantly inhibited following NFκB inhibition using PDTC in the intestine of grouper. The PDTC injection resulted in a significant reduction in the ß-glucan-induced increase in mucin levels. The ß-glucan-induced elevation of alkaline phosphatase (AKP) activity, component 3 (C3) content, and inflammatory factors were significantly suppressed following NFκB inhibition. The ßG + PDTC treatment resulted in a restoration of catalase (CAT) enzyme activity to the level observed in the CD treatment, while total antioxidant capacity (T-AOC) was decreased to the level of the ßG treatment. The ß-glucan-induced downregulation of caspase8 (casp8) was reversed following NFκB inhibition, as well as the mRNA levels of casp3 and casp9 being elevated to a greater extent. In conclusion, the ß-glucan-regulated intestinal immunity in grouper may be mediated by the NFκB pathway. Furthermore, the inhibitory effect of ß-glucan on apoptosis and oxidative stress may not be related to the NFκB signaling pathway.

The microbial communities of the rust layer were influenced by seawater microbial communities.

To reveal the responsible microorganisms of microbiologically-influenced-corrosion (MIC), using 16S rRNA and ITS sequencing techniques, we investigated the bacterial and fungal communities in rust layer and seawater. Results show that the corrosion-related genera of Erythrobacter, norank_f__Rhodothermaceae, and Acinetobacter bacteria, as well as Aspergillus fungi, were overrepresented in the rust layer, along with the Pseudoalteromonas and Marinobacterium bacteria in seawater, and Ramlibacter, Aquimarina, and Williamsia bacteria were first detected in the rust layer. SourceTracker analysis revealed that approximately 23.08% of bacteria and 21.48% of fungi originated from seawater. Stochastic processes governed the rust layer and seawater microbial communities, and network analysis showed coexistence and interaction among bacterial and fungal communities. These results indicate that the composition of microbial communities in the rust layer was influenced by the marine environmental microbial communities, which can provide basic data support for the control of MIC in marine-related projects.

Comparative genomics analysis of Streptococcus iniae isolated from Trachinotus ovatus: novel insight into antimicrobial resistance and virulence differentiation.

BACKGROUND: Streptococcus iniae is an important fish pathogen that cause significant economic losses to the global aquaculture industry every year. Although there have some reports on the genotype of S.iniae and its relationship with virulence, no genome-scale comparative analysis has been performed so far. In our previous work, we characterized 17 isolates of S.iniae from Trachinotus ovatus and divided them into two genotypes using RAPD and rep-PCR methods. Among them, BH15-2 was classified as designated genotype A (in RAPD) and genotype 1 (in rep-PCR), while BH16-24 was classified as genotype B and genotype 2. Herein, we compared the differences in growth, drug resistance, virulence, and genome between BH15-2 and BH16-24. RESULTS: The results showed that the growth ability of BH16-24 was significantly faster than that of BH15-2 at the exponential stage. Antimicrobial tests revealed that BH15-2 was susceptible to most of the tested antibiotics except neomycin and gentamycin. In contrast, BH16-24 was resistant to 7 antibiotics including penicillin, sulfasomizole, compound sulfamethoxazole tablets, polymyxin B, spectinomycin, rifampin and ceftazidime. Intraperitoneal challenge of T.ovatus, showed that the LD50 value of BH15-2 was 4.0 × 102 CFU/g, while that of BH16-24 was 1.2 × 105 CFU/g. The genome of S.iniae BH15-2 was 2,175,659 bp with a GC content of 36.80%. Meanwhile, the genome of BH16-24 was 2,153,918 bp with a GC content of 36.83%. Comparative genome analysis indicated that compared with BH15-2, BH16-24 genome had a large-scale genomic inversion fragment, at the location from 502,513 bp to 1,788,813 bp, resulting in many of virulence and resistance genes differentially expression. In addition, there was a 46 kb length, intact phage sequence in BH15-2 genome, which was absent in BH16-24. CONCLUSION: Comparative genomic studies of BH15-2 and BH16-24 showed that the main difference is a 1.28 Mbp inversion fragment. The inversion fragment may lead to abnormal expression of drug resistant and virulence genes, which is believed to be the main reason for the multiple resistance and weakened virulence of BH16-24. Our study revealed the potential mechanisms in underlying the differences of multidrug resistance and virulence among different genotypes of S.iniae.

Parvularcula maris sp. nov., an algicidal bacterium isolated from seawater.

A taxonomic study was carried out on strain BGMRC 0090T, which was isolated from seawater. The isolate was a Gram-negative, aerobic, flagellated, rod-shaped bacterium with algicidal activity. Optimal growth was observed at 30 °C, pH 6.0 and with 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain BGMRC 0090T belonged to the genus Parvularcula, with highest sequence similarity to Parvularcula lutaonensis CC-MMS-1T (98.4 %). Average nucleotide identity, amino acid identity and digital DNA-DNA hybridization values between strain BGMRC 0090T and five strains of the genus Parvularcula with publicly available genomes were below 84.0, 69.2 and 21.4 %, respectively. The genome of strain BGMRC 0090T was 3.2 Mb with 64.8 mol% DNA G+C content and encoded 2905 predicted proteins, three rRNA, 42 tRNA and four ncRNA genes. Some algicidal biosynthesis-associated genes were detected in the genome. Strain BGMRC 0090T contained Q-10 as the major quinone. The predominant fatty acids were identified as summed feature 8 (C18 : 1ω7c/ω6c) and C16 : 0. Based on the polyphasic evidence presented in this paper, strain BGMRC 0090T is concluded to represent a novel species of the genus Parvularcula, for which the name Parvularcula maris sp. nov. is proposed. The type strain is BGMRC 0090T (= KCTC 92591T=MCCC 1K08100T).

Planothidiumpseudolinkei sp. nov. (Bacillariophyta), a new marine monoraphid diatom species from the coast of Guangxi, China.

A new marine monoraphid diatom species, Planothidiumpseudolinkei sp. nov., is described from the coast of Guangxi, China. The detailed morphology of this epipsammic diatom is studied by using both light and scanning electron microscopy. P.pseudolinkei differs from congeners by a combination of morphological features including capitate apices, multiseriate striae, a small central area on the raphe valve and an oblong sinus on the rapheless valve. Ecological preferences of Planothidium are also briefly discussed.

Two new diatom species of the genus Gomphonemopsis (Bacillariophyceae) from the coast of China and two new combinations for the genus.

Two new diatom species belonging to the genus Gomphonemopsis are described, Gomphonemopsisnanasp. nov. and Gomphonemopsisgaoisp. nov. These two species were compared in detail with congeners. Gomphonemopsisnana is distinguished by its high stria density and small size. This species was found so far to be epiphytic only on the eelgrass collected from Qingdao Bay (Yellow Sea). Gomphonemopsisgaoi is characterized by its isopolar valves, simple proximal raphe endings and acutely rounded apices. This taxon was separated from the exoskeleton of marine copepods sampled from the Futian Mangrove Nature Reserve (South China Sea). In addition, two new combinations, Gomphonemopsisoahuensis (Hustedt) Lang Li, Yuhang Li & Changping Chen, comb. nov. and Gomphonemopsisplatypus (Østrup) Lang Li, Yuhang Li & Junxiang Lai, comb. nov. are proposed. This study increases the records and knowledge of Gomphonemopsis along the coast of China.

UV-ARTP compound mutagenesis breeding improves macrolactins production of Bacillus siamensis and reveals metabolism changes by proteomic.

Macrolactins are a type of compound with complex macrolide structure which mainly be obtained through microbiological fermentation now. They have excellent antifungal, antibacterial and antitumor activity. In order to improve macrolactins production, Bacillus siamensis YB304 was used as the research object, and a mutant Mut-K53 with stable genetic characters was selected by UV-ARTP compound mutagenesis. The yield of macrolactins was 156.46 mg/L, 3.95 times higher than original strain. The metabolic pathway changes and regulatory mechanism of macrolactins were analyzed by quantitative proteomics combined with parallel reaction monitoring. This study revealed that 1794 proteins were extracted from strain YB304 and strain Mut-K53, most of them were related to metabolism. After UV-ARTP compound mutagenesis treatment, the expression of 628 proteins were significantly changed, of which 299 proteins were significantly up-regulated. KEGG pathway analysis showed that differentially expression proteins mainly distributed in biological process, cellular component, and molecular function processing pathways. Such as utilization of carbon sources, glycolysis pathway, and amino acid metabolism pathway. Furthermore, key precursor substances such as acyl-CoA and amino acids of macrolactin biosynthesis are mostly up-regulated, which are one of the main reasons for increased production of macrolactin.This study will provide a new way to increase the yield of macrolactins through mutagenesis breeding and proteomics.

Quantitative law and scenario-based forecasting of different land use expansion, based on reliability analysis in mountainous areas.

The continuous high-intensity and disorderly expansion of construction land in mountainous areas threatens city development; consequently, the scientific guidance of its sustainable development has become a research hotspot. This work aimed to develop a new theoretical framework for predicting land expansion. Based on DMSP/OLS-Landsat 7 data correction from 2000 to 2019, to ensure data reliability, this study quantitatively analysed the expansion law of land-use and land-cover (LULC) in Huayuan, a typical mountainous area in China. Based on the land expansion law, the patch-generating land use simulation (PLUS) model was used to predict various types of LULCs in different scenarios. The results showed that (1) the reliability of LULC under multi-source spatio-temporal data correction reached more than 0.97. (2) The expansion law of industrial and mining land, urban living land, and traffic land is sprawl, while rural living land is enclaved and the expansion direction and intensity are obviously different. (3) The scale of land expansion in the natural-oriented scenario was significantly higher than that in the humanism-oriented scenario, with a higher value of 199.33 hm2. This study expands the case study of land use analysis and prediction, and provides scientific guidance for different land expansion planning, which can avoid the mismatch and waste of land resources. Furthermore, it also deepens the exploration of LULC identification reliability method and enriches the theory of different land use prediction in mountainous areas.

Improvement and application of qPCR assay revealed new insight on early warning of Phaeocystis globosa bloom.

Phaeocystis globosa bloom develops from its early solitary cells, providing clues for early warning of its bloom and timely responding to possible consequences. However, the early prediction requires quantification of the solitary cells for a thorough understanding of bloom formation. Therefore, we developed an accurate, sensitive, and specific qPCR assay for this need. Results show that the accuracy of qPCR was significantly enhanced by ameliorating DNA barcode design, improving genomic DNA extraction, and introducing a strategy of internal amplification control (IAC). This approach reached a quantification limit of 1 cell/reaction, making low-abundance cells (101-103 cells/L) detection possible, and we also observed a plunge in the abundance of the solitary cells before the bloom outbreak in two winters in 2019 and 2020 for the first time, which is quite unique from laboratory results showing an increase instead. The plunge in solitary-cell abundance might be associated with the attachment of solitary cells to solid matrices to form non-solitary attached aggregate, the precursor of colonies, which gains supports from other studies and needs more investigations in the future. Therefore, as the plunge in solitary-cell abundance is a sign of colony formation, it can be used as an early warning indicator to P. globosa bloom.

Effects of nutrient limitation on cell growth, exopolysaccharide secretion and TEP production of Phaeocystis globosa.

Phaeocystis globosa (P. globosa) often colonizes and produces mucus, which may cause massive blooms in coastal areas. To understand mechanism of the growth and the impact factors for better control of the bloom, we conducted a laboratory experiment on the effect of nitrogen (N) or phosphorus (P) limitation on the cell growth, production of exopolysaccharide (EPS), and transparent exopolymeric particles (TEP) of P. globosa. Results show no obvious differences in the N- and/or P-limitation in TEP production, polysaccharide secretion, and colony growth of P. globosa. Particularly in the death phase of the algae growth, the TEP production level in the experiment differed significantly, and was higher in the P-limitation group than that in the N-limitation group; additionally, the P-limitation group produced a relatively higher amount of EPS than N-limitation group, with greater cellular chlorophyll-a content, and in greater photosynthetic reaction rate of P. globosa cells, than those of the N-limitation group. However, under N-limited conditions, the algae colony survived longer. Under P-limited condition, P. globosa cells spend the photosynthesis-produced substances and energy for the secretion of extracellular substances but for cell reproduction, which was indicated by P. globosa cell growth and carbon content ratio between TEP and biomass.

1,5-anhydroglucitol biosensor based on light-addressable potentiometric sensor with RGO-CS-Fc/Au NPs nanohybrids.

In this paper, a novel silicon-based light-addressable potentiometric sensor (LAPS) has been designed for the detection of 1,5-anhydroglucitol (1,5-AG) in human serum. Reduced graphene oxide-chitosan-ferrocene (RGO-CS-Fc)/AuNPs nanohybrids and pyranose oxidase (PROD) enzyme is used to fabricate biological sensitive membrane unit by layer-by-layer assembly technology. When a bias voltage is provided to the LAPS system, the catalytic oxidation reaction between 1,5-AG and PROD to produce H2O2. The by-product H2O2 can oxidize Fc(Fe2+) ions in RGO-CS-Fc nanohybrids into Fc(Fe3+) ions, which cause the potential of the sensitive membrane surface to change and the potential shift of I-V curve will generate a corresponding offset response. Under the optimal conditions, the potential shift of the LAPS is linearly related to the concentration of 1,5-AG at 10 µg·mL-1 -350 µg·mL-1 with the correlation coefficient of 0.97414. The sensitivity is 0.44273 mV/µg·mL-1 and the lowest detection limit is 10 µg·mL-1. In addition, the biosensor showed good specificity, acceptable stability and satisfactory recovery rates (91.28%-107.66%), which would be a potential testing methods in actual clinical samples.

Spatiotemporal distribution of nitrogen biogeochemical processes in the coastal regions of northern Beibu Gulf, south China sea.

Nitrogen biogeochemistry occupies a central role in nitrogen cycles and exerts a significant influence on primary productivity and global carbon cycles. In order to better understand the nitrogen biogeochemistry in coastal regions, spatiotemporal nitrogen fixation, denitrification and anammox were investigated in the coastal regions of northern Beibu Gulf (NBG), South China Sea (SCS). Nitrogen fixation was mainly detected in the water column of outer bays, attributed to the low nitrate concentration and low N/P (N/P < 16). Comparisons of the nitrogen fixation rates between unicellular (<10 mm) and the filamentous diazotrophs (>10 mm) indicated that the contribution of unicellular diazotrophs was more important than filamentous diazotrophs. Besides, field investigation revealed that Richelia was the dominant species in filamentous diazotrophs. On the other hand, both denitrification and anammox were found in the surface sediment and denitrification dominated the nitrogen loss process. Denitrification was mainly related to the nitrate concentration in pore water and organic matter in the sediment, while anammox was mainly regulated by the concentration of nitrate and nitrite in pore water. Additionally, temperature, dissolved oxygen (DO) and salinity also had an impact on denitrification and anammox. The net areal yield of nitrogen biogeochemical processes was estimated to be -1079t/a, as an important pathway of nitrogen removal. This study adds to the knowledge of nitrogen biogeochemistry in the nutrient-replete coastal region and highlights its significance in such an environment.

[Phytoplankton pigment patterns and community structure in the Yangtze Estuary and its adjacent areas].

Three cruises were carried out in the Yangtze Estuary and its adjacent areas in May, November, June during 2009-2010. The spatial variations of phytoplankton community structure were investigated based on RP-HPLC analysis of pigments and CHEMTAX processing of the pigment data. 21 kinds of pigments were detected, among which chlorophyll a, peridinin, fucoxanthin, 19'-butanoyloxyfucoxanthin, 19'-hexanoyloxyfucoxanthin, chlorophyll b, diadinoxanthin, alloxanthin and zeaxanthin were the major pigments in the Yangtze Estuary and its adjacent areas. Chlorophyll a was the most abundant in all pigments, followed by fuxoxanthin. Other pigments generally contributed a minor proportion to the total pigments. High concentrations of fucoxanthin and peridinin were observed in May 2009 and June 2010, indicating blooms of diatoms and dinoflagellates. The results showed that the composition and distribution of phytoplankton pigments were influenced by environmental factors. The phytoplankton community, as determined by biomarker pigment concentration using HPLC and CHEMTAX, was composed mainly of diatoms, dinoflagellates, cryptophytes, chlorophytes, cyanobacteria, prymnesiophytes, chrysophytes and prasinophytes. The dominant algal groups were diatoms, dinoflagellates and chlorophytes in May 2009. The phytoplankton community was characterized by high contribution of diatoms in November 2009. Diatoms, dinoflagellates and cryptophytes accounted for 62.5% of chlorophyll a in June 2010, and the relative abundance of cyanobacteria was higher in this cruise. The spatial variations of phytoplankton community structure featured distinct regionality. Diatoms, chlorophytes and cryptophytes were the main groups in the inshore waters, and the abundances of prymnesiophytes, chrysophytes and cyanobacteria were increasing from inshore to the open sea.