Acid Phosphatase Activity Research Articles

Accumulation of coumaric acid is a key factor in tobacco continuous cropping obstacles

IntroductionPhenolic acids are believed to play a significant role in tobacco continuous cropping obstacles, but the strength and potential mechanisms of different phenolic acids remain unclear.MethodsThis study evaluated the allelopathic effects of six phenolic acids that exhibited cumulative effects in our previous research. Different concentrations of phenolic acids with the strongest allelopathic effects were added to potting soil to explore their impacts on tobacco growth and physiological characteristics, as well as on soil chemical properties and microbial community structure. ResultsThe results showed that coumaric acid exhibited the strongest direct allelopathic effect. Exogenous coumaric acid significantly reduced soil pH and shifted the soil microbial community from bacteria-dominated to fungi-dominated. Simultaneously, the abundance of bacteria related to nutrient utilization (e.g., Flavisolibacter, Methylobacterium) and fungi related to disease resistance (e.g., Fusicolla, Clonostachys) gradually decreased, along with a reduction in soil catalase, urease, invertase, and acid phosphatase activities. Leaf MDA levels increased continuously with higher concentrations of coumaric acid, while the root resistance hormone (jasmonic acid and the jasmonate-isoleucine complex) levels show the opposite trend.DiscussionCoumaric acid may inhibit tobacco growth by influencing the physiological processes in tobacco plants directly and the broader soil microecological balance indirectly. This study provides theoretical guidance for precise mitigation of continuous cropping obstacles in future tobacco cultivation.

Arbuscular mycorrhizal fungi and intercropping Vicia villosa mediate plant biomass, soil properties, and rhizosphere metabolite profiles of walnuts

Intercropping is a prevalent soil management strategy within orchards, whereas it is unclear how inoculation with arbuscular mycorrhizal (AM) fungi and intercropping affect tree growth, soil properties, and rhizosphere metabolite profiles. This study investigated the effects of inoculation with Diversispora spurca and intercropping with hairy vetch (Vicia villosa) on biomass production, soil available nutrients, water-stable aggregate (WSA) distribution, phosphatase activity, and secondary metabolite profiles in walnuts (Juglans regia). The intercropping only elevated soil nitrate N levels and WSA distribution at the 0.5–2 mm size, and also triggered 2159 differential metabolites (1378 up-regulated and 781 down-regulated), with armillaramide as the most prominently up-regulated metabolite, followed by the substance diminished upon D. spurca inoculation. Conversely, D. spurca inoculation increased walnut biomass, WSA distribution across the 0.25 − 2 mm size, and acid and neutral phosphatase activities, as well as triggered 2489 differential metabolites (897 up-regulated and 1592 down-regulated), with pteroside D being highest up-regulated differential metabolite, allowing a competitive advantage to AM plants in combating soil pathogens. Despite significantly suppressing root AM fungal colonization and biomass production in AM walnuts, intercropping significantly increased soil ammonium and nitrate N levels in AM walnuts as well as WSAs at the 1–4 mm size, exhibiting a synergistic effect. Flavone and flavonol biosynthesis and pyruvate metabolism were simultaneously involved following AM inoculation or intercropping. Co-application of AM inoculation and intercropping triggered 1006 differential metabolites, with urocanic acid being the most up-regulated metabolite, although it decreased following AM inoculation, suggesting the involvement of mycorrhizal hyphae in soil histidine uptake. Under intercropping, AM inoculation elicited 418 differential metabolites, with the most up-regulated metabolite being implicated in flavonoid pathways. AM inoculation primarily triggered the biosynthesis of unsaturated fatty acids, regardless of intercropping or not, implying a potential increase in unsaturated fatty acid contents of walnut kernels. It concluded that AM inoculation and intercropping interactively affected walnut growth, soil attributes, and soil microenvironment.Graphical

The Impact of Acute Ammonia Nitrogen Stress on Serum Biochemical Indicators and Spleen Gene Expression in Juvenile Yellowfin Tuna (Thunnus albacares)

The presence of ammonia nitrogen in water has a significant impact on the serum and spleen of fish, potentially leading to changes in substances such as proteins in the serum while also causing damage to the immune function of the spleen. To investigate the effects of ammonia nitrogen (NH3-N) stress on juvenile yellowfin tuna (Thunnus albacares), this study established three NH3-N concentrations, 0, 5, and 10 mg/L, denoted as L0, L1, and L2, respectively. Serum and spleen samples were collected at 6, 24, and 36 h. The effects of different NH3-N concentrations and exposure times on the physiological status of juvenile fish were investigated by analyzing serum biochemical indices and splenic gene expression. The results indicate that in the L1 group, the serum high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), complement 3 (C3) and complement 4 (C4) levels, and acid phosphatase (ACP) activity showed a trend of initially increasing and then decreasing. In the L2 group, the serum low-density lipoprotein cholesterol (LDL-C), HDL-C, and C4 levels and ACP activity also displayed an initially rising and then declining trend, while TG, C3, and creatinine (CRE) levels and alkaline phosphatase (AKP) activity showed an upward trend. In the L1 group, glutathione peroxidase 1b (GPX1b), interleukin 10 (IL-10), interleukin 6 receptor (IL-6r), and tumor necrosis factor alpha (TNF-α) gene expression levels in the spleen exhibited a trend of first increasing and then decreasing. In the L2 group, IL-10, IL-6r, tumor necrosis factor beta (TNF-β), caspase 2 (casp2), and caspase 9 (casp9) gene expression levels in the spleen also showed an initial increase followed by a decrease. When NH3-N levels are below 5 mg/L, it is recommended to limit stress exposure to no more than 36 h for the juvenile fish. For concentrations ranging from 5 to 10 mg/L, stress should be strictly controlled to within 24 h. Exposure to high NH3-N levels may affect biochemical indicators such as serum lipid metabolism, immunity, and metabolism in juvenile fish, and may damage the expression of antioxidant, immune gene, and apoptosis factors in the spleen. This study aims to deepen our understanding of the effects of NH3-N on juvenile tuna, with the goal of establishing effective water quality monitoring and management strategies. This will ensure the quality of aquaculture water, reduce the harm caused by NH3-N to juvenile yellowfin tuna, and enhance aquaculture efficiency and product quality.

Geranylgeranyl diphosphate synthase inhibition impairs osteoclast differentiation, morphology and resorptive activity

Abstract Nitrogen bisphosphonates (NBPs), such as zoledronic acid, target the enzyme farnesyl diphosphate synthase (FDPS) in the isoprenoid biosynthetic pathway (IBP), and are the frontline treatment for osteolytic bone diseases. The strong affinity of these agents for bone limits their distribution out of the skeleton. Geranylgeranyl diphosphate synthase (GGDPS) is directly downstream to FDPS in the IBP and novel GGDPS inhibitors such as RAM2061 have been shown to have key drug-like features including prolonged half-life, metabolic stability, and systemic distribution. Furthermore, RAM2061 exerts anti-neoplastic benefits in mouse models of multiple myeloma and Ewing sarcoma. Therefore, we are interested in determining the potential impact of RAM2061 on osteoclast biology and bone remodeling. Studies utilizing undifferentiated RAW264.7 cells demonstrated that treatment with RAM2061 depletes cells of GGPP, impairs protein geranylgeranylation and induces markers of the unfolded protein response pathway and apoptosis. Differentiation of RAW264.7 cells to mature osteoclasts is disrupted by RAM2061, resulting in decreased numbers of mature osteoclasts, altered morphology and decreased tartrate resistant acid phosphatase activity. Treatment of fully differentiated RAW264.7 cells with RAM2061 led to decreased resorptive activity. Confocal microscopy studies revealed that RAM2061 disrupts Cdc42 localization, inhibiting proper actin ring formation in osteoclasts. No significant impact on bone turnover markers or bone histomorphology was observed following a 3-week treatment of CD-1 mice with RAM2061, although decreased numbers of osteoclasts were observed. Liquid chromatography–tandem mass spectrometry studies confirmed accumulation of RAM2061 in bone from the in vivo studies as well as hydroxyapatite binding in vitro. In conclusion, these studies are the first to demonstrate the anti-osteoclastic activity of GGDPS inhibitor treatment and support future studies exploring the therapeutic benefit of this novel therapy in the setting of pathological bone remodeling.

Viscoelastic Hydrogel Modulates Phenotype of Macrophage-Derived Multinucleated Cells and Macrophage Differentiation in Foreign Body Reactions.

Biomaterial-induced macrophage-derived multinucleated cells (MNCs) are often observed at or near material implantation sites, yet their subtypes and roles in tissue repair and wound healing remain unclear. This study compares material-induced MNCs to cytokine-induced MNCs using both in vitro and in vivo models. 3D-embedded Raw264.7 cells and rat bone marrow-derived monocytes (BMDMs), with or without cytokines such as IL-4 and RANKL, were characterized for their MNC morphologies and subtypes via in situ immunocytochemistry and flow cytometry. Macrophage polarization and osteoclastic differentiation were assessed through NO production, arginase activity, and tartrate-resistant acid phosphatase levels. 3D matrix-induced MNCs expressed the same phenotypic heterogeneity as the IL-4 and RANK-treated ones. 3D matrix-induced MNCs displayed the same phenotypic heterogeneity as those treated with IL-4 and RANKL. A high viscoelastic matrix (1006.48 ± 92.29 Pa) induced larger populations of proinflammatory and osteoclast-like MNCs, whereas a low viscoelastic matrix (38.61 ± 7.56 Pa) supported active differentiation and gene expression across pro-, anti-inflammatory, and osteoclast-like macrophages. Matrix viscoelasticity also influenced the effects of IL-4 and RANKL on macrophage-derived MNC polarization. In an in vivo subcutaneous implantation model, medium to high viscoelastic matrices exhibited higher populations of CD86+ and RANK+ MNCs, while low viscoelastic matrices showed higher populations of CD206+ MNCs. These findings suggest that matrix viscoelasticity modulates macrophage differentiation and MNC phenotype, with low viscoelastic matrices potentially favoring anti-inflammatory MNCs and macrophage differentiation suitable for subcutaneous implantation.

Remediation of PBDE-contaminated soil using biochar-based bacterial consortium QY2Y

Biochar-immobilized microbial technologies hold substantial promise for the remediation of environmental contaminants. However, the understanding of remediation efficiency and the operation of microbial consortia in complex soil environments using biochar as a carrier remains limited. In this study, we explored the impacts of biochar combined with bacterial consortium QY2Y (consisting of Chitinophaga sp. MH-1, Achromobacter sp. YH-1, Methylobacterium sp. ZY-1, and Sphingomonas sp. GY-1) on BDE-47 dissipation in contaminated soil, as well as its effects on soil-plant systems. The findings indicated that the immobilization of biochar and QY2Y not only stimulated the biodegradation of BDE-47 (61.50%) but also decreased the bioavailability of BDE-47, subsequently reducing the concentration of BDE-47 in the edible portion of rapeseed (Brassica napus L.) by 82.00%. In addition, the combined treatment notably increased soil pH, enhanced the physicochemical properties and nutrient conditions of the soil, and amplified soil dehydrogenase, catalase, urease, and acid phosphatase activities by 58.72%, 69.25%, 54.24%, and 74.74%, respectively. The application of biochar-based QY2Y improved the soil microbial community structure, bolstered the interspecific symbiotic and cooperative relationships, and restructured the keystone taxa. The key species microbes had direct and significant positive effects on BDE-47 degradation, soil nutrients, and soil enzyme activity. This study deepens our understanding of the potential applications of immobilized microbial consortia in treating PBDE-contaminated soils and offers guidance for the management and remediation of contaminated soils.

Faba bean-wheat intercropping controls the occurrence of faba bean Fusarium wilt by improving the microecological environment of rhizosphere soil

BackgroundFusarium wilt is a severe soil-borne disease that affects faba bean production. Faba bean-wheat intercropping is often used to control the occurrence of Fusarium wilt in faba bean. AimsTo evaluate the effects of faba bean-wheat intercropping on the occurrence of faba bean Fusarium wilt and soil microecology. MethodsWe established two planting patterns, faba bean monocropping (M) and faba bean-wheat intercropping (I), to investigate Fusarium wilt occurrence and plant dry weight and assess changes in soil enzyme activities, microbial diversity, and community composition during different stages of disease onset. ResultsIntercropping effectively controlled faba bean Fusarium wilt at the three disease stages and increased the dry weight of faba bean plants. Intercropping promoted the activities of catalase (CAT), urease, sucrase, and acid phosphatase in the rhizosphere soil of faba bean at three disease stages. Bacterial and fungal diversity decreased with disease progression, and intercropping mitigated this trend. Compared with monocropping, intercropping increased the abundance of beneficial bacteria such as Proteobacteria, Actinobacteriota, Gemmatimonadota, Gemmatimonas, Conexibacter, and Sphingomonas, while reducing the abundance of pathogenic fungi such as Alternaria, Cladosporium, and Fusarium. Intercropping also increased the abundance of arbuscular mycorrhiza, soil saprophytes, and undefined saprophytes while decreasing the abundance of plant pathogens. ConclusionFaba bean-wheat intercropping enhanced soil enzyme activities, effective nutrient content, and alpha diversity indices of bacteria and fungi in the rhizosphere soil of faba bean, while promoting the abundance of beneficial bacteria, arbuscular mycorrhizal fungi, as well as both soil and undefined humus. Simultaneously, intercropping reduced the abundance of plant pathogens, facilitated nutrient cycling in the soil, provided sufficient nutrients for crop uptake, and mitigated the toxic effects of hydrogen peroxide on cells. Ultimately, this resulted in a reduced occurrence of Fusarium wilt.

Nutritional Innovation Using Green Seaweed (Ulva sp.) and Garlic Powder Extracts for White-Leg Shrimp (Litopenaeus vannamei) Challenged by Vibrio harveyi.

This study aimed to determine the antimicrobial effects of ethanolic extracts of Ulva sp. and garlic (Allium sativum) powder ethanolic extracts against Vibrio harveyi in vitro. The stimulatory effects of Ulva sp. extract (UE) and garlic powder extract (GPE) on the growth performance and innate immune responses of white-leg shrimp, Litopenaeus vannamei, and their challenge against V. harveyi infection were also investigated. A commercial shrimp diet (36.1% protein) was enriched with 0.5, 1.0 and 2.0gUE/kg diet and 2, 4 and 6gGPE/kg diet, whereas the control group was free of any supplement. Health juveniles of L. vannamei (average weight 2-3g) were distributed in 21 fiberglass reinforced plastic (FRP) tanks (500-L capacity) at a stocking density of 300 animals/tank to represent each treatment in triplicate. The animals were fed ad libitum on the experimental diets up to satiety four times daily for 60 days. The phytochemical analysis of ethanolic extracts of Ulva sp. and garlic powder evoked their richness of several bioactive compounds showing significant antibacterial activity against V. harveyi. The GPE exhibited a higher inhibition zone than that of the UE. The supplemented diets did not significantly affect weight gain %, final weight, feed conversion ratio, specific growth rate and survival rates of white shrimp compared to those fed on the control diet. Significant increases were observed in total haemocyte count, phagocytosis and phagocytic index of all treatments compared with the control group. There were significant increases in serum total protein, acid phosphatase activity, alkaline phosphatase, lysosomal enzyme activity, phenoloxidase activity and superoxide dismutase activity with offered diets with increasing the levels of ethanolic extracts of Ulva sp. and garlic powder up to 2.0gUE/kg diet and 6gGPE/kg diet, respectively. The ethanolic extraction of Ulva sp. and garlic powder-supplemented diet groups, particularly at treatments of 2.0 and 6gGPE/kg diet, respectively, significantly reduced the shrimp mortality induced by V. harveyi infection when compared with the control group. The net results evoked that ethanolic extraction of Ulva sp. (2.0gUE/kg) and garlic powder (6gGPE/kg diet) enhanced the immune response and disease resistance of the white-leg shrimp, L. vannamei. It is also noted that the GPE is more efficient than the UE in vitro and in vivo investigations.

Intercropping of tobacco and maize at seedling stage promotes crop growth through manipulating rhizosphere microenvironment.

Changes in the rhizosphere microbiome and metabolites resulting from crop intercropping can significantly enhance crop growth. While there has been an increasing number of studies on various crop combinations, research on the intercropping of tobacco and maize at seedling stage remains limited. This study is the first to explore rhizosphere effects of intercropping between tobacco and maize seedling stages, we analyzed the nitrogen, phosphorus and potassium nutrients in the soil, and revealed the important effects on soil microbial community composition and metabolite profiles, thereby regulating crop growth and improving soil balance. Compared with mono-cropping, intercropping increased the biomass of the two crops and promoted the nutrient absorption of nitrogen, phosphorus and potassium. Under intercropping conditions, the activities of sucrase, catalase and nitrate reductase in tobacco rhizosphere soil and the content of available potassium, the activities of nitrate reductase and acid phosphatase in maize rhizosphere soil were significantly increasing. Rhizosphere soil bacterial and fungal communities such as Sphingomonas, Massilia, Humicola and Penicillium respond differently to crop planting patterns, and soil dominant microbial communities are regulated by environmental factors such as pH, Organic Matter, Available Potassium, Nitrate Reductase, and Urease Enzyme. Network analysis showed that soil microbial communities were more complex after intercropping, and the reciprocal relationship between bacteria and fungi was enhanced. The difference of metabolites in soil between intercropping and monocropping system was mainly concentrated in galactose metabolism, starch and sucrose metabolism pathway, and the content of carbohydrate metabolites was significantly higher than that of monocropping soil. Key metabolites such as D-Sucrose, D-Fructose-6-Phosphate, D-Glucose-1-Phosphatel significantly influence the composition of dominant microbial communities such as Sphingomonas and Penicillium. This study explained the effects of intercropping between flue-cured tobacco and maize on the content of soil metabolites and soil microbial composition in rhizosphere soil, and deepened the understanding that intercropping system can improve the growth of flue-cured crops seedlings through rhizosphere effects.

BIOCHEMICAL PARAMETERS OF THE BLOOD OF RATS OF DIFFERENT AGES AFTER FILLING THE METAPHYSEAL DEFECT OF THE FEMUR WITH ALLOGENEIC BONE IMPLANTS WITH LOCAL ADMINISTRATION OF BLOOD PLASMA ENRICHED WITH PLATELETS

A promising method of regenerative medicine is the saturation of allografts with platelet-rich plasma (PRP). Objective. To evaluate the course of metabolic processes after filling a hole defect in the distal metaphysis of the femur with allogeneic bone implants in conditions of additional local administration of allogeneic PRP. Methods. In the model of a hole defect in white rats with the filling of the defect with an allograft, as well as with additional local stimulation of PRP on the 7th day; on the 3rd and 7th days and on the 1st, 3rd and 7th days in blood serum, the content of glycoproteins (GP), chondroitin sulfates (CST), total protein (TP), calcium (Ca), activity alkaline phosphatase (AlP) and acid phosphatase (AcP). The results. 14 day. In the 3-month rats, under one stimulation, an increase in TP and Ca, a decrease in AcP was observed, with two stimulations there was a 1.11 times smaller GP, 1.13 times larger TP and 1.43 times Ca compared to those in rats without stimulation. During three stimulations, the GP was 1.24 times lower than that in animals without stimulation. In the 12-month rats in comparison with the data of rats without stimulation, 1.15 times higher GP, Ca, AlP activity, and 1.44 times more AcP were noted. 28 days of the 3-month rats for one injection exceeded the data of animals without stimulation by 1.34 times for GP, and were inferior to them by 1.31 times for AcP. In the 12-month rats, compared to these animals without stimulation, with three injections, a 1.19 times greater TP was noted. 90 d. In the 3-month rats for one injection showed 1.24 times less CST with 1.28 times lower AlP compared to data from rats without stimulation. 12-month rats exceeded the data of the group without stimulation by 1.43 times for AlP. Conclusions. In rats with an alloimplant (especially at 12 months), an increase in connective tissue formation markers and a decrease in LF activity were observed. Filling the defect with an alloimplant led to an increase in inflammation indicators and an increase in markers of bone tissue formation.

Facilitating effects of plant extracts on soil health and replanted Panax ginseng growth in recession soil.

Plant extracts have been shown to be effective agricultural strategies for improving soil fertility and quality, and promoting plant growth in soil degradation remediation. The application of plant extracts improves the material cycle of soil microecology, such as the decomposition of nitrogen, phosphorus, and potassium, while increasing plant resistance. However, there is currently no experiment to demonstrate whether plant extracts have a promoting effect on the growth of ginseng and the mechanism of action. Pot experiments were carried out to investigate the effects of extracts, namely Rubia cordifolia (RC), Schisandra chinensis (SC), and Euphorbia humifusa (EH) on soil properties, enzyme activities, and plant physiological characteristics were evaluated. Results showed that compared with CK, plant extract-related treatments increased soil Organic carbon (OC), Available nitrogen (AN), Available phosphorus (AP) contents, and Soil urease activity. (S-UE), Soil sucrase activity (Soil sucrase), Soil acid phosphatase activity. (S-ACP). Meanwhile, plant extract-related treatments significantly increased plant physiological properties and TP (Total protein) content, and decreased the content of MDA (malondialdehyde) by 15.70% -36.59% and PRO (proline) by 30.13% -148.44%. Furthermore, plant extract-related treatments also significantly promote plant growth and reduce plant incidence, the fresh weight of ginseng increased by 27.80% -52.08%, ginseng root activity increased by 45.13% -90.07%, and ginseng incidence rate decreased by 20.00% -46.67%. Through correlation analysis between fresh weight of ginseng and root parameters and soil index, fresh weight is significantly positively correlated with root diameter, fiber root number, root activity, total protein (TP), catalytic activity (CAT) and superoxide dismutase activity (SOD), H, soil urea activity (S-UE), soil sucrose activity (S-SC), soil acid phosphate activity (S-ACP), and soil laccase activity (SL); The fresh weight was significantly negatively correlated with incidence rate, disease severity index, and malondialdehyde content (MDA). In summary, plant extract-related treatments improve soil quality and promote ginseng growth, further enhancing soil health and plant disease resistance. These findings provide new insights into ginseng cultivation and soil health management and highlight a new approach that can be applied to a wider range of agricultural practices and environmental sustainability.

Polystyrene nanoplastics induce apoptosis, histopathological damage, and glutathione metabolism disorder in the intestine of juvenile East Asian river prawns (Macrobrachium nipponense)

Nanoplastics (NPs) are widely distributed in the aquatic environment and have become a global concern as a new type of pollutant. Many researchers have studied the physiological effects of NPs on aquatic organisms, but relatively little is known about their effects on intestinal immune function in crustaceans. Therefore, we used NPs concentrations of 0, 5, 10, 20, 40 mg/L for 28 days of stress, evaluated the effects of NPs exposure on intestinal cell apoptosis, histopathological damage, and glutathione (GSH) metabolism of juvenile East Asian river prawns (Macrobrachium nipponense). As NPs concentration increased, the contents of total GSH and oxidized glutathione decreased gradually (P < 0.05), the concentration of GSH first increased and then decreased (P < 0.05), and the activities of lysozyme, acid phosphatase, phenoloxidase, and alkaline phosphatase first increased and then decreased (P < 0.05). Additionally, intestinal tissue structure was damaged, and the apoptosis rate significantly increased (P < 0.05). The expression of intestinal autophagy genes (CTL, ALF, Crustin, ATG8, and BCL-2) increased at first and then decreased, the expression levels of TNF and Wnt4 significantly decreased, and the expression of Beclin significantly increased with increasing NPs concentration. We also found that AP-1 and PTEN were highly expressed in the hepatopancreas and were involved in intestinal immune responses. Our results showed that exposure to NPs may induce apoptosis of intestinal tissue cells, induce autophagy, and inhibit GSH metabolism, thereby reducing intestinal immune function of M. nipponense. These findings provide a reference for healthy aquaculture and ecological risk assessment of prawns.

Replacing sea mud with attachment of suspension cage can improve growth and gut health for sea cucumber Apostichopus japonicus

During the suspension cage culture procedure, a substantial quantity of attachments from the suspension cage will amass in the tidal flats, resulting in environmental pollution and resource wastage. The aim of the present study was to explore the feasibility of using attachments of suspension cage (ASC) as a raw feed material for sea cucumbers. Different ASC concentrations (0, 25, 50, 75, and 100%) were used in sea cucumber (Apostichopus japonicus) fed for 60 d. Replacing sea mud with 50% ASC significantly improved sea cucumber growth, reduced the feed coefficient, significantly increased the activity of intestinal digestive enzymes, improved the activities of acid phosphatase and alkaline phosphatase, improved the intestinal structure, and enriched intestinal microbiome diversity (P&amp;lt;0.05). Replacing sea mud with 75% and 100% ASC significantly improved sea cucumber total antioxidant capacity and total superoxide dismutase (P&amp;lt;0.05).Furthermore, expressions of c-myc gene, mapk-7 gene and fgfr-1 gene, which all three growth genes, significantly increased in the 50% ASC treatment compared to that in the control; whereas, that of gdf-8 gene, a negative growth regulator, was inhibited (P&amp;lt;0.05). Based on the quadratic regression analysis for the weight gain rate, the appropriate levels of ASC substitution were estimated as to be 44.28%. The results suggested the potential of ASC as a new raw feed material for sea cucumbers.

Antibacterial activity and mechanism of combination natural essential oils from oregano and cinnamon against enterotoxigenic Escherichia coli K99

Essential oils from oregano (OEO) and cinnamon (CEO) have been reported an excellent antibacterial activity, but their synergistic antibacterial effect is rarely studied. In this study, the chemical compositions of OEO, CEO and mixed OEO with CEO (MEO) and their antibacterial activities were analyzed, the mode of MEO action against enterotoxigenic Escherichia coli K99 (ETEC K99) were investigated. Results showed that MEO had the synergistic antibacterial effect, which was proven by fractional inhibitory concentration index of 0.375. The inhibition zone diameter measured varying with volume ratio of OEO and CEO showed that the best synergistic effect was at a ratio 2:8. Meanwhile, the antibacterial mechanism of MEO demonstrated that the MEO changed the morphology of ETEC K99 by scanning and transmission electron microscopy, and disrupted the cell membrane integrity that were proved by the leakage of intracellular nucleic acid and lower alkaline phosphatase activity of ETEC K99. In addition, the MEO also inhibited DNA biosynthesis, biofilm formation and scavenged mature biofilm of ETEC K99. These results confirm the synergistic effect on improvement of antibacterial activity and elucidate a variety of mechanisms through which the ETEC K99 was inactivated, and support the application of MEO to against ETEC K99.

Bone regeneration-enhancing effects of extremely low-frequency electromag- netic fields: Analysis using fish scales as a bone model.

Electromagnetic fields (EMFs) noninvasively promote fracture healing, prevent osteoporosis, promote diaphyseal growth, enhance differentiation, and stimulate cell division. However, no good model systems for analyzing bone regeneration have been reported. In this study, we examined the in vivo regeneration of scales having osteoblasts and osteoclasts using a new magnetic field generator for exposing aquatic animals to EMFs at a sine-wave frequency of 60 Hz. Goldfish scales were put into a fish-breeding space with the developed magnetic field generator and exposed to extremely low-frequency electromagnetic fields (ELF-EMFs) of 60 Hz at an intensity of 1, 3, and 5 mT for 10 days while being regenerated the scales. After exposure, alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) activities in the goldfish scales were measured as markers of osteoblasts and osteoclasts, respectively. As a result, both ALP and TRAP activities in regenerating scales exposed to 3 mT ELF-EMFs were higher than those in regenerating scales exposed to 1 and 5 mT ELF-EMFs. Exposure of scales to 3 mT ELF-EMFs significantly enhanced the scale regeneration rate. Exposure of rat calvaria to 3 mT ELF-EMFs also increased both ALP and TRAP activities like in goldfish scales. Thus, we concluded that 3 mT ELF-EMFs contribute to the medical treatment of bone diseases.

Histochemical Analysis and Distribution of Digestive Enzymes in the Gastrointestinal System of the European Barracuda Sphyraena sphyraena (Linnaeus, 1758).

In this study, we examined the gastrointestinal tract of the European barracuda (Sphyraena sphyraena) living in the Adriatic Sea near Dubrovnik, Croatia. The study aimed to identify the presence and distribution of four digestive enzymes: alkaline phosphatase, aminopeptidase, acid phosphatase, and non-specific esterase. We found that alkaline phosphatase activity was present in the brush border of the enterocytes in all the investigated intestinal segments. The activity of the alkaline phosphatase was the strongest in the pyloric caeca but strong only in the basal part of the intestinal villi in the anterior and middle intestinal segments. In the posterior intestinal segment, alkaline phosphatase had strong activity along the entire villi. The activity of acid phosphatase was weak in all the investigated parts of the intestine, except in the posterior part, where it was moderate. Aminopeptidase was detected in the brush border of the intestinal epithelium, with stronger activity in the pyloric caeca and anterior part of the intestine and weaker activity posteriorly. The activity of the non-specific esterase was moderate in the pyloric caeca and anterior part of the intestine, while it was weak in the posterior segment and the lamina propria in all parts of the digestive tract. Weak acid phosphatase activity was observed only in the lamina propria of the posterior part of the intestine. This study is the first to evaluate the activity of digestive enzymes in the European barracuda.

Osteogenic and osteoclastic differentiation by control release of adenosine within PEI/HA/Collagen composite coatings

In virtue of the rapid metabolism and short half-life of adenosine (AD) in the human body, it is highly desired to explore new adenosine delivery system with controlled and steady release capacity. In this study, composite coatings consisting of polyethyleneimine (PEI), hyaluronic acid (HA) and collagen type I (COL-1) on titanium substrate surface are developed by layer-by-layer self-assembly technique, in which different concentrations of AD are encapsulated. In-vitro release study demonstrated a steady and sustained release of AD from the composite coating system. Adenosine within composite coatings depicted a concentration-dependent regulatory effect on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and osteoclastic differentiation of mouse leukemia cells of monocyte macrophage (RAW264.7) cells. All of these coatings exhibited good cell viability and promoted osteoblast cell proliferation. Among various composite coatings, the optimized coating specimen with loading of 65 μg AD (AD2) exhibits the best osteoconductive and osteoinductive activities with significantly enhanced osteoblast-associated gene expressions, and increased alkaline phosphatase (ALP) activity and calcium deposits. On the other hand, AD2 specimen shows the activity of inhibiting osteoclast differentiation and osteobclast-associated gene expressions as well as suppressing tartrate-resistant acid phosphatase (TRAP) activity. The superior performance of AD/PEI/HA/COL composite coatings indicated that this coating system could be useful for bone tissue engineering.

A superior chitin product: Black soldier fly larvae chitin, beneficial to growth performance, muscle quality and health status of largemouth bass Micropterus salmoides, in comparison to shrimp chitin

We appraised the effects of black soldier fly larvae chitin (BSFC) with four levels (0.4 %, 0.8 %, 1.2 % and 1.6 %) and shrimp chitin (SC) with three levels (0.4 %, 0.8 % and 1.6 %) in diets on the growth performance, nutrient composition and health status of largemouth bass (8.00 ± 0.29 g) fed for 56 days. The results showed that the dietary 0.4 % BSFC distinctly increased the specific growth rate and weight gain rate of fish (P < 0.05), and evidently promoted the deposition of C22:1n-9, C24:1n-9 and increased total monounsaturated fatty acid content in the muscle (P < 0.05). BSFC and SC distinctly stimulated the secretion of β-N-acetylglucosaminidase in the hindgut (P < 0.05). The antioxidant status of fish was improved by BSFC and SC, which was attributed to the decreased malonaldehyde contents in the muscle and midgut and the increased total superoxide dismutase and catalase activities in the midgut. The 1.2 %–1.6 % BSFC and 0.8 %–1.6 % SC enhanced the non-specific immunity of fish, which was attributed to the marked promotion of lysozyme, acid phosphatase and alkaline phosphatase activities by BSFC and the distinct enhancement of lysozyme activity by SC in the serum or midgut (P < 0.05). BSFC and SC notably reduced the abundance of potential pathogenic bacterium Vibrio and Photobacterium (P < 0.05), and 0.4 %, 1.2 %, 1.6 % BSFC and 0.4 % SC markedly increased the abundance of probiotic bacterium Cetobacterium (P < 0.05), and the BSFC and SC of 0.8 % significantly increased the abundance of the probiotics Bifidobacterium and Bacillus respectively (P < 0.05). The increase of acetic acid, propionic acid, butyric acid and valeric acid concentrations in the hindgut might be related to the increase of intestinal probiotics, and 0.8 %–1.6 % BSFC and 0.4 % SC obviously increased the total short-chain fatty acid levels in the hindgut (P < 0.05). BSFC and SC remarkably reduced muscle fiber area and diameter (P < 0.05), while fiber densities were significantly increased under 0.4 %, 0.8 % BSFC and 0.4 % SC supplementation (P < 0.05), which was accordant with increased mRNA expression levels of myod, myf-5, smad-2, tgf-β1 and decreased mRNA expression levels of myos in the muscle. Collectively, BSFC and SC had no negative effect on the growth of largemouth bass, and improved the health status of fish. The dietary 0.4 % BSFC supplementation was beneficial to fish growth and muscle quality, and BSFC was a superior chitin product in comparison to SC.

Screening high-efficiency promoter to construct trans-vp28 gene Anabaena sp. PCC7120 against white spot syndrome virus of Litopenaeus vannamei

This study aimed to select high-quality promoters to construct trans-vp28 gene Anabaena sp. PCC7120 and feed Litopenaeus vannamei to assess the effect of L.vannamei against white spot syndrome virus (WSSV). Transgenic algae were created using five plasmids containing PrbcL, Pcpc560, Ptrc, Ptac, and PpsbA. According to the gene expression efficiency and the growth index of transgenic algae, Pcpc560 was determined to be the most efficient promoter. Shrimps were continuously fed trans-vp28 gene Anabaena sp. PCC7120 for one week and then challenged with WSSV. After the challenge, the transgenic algae group (vp28-7120 group) was continuously immunized [continuous immunization for 0 days (vp28-7120-0d); continuous immunization for 2 days (vp28-7120-2d); continuous immunization for 4 days (vp28-7120-4d)]. After seven days, the daily survival rate of each experimental group was continuously tracked. Following the viral challenge, the hepatopancreas samples were assayed for their levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), thioredoxin peroxidase (TPX), acid phosphatase (ACP), and alkaline phosphatase (AKP) at varying time intervals. In comparison to the positive control group (challenge and no vaccination) and the wild-type group (challenge, fed wild-type Anabaena sp. PCC7120), the vp28-7120 group (challenge, fed trans-vp28 gene Anabaena sp. PCC7120) exhibited a remarkable increase in survival rates, reaching 50 % (vp28-7120-0d), 76.67 % (vp28-7120-2d), and 80 % (vp28-7120-4d). Furthermore, the vp28-7120 group consistently displayed significantly higher activities of SOD, CAT, GSH-Px, ACP, and AKP, while exhibiting notably lower TPX activity, when compared to the control group. These results indicate that the Pcpc560 promoter effectively elevated the expression level of the exogenous vp28 gene and spurred the growth of the trans-vp28 gene Anabaena sp. PCC7120. Consequently, trans-vp28 gene Anabaena sp. PCC7120 significantly bolstered the immunity of L.vannamei. Therefore, utilizing the Pcpc560 promoter to develop trans-vp28 gene Anabaena sp. PCC7120 based oral vaccine is highly beneficial for industrial-scale cultivation, advancing its commercialization prospects.

Biochar and wood vinegar altered the composition of inorganic phosphorus bacteria community in saline-alkali soils and promoted the bioavailability of phosphorus

As an important part of the ecosystem, saline-alkali soils are in urgent need of efficient and environmentally friendly soil conditioners. Biochar and wood vinegar are regarded as organic soil improvement and plant growth regulators to improve soil physicochemical properties and promote crop growth. However, the mechanism of how inorganic phosphorus bacteria increase phosphorus when biochar and wood vinegar applied to saline-alkali soils is not clear. Herein, the present study was designed to investigate the effects of biochar and wood vinegar with different rates on physicochemical properties of saline-alkali soils and inorganic phosphorus bacteria diversities and to discuss the mechanism of biochar and wood vinegar on available phosphorus by pot experiments. The application of biochar and wood vinegar exhibited an effect on the decrease in pH and salt contents and the increase in soil porosity, soil nutrients, and hundred-grain weight of rice. The 600 kg ha−1 biochar and 1800 kg ha−1 wood vinegar group showed the most significant increment in available phosphorus, alkaline phosphatase, acid phosphatase, and neutral phosphatase activities, with the increases of 49.24%, 40.35%, 48%, and 149%, respectively. The 600 kg ha−1 biochar and 1200 kg ha−1 wood vinegar group significantly enhanced microbial biomass phosphorus concentrations by 41.29%. Moreover, biochar and wood vinegar shifted inorganic phosphorus bacteria composition structure and promoted its diversities, more so at a higher rate of wood vinegar application. The dominant species of inorganic phosphorus bacteria were Proteobacteria, Gammaproteobacteria, Alphaproteobacteria, Pseudomonas, and Rhizobium in saline-alkali soils. The Alphaproteobacteria and Hydrogenophaga were the key microorganisms reducing pH and salt contents and increasing available phosphorus contents in saline-alkali soils. In conclusion, the application of biochar and wood vinegar was a useful strategy to improve saline-alkali soils.