Enzymes Acid Phosphatase Research Articles

Biochar, Organic Fertilizer, and Bio-Organic Fertilizer Improve Soil Fertility and Tea Quality

Tea, the world’s second most traded commodity, significantly impacts the economies of producing countries. However, prolonged cultivation leads to soil degradation, particularly through acidification and the depletion of essential nutrients, which adversely affects tea quality. This study investigates the effects of biomass materials—biochar, organic fertilizer, and bio-organic fertilizer—on both tea quality and soil properties. The results revealed that all biomass treatments improved the catechin quality index (CQI) of tea, with bio-organic fertilizer (BOF) yielding the highest CQI at 629.41, followed closely by biochar (624.16) and organic fertilizer (581.34). Soil analysis indicated that biochar increased soil pH from 4.53 to 5.54, total carbon by 194.6% (from 12.61 g kg−1 to 24.42 g kg−1), and nitrogen levels by 11.7% (from 14.91 mg kg−1 to 16.17 mg kg−1), while reducing soluble salts significantly. Furthermore, biomass treatments enhanced enzyme activities, with urease and acid phosphatase increasing by up to 136.6% and 149.5%, respectively. Correlation analysis revealed significant positive relationships, with tea amino acid content correlating with soil total nitrogen (r = 0.62, p < 0.05) and tea polyphenols positively correlating with available potassium (r = 0.60, p < 0.05). This study demonstrates that integrating biomass materials into tea cultivation not only enhances tea quality but also contributes to soil health, supporting sustainable tea garden management practices.

Biomarkers of oxidative stress, biochemical changes, and the activity of lysosomal enzymes in the livers of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against yersiniosis before a Yersinia ruckeri challenge

Abstract Introduction This study aimed to evaluate biomarkers of oxidative stress (2-thiobarbituric acid reactive substances, aldehyde and ketone derivatives of oxidatively modified proteins and total antioxidant capacity), the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase), that of lysosomal enzymes (alanyl aminopeptidase, leucyl aminopeptidase, β-N-acetylglucosaminidase and acid phosphatase) and changes in biochemical parameters (alanine aminotransferase, aspartate aminotransferase, de Ritis ratio, lactate dehydrogenase activity, lactate and pyruvate levels and their ratio) in the liver tissue of fish that were vaccinated against enteric redmouth disease and challenged with its causative agent, the bacterium Yersinia ruckeri. Material and Methods The vaccine was administered orally to trout, some of which were challenged with Y. ruckeri 61 days later. For comparison, unvaccinated and unchallenged trout and unvaccinated and challenged trout were also evaluated. Results In the unvaccinated fish, infection with Y. ruckeri disrupted the pro-oxidant/antioxidant balance, led to a significant increase in lipid peroxidation and oxidative modification of proteins, decreased total antioxidant capacity and significantly increased the activity of lysosomal enzymes. In vaccinated fish, the Y. ruckeri challenge increased the activity of glutathione-related enzymes and decreased lipid peroxidation, anaerobic metabolism and the activity of lysosomal enzymes in fish livers relative to the unvaccinated and challenged group. In contrast, these parameters increased after the Y. ruckeri challenge in unvaccinated trout relative to those in the untreated group. Conclusion Vaccination exerted a protective effect during the Y. ruckeri challenge and had no adverse effect on fish livers.

Impact of Intercropping Five Medicinal Plants on Soil Nutrients, Enzyme Activity, and Microbial Community Structure in Camellia oleifera Plantations.

Intercropping medicinal plants plays an important role in agroforestry that can improve the physical, chemical, and biological fertility of soil. However, the influence of intercropping medicinal plants on the Camellia oleifera soil properties and bacterial communities remains elusive. In this study, five intercropping treatment groups were set as follows: Curcuma zedoaria/C. oleifera (EZ), Curcuma longa/C. oleifera (JH), Clinacanthus nutans/C. oleifera (YDC), Fructus Galangae/C. oleifera (HDK), and Ficus simplicissima/C. oleifera (WZMT). The soil chemical properties, enzyme activities, and bacterial communities were measured and analyzed to evaluate the effects of different intercropping systems. The results indicated that, compared to the C. oleifera monoculture group, YDC and EZ showed noticeable impacts on the soil chemical properties with a significant increase in total nitrogen (TN), nitrate nitrogen (NN), available nitrogen (AN), available phosphorus (AP), and available potassium (AK). Among them, the content of TN and AK in the rhizosphere soil of Camellia oleifera in the YDC intercropping system was the highest, which was 7.82 g/kg and 21.94 mg/kg higher than CK. Similarly, in the EZ intercropping system, the content of NN and OM in the rhizosphere soil of Camellia oleifera was the highest, which was higher than that of CK at 722.33 mg/kg and 2.36 g/kg, respectively. Curcuma longa/C. oleifera (JH) and Clinacanthus nutans/C. oleifera (YDC) had the most effect on soil enzyme activities. Furthermore, YDC extensively increased the activities of hydrogen peroxide and acid phosphatase enzymes; the increase was 2.27 mg/g and 3.21 mg/g, respectively. While JH obviously increased the urease activity, the diversity of bacterial populations in the rhizosphere soil of the intercropping plants decreased, especially the Shannon index of YDC and HDK. Compared with the monoculture group, the bacterial community abundance and structure of JH and YDC were quite different. The relative abundance of Actinobacteriota and Firmicutes was increased in YDC, and that of Acidobacteriota and Myxococcota was increased in JH. According to the redundancy analysis (RDA), pH, total potassium, and soil catalase activity were identified as the main factors influencing the microbial community structure of the intercropping systems. In conclusion, intercropping with JH and YDC increased the relative abundance of the dominant bacterial communities, improved the microbial community structure, and enhanced the soil nutrients and enzyme activities. Therefore, in the future, these two medicinal plants can be used for intercropping with C. oleifera.

Phosphorus addition stimulates overall carbon acquisition enzymes but suppresses overall phosphorus acquisition enzymes: A global meta-analysis

Of the total land worldwide, 43 % is phosphorus (P)-constrained, which is significantly greater than the proportion of nitrogen-constrained land (18 %). However, there are some uncertainties regarding the responses of enzyme activities to P addition on a global scale. We collected data from 99 publications and performed a meta-analysis to assess the responses of 11 extracellular enzymes, including carbon, nitrogen, and phosphorus acquisition enzymes, and oxidases. P addition significantly increased the activities of β-1, 4-glucosidase, sucrase, cellobiohydrolase, and alkaline phosphatase, while lowering the activities of acid phosphatase and phosphodiesterase. The responses of enzyme activities to P addition differed considerably across the three ecosystems (forest, grassland, and cropland). P addition significantly decreased the activities of acid phosphatase, phosphodiesterase, and P-acquisition enzymes in forests, but increased the activities of leucine aminopeptidase, carbon acquisition enzymes, and nitrogen acquisition enzymes in grasslands. Sucrase, β-1,4-glucosidase, N-acetyl-β-glucosaminidase, alkaline phosphatase, and carbon acquisition enzymes were significantly stimulated in croplands. In addition, all P addition rates significantly increased the activity of alkaline phosphatase, a P-acquisition enzyme, in croplands. Ca(H2PO4)2 had a greater effect on enzyme activity than other fertilizer types, possibly because Ca2+ can improve the soil structure and enhance microbial metabolic activities. Furthermore, structural equation models revealed that mean annual precipitation and ecosystem type had significant impacts on the response of P-acquisition enzymes, but no factor could directly influence the response of carbon-acquisition enzymes. Our investigation of enzyme activities in response to P addition complements the biogeochemical model and helps forecast soil nutrient dynamics. We propose that rather than focusing on a single factor, P fertilizer addition programs should take into account the effects of mean annual precipitation, ecosystem type, and nutrient interactions, which could also reduce P pollution.

Effects of leguminous green manure-crop rotation on soil enzyme activities and stoichiometry

Abstract As a crucial strategy for sustainable agricultural production, green manure-crop rotation can regulate soil nutrient cycling and decrease the reliance on nitrogen fertilizers. However, we still lack a comprehensive understanding of the changes in soil eco-enzyme activities, microbial metabolism, and nutrient limitations caused by leguminous green manure crop rotation. Here, we conducted field experiments of leguminous green manure-crop rotation across China to analyze soil extracellular enzyme activities, specifically β-glucosidase (BG), N-acetyl-β-D-glucosaminidase (NAG), leucine aminopeptidase (LAP), and acid phosphatase (AP). The study revealed that long-term green manure-crop rotation increased carbon and nitrogen accumulation in farmland, with a significant average increase of 20.1% and 36.4% in BG, AP enzyme activities in topsoil, while showing a decrease in ln(NAG+LAP):ln(AP) ratios. The ratios of ln(BG):ln(NAG+LAP) and ln(NAG+LAP):ln(AP) in soil across various regions were typically below 1:1, indicating that soil microbial activity is more constrained by nitrogen and phosphorus nutrients rather than by carbon. Precipitation, temperature, soil total carbon, and total nitrogen were identified as key environmental factors for extracellular enzyme activities and stoichiometric ratios. Our study highlights that the green manure-crop rotation alleviates nitrogen limitation while enhancing phosphorus limitation, and is closely related to the accumulation of total carbon and nitrogen in the soil.

The Short-term Exposure Effect of Chlorpyrifos (20% EC) on Haematological, Biochemical and Histopathological Response of Striped Catfish Pangasianodon hypophthalmus

An experiment of acute toxicity was conducted to find out the lethal concentration 50 (LC50) of chloropyrifos (CPF) for Pangasianodon hypophthalmus. Our findings indicated that the 96-h LC50 value of CPF for P. hypophthalmus was 0.106 mg L-1. Effects of different concentrations of CPF (0.09, 0.1, 0.11, 0.12, and 0.13 mg L-1) for 96-h on hemato-biochemical, enzyme and histological effects were evaluated in this study. Acute exposure induced significant differences in enzymes acid phosphatase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and acetylcholine of blood serum of the treated group as compared to control group. However, significant decreases in serum total protein, globulin, albumin, and triglycerides levels while serum glucose enhanced with consequent increasing doses of CPF exposure. Also, CPF induced significant declines in RBC count, haemoglobin and hematocrit level whereas the exposure groups showed significant rises in WBC and blood indices levels as compared to control group. Furthermore, CPF exposure causes gill curling, epithelium lifting, necrosis, and lamellae degeneration. Hemosiderin, sinusoid dilations, necrosis, malanomacrophage, hepatocyte hypertrophy were observed in liver histology. Reduction of lumen tubule, hyperplasia, increased bowman space and degenerations were seen in kidney. Overall findings indicates that the acute CPF exposure to P. hypophthalmus resulted in significant adverse changes in biochemical, hematological, and histopathological response.

Field-scale assessment of vermicompost amendments for diuron-contaminated soil: Implications for soil quality and pesticide fate

This comprehensive study investigated the distribution, dissipation, and environmental impact of diuron and its metabolites (DPMU, DPU, and DCA), as well as its effects on enzyme activities, bacterial abundance, and community composition in response to different soil treatments using two vermicomposts. One vermicompost was derived from vine shoot waste, and the other from wet olive cake (alperujo). The study was conducted under field conditions in a semi-arid region. The results indicated that diuron concentrations decreased with soil depth, with the highest levels in the upper 0–5 cm layer. The application of both vermicomposts reduced diuron leaching into the soil, facilitating its dissipation compared to the organically unamended soil. DPMU was the main metabolite detected in the soil, especially at the end of the experiment. Enzyme activities, including dehydrogenase, β-glucosidase, protease, and acid phosphatase, showed variations with treatment and time, indicating microbial adaptation. In general, both vermicomposts increased soil enzyme activities, even when diuron was co-applied. Bacterial communities were affected by diuron and vermicomposts, with Actinobacteria and Alphaproteobacteria showing significant shifts in relative abundance. Principal Component Analysis (PCA) confirmed structural changes in bacterial communities due to diuron and vermicomposts. Warmer temperatures and evapotranspiration may contribute to the distribution of diuron in the different layers of soil and affect microbial activity. Overall, vermicomposts from wet olive cake proved effective in mitigating diuron in the different soil layers, promoting enzyme activities, and influencing bacterial communities. These findings underscore the complexity of interactions between diuron, organic amendments, and soil microorganisms, highlighting the potential for enhanced herbicide dissipation and microbial resilience and adaptation.

Combined effects of polyethylene terephthalate and abamectin on enzymatic activity and histopathology response in juvenile zebrafish (Danio rerio).

One of the most pressing global environmental issues is the widespread abundance and distribution of microplastics (MPs). MPs can act as vectors for other contaminants in the environment making these small plastic particles hazardous for ecosystems. The presence of MPs in aquatic environments may pose threats to aquatic organisms that ingest them. This study examined effects of abamectin (ABM) and polyethylene terephthalate (PET) MP fragments on histopathological and enzymatic biomarkers in zebrafish (Danio rerio). Zebrafish were exposed for 96 h to pristine PET-MPs at concentrations of 5 mg/L and 10 mg/L, ABM alone at 0.006 mg/L, and the same concentration of ABM in the presence of PET-MPs in aquaria. Histopathological analysis revealed tissue content changes in liver and kidney in the presence of ABM individually and in combination with MPs. Results of enzymatic analysis showed that MPs increased the bioavailability and toxicity of pesticides due to inhibition of catalase (CAT) and acid phosphatase (ACP) enzymes. However, MPs did not affect the toxicity of ABM for glutathione s-transferase (GST) enzyme. Despite the inhibition of acetylcholinesterase (AChE) in MPs or ABM treatments, and some neurotoxicity, no change in activity of this enzyme and neurotoxicity was observed in the combined MPs and ABM treatments, although toxicity effects of MPs and ABM on zebrafish require more detailed studies.

Impact of soil moisture regimes on greenhouse gas emissions, soil microbial biomass, and enzymatic activity in long-term fertilized paddy soil

Two potent greenhouse gases that are mostly found in agricultural soils are methane and nitrous oxide. Therefore, we investigated the effect of different moisture regimes on microbial stoichiometry, enzymatic activity, and greenhouse gas emissions in long-term paddy soils. The treatments included a control (CK; no addition), chemical fertilizer (NPK), and NPK + cattle manure (NPKM) and two moisture regimes such as 60% water-filled pore spaces (WFPS) and flooding. The results revealed that 60% water-filled pore spaces (WFPS) emit higher amounts of N2O than flooded soil, while in the case of CH4 the flooded soil emits more CH4 emission compared to 60% WFPS. At 60% WFPS higher N2O flux values were recorded for control, NPK, and NPKM which are 2.3, 3.1, and 3.5 µg kg−1, respectively. In flooded soil, the CH4 flux emission was higher, and the NPKM treatment recorded the maximum CH4 emissions (3.8 µg kg−1) followed by NPK (3.2 µg kg−1) and CK (1.7 µg kg−1). The dissolved organic carbon (DOC) was increased by 15–27% under all flooded treatments as compared to 60% WPFS treatments. The microbial biomass carbon, nitrogen, and phosphorus (MBC, MBN, and MBP) significantly increased in the flooded treatments by 8–12%, 14–21%, and 4–22%, respectively when compared to 60% WFPS. The urease enzyme was influenced by moisture conditions, and significantly increased by 42–54% in flooded soil compared with 60% WFPS while having little effect on the β-glucosidase (BG) and acid phosphatase (AcP) enzymes. Moreover DOC, MBC, and pH showed a significant positive relationship with cumulative CH4, while DOC showed a significant relationship with cumulative N2O. In the random forest model, soil moisture, MBC, DOC, pH, and enzymatic activities were the most important factors for GHG emissions. The PLS-PM analysis showed that soil properties and enzymes possessed significantly directly impacted on CH4 and N2O emissions, while SMB had indirect positive effect on CH4 and N2O emissions.

The impact on Cd bioavailability and accumulation in rice (Oryza sativa L.) induced by dry direct-seeding cultivation method in field-scale experiments

Dry direct-seeded rice cultivation has gained popularity and expanded its cultivated area due to reduced labor requirements and water consumption. However, the impact of this cultivation method on cadmium (Cd) bioavailability in soil and the accumulation levels in grains remains uncertain. Field experiments were conducted in acidic soils at two locations in southern China to compare rice varieties and evaluate the dry direct-seeding method alongside the wet direct-seeding and traditional transplanting methods. Dry direct-seeded rice reached significantly higher Cd concentrations in its tissues starting from the heading stage than transplanted rice. Cd accumulation levels by the maturation stage in the brown rice of dry direct-seeded rice were 18.33 %–150.69 % higher than those of wet direct-seeded and transplanted rice, with a considerable ability to translocate Cd into brown rice. Furthermore, dry direct seeding decreased iron plaque formation, particularly in the amorphous Fe form; it resulted in high soil temperature and low moisture content during tillering, elevating Cd availability in the soil. Additionally, the proportion of ions and more labile forms of Cd in the soil solution was high. Moreover, the soil under dry direct seeding had high urease and acid phosphatase enzyme activities. However, low richness and diversity in the bacterial community were characterized by a significant increase in the relative abundance of Actinobacteria and Gammaproteobacteria at the class level, while exhibiting decreased relative abundances of Alphaproteobacteria, Bacilli, and KD4-96, along with fewer biomarkers. Nonetheless, these differences are gradually reduced during the maturation stage. Overall, although dry direct seeding offers several advantages, it is crucial to implement additional measures to mitigate the increased health risks linked to rice cultivation through this approach in Cd-contaminated areas.

Broad-spectrum pH functional chitosan-phosphatase beads for the generation of plant-available phosphorus: utilizing the insoluble P pool.

Utilization of organic phosphates and insoluble phosphates for the gradual generation of plant-available phosphorus (P) is the only sustainable solution for P fertilization. Enzymatic conversions are one of the best sustainable routes for releasing P to soil. Phosphatase enzyme aids in solubilizing organic and insoluble phosphates to plant-available P. We herein report the preparation of highly functional chitosan beads co-immobilized with acid phosphatase and alkaline phosphatase enzymes via a glutaraldehyde linkage. The dual enzyme co-immobilized chitosan beads were characterized using Fourier-transform infrared (FTIR), thermogravimetric (TGA), and scanning electron microscopy-energy dispersive x-ray (SEM-EDX) analyses to confirm the immobilization. The co-immobilized system was found to be active for a broader pH range of ∼4-10 than the individually bound enzymes and mixed soluble enzymes. The bound matrix exhibited pH optima at 6 and 9, respectively, for acid and alkaline phosphatase and a temperature optimum at 50°C. The phosphate-solubilizing abilities of the chitosan-enzyme derivatives were examined using insoluble tri-calcium phosphate (TCP) for wide pH conditions of 5.5, 7, and 8.5 up to 25 days. The liberation of phosphate was highest (27.20mg/mL) at pH 5.5 after the defined period. The residual soil phosphatase activity was also monitored after 7days of incubation with CBE for three different soils of pH ∼5.5, 7, and 8.5. The residual phosphatase activity increased for all the soils after applying the CBE. The germination index of the Oryza sativa (rice) plant was studied using different pH buffer media upon the application of the CBE in the presence of tri-calcium phosphate as a phosphate source. Overall, the dual-enzyme co-immobilized chitosan beads were highly effective over a wide pH range for generating plant-available phosphates from insoluble phosphates. The chitosan-enzyme derivative holds the potential to be used for sustainable phosphorus fertilization with different insoluble and organic phosphorus sources.

RHIZOSPHERE MICROBIOME, SOIL ENZYMES AND GROUNDNUT YIELD AS INFLUENCED BY ADDITION OF BIOCHAR AND FERTILIZERS IN IRRIGATED ALFISOL OF NORTH COASTAL ANDHRA PRADESH

The field experiment was conducted in irrigated Alfisols of North coastal Andhra Pradesh tostudy the effect of biochar and chemical fertilizers on rhizosphere microbiome, soil enzymes andyield performance of groundnut crop (variety K-6) during rabi, 2018-19. Biochar application tosoil significantly increased soil bacterial, fungal and actinomycetes population. At pod developmentstage, the highest number of bacterial count (39.0 x106 CFU g-1 soil) was observed in T5 (100%RDF + biochar @ 6 t ha-1) which was on par with all the biochar applied treatments (T3, T4, T6, T7,T8). The Biochar applied @ 6 t ha-1 (T5 & T8) significantly increased the bacterial population ascompared to non-biochar applied treatments (T1 & T2). Fungal and actinomycetes populationfollowed the similar trend of bacterial count. Soil urease activity was significantly superior inbiochar applied treatments (T3, T4, T5, T6, T7, T8) compared to non biochar applied treatments (T1and T2). With increased rates of biochar application the urease activity markedly increased insoil. Similar trend was noticed with respect to dehydrogenase, acid phosphatase and alkalinephosphatase enzymes in soil in response to addition of biochar. At pod development stage, thehighest leaf area index (3.16) was recorded with 100% RDF + biochar @ 6 t ha-1(T5) which wassignificantly higher than T1 (control), T2 (100% RDF) and T6 (75% RDF + biochar @ 2 t ha-1). Thehighest drymatter accumulation of 2951 kg ha-1 at peg penetration and 6428 kg ha-1 at poddevelopment was observed in T5 (100% RDF + biochar @ 6 t ha-1) which was on par with T3(100% RDF + biochar @ 2 t ha-1), T4 (100% RDF + biochar @ 4 t ha-1), T8 (75% RDF + biochar @6 t ha-1) treatments. Groundnut pod yield was highest (4019.58 kg ha-1) in treatment received100% RDF + biochar @ 6 t ha-1, which was on par with T4 (100% RDF + biochar @ 6 t ha-1) and T8(75% RDF + biochar @ 6 t ha-1).

Effects of Clothianidin on Biochemical Parameters of Adult Zebrafish

The ubiquitous use of insecticides leads to detrimental effects on non-target organisms due to accidental exposure. Neonicotinoid insecticides are popularly used worldwide for their high affinity for arthropod nicotinic receptors which effectively kill insect pests. Low affinity towards vertebrate nicotinic receptors, make them safer as compared to traditional insecticides. Recent studies demonstrated that neonicotinoid exposure can cause some toxicity in vertebrates including humans. Zebrafish is one of the popular model organisms to study ecotoxicity. This is the first study on adult zebrafish to report the effect of novel neonicotinoid, Clothianidin on mortality, liver antioxidant stress profile, liver function profile and brain acetylcholinesterase (AChE). Observations were made over two treatment periods, 96 hours, and 21 days, in five groups exposed to varying concentrations of Clothianidin viz. 30mg/L, 50mg/L, 70mg/L, 90mg/L, 110mg/L and a control. Although no mortality was observed, Clothianidin exposure led to increased activity of superoxide dismutase (SOD) enzyme, lipid peroxidation and decreased catalase (CAT), glutathione-S-transferase (GST). Treated groups also showed increased concentrations of liver enzyme alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and acid phosphatase (ACP). These results indicate that Clothianidin causes disturbances in the hepatic function. A prominent decreasing trend observed in brain AChE shows that Clothianidin inhibits AChE in adult zebrafish. Further investigations on DNA damage and gene expression studies could be conducted to understand the exact mechanism of action of Clothianidin

Warming enhances the negative effects of shrub removal on phosphorus mineralization potential

Shrubs have developed various mechanisms for soil phosphorus utilization. Shrub encroachment caused by climate warming alters organic phosphorus mineralization capability by promoting available phosphorus absorption and mediating root exudates. However, few studies have explored how warming regulates the effects of dominant shrubs on soil organic phosphorus mineralization capability. We provide insights into warming, dominant shrub removal, and their interactive effects on the soil organic phosphorus mineralization potential in the Qinghai-Tibetan Plateau. Real-time polymerase chain reaction was used to quantify the soil microbial phosphatase genes (phoC and phoD), which can characterize the soil organic phosphate mineralization potential. We found that warming had no significant effect on the soil organic phosphate-mineralized components (total phosphate, organic phosphate, and available phosphate), genes (phoC and phoD), or enzymes (acid and alkaline phosphatases). Shrub removal negatively influenced the organic phosphate-mineralized components and genes. It significantly decreased soil organic phosphate mineralization gene copy numbers only under warming conditions. Warming increased fungal richness and buffered the effects of shrub removal on bacterial richness and gene copy numbers. However, the change in the microbial community was not the main factor affecting organic phosphate mineralization. We found only phoC copy number had significant correlation to AP. Structural equation modelling revealed that shrub removal and the interaction between warming and shrub removal had a negative direct effect on phoC copy numbers. We concluded that warming increases the negative effect of shrub removal on phosphorus mineralization potential, providing a theoretical basis for shrub encroachment on soil phosphate mineralization under warming conditions.

Effect of Chitosan Degradation Products, Glucosamine and Chitosan Oligosaccharide, on Osteoclastic Differentiation.

Chitosan, a natural cationic polysaccharide derived from crustaceans and shellfish shells, is known for its advantageous biological properties, including biodegradability, biocompatibility, and antibacterial activity. Chitosan and its composite materials are studied for their potential for bone tissue repair. However, the effects of chitosan degradation products, glucosamine (GlcN) and chitosan oligosaccharide (COS), on osteoclasts remain unclear. If these chitosan degradation products promote osteoclastic differentiation, careful consideration is required for the use of chitosan and related materials in bone repair applications. Here, we assessed the effects of high (500 μg/mL) and low (0.5 μg/mL) concentrations of GlcN and COS on osteoclastic differentiation in human peripheral blood mononuclear cells (PBMCs) and murine macrophage-like RAW264 cells. A tartrate-resistant acid phosphatase (TRAP) enzyme activity assay, TRAP staining, and actin staining were used to assess osteoclastic differentiation. High concentrations of GlcN and COS, but not low concentrations, suppressed macrophage colony-stimulating factor (M-CSF)- and RANKL-dependent increases in TRAP enzyme activity, TRAP-positive multinuclear osteoclast formation, and actin ring formation in PBMCs without cytotoxicity. Similar effects were observed in the RANKL-dependent osteoclastic differentiation of RAW264 cells. In conclusion, chitosan degradation products do not possess osteoclast-inducing properties, suggesting that chitosan and its composite materials can be safely used for bone tissue repair.

CRITICAL LEVELS AND DOSES OF MAXIMUM TECHNICAL EFFICIENCY OF PHOSPHORUS IN Handroanthus heptaphyllus SUBJECTED TO PHOSPHORUS FERTILIZATION

Tropical and subtropical soils do not always provide the necessary amount of phosphorus (P) to supply the nutrient demand of native trees, such as Handroanthus heptaphyllus (Mart.) Mattos (Ipê-roxo). Therefore, it becomes necessary to apply phosphate fertilizers. However, decision-making and doses to be applied will be more assertive if P's critical levels (NC) in the soil and leaves and doses of maximum technical efficiency (MET) are defined to improve the fertilizer recommendation system for native trees. The study aimed to propose NC and MET of P in H. heptaphyllus submitted to P application. The experiment was conducted in Red Argisol submitted to the application of five doses of P (0, 20, 40, 80, and 160 kg ha-1). After 24 months of planting, the P content in the soil and its concentration in the leaf tissue, height growth (H) and stem diameter (DC), chlorophyll A fluorescence, and acid phosphatase enzyme activity were determined. The NC in soil and leaves were calculated. The addition of phosphorus fertilization increased the P levels in the soil and leaves, promoting higher H and DC in the H. heptaphyllus plants. The greater availability of P reduced photochemical energy losses by fluorescence. The NC in the soil for growth variables height and stem diameter was 20.0 and 21.2 mg dm-3. The CN in leaves for growth and physiological parameters was, on average, 1.6 g kg-1.

Lysosomal enzymes and the oxygen burst capability of monocyte-derived macrophages in active drug-resistant tuberculosis patients in relation to cell attachment

Drug resistance to tuberculosis (TB) has become an obstacle in eliminating tuberculosis. The transmission of drug-resistant TB from patients increases the incidence of primary drug-resistant (DR) TB in individuals who are in close contact. Therefore, it is necessary to incorporate an immunological approach into preventive therapy. This study focuses on the activity of lysosomal enzymes, oxygen bursts, and the attachment ability of macrophages among individuals diagnosed with active drug-resistant TB compared with close contacts with latent TB or healthy cases. We measured macrophage oxygen burst ability (Water-soluble tetrazolium salt (WST) test, Nitric Oxide production, and myeloperoxidase activity) and the degradative ability of lysosomes (activity of the β-glucuronidase and acid phosphatase enzymes). Six active DR-TB patients and 18 close-contact cases (8 Latent Tuberculosis Infection (LTBI); 10 healthy) were recruited at Universitas Indonesia Hospital. The macrophage attachment of the LTBI group was higher than in the other groups. NO production, myeloperoxidase activity, β-glucuronidase, and acid phosphatase were higher in the active DR-TB group. A negative correlation was uncovered between phagocytosis and NO production, myeloperoxidase activity, and lysosomal enzymes. The difference in macrophage function is expected to be a further reference in active DR-TB treatment or preventive therapy.

Soil properties and quinoa yield as affected by irrigation and vermicompost application under a drip-tape irrigation system

IntroductionQuinoa a crop with high nutritional value has the widespread adaptability to different climates and there is an increased demand for this product in the world. Characterized by roots with extensive penetration capabilities and wide, alternate leaves giving rise to diverse inflorescences, quinoa plants exhibit variable heights ranging from 20 to 300 cm. The duration of their growth phase spans 95 to 125 days’ contingent upon cultivar selection and prevailing climatic conditions.Materials and methodsIn order to reduce chemical fertilizers consumption through using organic fertilizers a two-year study was conducted at the Research Farm of the University of Kurdistan, Iran, during the 2021–2022 growing seasons. In this study the effects of varying levels of irrigation and vermicompost application on soil characteristics and quinoa (Chenopodium quinoa) yield were investigated. The experiments were arranged as split plots within a randomized complete block design. The main factor consisted of four irrigation levels (50%, 75%, 100%, and 125% of quinoa’s water requirement), while the sub-factor encompassed four vermicompost application rates (0, 5, 10, and 15 tons per hectare). The irrigation method employed was a drip-tape irrigation system with a seven-day interval between irrigations, and water quantities were measured using a volumetric meter on the main pipeline.ResultsThe results demonstrated that the highest levels of available soil phosphorus (21.83 mg kg−1) and soil ammonium (36.08 mg kg−1) were observed in the treatment receiving 50% of quinoa’s water requirement combined with 15 tons per hectare of vermicompost. Additionally, the application of 15 tons per hectare of vermicompost led to the highest concentrations of soil nitrate (14.16 mg kg−1), available potassium (144.62 mg kg−1), and quinoa seed yield (1784.01 kg ha−1)). Over both years, the greatest activity of alkaline and acid phosphatase enzymes was noted in the treatment receiving 125% of quinoa’s water requirement in combination with 15 tons per hectare of vermicompost. Vermicompost application was found to enhance yield and ameliorate drought stress by enhancing soil physical and chemical properties and improving soil moisture retention.

Identifying the in vivo-induced antigenic genes is a strategy to develop DNA vaccine against Nocardia seriolae in hybrid snakehead (Channa maculata ♀ × Channa argus ♂)

Nocardia seriolae has been identified as the causative agent of fish nocardiosis, resulting in serious economic losses in aquaculture. With an aim to screen potential candidates for vaccine development against N. seriolae, the in vivo-induced genes of N. seriolae in hybrid snakehead (Channa maculate ♀ × Channa argus ♂) model were profiled via in vivo-induced antigen technology (IVIAT) in the present study, and 6 in vivo-induced genes were identified as follows: IS701 family transposase (is701), membrane protein insertase YidC (yidC), ergothioneine biosynthesis glutamate-cysteine ligase (egtA), molybdopterin respectively-dependent oxidoreductase (mol), phosphoketolase family protein (Ppl), hypothetical protein 6747 (hp6747). Additionally, the yidC was inserted into eukaryotic expression vector pcDNA3.1-myc-his-A to construct a DNA vaccine named as pcDNA-YidC to evaluate immunoprotection in hybrid snakehead after artificial challenge with N. serioale. Results showed that the transcription of yidC was detected in spleen, trunk kidney, muscle and liver in vaccinated fish, suggesting that this antigenic gene can be recombinantly expressed in fish. Meanwhile, indexes of humoral immunity were evaluated in the vaccinated fish through assessing specific-antibody IgM and serum enzyme activities, including lysozyme (LZM), superoxide dismutase (SOD), acid phosphatase (ACP) and alkaline phosphatase (AKP). Quantitative real-time PCR analysis indicated that pcDNA-YidC DNA vaccine could notably enhance the expression of immune-related genes (CD4、CD8α、MHCIIα、TNFα、IL-1β and MHCIα) in 4 tissues (spleen, trunk kidney, muscle and liver) of the vaccinated fish. Finally, an immuno-protection with a relative survival rate of 65.71 % was displayed in vaccinated fish in comparison to the control groups. Taken together, these results indicate that pcDNA-YidC DNA vaccine could boost strong immune responses in hybrid snakehead and show preferably protective efficacy against N. seriolae, indicating that IVIAT is a helpful strategy to screen the highly immunogenic antigens for vaccine development against fish nocardiosis.

Effects of Photoperiod on Survival, Growth, Physiological, and Biochemical Indices of Redclaw Crayfish (Cherax quadricarinatus) Juveniles

Through a 30-day experiment, this study investigated the effects of five photoperiods (0L:24D, 6L:18D, 12L:12D, 18L:6D, and 24L:0D) on the survival, enzyme activity, body color, and growth-related gene expression of redclaw crayfish (Cherax quadricarinatus) juveniles. The results showed that C. quadricarinatus juveniles under 18L:6D and 24L:0D photoperiods exhibited the highest survival rate, which was significantly higher than the survival rates of juveniles under the other three photoperiods (p < 0.05). However, the 0L:24D group had the highest final body weight and weight gain rate, significantly surpassing those of the 12L:12D, 18L:6D, and 24L:0D groups (p < 0.05). Regarding enzyme activity and hormone levels, juveniles under the 18L:6D photoperiod exhibited relatively higher activity of superoxide dismutase (SOD), acid phosphatase (ACP), and lysozyme (LZM) enzymes than those under other photoperiods, but their levels of melatonin and cortisol were relatively low. In addition, the 24L:0D group showed the highest malondialdehyde (MDA) content. Analysis of gene expression levels revealed that retinoid X receptor (RXR) and α-amylase (α-AMY) genes in C. quadricarinatus juveniles exhibited significantly higher expression levels under the 18L:6D photoperiod than those under the other four photoperiods (p < 0.05). With increasing daylight exposure, the body color of C. quadricarinatus changed from pale blue to yellow–brown. In summary, C. quadricarinatus juveniles achieved high survival rates, good growth performance, strong antioxidant stress response, and immune defense capabilities under an 18 h photoperiod. Therefore, in the industrial seedling cultivation of redclaw crayfish, it is recommended to provide 18 h of daily light. Further, the study demonstrated the ability to manipulate the body color of C. quadricarinatus through controlled artificial photoperiods. These findings provide essential technical parameters needed for the industrial cultivation of C. quadricarinatus juveniles.