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1.
Heliyon ; 10(13): e33995, 2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-39091955

RESUMO

Globally, breast cancer is a primary contributor to cancer-related fatalities and illnesses among women. Consequently, there is a pressing need for safe and effective treatments for breast cancer. Bioactive compounds from endophytic fungi that live in symbiosis with medicinal plants have garnered significant interest in pharmaceutical research due to their extensive chemical composition and prospective medicinal attributes. This review underscores the potentiality of fungal endophytes as a promising resource for the development of innovative anticancer agents specifically tailored for breast cancer therapy. The diversity of endophytic fungi residing in medicinal plants, success stories of key endophytic bioactive metabolites tested against breast cancer and the current progress with regards to in vivo studies and clinical trials on endophytic fungal metabolites in breast cancer research forms the underlying theme of this article. A thorough compilation of putative anticancer compounds sourced from endophytic fungi that have demonstrated therapeutic potential against breast cancer, spanning the period from 1990 to 2022, has been presented. This review article also outlines the latest trends in endophyte-based drug discovery, including the use of artificial intelligence, machine learning, multi-omics approaches, and high-throughput strategies. The challenges and future prospects associated with fungal endophytes as substitutive sources for developing anticancer drugs targeting breast cancer are also being highlighted.

2.
BMC Plant Biol ; 24(1): 642, 2024 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-38972980

RESUMO

Among the several threats to humanity by anthropogenic activities, contamination of the environment by heavy metals is of great concern. Upon entry into the food chain, these metals cause serious hazards to plants and other organisms including humans. Use of microbes for bioremediation of the soil and stress mitigation in plants are among the preferred strategies to provide an efficient, cost-effective, eco-friendly solution of the problem. The current investigation is an attempt in this direction where fungal strain PH1 was isolated from the rhizosphere of Parthenium hysterophorus which was identified as Aspergillus niger by sequence homology of the ITS 1 and ITS 4 regions of the rRNA. The strain was tested for its effect on growth and biochemical parameters as reflection of its potential to mitigate Pb stress in Zea mays exposed to 100, 200 and 500 µg of Pb/g of soil. In the initial screening, it was revealed that the strain has the ability to tolerate lead stress, solubilize insoluble phosphate and produce plant growth promoting hormones (IAA and SA) and other metabolites like phenolics, flavonoids, sugar, protein and lipids. Under 500 µg of Pb/g of soil, Z. mays exhibited significant growth retardation with a reduction of 31% in root length, 30.5% in shoot length, 57.5% in fresh weight and 45.2% in dry weight as compared to control plants. Inoculation of A. niger to Pb treated plants not only restored root and shoot length, rather promoted it to a level significantly higher than the control plants. Association of the strain modulated the physio-hormonal attributes of maize plants that resulted in their better growth which indicated a state of low stress. Additionally, the strain boosted the antioxidant defence system of the maize there by causing a significant reduction in the ascorbic acid peroxidase (1.5%), catalase (19%) and 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging activity (33.3%), indicating a lower stress condition as compared to their non-inoculated stressed plants. Based on current evidence, this strain can potentially be used as a biofertilizer for Pb-contaminated sites where it will improve overall plant health with the hope of achieving better biological and agricultural yields.


Assuntos
Antioxidantes , Aspergillus niger , Chumbo , Fosfatos , Fotossíntese , Zea mays , Zea mays/crescimento & desenvolvimento , Zea mays/microbiologia , Zea mays/efeitos dos fármacos , Zea mays/metabolismo , Aspergillus niger/metabolismo , Chumbo/metabolismo , Antioxidantes/metabolismo , Fotossíntese/efeitos dos fármacos , Fosfatos/metabolismo , Poluentes do Solo/metabolismo , Estresse Fisiológico , Biodegradação Ambiental
3.
Sci Rep ; 14(1): 2375, 2024 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-38287143

RESUMO

Potato (Solanum tuberosum) is the third crucial global crop facing threats from Alternaria solani, a necrotrophic fungal pathogen causing early blight disease. Beyond crop impact, it leads to substantial production reduction and economic losses worldwide. This study introduces a green synthesis method for producing Ferric Oxide nanoparticles (FNPs) using dried Guava (Psidium guajava) leaves. Guava leaf extract acts as a reducing agent, with iron (III) chloride hexahydrate (FeCl3·6H2O) as the oxidizing agent. This study employed various characterization techniques for Ferric Oxide nanoparticles (FNPs). Fourier Transform Infrared Spectroscopy (FTIR) revealed peaks at 877 cm-1, 1180 cm-1, 1630 cm-1, 1833 cm-1, 2344 cm-1, and 3614 cm-1, associated with Maghemite vibrations, polyphenol compounds, and amino acids. UV-Vis spectroscopy exhibited a characteristic absorbance peak at 252 nm for FNPs. Scanning Electron Microscope (SEM) images illustrated particle sizes of 29-41 nm, and Energy Dispersive Spectroscopy (EDS) indicated elemental composition. X-ray diffraction (XRD) confirmed crystalline FNPs with peaks at 26.78, 30.64, 36.06, 38.21, 43.64, 53.52, 57.42, 63.14 and 78.32. Disease resistance assays demonstrated FNPs' effectiveness against A. solani, reducing disease incidence and severity. In the leaf detach assay, concentrations of 15, 10 and 5 mg/L showed a dose-dependent reduction in disease severity and incidence. The Greenhouse Assay confirmed FNPs' concentration-dependent effect on disease incidence and severity. The study also explored FNPs' potential as biocontrol agents showing no adverse effects on overall plant development. Additionally, the study highlighted the agronomic potential of FNPs in enhancing plant growth and development emphasizing their role as micronutrients in biofortification. The findings suggest the promising application of FNPs in plant protection and biofortification strategies.


Assuntos
Alternaria , Nanopartículas Metálicas , Nanopartículas , Solanum tuberosum , Nanopartículas/química , Compostos Férricos/química , Espectroscopia de Infravermelho com Transformada de Fourier , Nanopartículas Metálicas/química , Extratos Vegetais/química , Difração de Raios X , Antibacterianos/química
4.
Curr Pharm Des ; 29(41): 3324-3339, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38111115

RESUMO

INTRODUCTION: In the present study, we aimed to investigate the extraction and identification of the potential phytochemicals from the Methanolic Extract of Dryopteris ramosa (MEDR) using GC-MS profiling for validating the traditional uses of MEDR its efficacy in inflammations by using in-vitro, in-vivo and in silico approaches in anti-inflammatory models. METHODS: GC-MS analysis confirmed the presence of a total of 59 phytochemical compounds. The human red blood cells (HRBC) membrane stabilization assay and heat-induced hemolysis method were used as in-vitro anti-inflammatory activity of the extract. The in-vivo analysis was carried out through the Xylene-induced mice ear oedema method. It was found that MEDR at a concentration of 20 µg, 30 µg, and 40 µg showed 35.45%, 36.01%, and 36.33% protection to HRBC in a hypotonic solution, respectively. At the same time, standard Diclofenac at 30 µg showed 45.31% protection of HRBC in a hypotonic solution. RESULTS: The extract showed inhibition of 25.32%, 26.53%, and 33.31% cell membrane lysis at heating at 20 µg, 30 µg, and 40 µg, respectively. In comparison, standard Diclofenac at 30 µg showed 50.49% inhibition of denaturation to heat. Methanolic extract of the plant exhibited momentous inhibition in xylene-induced ear oedema in mice treated with 30 µg extract were 47.2%, 63.4%, and 78.8%, while inhibition in mice ear oedema treated with 60 µg extract was 34.7%, 43.05%, 63.21% and reduction in ear thickness of standard drug were 57.3%, 59.54%, 60.42% recorded at the duration of 1, 4 and 24 hours of inflammation. Molecular docking and simulations were performed to validate the anti-inflammatory role of the phytochemicals that revealed five potential phytochemicals i.e. Stigmasterol,22,23dihydro, Heptadecane,8methyl, Pimaricacid, Germacrene and 1,3Cyclohexadiene,_5(1,5dimethyl4hexenyl)-2methyl which revealed potential or significant inhibitory effects on cyclooxygenase-2 (COX-2), tumour necrosis factor (TNF-α), and interleukin (IL-6) in the docking analysis. CONCLUSION: The outcome of the study signifies that MEDR can offer a new prospect in the discovery of a harmonizing and alternative therapy for inflammatory disease conditions.


Assuntos
Dryopteris , Camundongos , Humanos , Animais , Xilenos/efeitos adversos , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Extratos Vegetais/química , Simulação de Acoplamento Molecular , Diclofenaco/efeitos adversos , Soluções Hipotônicas/efeitos adversos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Inflamação/tratamento farmacológico , Edema/induzido quimicamente , Edema/tratamento farmacológico , Metanol/efeitos adversos , Fator de Necrose Tumoral alfa
5.
MycoKeys ; 100: 171-204, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38098977

RESUMO

Tetraplosphaeriaceae (Pleosporales, Ascomycota) is a family with many saprobes recorded from various hosts, especially bamboo and grasses. During a taxonomic investigation of microfungi in tropical and subtropical forest regions of Guizhou, Hainan and Yunnan provinces, China, several plant samples were collected and examined for fungi. Four newly discovered species are described based on morphology and evolutionary relationships with their allies inferred from phylogenetic analyses derived from a combined dataset of LSU, ITS, SSU, and tub2 DNA sequence data. Detailed illustrations, descriptions and taxonomic notes are provided for each species. The four new species of Tetraplosphaeriaceae reported herein are Polyplosphaeriaguizhouensis, Polyplosphaeriahainanensis, Pseudotetraploayunnanensis, and Tetraploahainanensis. A checklist of Tetraplosphaeriaceae species with available details on their ecology is also provided.

6.
Metabolites ; 13(11)2023 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-37999244

RESUMO

Salicylic acid (SA), benzothiadiazole (BTH), and methyl jasmonate (MeJA) are potential elicitors found in plants, playing a crucial role against various biotic and abiotic stresses. The systemic acquired resistance (SAR) mechanism was evaluated in cotton plants for the suppression of Cotton leaf curl Multan Virus (CLCuMuV) by the exogenous application of different elicitors. Seven different treatments of SA, MeJA, and BTH were applied exogenously at different concentrations and combinations. In response to elicitors treatment, enzymatic activities such as SOD, POD, CAT, PPO, PAL, ß-1,3 glucanse, and chitinase as biochemical markers for resistance were determined from virus-inoculated and uninoculated cotton plants of susceptible and tolerant varieties, respectively. CLCuMuV was inoculated on cotton plants by whitefly (Bemesia tabaci biotype Asia II-1) and detected by PCR using specific primers for the coat protein region and the Cotton leaf curl betasatellite (CLCuMuBV)-associated component of CLCuMuV. The development of disease symptoms was observed and recorded on treated and control plants. The results revealed that BTH applied at a concentration of 1.1 mM appeared to be the most effective treatment for suppressing CLCuMuV disease in both varieties. The enzymatic activities in both varieties were not significantly different, and the disease was almost equally suppressed in BTH-treated cotton plants following virus inoculation. The beta satellite and coat protein regions of CLCuMuV were not detected by PCR in the cotton plants treated with BTH at either concentration. Among all elicitors, 1.1 mM BTH was proven to be the best option for inducing resistance after the onset of CLCuMuV infection and hence it could be part of the integrated disease management program against Cotton leaf curl virus.

7.
Chemosphere ; 343: 140075, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37678601

RESUMO

In this study, a trimetallic nanocomposite comprising Silver Copper-Nickel (Ag-Cu-Ni TNC) was synthesized and analysed for its efficiency in degrading benzene and benzopyrene, which has five fused benzene rings. Fabrication of trimetallic nanocomposites were characterized using UV spectroscopy, FTIR studies, SEM EDAX, and DLS results. XRD confirmed the cubic crystalline Fcc structure of Ag-Cu-Ni TNC. Photocatalytic degradation analysis revealed that Ag-Cu-Ni TNC has the efficient photocatalytic ability, and the optimum condition required for efficient degradation of benzene and benzopyrene was identified as 2 µg/mL of PAH molecule, 10 µg/mL of Ag-Cu-Ni TNC at pH 5, stirring time of 2 h placed under UV light. Based on these optimum conditions, kinetic and isotherm studies were performed, revealing that the adsorption of benzene and benzopyrene by Ag-Cu-Ni TNC fits well with the Pseudo-second order kinetic model and Freundlich isotherm model. Thus, our study's adsorption of PAH molecule from aqueous solution takes place through chemisorption and involves heterogeneous adsorption phenomena.

8.
Environ Res ; 235: 116672, 2023 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-37453502

RESUMO

The heavy metals (HMs) in seafood are alarming due to their biomagnification in the food chain. The concentrations of As, Cd, Hg, Pb, Cr, and Ni in both fresh and dried fish were quantified, and the potential exposure and safe intake levels for human consumption were assessed by the European Commission (EC) and the Food Safety Standard Authority of India (FSSAI). HMs concentrations ranged from 0.003 mg/kg (Cr) to 2.08 mg/kg for (As) and 0.007 mg/kg (Hg) to 2.76 mg/kg (As). Cd, Hg, and Pb levels in fresh and dried fish were below the maximum residue limits (MRLs) set by the EC and FSSAI, which were 0.1 mg/kg, 0.5 mg/kg, and 0.3 mg/kg, respectively. Cr and As concentrations were also below the MRLs of 12 mg/kg and 76 mg/kg for aquatic products specified by FSSAI. The concentration of HMs in fresh and dried fish was found in the order of As > Cr > Ni > Pb > Cd > Hg and As > Cd > Cr > Ni > Pb > Hg, while the fresh and dried fishes contained HMs in the following order: E. areolatus > S. longiceps > L.lentjen > S. barracuda > E. affinis > S. javus and DA > DS > DR > DB > DSF. The metal pollution index (MPI) validates seafood is HMs free, while the single (Pi) and Nemerow integrated pollution index (Pnw) indicate that concentrations of Cd and As in fresh and dried fish have exceeded the threshold value. The target hazard quotient (THQ<1), hazard index (HI < 1), and target cancer risk (TCR<10-4) indicate that there are no non-carcinogenic and carcinogenic risks through the consumption of seafood and seafood products collected from the Tuticorin coast and marketed at the domestic and international levels. The preliminary findings emphasize the importance of formulating domestic legislation/government initiatives to promote seafood and its consumption. The attainment of this objective shall be facilitated by examining the levels of persistent organic pollutants (POPs) in seafood and evaluating its potential risk to consumers.


Assuntos
Mercúrio , Metais Pesados , Poluentes Químicos da Água , Animais , Humanos , Cádmio/análise , Chumbo , Alimentos Marinhos/análise , Contaminação de Alimentos/análise , Medição de Risco , Metais Pesados/análise , Mercúrio/análise , Monitoramento Ambiental , Poluentes Químicos da Água/análise
9.
Heliyon ; 9(5): e15909, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37206037

RESUMO

The present study examined the biological potential and phytochemicals of Sophora mollis, Mucuna pruriens, and Indigofera atropurpurea methanolic leaf extracts. In vitro anti-acetylcholinesterase and anti-lipase assays were performed using different concentrations of plant extracts, and the IC50 values were determined. The cytotoxic potential of the selected plant extracts was assessed against HeLa, PC3, and 3T3 cell lines using an MTT assay. S. mollis leaf extract displayed the highest inhibition percentage (114.60% ± 19.95 at 1000 µg/mL) for the anti-acetylcholinesterase activity with a prominent IC50 value of 75.9 µg/mL. The anti-lipase potential was highest with the M. pruriens leaf extract (355.5 µg/mL IC50), followed by the S. mollis extract (862.7 µg/mL IC50). Among the cell lines tested, the cytotoxic potential of the I. atropurpurea extract (91.1 ppm IC50) against the PC3 cell line was promising. High-performance liquid chromatography revealed gallic acid, chlorogenic acid, caffeic acid, vanillic acid, rutin trihydrate, and quercetin dihydrate in varying concentrations in all plant species. The concentration of chlorogenic acid (69.09 ppm) was highest in M. pruriens, and the caffeic acid concentration (45.20 ppm) was higher in S. mollis. This paper reports the presence of bioactive therapeutic compounds in selected species of the Fabaceae family that could be micro-propagated, isolated, and utilized in pharmaceutical industries.

10.
Environ Res ; 231(Pt 1): 116033, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37142082

RESUMO

The current study aimed to investigate the effects of lead nitrate exposure on the enzymatical, haematological, and histological changes in the gill, liver, and kidney of Pangasius hypophthalmus. The fish were divided into six groups and treated with different Pb concentrations. The LC50 value of Pb was 55.57 mg/L at 96 h for P. hypophthalmus, and sublethal toxicity was assessed for 45 days at 1/5th (11.47 mg/L) and 1/10th (5.57 mg/L) of LC50 concentration. Enzymes such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) levels, alkaline phosphate (ALP), and lactate dehydrogenase (LDH) content increased significantly during sublethal toxicity of Pb. The reduction of HCT and PCV indicates an anemic condition due to the toxicity of Pb. Differential leucocytes, lymphocytes, and monocytes and their % values significantly decreased, indicating Pb exposure. The main histological changes observed in the gills were the destruction of secondary lamellae, the fusion of adjacent gill lamellae, primary lamellae hypertrophy, and severe hyperplasia, while in kidney exposed to Pb showed melanomacrophages, increased periglomerular, peritubular space, vacuolation, shrunken glomerulus, destruction of tubular epithelium, and hypertrophy of distal convoluted segment. The liver showed severe necrosis and rupture of hepatic cells, hyper trepheoid bile duct, shifting of nuclei, and vascular hemorrhage, while in the brain, binucleus, mesoglea cells, vacuole, and ruptured nucleus were observed. In conclusion, P. hypophthalmus, which has been exposed to Pb has developed a number of toxicity markers. Consequently, prolonged exposure to higher Pb concentrations may be harmful to fish health. The findings strongly suggest that the lead had a detrimental impact on the P. hypophthalmus population, as well as on the water quality and non-target aquatic organisms.


Assuntos
Peixes-Gato , Metais Pesados , Poluentes Químicos da Água , Animais , Chumbo/toxicidade , Metais Pesados/toxicidade , Antioxidantes/farmacologia , Fígado , Brânquias , Poluentes Químicos da Água/toxicidade
11.
Plant J ; 114(6): 1227-1242, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36917083

RESUMO

Anthropogenic global change is driving an increase in the frequency and intensity of drought and flood events, along with associated imbalances and limitation of several soil nutrients. In the context of an increasing human population, these impacts represent a global-scale challenge for biodiversity conservation and sustainable crop production to ensure food security. Plants have evolved strategies to enhance uptake of soil nutrients under environmental stress conditions; for example, symbioses with fungi (mycorrhization) in the rhizosphere and the release of exudates from roots. Although crop cultivation is managed for the effects of limited availability of nitrogen (N) and phosphorus (P), there is increasing evidence for limitation of plant growth and fitness because of the low availability of other soil nutrients such as the metals potassium (K), calcium (Ca), magnesium (Mg), and iron (Fe), which may become increasingly limiting for plant productivity under global change. The roles of mycorrhizas and plant exudates on N and P uptake have been studied intensively; however, our understanding of the effects on metal nutrients is less clear and still inconsistent. Here, we review the literature on the role of mycorrhizas and root exudates in plant uptake of key nutrients (N, P, K, Ca, Mg, and Fe) in the context of potential nutrient deficiencies in crop and non-crop terrestrial ecosystems, and identify knowledge gaps for future research to improve nutrient-uptake capacity in food crop plants.


Assuntos
Micorrizas , Humanos , Solo , Cálcio , Ecossistema , Magnésio , Potássio , Ferro , Plantas/microbiologia , Exsudatos e Transudatos , Raízes de Plantas , Fósforo
12.
Environ Res ; 233: 115601, 2023 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-36863657

RESUMO

The advancement of effective and long-lasting electrocatalysts for energy storage devices is crucial to reduce the impact of the energy crisis. In this study, a two-stage reduction process was used to synthesize carbon-supported cobalt alloy nanocatalysts with varying atomic ratios of cobalt, nickel and iron. The formed alloy nanocatalysts were investigated using energy-dispersive X-ray spectroscopy, X-ray diffraction, and transmission electron microscopy to determine their physicochemical characterization. According to XRD results, Cobalt-based alloy nanocatalysts form a face-centered cubic solid solution pattern, illustrating thoroughly mixed ternary metal solid solutions. Transmission electron micrographs also demonstrated that samples of carbon-based cobalt alloys displayed homogeneous dispersion at particle sizes ranging from 18 to 37 nm. Measurements of cyclic voltammetry, linear sweep voltammetry, and chronoamperometry revealed that iron alloy samples exhibited much greater electrochemical activity than non-iron alloy samples. The alloy nanocatalysts were evaluated as anodes for the electrooxidation of ethylene glycol in a single membraneless fuel cell to assess their robustness and efficiency at ambient temperature. Remarkably, in line with the results of cyclic voltammetry and chronoamperometry, the single-cell test showed that the ternary anode works better than its counterparts. The significantly higher electrochemical activity was observed for alloy nanocatalysts containing iron than for non-iron alloy catalysts. Iron stimulates nickel sites to oxidize cobalt to cobalt oxyhydroxides at lower over-potentials, which contributes to the improved performance of ternary alloy catalysts containing iron.


Assuntos
Etilenoglicol , Níquel , Cobalto , Ligas/química , Carbono/química
13.
ACS Omega ; 6(8): 5548-5559, 2021 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-33681595

RESUMO

An in vitro study was conducted to assess the impact of organochlorine pesticides (OCPs) on cellular growth, morphology, cell viability, biofilm-formation activity, and growth-regulating substances of a soil bacterium. Phosphate-solubilizing EAM 35 isolated from rhizosphere soil was molecularly identified as Enterobacter cloacae (accession number MT672578.1). Strain EAM 35 tolerated varying levels of OCPs, viz., benzene hexachloride (BHC), chlorpyrifos (CP), dieldrin (DE), and endosulfan (ES). The toxicity of OCPs to strain EAM 35 was displayed in a concentration-dependent manner. Among the OCPs, ES at a concentration of 200 µM showed a higher toxicity, where it maximally reduced the bacterial synthesis of indole-3-acetic acid (IAA), salicylic acid (SA), and 2,3-dihydroxy-benzoic acid (DHBA) by 73% (p ≤ 0.001), 85% (p ≤ 0.005), and 83% (p ≤ 0.001), respectively, over the control. While comparing the toxicity of OCPs to P-solubilizing activity of E. cloacae after 10 days of growth, the toxicity pattern followed the order ES (mean value = 82.6 µg mL-1) > CP (mean value = 93.2 µg mL-1) > DE (mean value = 113.6 µg mL-1) > BHC (mean value = 127 µg mL-1). Furthermore, OCP-induced surface morphological distortion in E. cloacae EAM 35 was observed as gaps, pits on both cellular facets, and fragmented and disorganized cell structure under a scanning electron microscope (SEM). The membrane-compromised cells increased as the concentrations of OC pesticides increased from 25 to 200 µM. Additionally, microbial counts (log10 CFU/mL) were also affected after pesticide exposure and decreased with increasing concentrations. While assessing the impact of OCPs on inhibition (%) of log10 CFU/mL, 150, 175, and 200 µM concentrations of ES completely reduced the growth of E. cloacae. Similarly, while comparing the toxicity of higher concentrations of OCPs to bacterial growth, sensitivity followed the order ES > DE > CP > BHC. In addition, the biofilm-formation ability of strain EAM 35 was inhibited in a pesticide-dose-dependent manner, and it was statistically (p ≤ 0.05, p ≤ 0.005, and p ≤ 0.001) significant. Conclusively, the present study clearly suggests that before applying pesticides to soil, their recommended dose should carefully be monitored.

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