Search Results (4,172)

Municipal wastewaters pose a severe risk to the environment and human health when discharged untreated. This is due to their high content of pathogens, such as viruses and bacteria, which can cause diseases like cholera. Herein, the research and development of porphyrin-modified polyethersulfone (PES) ultrafiltration (UF) membranes was conducted to improve bacterial inactivation in complex municipal wastewater and enhance the fouling resistance and filtration performance. The synthesis and fabrication of porphyrin nanofillers and the resultant membrane characteristics were studied. The incorporation of porphyrin-based nanofillers improved the membrane’s hydrophilicity, morphology, and flux (247 Lm⁻² h⁻¹), with the membrane contact angle (CA) decreasing from 90° to ranging between 58° and 50°. The membrane performance was monitored for its flux, antifouling properties, reusability potential, municipal wastewater, and humic acid. The modified membranes demonstrated an effective application in wastewater treatment, achieving notable antibacterial activity, particularly under light exposure. The In-BP@SW/PES membrane demonstrated effective antimicrobial photodynamic effects against both Gram-positive S. aureus and Gram-negative E. coli. It achieved at least a 3-log reduction in bacterial viability, meeting Food and Drug Administration (FDA) standards for efficient antimicrobial materials. Among the variants tested, membranes modified with In-PB@SW nanofillers exhibited superior antifouling properties with flux recovery ratios (FRRs) of 78.9% for the humic acid (HA) solution and 85% for the municipal wastewater (MWW), suggesting a strong potential for long-term filtration use. These results highlight the promise of porphyrin-functionalized membranes as multifunctional tools in advanced water treatment technologies. Full article

Background/Objectives: Hemodialysis catheter-related bloodstream infection (HD-CRBSIs) is a main cause of morbidity in hemodialysis. New preventive strategies have emerged, such as using lock solutions with antiseptic or antibiotic capacity. In this study, the antimicrobial effect was analyzed in vitro and with a catheter model of lock solutions of gentamicin (LSG), gentamicin/heparin (LSG/H), and gentamicin/citrate (LSG/C) in clinical and ATCC strains of Pseudomonas aeruginosa and Staphylococcus aureus. Methods: The formation, minimum inhibitory concentration, and minimum inhibitory concentration of the biofilm and minimum biofilm eradication concentration of the lock solutions were determined. Additionally, colony-forming unit assays were performed to evaluate the antimicrobial efficacy of the lock solutions in a hemodialysis catheter inoculation model. Results: The minimum inhibitory concentration (MIC) of planktonic cells of both P. aeruginosa and S. aureus for LSG/H and LSG/C was 4 µg/mL. In the minimum biofilm inhibitory concentration (MBIC) tests, the LSG/H was less effective than LSG/C, requiring higher concentrations for inhibition, contrary to the minimum biofilm eradication concentration (MBEC), where LSG/H was more effective. All lock solutions eradicated P. aeruginosa biofilms in the HD catheter model under standard conditions. Nevertheless, under modified conditions, the lock solutions were not as effective versus ATCC and clinical strains of S. aureus. Conclusions: Our analysis shows that the lock solutions studied managed to eradicate intraluminal mature P. aeruginosa in non-tunneled HD catheters under standard conditions. Biofilm inhibition and eradication were observed at low gentamicin concentrations, which could optimize the gentamicin concentration in lock solutions used in HD catheters. Full article

Background/Objective: Staphylococcus aureus (S. aureus) is a clinically significant pathogenic bacterium. Daptomycin (DAP) is a cyclic lipopeptide antibiotic used to treat infections caused by multidrug-resistant Gram-positive bacteria, including S. aureus. However, DAP currently faces clinical limitations due to its short half-life, toxic side effects, and increasingly severe drug resistance issues. This study aimed to develop a biomimetic nano-drug delivery system to enhance targeting ability, prolong blood circulation, and mitigate resistance of DAP. Methods: DAP-loaded chitosan nanocomposite particles (DAP-CS) were prepared by electrostatic self-assembly. Macrophage membrane vesicles (MM) were prepared by fusion of M1-type macrophage membranes with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). A biomimetic nano-drug delivery system (DAP-CS@MM) was constructed by the coextrusion process of DAP-CS and MM. Key physicochemical parameters, including particle diameter, zeta potential, encapsulation efficiency, and membrane protein retention, were systematically characterized. In vitro immune escape studies and in vivo zebrafish infection models were employed to assess the ability of immune escape and antibacterial performance, respectively. Results: The particle size of DAP-CS@MM was 110.9 ± 13.72 nm, with zeta potential +11.90 ± 1.90 mV, and encapsulation efficiency 70.43 ± 1.29%. DAP-CS@MM retained macrophage membrane proteins, including functional TLR2 receptors. In vitro immune escape assays, DAP-CS@MM demonstrated significantly enhanced immune escape compared with DAP-CS (p < 0.05). In the zebrafish infection model, DAP-CS@MM showed superior antibacterial efficacy over both DAP and DAP-CS (p < 0.05). Conclusions: The DAP-CS@MM biomimetic nano-drug delivery system exhibits excellent immune evasion and antibacterial performance, offering a novel strategy to overcome the clinical limitations of DAP. Full article

This study evaluated the effects of different storage conditions, varying in light exposure, relative humidity (RH), and packaging materials, on the physicochemical stability of Lobosphaera sp. biomass, the retention of bioactive compounds, and the bioactivity of its extracts. Under light and 75% RH, the biomass absorbed moisture over time, reaching 0.779 ± 0.003 g/g dry weight (DW) after three months. This was accompanied by a decline in luminosity, chroma, and hue values. In contrast, samples stored under other conditions showed minimal changes, indicating that high humidity, combined with light exposure, compromises biomass stability. Packaging in metalized polyethylene terephthalate (PETmet/PE) effectively preserved the water content, color, and carotenoid levels during a two-month storage period. Bioactive compounds extracted via hydroethanolic ultrasound-assisted extraction yielded 15.48 ± 1.35% DW. Total phenolic content (TPC) of the extracts declined over time in both PETmet/PE and low-density polyethylene (LDPE) packaging, though the decrease was less pronounced in PETmet/PE. Antioxidant activity, assessed via the ABTS assay, remained stable, regardless of storage duration or packaging. Antimicrobial activity of the extract decreased over time but remained more effective against Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes), with PETmet/PE packaging better preserving antimicrobial efficacy than LDPE. These findings underscore the importance of optimized storage conditions and packaging for maintaining the quality and bioactivity of Lobosphaera sp. biomass and its extracts. Full article

In response to the growing demand for clean-label preservatives, this study investigates the potential of Laetiporus sulphureus, an edible polypore mushroom, as a multifunctional additive in cooked sausages. The ethanolic extract of L. sulphureus (LsEtOH) was evaluated for its chemical composition, antioxidant capacity, and antimicrobial activity. Leucine (12.4 ± 0.31 mg/g d.w.) and linoleic acid (68.6%) were identified as the dominant essential amino acid and fatty acid. LsEtOH exhibited strong antioxidant activity, with IC₅₀ values of 215 ± 0.05 µg/mL (DPPH•), 182 ± 0.40 µg/mL (NO•), and 11.4 ± 0.01 µg/mL (OH•), and showed a selective inhibition of Gram-positive bacteria, particularly Staphylococcus aureus (MIC/MBC: 0.31/0.62 mg/mL). In cooked sausages treated with 0.05 mg/kg of LsEtOH, lipid peroxidation was reduced (TBARS: 0.26 mg MDA/kg compared to 0.36 mg MDA/kg in the control), microbial growth was suppressed (33.3 ± 15.2 CFU/g in the treated sample compared to 43.3 ± 5.7 CFU/g in the control group), and color and pH were stabilized over 30 days. A sensory evaluation revealed minor flavor deviations due to the extract’s inherent aroma. Encapsulation and consumer education are recommended to enhance acceptance. This is the first study to demonstrate the efficacy of L. sulphureus extract as a natural preservative in a meat matrix, supporting its application as a clean-label additive for shelf life and safety improvement. Full article

Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have compounding effects on the color, texture, flavor, and nutritional quality of the product or do not effectively reduce the pathogens that food can be exposed to. Some bacterial pathogens particularly possess traits and tactics that make them even more difficult to mitigate such as biofilm formation. Non-thermal plasma sanitation techniques, including plasma-treated water (PTW), have proven to be promising methods that significantly reduce pathogenic bacteria that food is exposed to. Published work reveals that PTW can effectively mitigate both gram-positive and gram-negative bacterial biofilms. This study presents a novel analysis of the differences in antimicrobial effects of PTW treatment between biofilm-forming gram-positive bacteria, commonly associated with foodborne illness, that are sporulating (Bacillus cereus) and non-sporulating (Listeria monocytogenes). After treatment with PTW, the results suggest the following hypotheses: (1) that the non-sporulating species experiences less membrane damage but a greater reduction in metabolic activity, leading to a possible viable but non-culturable (VBNC) state, and (2) that the sporulating species undergoes spore formation, which may subsequently convert into vegetative cells over time. PTW treatment on gram-positive bacterial biofilms that persist in food processing environments proves to be effective in reducing the proliferating abilities of the bacteria. However, the variance in PTW’s effects on metabolic activity and cell vitality between sporulating and non-sporulating species suggest that other survival tactics might be induced. This analysis further informs the application of PTW in food processing as an effective sanitation method. Full article

Pressure therapy combined with silicone has a significant effect on scar hyperplasia, but limitations such as long-term wearing of compression garments (CGs) can easily cause bacterial infection, cleanliness, and lifespan problems of CGs caused by the tedious operation of applying silicone. In this study, a compression garment fabric (CGF) with both inhibition of scar hyperplasia and antibacterial function was prepared. A polydimethylsiloxane (PDMS)-loaded microcapsule (PDMS-M) was prepared with chitosan quaternary ammonium salt (HACC) and sodium alginate (SA) as wall materials and PDMS as core materials by the complex coagulation method. The PDMS-Ms were finished on CGF and modified with (3-aminopropyl)triethoxysilane (APTES) to obtain PDMS-M CGF, which was further treated with HACC to produce PDMS-M-HACC CGF. X-ray Photoelectron Spectroscopy(XPS) and Fourier transform infrared spectroscopy (FTIR) analysis confirmed the formation of covalent bonding between PDMS-M and CGF. The PDMS-M CGF exhibited antibacterial rates of 94.2% against Gram-negative bacteria Escherichia coli (E. coli, AATCC 6538) and of 83.1% against Gram-positive bacteria Staphylococcus aureus (S. aureus, AATCC 25922). The antibacterial rate of PDMS-M-HACC CGF against both E. coli and S. aureus reached 99.9%, with wash durability reaching grade AA for E. coli and approaching grade A for S. aureus. The finished CGF maintained good biocompatibility and showed minimal reduction in moisture permeability compared to unfinished CGF, though with decreased elastic recovery, air permeability and softness. The finished CGF of this study is expected to improve the therapeutic effect of hypertrophic scars and improve the quality of life of patients with hypertrophic scars. Full article

Soil microbial communities are crucial for ecosystem services, soil fertility, and the resilience of agroecosystems. This study investigated how long-term (31 years) agronomic practices—tillage, NPK fertilization, and cropping system—along with measured environmental variables influence the microbial biomass and its community composition in Chernozem soil under corn cultivation. The polyfactorial field experiment included three tillage treatments ((moldboard (MT), ripped (RT), strip (ST)), two fertilization regimes (NPK (N: 160; P: 26; K: 74 kg/ha), and unfertilized control) and two cropping systems (corn monoculture and corn–wheat biculture). The soil samples (0–30 cm) were collected in June and September 2023. Microbial biomass and community structure were quantified using phospholipid fatty acid (PLFA) analysis, which allowed the estimation of total microbial biomass and community composition (arbuscular mycorrhizal (AM) fungi, fungi, Gram-negative (GN) and Gram-positive (GP) bacteria, actinomycetes). Our results showed that microbial biomass increased from June to September, rising by 270% in unfertilized plots and by 135% in NPK-fertilized plots, due to higher soil moisture. Reduced tillage, especially ST, promoted significantly higher microbial biomass, with biomass reaching 290% and 182% of that in MT plots in June and September, respectively. MT had a higher ratio of bacteria-to-fungi compared to RT and ST, indicating a greater sensitivity of fungi to disturbance. NPK fertilization lowered soil pH by about one unit (to 4.1–4.8) and reduced microbial biomass—by 2% in June and 48% in September—compared to the control, with the particular suppression of AM fungi. The cropping system had a smaller overall effect on microbial biomass. Full article

The emergence and spread of antimicrobial resistance among pathogens are significantly reducing available therapeutic options, highlighting the urgent need for novel and complementary treatment strategies. Antimicrobial photodynamic therapy (aPDT) is a promising alternative approach that can overcome antimicrobial resistance through a multitarget mechanism of action, exerting direct bactericidal and fungicidal effects with minimal risk of resistance development. Although aPDT has shown efficacy against a variety of pathogens, data on its activity against large collections of clinical multidrug-resistant strains are still limited. In this study, we assessed the antimicrobial activity of the photosensitizer RLP068/Cl combined with a red light-emitting LED source at 630 nm (Molteni Farmaceutici, Italy) against a large panel of Gram-negative and Gram-positive bacterial strains harboring relevant resistance traits and Candida species. Our results demonstrated the significant microbicidal activity of RLP068/Cl against all of the tested strains regardless of their resistance phenotype, with particularly prominent activity against Gram-positive bacteria (range of bactericidal concentrations 0.05–0.1 µM), which required significantly lower exposure to photosensitizer compared to Candida and Gram-negative species (range 5–20 µM). Overall, these findings support the potential use of RLP068/Cl-mediated aPDT as an effective therapeutic strategy for the management of localized infections caused by MDR organisms, particularly when conventional therapeutic options are limited. Full article

Cancer and infectious diseases are major causes of global morbidity and mortality stressing the need to find novel drugs with promising dual anticancer and antimicrobial efficacy. Gold complexes have been studied for the past years due to their anticancer properties, with a few of them displaying antimicrobial properties, which support their pharmacological interest. Within this scope, we investigated six gold bisdithiolate complexes [Au (bdt)₂]⁻ (1), [Au (dcbdt)₂]⁻ (2), [Au (3-cbdt)₂]⁻ (3), [Au (4-cbdt)₂]⁻ (4), [Au (pdt)₂]⁻ (5) and [Au (dcdmp)₂]⁻ (6), and) against the ovarian cancer cell lines A2780 and A2780cisR, the Gram-positive bacteria Staphylococcus aureus Newman, the Gram-negative bacteria Escherichia coli ATCC25922 and Burkholderia contaminans IST408, and the pathogenic yeasts Candida glabrata CBS138 and Candida albicans SC5134. Complexes 2 and 6, with ligands containing aromatic pyrazine or phenyl rings, substituted with two cyanonitrile groups, showed after 24 h of incubation high anticancer activities against A2780 ovarian cancer cells (IC₅₀~5 µM), being also able to overcome cisplatin resistance in A2780cisR cells. Both complexes induced the formation of ROS, activated caspase-3/7, and induced necrosis (LDH release) in a dose-dependent way, in a greater extent in the case of 6. Among the bacterial and fungal strains tested, only complex 6 presented antimicrobial activity against S. aureus Newman, indicating that this complex is a potential novel anticancer and antibacterial agent. These results delve into the structure-activity relationship of the complexes, considering molecular alterations such as replacing a phenyl group for a pyrazine group, and the inclusion of one or two cyanonitrile appendage groups, and their effects on biological activity. Overall, both complexes were found to be promising leads for the development of future anticancer drugs against low sensitive or cisplatin resistant tumors. Full article

Background: Unambiguous clinical interpretation of PCR results for urinary tract infections (UTIs) remains a challenge. Here we compare and correlate multiplex qPCR results (quantification cycle values) with traditional microbial culture results (colony forming units) for clinical samples. Methods: Serial dilutions [10⁸ to 10⁰ colony forming units (CFU)/mL] were performed on five Gram-negative and two Gram-positive UTI-causing bacterial pathogens. For each dilution, quantitative cultures on solid media to confirm CFU/mL values and a real-time PCR UTI panel employing a nanofluidic Open Array^TM platform producing quantification cycle (Cq) values were performed. Cq values were correlated with CFU/mL values, generating a semi-quantitative interpretive scale for clinical samples. The clinical utility of the scale was then assessed using PCR and culture data from 168 clinical urine samples. Results: For Gram-negative bacteria, Cq values of <23, 23 to 28, and >28 corresponded with ≥10⁵ CFU/mL, <10⁵ CFU/mL and negative cultures, respectively. For Gram-positive bacteria, Cq values of <26, 26 to 30, and >30 corresponded with ≥10⁵ CFU/mL, <10⁵ CFU/mL and negative cultures, respectively. Among 168 urine specimens (including 138 Gram-negative and 30 Gram-positive bacteria), there was 83.3% agreement (n = 140/168) and 16.6% non-agreement (n = 28/168) between culture CFU/mL and qPCR Cq. Gram-negative bacteria had higher agreement (87.6%, 121/138) than Gram-positive bacteria (63.3%, 19/30). Conclusions: This study demonstrates that qPCR Cq results can be directly correlated with traditional urine quantitative culture results and reliably identify the clinically relevant cutoff of 10⁵ CFU/mL for detected uropathogens. Full article

Background/Objectives: The clinical potential of antimicrobial peptides (AMPs) against dual threats like antimicrobial resistance (AMR) and cancer is often limited by their high host cell toxicity. Here, we focused on brevinin-2OS (B2OS), a novel peptide from the skin of Odorrana schmackeri with potent haemolytic activity. The objective was to study the structure–activity relationship and optimise the safety via targeted modifications. Methods: A dual-modification strategy involving C-terminal truncation and subsequent N-terminal D-amino acid substitution was employed. The bioactivities and safety profiles of the resulting analogues were evaluated using antimicrobial, haemolysis, and cytotoxicity assays. Result: Removal of the rana box in B2OS(1-22)-NH₂ substantially reduced haemolysis while maintaining bioactivities. Remarkably, the D-leucine substitution in [D-Leu²]B2OS(1-22)-NH₂ displayed a superior HC₅₀ value of 118.1 µM, representing a more than ten-fold improvement compared to its parent peptide (HC₅₀ of 10.44 µM). This optimised analogue also demonstrated faster bactericidal kinetics and enhanced membrane permeabilisation, leading to a greater than 22-fold improvement in its therapeutic index against Gram-positive bacteria. Conclusions: The C-terminal rana box is a primary determinant of toxicity rather than a requirement for activity in the B2OS scaffold. The engineered peptide [D-Leu²]B2OS(1-22)-NH₂ emerges as a promising lead compound, and this dual-modification strategy provides a powerful design principle for developing safer, more effective peptide-based therapeutics. Full article

Soil salinization poses a significant constraint to agricultural productivity. However, certain plant growth-promoting bacteria (PGPB) can mitigate salinity stress and enhance crop performance. In this study, a bacterial isolate, R-18, isolated from saline-alkali soil in Ningxia, China, was identified as Enterobacter bugandensis based on 16S rRNA gene sequencing. The isolate was characterized for its morphological, biochemical, and plant growth-promoting traits and was evaluated for its potential to alleviate NaCl-induced stress in maize (Zea mays L.) under hydroponic conditions. Isolate R-18 exhibited halotolerance, surviving at NaCl concentrations ranging from 2.0% to 10.0%, and alkaliphilic adaptation, growing at pH 8.0–11.0. Biochemical assays confirmed it as a Gram-negative bacterium, displaying positive reactions in the Voges–Proskauer (V–P) tests, catalase activity, citrate utilization, fluorescent pigment production, starch hydrolysis, gelatin liquefaction, and ammonia production, while testing negative for the methyl red and cellulose hydrolysis. Notably, isolate R-18 demonstrated multiple plant growth-promoting attributes, including nitrogen fixation, phosphate and potassium solubilization, ACC deaminase activity, and indole-3-acetic acid (IAA) biosynthesis. Under 100 mM NaCl stress, inoculation with isolate R-18 significantly enhanced maize growth, increasing plant height, stem dry weight, root fresh weight, and root dry weight by 20.64%, 47.06%, 34.52%, and 31.25%, respectively. Furthermore, isolate R-18 improved ion homeostasis by elevating the K⁺/Na⁺ ratio in maize tissues. Physiological analyses revealed increased chlorophyll and proline content, alongside reduced malondialdehyde (MDA) levels, indicating mitigated oxidative damage. Antioxidant enzyme activity was modulated, with decreased superoxide dismutase (SOD) and peroxidase (POD) activities but increased catalase (CAT) activity. These findings demonstrated that Enterobacter bugandensis R-18 effectively alleviated NaCl-induced growth inhibition in maize by enhancing osmotic adjustment, reducing oxidative stress, and improving ion balance. Full article

Background/Objectives: Antimicrobial resistance (AMR) is a health related threat world-wide. Biosynthesized gold nanoparticles (AuNPs) using plant extracts have been reported to exhibit certain biological activity. This study aimed to biosynthesize AuNPs using an aqueous extract of Eruca sativa leaves and to evaluate their biocompatibility, antimicrobial activity, and antioxidant properties. Methods: AuNPs were biosynthesized using an aqueous extract of Eruca sativa leaves. Their biocompatibility was evaluated through hemolytic activity and assessments of hepatic and renal functions in rats. AuNPs were biologically evaluated as antimicrobial and antioxidant agents. Results: The AuNPs exhibited particle sizes of 27.78 nm (XRD) and 69.41 nm (AFM). Hemolysis assays on red blood cells revealed negligible hemolytic activity (<1%). Hepatic enzyme levels, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were studied. ALT, AST, and ALP levels showed no significant changes compared to the negative control. However, LDH levels were elevated at higher concentration (52.8 µg/mL), while the lower concentration (26.4 µg/mL) appeared to be safer. Renal biomarkers, urea and creatinine, showed no significant changes at either concentration, indicating minimal nephrotoxicity. The antimicrobial activity of AuNPs, plant extract, and gold salt was tested against five microorganisms: two Gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumoniae), two Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), and a fungal strain (Candida albicans). The AuNPs exhibited minimum inhibition concentrations (MICs) of 13.2 µg/mL against S. aureus and S. pneumoniae, 26.4 µg/mL against E. coli and C. albicans, and 39.6 µg/mL against P. aeruginosa, suggesting selectivity towards Gram-positive bacteria. Furthermore, the AuNPs demonstrated strong antioxidant activity, surpassing that of vitamin C. Conclusions: The biosynthesized AuNPs exhibited promising biocompatibility, selective antimicrobial properties, and potent antioxidant activity, supporting their potential application in combating the AMR. Full article

This study analyzed the heavy metal tolerance and chromium reduction and the potential of plant growth to promote Rhizobium sp. OS-1. By genetic makeup, the Rhizobium strain is nitrogen-fixing and phosphate-solubilizing in metal-contaminated agricultural soil. Among the Rhizobium group, bacterial strain OS-1 showed a significant tolerance to heavy metals, particularly chromium (900 µg/mL), zinc (700 µg/mL), and copper. In the initial investigation, the bacteria strains were morphologically short-rod, Gram-negative, appeared as light pink colonies on media plates, and were biochemically positive for catalase reaction and the ability to ferment glucose, sucrose, and mannitol. Further, bacterial genomic DNA was isolated and amplified with the 16SrRNA gene and sequencing; the obtained 16S rRNA sequence achieved accession no. HE663761.1 from the NCBI GenBank, and it was confirmed that the strain belongs to the Rhizobium genus by phylogenetic analysis. The strain’s performance was best for high hexavalent chromium [Cr(VI)] reduction at 7–8 pH and a temperature of 30 °C, resulting in a total decrease in 96 h. Additionally, the adsorption isotherm Freundlich and Langmuir models fit best for this study, revealing a large biosorption capacity, with Cr(VI) having the highest affinity. Further bacterial chromium reduction was confirmed by an enzymatic test of nitro reductase and chromate reductase activity in bacterial extract. Further, from the metal biosorption study, an Artificial Neural Network (ANN) model was built to assess the metal reduction capability, considering the variables of pH, temperature, incubation duration, and initial metal concentration. The model attained an excellent expected accuracy (R² > 0.90). With these features, this bacterial strain is excellent for bioremediation and use for industrial purposes and agricultural sustainability in metal-contaminated agricultural fields. Full article