Green Synthesis of Iron Nanoparticles from Spinach Leaf and Banana Peel Aqueous Extracts and Evaluation of Antibacterial Potential

Spinacia oleracea (spinach) and Musa acuminata (banana) were chosen for the study, and aqueous extracts of spinach leaf extract (SLE) and banana peel extract (BPE) were prepared for the synthesis of iron nanoparticles (FeNPs), and their antibacterial potential against pathogenic bacteria Bacillus subtilis (MTTC 1133) and Escherichia coli (MTTC 62) was evaluated. In 10 minutes at 60°C, the color of the mixture (FeCl3+SLE) changed from light green to dark blackish-brown, and the color of the mix (FeCl3+BPE) changed from transparent yellow to dark black, confirming the synthesis of FeNPs from SLE and BPE, respectively. The UV-Vis spectra of spinach- and banana-derived FeNPs revealed two peaks ranging from 240 to 430 nm and multiple peaks at 240, 270, and 395 nm, respectively. FTIR spectroscopy was used to show different functional groups on BPE and SLE, and their role in FeNP synthesis was predicted. TEM micrographs showed that the particles were in nanoscale, ranging in size from 20 to 50 nm for BPE-derived FeNPs and 10 to 70 nm for SLE-derived FeNPs. The FeNP (BPE and SLE) XRD analysis revealed amorphism, with a weak iron characteristic peak, indicating noncrystallinity. The antibacterial potential of BPE- and SLE-FeNPs was investigated, and inhibition zones (mm) against B. subtilis ( 22.70 ± 0.4 ) and E. coli ( 20.45 ± 1.66 ) were observed, as well as SLE-FeNPs against B. subtilis ( 23.56 ± 1.00 ) and E. coli ( 20.33 ± 0.58 ). There were no significant differences in antibacterial activities between BPE-FeNPs and SLE-FeNPs. Positive controls were tetracycline and gentamicin, both standard antibiotics, at 5 μg/disk. SLE- and BPE-derived green FeNPs were also analysed in vivo of D. melanogaster life history traits, i.e., fecundity, hatchability, viability, and duration of development for toxicity evaluation. SLE- and BPE-derived green FeNPs at a concentration of 10 mg/L were fed flies compared to normal diet-fed flies (control sample), and no significant differences were observed between them. The findings suggest that FeNPs have a high antibacterial potential and could be used as antibacterial agents against pathogenic bacteria while being nontoxic in nature.

PENGARUH PEMBERIAN EKSTRAK DAUN BAYAM MERAH (Amaranthus tricolor L.) TERHADAP KADAR LDL DAN HDL PADA TIKUS PUTIH (Rattus norvegicus) GALUR WISTAR JANTAN YANG DIBERI DIET TINGGI LEMAK

Background: Hyperlipidemia is a condition where there is an increase or decrease in the lipid fraction in the blood due to a metabolic disorder of the lipid itself. The lipid fraction that experienced abnormalities was usually an increase in total cholesterol, triglycerides, LDL cholesterol and a decrease in HDL cholesterol (Perkeni, 2015). Anthocyanin compounds are one of the flavonoid compounds that have the potential as an alternative therapy for dyslipidemia. The leaves of red chicken (Amaranthus tricolor L.) contain anthocyanin compounds that can be used as an alternative therapy for hyperlipidemia.Objective: To determine the effect of red spinach leaf extract (Amaranthus tricolor L.) on LDL and HDL blood of white wistar rats induced by a high-fat diet.Methodology: Methods Reviewing articles according to the issue to be studied. It was carried out according to the PICOST analysis, namely population, intervention, comparison, output, study and times from 583 selected journals and 5 journals. Results: Literature from 5 journals contained 5 journals discussing red spinach leaf extract (Amaranthus tricolor L.) (100%), 5 journals discussing high-fat diets (100%), 4 journals discussing blood LDL (80%) and 3 journals. discussed blood HDL (60%). Conclusion: The administration of red spinach leaf extract (Amaranthus tricolor L.) affected the blood LDL of rats at a dose of 200 mg/kg BW to 800 mg/kg BW and HDL rat blood at a dose of 200 mg/kg BW to 400 mg/kg BW . ABSTRAK Latar Belakang : Hiperlipidemia adalah keadaan dimana terjadinya peningkatan ataupun penurunan fraksi lipid dalam darah karena adanya suatu kelainan metabolisme dari lipid itu sendiri. Fraksi lipid yang mengalami kelainan biasanya adalah kenaikan kadar kolesterol total, trigliserida Keywords: High Fat Diet, LDL, HDL, Red Spinach Leaf Extract , kolesterol LDL dan penurunan dari kolesterol HDL (Perkeni, 2015). Senyawa antosianin merupakan salah satu senyawa flavonoid yang dapat berpotensi sebagai terapi alternatif untuk dislipidemia. Pada daun ayam merah (Amaranthus tricolor L.) terdapat senyawaantosianin yang dapat digunakan sebagai terapi alternative hiperlipidemia.Tujuan : Mengetahui pengaruh pemberian ekstrak daun bayam merah (Amaranthus tricolor L.) terhadap LDL dan HDL darah tikus putih wistar yang diinduksi diet tinggi lemak.Metodologi : Melakukan telaah artikel sesuai dengan issue yang akan diteliti. Dilakukan sesuai dengan analisis PICOST yaitu population, intervention, comparation, output, studi dan times dari 583 jurnal terpilih 5 jurnal.Hasil : Literature dari 5 jurnal terdapat 5 jurnal yang membahas tentang ekstrak daun bayam merah (Amaranthus tricolor L.) (100%), 5 jurnal membahas diet tinggi lemak (100%), 4 jurnal membahas LDL darah (80%) dan 3 jurnal membahas HDL darah (60%).Kesimpulan : Pemberian ekstrak daun bayam merah (Amaranthus tricolor L.) berpengaruh terhadap LDL darah tikus dengan dosis pemberian 200 mg/kg BB hingga 800 mg/kg BB dan HDL darah tikus dengan dosis pemberian 200 mg/kg BB hingga 400 mg/kg BB. Kata kunci : Diet Tinggi Lemak, LDL, HDL, Ekstrak Daun Bayam Merah

STUDI LITERATUR PENGARUH PEMBERIAN EKSTRAK DAUN BAYAM MERAH (Amaranthus tricolor L.) TERHADAP KADAR SGOT DAN SGPT PADA TIKUS PUTIH (Rattus norvegicus) JANTAN GALUR WISTAR YANG DIBERI DIET TINGGI LEMAK

Background : The prevalence of hyperlipidemia in Indonesia is increasing from year to year, in 2008 it was recorded at 35.1%. Then in 2013 it increased to 35.9%(WHO, 2013). Increased hyperlipidemia triggers Non-Alcoholic Fatty Liver Disease (NAFLD)(Ghanaei et al., 2019). NAFLD conditions cause liver cell damage, causing increased levels of liver enzymes such as SGOT and SGPT (Prahastuti et al., 2020). According to Pradana et al. (2017) it is said that red spinach leaf extract has antioxidant activity that is able to inhibit the incision of the oxidation chain reaction because it contains flavonoid compounds, quercetin, tannins, and saponins which have the potential as a preventive therapy for fatty liver. Objective: To determine the effect of red spinach (Amaranthus tricolor L.) leaf extract on SGOT and SGPT levels in white rats (Rattus norvegicus) fed a high-fat diet. Research Methods: The research method uses a literature study. The database in the literature study was obtained from Google Scholar, PubMed and NCBI which was taken by Population, Intervention, Comparation, Output, Study, and Time (PICOST) from 5,166 journals and finally 8 journals were obtained. Results: Literature from 8 journals is known to have 8 that discuss red spinach leaf extract. Conclusion: From the literature obtained, there was an effect of giving red spinach leaf extract (Amaranthus tricolor L.) on the levels of SGOT and SGPT given a high-fat diet. The dose is 200mg/kgBB and 400mg/kgBB with a duration of administration of 7 to 29 days. Keyword : High fat diet, Red Spinach Extract, Hyperlipidemia, NAFLD, SGOT, SGPT, White Rat

Siklus hidup Spodoptera frugiperda J.E. Smith dengan pakan daun bayam cabut hijau dan daun bayam duri hijau di laboratorium

Spodoptera frugiperda merupakan hama baru di Indonesia yang menyerang tanaman jagung dan memiliki berbagai macam tanaman inang lain. Salah satu tanaman yang dimungkinkan menjadi inang dari hama ini di Indonesia adalah bayam. Tujuan penelitian ini untuk mengetahui lama siklus hidup S. frugiperda yang diberikan pakan daun bayam cabut hijau dan daun bayam duri hijau. Metode penelitian ini yaitu permeliharaan S. furgiperda dengan sumber pakan yang berbeda dan mengamati beberapa parameter. Analisis statistik berupa inferensial, data yang dilakukan dengan uji normalitas Shapiro-Wilk. Apabila data normal dilakukan uji homogenitas dengan dilanjutkan analisis ANNOVA. Hasil penelitian menunjukan bahwa S. frugiperda pada pakan kontrol (daun jagung) memiliki lama siklus hidup 38 hari, daun bayam cabut hijau 40 hari, dan daun bayam duri hijau 42 hari. Panjang larva S. frugiperda paling panjang didapatan pada pakan daun bayam duri hijau, sedangkan diameter kepala tidak terdapat perbedaan antara ketiga pakan. Bobot larva paling berat didapatkan pada pakan daun bayam cabut hijau. Panjang dan berat pupa S. frugiperda paling tinggi didapatkan pada pakan daun bayam duri hijau. Jumlah jantan paling banyak dihasilkan pada pakan daun bayam cabut hijau dan daun bayam duri hijau, sedangkan jumlah betina paling banyak dihasilkan pakan kontrol. Jumlah telur paling banyak dihasilkan pada pakan kontrol dan paling sedikit pada bayam duri hijau. Pakan bayam cabut dan bayam duri daun bayam cabut hijau dan bayam duri hijau dapat digunakan sebagai pakan alternatif bagi S. frugiperda di laboratorium. Kata kunci: pakan alternatif; perkembangan; pertumbuhan; Spodoptera frugiperda ABSTRACTLife cycle of Spodoptera frugiperda J.E. Smith with green spinach and green thorn spinach leaves in the laboratory. Spodoptera frugiperda is a new pest in Indonesia that attacks maize and has a wide variety of other host plants. One of the plants that is possible to host this pest in Indonesia is spinach. The purpose of this study was to determine the length of the life cycle of S. frugiperda which was fed with green spinach leaves and green thorn spinach leaves. This research method is the maintenance of S. furgiperda with different feed sources and observing several parameters. Statistical analysis in the form of inferential, the data was done by using the Shapiro-Wilk normality test. If the data is normal, the homogeneity test is carried out followed by ANNOVA analysis. The results showed that S. frugiperda in control diets (corn leaves) had a life cycle length of 38 days, green spinach leaves 40 days, and green thorn spinach leaves 42 days. Thelarvae of longestS. frugiperda were found in green thorn spinach leaf feed, while there was no difference in head diameter between the three diets. The heaviest larval weight was found in green pulled spinach feed. Thelength and weight ofpupa were S. frugiperda highestfound in green thorn spinach leaf feed. The highest number of males was produced in green spinach leaf and green spinach leaf feed, while the highest number of females was produced in control feed. The highest number of eggs was produced in the control feed and the least was the green thorn spinach. Spinach feed and spinach spines, green pulled spinach and green spiny spinach can be used as an alternative feed for S. frugiperda in the laboratory. Keywords: alternative food; development; growth; Spodoptera frugiperda

Antimicrobial-Loaded Polymeric Micelles Inhibit Enteric Bacterial Pathogens on Spinach Leaf Surfaces During Multiple Simulated Pathogen Contamination Events

Protecting fresh-packed produce microbiological safety against pre- and post-harvest microbial pathogen contamination requires innovative antimicrobial strategies. Although largely ignored in the scientific literature, there exists the potential for gross failure in food safety protection of fresh fruits and vegetables leading to opportunity for multiple produce contamination events to occur during production and post-harvest handling of food crops. The primary objective of this research was to determine the efficacy of plant-derived antimicrobial-loaded nanoparticles to reduce Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium on spinach leaf surfaces whilst simulating multiple pathogen contamination events (pre-harvest and post-harvest). Spinach samples were inoculated with a blend of E. coli O157:H7 and S. Typhimurium, each diluted to ~8.0 log10 CFU/mL. The inoculated samples were then submerged in solutions containing nanoparticles loaded with geraniol (GPN; 0.5 wt.% geraniol), unencapsulated geraniol (UG; 0.5 wt.%), or 200 ppm chlorine (HOCl; pH 7.0), with untreated samples serving for controls. Following antimicrobial treatment application, samples were collected for surviving pathogen enumeration or were placed under refrigeration (5°C) for up to 10 days, with periodic enumeration of pathogen loads. After 3 days of refrigerated storage, all samples were removed, aseptically opened and subjected to a second inoculation with both pathogens. Treatment of spinach surfaces with encapsulated geraniol reduced both pathogens to non-detectable numbers within 7 days of refrigerated storage, even with a second contamination event occurring 3 days after experiment initiation. Similar results were observed with the UG treatment, except that upon recontamination at day 3, a higher pathogen load was detected on UG-treated spinach vs. GPN-treated spinach. These data fill a research gap by providing a novel tool to reduce enteric bacterial pathogens on spinach surfaces despite multiple contamination events, a potential food safety risk for minimally processed edible produce.

Supercritical carbon dioxide decellularization of plant material to generate 3D biocompatible scaffolds

The use of plant-based biomaterials for tissue engineering has recently generated interest as plant decellularization produces biocompatible scaffolds which can be repopulated with human cells. The predominant approach for vegetal decellularization remains serial chemical processing. However, this technique is time-consuming and requires harsh compounds which damage the resulting scaffolds. The current study presents an alternative solution using supercritical carbon dioxide (scCO2). Protocols testing various solvents were assessed and results found that scCO2 in combination with 2% peracetic acid decellularized plant material in less than 4 h, while preserving plant microarchitecture and branching vascular network. The biophysical and biochemical cues of the scCO2 decellularized spinach leaf scaffolds were then compared to chemically generated scaffolds. Data showed that the scaffolds had a similar Young’s modulus, suggesting identical stiffness, and revealed that they contained the same elements, yet displayed disparate biochemical signatures as assessed by Fourier-transform infrared spectroscopy (FTIR). Finally, human fibroblast cells seeded on the spinach leaf surface were attached and alive after 14 days, demonstrating the biocompatibility of the scCO2 decellularized scaffolds. Thus, scCO2 was found to be an efficient method for plant material decellularization, scaffold structure preservation and recellularization with human cells, while performed in less time (36 h) than the standard chemical approach (170 h).

Three New Fungal Leaf Spot Diseases of Spinach in the United States and the Evaluation of Fungicide Efficacy for Disease Management

Leaf spot diseases of spinach, caused by Colletotrichum spinaciae, has become a major production constraint in several production areas, including Texas, in recent years. Leaf spot symptoms were observed in several fields in Texas in 2016 and 2017, with typical anthracnose-like symptoms and leaves with small, circular, and sunken lesions that appeared similar to injury from windblown sand. The lesions were plated on potato dextrose agar, from which fungal cultures were recovered. The fungi were identified based on morphology and sequence analysis of the introns of glutamate synthetase and glyceraldehyde-3-phosphate dehydrogenase (for isolates determined to be Colletotrichum spp.) and the internal transcribed spacer ribosomal DNA (for isolates determined to be Myrothecium spp.). Based on foliar symptoms, fungal colony and spore morphology, pathogenicity tests of fungal isolates on the spinach cultivar ‘Viroflay’, and DNA sequence analysis of the isolates, the symptoms on spinach leaves for two sets of samples were caused by Colletotrichum coccodes and Colletotrichum truncatum, and leaf spots resembling damage from windblown sand were caused by Myrothecium verrucaria. This is the first report of spinach leaf spot diseases caused by C. coccodes, C. truncatum, and M. verrucaria in the United States. C. coccodes and C. truncatum caused severe symptoms on the spinach cultivar ‘Viroflay’, whereas M. verrucaria caused symptoms of intermediate severity. Fungicide efficacy tests demonstrated that chlorothalonil, mancozeb, pyraclostrobin, fluxapyroxad + pyraclostrobin, and penthiopyrad were completely effective at preventing leaf spots caused by any of these pathogens when applied 24 h before inoculation of ‘Viroflay’ plants in greenhouse trials.

Spinach and Chive for Kidney Tubule Engineering: the Limitations of Decellularized Plant Scaffolds and Vasculature

Tissue decellularization yields complex scaffolds with retained composition and structure, and plants offer an inexhaustible natural source of numerous shapes. Plant tissue could be a solution for regenerative organ replacement strategies and advanced in vitro modeling, as biofunctionalization of decellularized tissue allows adhesion of various kinds of human cells that can grow into functional tissue. Here, we investigated the potential of spinach leaf vasculature and chive stems for kidney tubule engineering to apply in tubular transport studies. We successfully decellularized both plant tissues and confirmed general scaffold suitability for topical recellularization with renal cells. However, due to anatomical restrictions, we believe that spinach and chive vasculature themselves cannot be recellularized by current methods. Moreover, gradual tissue disintegration and deficient diffusion capacity make decellularized plant scaffolds unsuitable for kidney tubule engineering, which relies on transepithelial solute exchange between two compartments. We conclude that plant-derived structures and biomaterials need to be carefully considered and possibly integrated with other tissue engineering technologies for enhanced capabilities.