Bioassay-Guided Isolation and Identification of Cytotoxic Compounds from Melaleuca quinquenervia Fruits.

The fruit extract of Melaleuca quinquenervia yielded a total of 19 compounds, including two novel spiro-biflavonoid enantiomers (1a and 1b) and a chalcone derivative (3). Their structures were determined through spectroscopic analysis. The enantiomers of the racemic mixture of compound 1 were successfully resolved into (+)-1 and (-)-1 using chiral-phase HPLC. Single-crystal X-ray diffraction analysis was also used to confirm the structure of 1. The enantiomeric configurations of 1 and 2 were determined through a comparison of the calculated and experimental electronic circular dichroism spectra. Compounds 2 (melanervin), 14 (methyl betulinate), 15 (3-O-acetylbetulinic acid), and 16 (pyracrenic acid) were found to be highly cytotoxic, with compound 16 showing superior growth inhibition of nonsmall cell lung cancer cells (A549 cells) (IC50 2.8 ± 0.1 µM) compared to cisplatin (IC50 3.3 ± 0.0 µM), a positive control chemotherapeutic drug. Both compound 16 and cisplatin were significantly more cytotoxic toward A549 lung cancer cells compared to nontumorigenic Vero E6 cells.

Nutritional, physicochemical, and functional properties of Hawaiian taro (Colocasia esculenta) flours: A comparative study.

Taro (Colocasia esculenta) flour is a viable carbohydrate alternative and a functional additive for food formulation; however, different taro varieties may possess distinct characteristics that may influence their suitability for food production. This study evaluated the nutritional, physicochemical, and functional properties of flours from five Hawaiian taro varieties: Bun-Long, Mana Ulu, Moi, Kaua'i Lehua, and Tahitian. Tahitian, Bun-long, and Moi had high total starch contents of 40.8, 38.9, and 34.1 g/100 g, respectively. Additionally, Moi had the highest neutral detergent fiber (25.5 g/100 g), lignin (1.39 g/100 g), and cellulose (5.31 g/100 g). In terms of physicochemical properties, Tahitian showed the highest water solubility index (33.3 g/100 g), while Tahitian and Moi exhibited the two highest water absorption indices (5.81 g/g and 5.68 g/g, respectively). Regarding functional properties, Tahitian had the highest water absorption capacity (3.48 g/g), and Tahitian and Moi had the two highest oil absorption capacities (3.15 g/g and 2.68 g/g, respectively). Therefore, the flours from these Hawaiian taro varieties possess promising characteristics that could enhance food quality when used as alternative additives in food processing.

Physiochemical and Sensory Properties of a Turmeric, Ginger, and Pineapple Functional Beverage with Effects of Pulp Content.

Beverage mixtures based on pineapple juice (80-100%), with varying concentrations of turmeric (0-20%) and ginger (0-20%) juice were developed. The pineapple juice alone exhibited a total soluble solid (TSS) content of 15.90-16.03 °Brix. The total polyphenols content (TPC) varied between 0.32 and 1.79 mg GAE/mL, and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) inhibition was between 40.56% and 86.19% and correlated with the TPC and curcumin and other curcuminoids. The formulations with a high pulp content showed a significantly higher TPC and greater DPPH inhibition than those with a low pulp content. Turmeric and ginger with a high amount of pulp had a higher abundance of volatile compounds. Significant differences were observed by the panelists in the taste and mouthfeel attributes and the low-pulp juices were associated with increased palatability due to the better mouthfeel, higher sweetness, and decreased bitterness, pepperiness, pulpiness, and spiciness. The pineapple juice mixtures with 10% turmeric juice and 10% or less ginger juice were most preferred by sensory panelists.

Supplementation of papaya leaf juice has beneficial effects on glucose homeostasis in high fat/high sugar-induced obese and prediabetic adult mice.

Prediabetes is characterized by a cluster of glycemic parameters higher than normal but below the threshold of type 2 diabetes mellitus (T2DM). In recent years, phytochemical-rich plant extracts have gained popularity as therapeutic agents for metabolic disorders. This study investigated the effects of papaya leaf (PL) juice supplementation on blood glucose levels in diet-induced obese and prediabetic adult mice. B65JL F1 mice (n = 20) at 12-14 months old were fed a high fat/sugar diet (HFHS) for 120 days. Mice were switched to restricted rodent chow of 3 g feed/30 g body weight/day, supplemented with 3 g/100 mL PL juice for 30 days. HFHS diet remarkably increased fasting plasma glucose levels from 114 ± 6.54 mg/dL to 192.7 ± 10.1 mg/dL and body weight from 32.5 ± 1.6 to 50.3 ± 4.1 g. HFHS diet results in hyperglycemia, insulin resistance, hyperlipidemia, and liver steatosis. The combination of PL juice and restricted diet significantly reduced body weight and fasting blood glucose levels to 43.75 ± 1.4 g and 126.25 ± 3.2 mg/dl, respectively. Moreover, PL juice with a restricted diet significantly improved lipid profile: cholesterol from 204 to 150 mg/dL, LDL-c from 110.4 to 50 mg/dL, and triglyceride from 93.7 to 60 mg/dL. Additionally, PL juice combined with a restricted diet significantly reduced adiposity, reversed fatty liver, and restored skeletal muscle Glut4 and phosphorylated (p-AKT (ser473). This study demonstrated that supplementation of PL juice with a restricted diet was more effective than a restricted diet alone in reversing major symptoms related to prediabetic and obesity conditions.

Physical, Chemical, and Sensory Properties of a Turmeric-Fortified Pineapple Juice Beverage.

Beverage mixtures based on pineapple (Ananas comosus) and turmeric (Curcuma longa) juice as a ready-to-drink product were developed, and their physicochemical, nutritional, and sensory properties were evaluated. Four different concentrations of turmeric juice (5%, 10%, 15%, and 20% (v/v)) were added to pineapple juice to make turmeric-fortified pineapple (TFP) juice samples. Pineapple juice without turmeric was the control. The L*, a*, b*, titratable acidity (TA), total antioxidant capacity, and %DPPH scavenging values, as well as the concentrations of the phenolic compounds curcumin and demethoxycurcumin, were significantly increased with increasing turmeric concentration. Thirty volatile compounds were detected in the mixed juice samples with turmeric. Most of the turmeric-specific compounds, including monoterpenes, sesquiterpenes and turmerones, were detected in the TFP juice samples. While the antioxidant activity of the juice samples increased with increasing turmeric concentration, the pineapple juice fortified with 10% turmeric (10%T) had the best overall quality as determined by panelists. Greater concentrations of turmeric were associated with decreased palatability due to reduced mouthfeel and sweetness and increased aftertaste and sourness. These results suggest that the 10%T juice could be developed into a commercial functional beverage with increased overall flavor and nutritional quality.

Economic Benefits from Managing Coffee Berry Borer (Hypothenemus hampei) in Hawaii.

Coffee berry borer (CBB) is considered the most damaging insect pest of coffee worldwide. CBB was first detected on Hawai'i Island in 2010, and quickly spread throughout the state's coffee-growing regions. With the introduction of this pest, Hawaii's small yet economically important coffee industry was changed forever with growers facing significantly higher production and labor costs, as well as decreased yield and coffee quality. We assessed the economic benefits of managing CBB based on three strategies that emerged in Hawaii over the last decade: (1) the use of the entomopathogenic fungus Beauveria bassiana alone, (2) early integrated pest management (IPM), which combined monitoring and sanitation with spraying B. bassiana, and (3) research-based IPM, which focused on CBB biology in Hawaii, optimization of monitoring, B. bassiana applications, and cultural controls. From 2011 to 2021, the economic benefits from managing CBB were USD 52 million using B. bassiana alone, USD 69 million from early IPM, and USD 130 million from research-based IPM, for a total of USD 251 million from all management. Our findings suggest that all types of management provide economic benefits to Hawaii growers, but management strategies based on Hawaii-specific research have provided the greatest gains in coffee yield, price, and revenue.

Frequent and efficient harvesting as an economically viable strategy to regulate coffee berry borer on commercial farms in Hawaii.

Coffee is the second most economically important agricultural crop in Hawaii, valued at around $175M for green and roasted coffee in the 2021-2022 season. With the introduction of coffee berry borer (CBB, Hypothenemus hampei Ferrari) to Hawaii in 2010, growers have faced a significant challenge in producing the specialty coffee that the region is known for. This tiny beetle infests the coffee seed and reduces the yield and quality of coffee products. While field sanitation, frequent harvesting and strip-picking are known to be essential for controlling CBB, the associated costs and benefits of these cultural control practices have not been estimated for Hawaii. In the present study, we examined two CBB management strategies across 10 commercial coffee farms on Hawaii Island: (i) conventional management including frequent sprays of pesticides and few rounds of sanitation and harvesting, and (ii) cultural control-focused management consisting of few sprays of pesticides and frequent sanitation and harvesting. Cultural management resulted in significantly lower mean CBB infestation (4.6% vs. 9.0%), total defects (5.5% vs. 9.1%), and CBB damage to processed coffee (1.6% vs. 5.7%) compared to conventional management. Additionally, yields were higher (mean increase of 3,024 lbs of cherry/acre) and harvested more efficiently (4.8 vs. 7.9 raisins/tree) on culturally managed vs. conventional farms. Lastly, the cost of chemical controls was 55% lower and the net benefit of frequent harvesting was 48% higher on cultural vs. conventional farms. Our findings demonstrate that frequent and efficient harvesting is an effective and economically viable alternative to frequent pesticide applications.

Selective Detection of Escherichia coli K12 and Staphylococcus aureus in Mixed Bacterial Communities Using a Single-Walled Carbon Nanotube (SWCNT)-Functionalized Electrochemical Immunosensor with Dielectrophoretic Concentration.

An electrochemical immunosensor has been developed for the rapid detection and identification of potentially harmful bacteria in food and environmental samples. This study aimed to fabricate a microwire-based electrochemical immunosensor (MEI sensor) for selective detection of Escherichia coli and Staphylococcus aureus in microbial cocktail samples using dielectrophoresis (DEP)-based cell concentration. A gold-coated tungsten microwire was functionalized by coating polyethylenimine, single-walled carbon nanotube (SWCNT) suspension, streptavidin, biotinylated antibodies, and then bovine serum albumin (BSA) solutions. Double-layered SWCNTs and 5% BSA solution were found to be optimized for enhanced signal enhancement and nonspecific binding barrier. The selective capture of E. coli K12 or S. aureus cells was achieved when the electric field in the bacterial sample solution was generated at a frequency of 3 MHz and 20 Vpp. A linear trend of the change in the electron transfer resistance was observed as E. coli concentrations increased from 5.32 × 102 to 1.30 × 108 CFU/mL (R2 = 0.976). The S. aureus MEI sensor fabricated with the anti-S. aureus antibodies also showed an increase in resistance with concentrations of S. aureus (8.90 × 102-3.45 × 107 CFU/mL) with a correlation of R2 = 0.983. Salmonella typhimurium and Listeria monocytogenes were used to evaluate the specificity of the MEI sensors. The functionalization process developed for the MEI sensor is expected to contribute to the sensitive and selective detection of other harmful microorganisms in food and environmental industries.

Anti-inflammatory Quinoline Alkaloids from the Roots of Waltheria indica.

Sixteen new quinoline alkaloids (1a-7, 8a, 9, 10, 13-15, 17, and 21) and 10 known analogs (8b, 11, 12, 16, 18-20, and 22-24), along with three known cyclopeptide alkaloids (25-27), were isolated from the roots of Waltheria indica. The structures of the new compounds were elucidated by detailed NMR and circular dichroism with computational support and mass spectrometry data interpretation. Anti-inflammatory potential of isolates was evaluated based on inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production and tumor necrosis factor-alpha (TNF-α)-induced nuclear factor kappa B (NF-κB) activity with cell culture models. In the absence of cell growth inhibition, compounds 6, 8a, 9-11, 13, 21, and 24 reduced TNF-α-induced NF-κB activity with IC50 values ranging from 7.1 to 12.1 µM, comparable to the positive control (BAY 11-7082, IC50 = 9.7 µM). Compounds 6, 8a, 8b, and 11 showed significant NO-inhibitory activity with IC50 values ranging from 11.0 to 12.8 µM, being more active than the positive control (l-NMMA, IC50 = 22.7 µM). Structure-activity relationships indicated that NO inhibitory activity was significantly affected by C-8 substitution. Inhibition of LPS-induced nitric oxide synthase (iNOS) by 8b [(5S)-waltherione M, IC50 11.7 ± 0.8 µM] correlated with inhibition of iNOS mRNA expression. The biological potential of W. indica metabolites supports the traditional use of this plant for the treatment of inflammatory-related disorders.

First detection and complete genome sequence of a new tobamovirus naturally infecting Hibiscus rosa-sinensis in Hawaii.

High-throughput sequencing was used to analyze Hibiscus rosa-sinensis (family Malvaceae) plants with virus-like symptoms in Hawaii. Bioinformatic and phylogenetic analysis revealed the presence of two tobamoviruses, hibiscus latent Fort Pierce virus (HLFPV) and a new tobamovirus with the proposed name "hibiscus latent Hawaii virus" (HLHV). This is the first report of the complete sequence, genome organization, and phylogenetic characterization of a tobamovirus infecting hibiscus in Hawaii. RT-PCR with virus-specific primers and Sanger sequencing further confirmed the presence of these viruses. Inoculation experiments showed that HLFPV could be mechanically transmitted to Nicotiana benthamiana and N. tabacum, while HLHV could only be mechanically transmitted to N. benthamiana.

Survey of Viruses Infecting Basella alba in Hawaii.

Malabar spinach plants (Basella alba, Basellaceae) with leaves exhibiting symptoms of mosaic, rugosity, and malformation were found in a community garden on Oahu, HI in 2018. Preliminary studies using enzyme-linked immunosorbent assay and reverse-transcription (RT)-PCR identified Basella rugose mosaic virus (BaRMV) in symptomatic plants. However, nucleotide sequence analysis of RT-PCR amplicons indicated that additional potyviruses were also present in the symptomatic Malabar spinach. High-throughput sequencing (HTS) analysis was conducted on ribosomal RNA-depleted composite RNA samples of potyvirus-positive plants from three locations. Assembled contigs shared sequences similar to BaRMV, chilli veinal mottle virus (ChiVMV), Alternanthera mosaic virus (AltMV), Basella alba endornavirus (BaEV), broad bean wilt virus 2 (BBWV2), and Iresine viroid 1. Virus- and viroid-specific primers were designed based on HTS sequencing results and used in RT-PCR and Sanger sequencing to confirm the presence of these viruses and the viroid. We tested 63 additional samples from six community gardens for a survey of viruses in Malabar spinach and found that 21 of them were positive for BaRMV, 57 for ChiVMV, 21 for AltMV, 19 for BaEV, and 14 for BBWV2. This is the first characterization of the virome from B. alba.

Genetic Diversity of Viral Populations Associated with Ananas Germplasm and Improvement of Virus Diagnostic Protocols.

Pineapple (Ananas comosus L. [Merr.]) accessions from the U.S. Tropical Plant Genetic Resources and Disease Research (TPGRDR) in Hilo, Hawaii were subjected to RNA-sequencing to study the occurrence of viral populations associated with this vegetatively propagated crop. Analysis of high-throughput sequencing data obtained from 24 germplasm accessions and public domain transcriptome shotgun assembly (TSA) data identified two novel sadwaviruses, putatively named "pineapple secovirus C" (PSV-C) and "pineapple secovirus D" (PSV-D). They shared low amino acid sequence identity (from 34.8 to 41.3%) compared with their homologs in the Pro-pol region of the previously reported PSV-A and PSV-B. The complete genome (7485 bp) corresponding to a previously reported partial sequence of the badnavirus, pineapple bacilliform ER virus (PBERV), was retrieved from one of the datasets. Overall, we discovered a total of 69 viral sequences representing ten members within the Ampelovirus, Sadwavirus, and Badnavirus genera. Genetic diversity and recombination events were found in members of the pineapple mealybug wilt-associated virus (PMWaV) complex as well as PSVs. PMWaV-1, -3, and -6 presented recombination events across the quintuple gene block, while no recombination events were found for PMWaV-2. High recombination frequency of the RNA1 and RNA2 molecules from PSV-A and PSV-B were congruent with the diversity found by phylogenetic analyses. Here, we also report the development and improvement of RT-PCR diagnostic protocols for the specific identification and detection of viruses infecting pineapple based on the diverse viral populations characterized in this study. Given the high occurrence of recombination events, diversity, and discovery of viruses found in Ananas germplasm, the reported and validated RT-PCR assays represent an important advance for surveillance of viral infections of pineapple.

Genome sequence of pineapple secovirus B, a second sadwavirus reported infecting Ananas comosus.

The complete genome sequence of pineapple secovirus B (PSV-B), a new virus infecting pineapple (Ananas comosus) on the island of Oahu, Hawaii, was determined by high-throughput sequencing (HTS). The genome comprises two RNAs that are 5,956 and 3,808 nt long, excluding the 3'-end poly-A tails, both coding for a single large polyprotein. The RNA1 polyprotein contains five conserved domains associated with replication, while the RNA2 polyprotein is cleaved into the movement protein and coat protein. PSV-B is representative of a new species in the subgenus Cholivirus (genus Sadwavirus; family Secoviridae), as the level of amino acid sequence identity to recognized members of this subgenus in the Pro-Pol and coat protein regions is below currently valid species demarcation thresholds.

Preserving plant samples from remote locations for detection of RNA and DNA viruses.

Viral diseases in plants have a significant impact on agricultural productivity. Effective detection is needed to facilitate accurate diagnosis and characterization of virus infections essential for crop protection and disease management. For sensitive polymerase chain reaction (PCR)-based methods, it is important to preserve the integrity of nucleic acids in plant tissue samples. This is especially critical when samples are collected from isolated areas, regions distant from a laboratory, or in developing countries that lack appropriate facilities or equipment for diagnostic analyses. RNAlater ® provides effective, reliable sample storage by stabilizing both RNA and DNA in plant tissue samples. Our work indicated that total RNA or DNA extracted from virus-infected leaf samples preserved in RNAlater ® was suitable for reverse transcription polymerase chain reaction (RT-PCR), PCR, Sanger sequencing, high-throughput sequencing (HTS), and enzyme-linked immunosorbent assay (ELISA)-based diagnostic analyses. We demonstrated the effectiveness of this technology using leaf tissue samples from plants with virus symptoms grown in farmers' fields in Bangladesh. The results revealed that RNAlater ® technology was effective for detection and characterization of viruses from samples collected from remote areas and stored for extended periods. Adoption of this technology by developing countries with limited laboratory facilities could greatly increase their capacity to detect and diagnose viral infections in crop plants using modern analytical techniques.

Coumarinolignans with Reactive Oxygen Species (ROS) and NF-κB Inhibitory Activities from the Roots of Waltheria indica.

Seven new coumarinolignans, walthindicins A-F (1a, 1b, 2-5, 7), along with five known analogs (6, 8-11), were isolated from the roots of Waltheria indica. The structures of the new compounds are determined by detailed nuclear magnetic resonance (NMR), circular dichroism (CD) with extensive computational support, and mass spectroscopic data interpretation. Compounds were tested for their antioxidant activity in Human Cervical Cancer cells (HeLa cells). Compounds 1a and 6 showed higher reactive oxygen species (ROS) inhibitory activity at 20 µg/mL when compared with other natural compound-based antioxidants such as ascorbic acid. Considering the role of ROS in nuclear-factor kappa B (NF-κB) activation, compounds 1a and 6 were evaluated for NF-κB inhibitory activity and showed a concentration-dependent inhibition in Human Embryonic Kidney 293 cells (Luc-HEK-293).

Complete genome organization and characterization of Hippeastrum latent virus.

The complete genome sequences of two carlaviruses were determined by high-throughput sequencing of RNA extracted from ringspot and mosaic, disease symptoms on leaves of spider lily plants (Crinum asiaticum, family Amaryllidaceae) growing as landscape plants in Hawaii. One, named Nerine latent virus (NeLV)-Hawaii with a genome of 8281 nucleotide exhibited the highest nucleotide identity and amino acid similarity of 95.5% and 96.0%, respectively, to the genome sequence of an isolate of NeLV from Narcissus sp. in Australia (JQ395044). The second, named Hippeastrum latent virus (HiLV)-Hawaii with a genome of 8497 nucleotides exhibited the highest nucleotide identity and amino acid similarity, 84.3% and 88.7%, respectively, to the sequence of a previously uncharacterized HiLV isolate from a potted flowering plant, Amaryllis (Hippeastrum hybridum Hort) in Taiwan (DQ098905). The amino acid sequence similarities of replicase (Rep) and coat protein (CP) between HiLV-Hawaii and NeLV-Hawaii were 44.8% and 38.4%, respectively. Results of viral protein Rep and CP amino acid sequence comparisons from various carlaviruses provide evidence that HiLV and NeLV, previously classified as synonymous viruses are in fact unique viruses. This is the first report for the complete sequence, organization, and phylogenetic characterization of HiLV and the first detection of HiLV both in C. asiaticum and in the USA.

Antimicrobial Activity of Ohelo Berry (Vaccinium calycinum) Juice against Listeria monocytogenes and Its Potential for Milk Preservation.

Listeria monocytogenes is a foodborne pathogen and causes illnesses with a high mortality rate in susceptible populations. Several dairy-related outbreaks have been attributed to contamination by L. monocytogenes, which requires antimicrobial interventions to enhance the safety of these products. This study aimed to determine the antimicrobial activity of the ohelo berry (Vaccinium calycinum), a Hawaiian wild relative of cranberry, against L. monocytogenes in culture media and milk products. The effect of ohelo berry juice at its sub-inhibitory concentrations on the physicochemical properties, biofilm formation, and gene expression of L. monocytogenes was also investigated. The minimum inhibitory concentration of ohelo berry juice against L. monocytogenes was 12.5%. The sub-inhibitory concentration of ohelo berry juice (6.25%) significantly increased the auto-aggregation and decreased the hydrophobicity, swimming motility, swarming motility, and biofilm formation capability of L. monocytogenes. The relative expression of genes for motility (flaA), biofilm formation and disinfectant resistance (sigB), invasion (iap), listeriolysin (hly), and phospholipase (plcA) was significantly downregulated in L. monocytogenes treated by the 6.25% juice. L. monocytogenes was significantly inhibited in whole and skim milk supplemented with 50% ohelo berry juice, regardless of the fat content. These findings highlight the potential of ohelo berry as a natural preservative and functional food to prevent L. monocytogenes infection.

Aqueous chlorine dioxide generated with organic acids have higher antimicrobial efficacy than those generated with inorganic acids.

Chlorine dioxide (ClO2) is commonly generated by mixing sodium chlorite and acid. This study aimed to evaluate how acid affects the release kinetics and antimicrobial property of ClO2. Solutions made with weak acids released ClO2 more slowly and had higher stability than those made with hydrochloric acid. Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes were treated with 1, 2.5, and 5 ppm ClO2 for 3 or 5 min. Lettuce inoculated with the pathogenic bacteria were treated with 2.5 and 5 ppm ClO2 for 5 min. The effects of peptone load at 0.01% and 0.02% on the antimicrobial efficacy of ClO2 were investigated in S. Typhimurium cell suspensions. The contribution of acids alone at the pH of the ClO2 solutions to bacterial reduction was also evaluated. The 2.5 ppm ClO2 solutions made with citric acid, lactic acid, and malic acid showed higher reductions in all three bacteria than ClO2 made with hydrochloric acid and sodium bisulfate. The 5 ppm ClO2 solutions produced with organic acids reduced populations of all bacterial strains from 7 log CFU/mL to undetectable level in 3 min, except S. Typhimurium treated by ClO2 produced with lactic acid. On inoculated Romaine lettuce model, 5 ppm ClO2 produced with lactic acid and malic acid resulted in the highest reduction of E. coli O157:H7, S. Typhimurium, and L. monocytogenes of approximately 1.4, 1.7, and 2.4 log CFU/g, respectively. The antimicrobial efficacy of ClO2 made with HCl and NaHSO4 were affected by 0.01% and 0.02% peptone load, respectively. Food-grade organic acids produced aqueous ClO2 solutions with stronger antimicrobial properties than inorganic acids. The acids alone at the pH of ClO2 did not show significant bacterial reductions.

A single-walled carbon nanotubes-based electrochemical impedance immunosensor for on-site detection of Listeria monocytogenes.

Real-time and sensitive detection of pathogenic bacteria in food is in high demand to ensure food safety. In this study, a single-walled carbon nanotubes (SWCNTs)-based electrochemical impedance immunosensor for on-site detection of Listeria monocytogenes (L. monocytogenes) was developed. A gold-plated wire was functionalized using polyethylenimine (PEI), SWCNTs, streptavidin, biotinylated L. monocytogenes antibodies, and bovine serum albumin (BSA). A linear relationship (R2  = 0.982) between the electron transfer resistance measurements and concentrations of L. monocytogenes within the range of 103 -108 CFU/ml was observed. In addition, the sensor demonstrated high selectivity towards the target in the presence of other bacterial cells such as Salmonella Typhimurium and Escherichia coli O157:H7. To facilitate the demand for on-site detection, the sensor was integrated into a smartphone-controlled biosensor platform, consisting of a compact potentiostat device and a smartphone. The signals from the proposed platform were compared with a conventional potentiostat using the immunosensor interacted with L. monocytogenes (103 -105 CFU/ml). The signals obtained with both instruments showed high consistency. Recovery percentages of lettuce homogenate spiked with 103 , 104 , and 105 CFU/ml of L. monocytogenes obtained with the portable platform were 90.21, 90.44, and 93.69, respectively. The presented on-site applicable SWCNT-based immunosensor platform was shown to have a high potential to be used in field settings for food and agricultural applications. PRACTICAL APPLICATION: The developed immunosensor was developed for on-site detection of L. monocytogenes. The limit of detection of the sensor was 103 CFU/ml with a detection time of 10 min. In order to facilitate the requirements for effective on-site screening for food safety, the sensor was integrated into a smartphone-controlled platform, so that the bio-molecular interactions were converted into impedance signals and transmitted wirelessly to a smartphone by a hand-held EIS transducer.

First Detection and Genome Characterization of a New RNA Virus, Hibiscus Betacarmovirus, and a New DNA Virus, Hibiscus Soymovirus, Naturally Infecting Hibiscus spp. in Hawaii.

Hibiscus (Hibiscus spp., family Malvaceae) leaves exhibiting symptoms of mosaic, ringspot, and chlorotic spots were collected in 2020 on Oahu, HI. High-throughput sequencing analysis was conducted on ribosomal RNA-depleted composite RNA samples extracted from symptomatic leaves. About 77 million paired-end reads and 161,970 contigs were generated after quality control, trimming, and de novo assembly. Contig annotation with BLASTX/BLASTN searches revealed a sequence (contig 1) resembling the RNA virus, hibiscus chlorotic ringspot virus (genus Betacarmovirus), and one (contig 2) resembling the DNA virus, peanut chlorotic streak virus (genus Soymovirus). Further bioinformatic analyses of the complete viral genome sequences indicated that these viruses, with proposed names of hibiscus betacarmovirus and hibiscus soymovirus, putatively represent new species in the genera Betacarmovirus and Soymovirus, respectively. RT-PCR using specific primers, designed based on the retrieved contigs, coupled with Sanger sequencing, further confirmed the presence of these viruses. An additional 54 hibiscus leaf samples from other locations on Oahu were examined to determine the incidence and distribution of these viruses.