This study examined an aquaponic approach of circulating water containing ammonia excretions from African catfish grown in an aquaculture tank for bacterial conversion into nitrates, which then acted as a nutrient substance to cultivate lettuce in hydroponic tank. We found that microwave pyrolysis biochar (450 g) having microporous (1.803 nm) and high BET surface area (419 m2/g) was suitable for use as biological carrier to grow nitrifying bacteria (63 g of biofilm mass) that treated the water quality through removing the ammonia (67%) and total suspended solids (68%), resulting in low concentration of remaining ammonia (0.42 mg/L) and total suspended solid (59.40 mg/L). It also increased the pH (6.8), converted the ammonia into nitrate (29.7 mg/L), and increased the nitrogen uptake by the lettuce (110 mg of nitrogen per plant), resulting in higher growth in lettuce (0.0562 %/day) while maintaining BOD5 level (3.94 mg/L) at acceptable level and 100% of catfish survival rate. Our results demonstrated that microwave pyrolysis biochar can be a promising solution for growing nitrifying bacteria in aquaponic system for simultaneous toxic ammonia remediation and generation of nitrate for growing vegetable in aquaculture industry.
Food security and sustainable development of agriculture has been a key challenge for decades. To support this, nanotechnology in the agricultural sectors increases productivity and food security, while leaving complex environmental negative impacts including pollution of the human food chains by nanoparticles. Here we model the effects of silver nanoparticles (Ag-NPs) in a food chain consisting of soil-grown lettuce Lactuca sativa and snail Achatina fulica. Soil-grown lettuce were exposed to sulfurized Ag-NPs via root or metallic Ag-NPs via leaves before fed to snails. We discover an important biomagnification of silver in snails sourced from plant root uptake, with trophic transfer factors of 2.0-5.9 in soft tissues. NPs shifts from original size (55-68 nm) toward much smaller size (17-26 nm) in snails. Trophic transfer of Ag-NPs reprograms the global metabolic profile by down-regulating or up-regulating metabolites for up to 0.25- or 4.20- fold, respectively, relative to the control. These metabolites control osmoregulation, phospholipid, energy, and amino acid metabolism in snails, reflecting molecular pathways of biomagnification and pontential adverse biological effects on lower trophic levels. Consumption of these Ag-NP contaminated snails causes non-carcinogenic effects on human health. Global public health risks decrease by 72% under foliar Ag-NP application in agriculture or through a reduction in the consumption of snails sourced from root application. The latter strategy is at the expense of domestic economic losses in food security of $177.3 and $58.3 million annually for countries such as Nigeria and Cameroon. Foliar Ag-NP application in nano-agriculture has lower hazard quotient risks on public health than root application to ensure global food safety, as brought forward by the United Nations Sustainable Development Goals.
Understanding metabolite changes and underlying metabolic pathways that may be affected in target plants following essential oils (EOs) exposure is of great importance. In this study, a gas chromatography-mass spectrometry (GC/MS) based metabolomics approach was used to determine the metabolite changes in lettuce (Lactuca sativa L.) shoot and root after exposure to different concentrations of W. trilobata EO. Multivariate analyses of principal component analysis (PCA) and orthogonal partial least-discriminant analysis (OPLS-DA) corroborated that shoot and root of lettuce responded differently to W. trilobata EO. In EO-exposed shoot samples, an increase in the levels of malic acid, glutamine, serine, lactose and α-glucopyranose affected important metabolism pathways such as glycolysis, fructose and mannose metabolism and galactose metabolism. The findings suggest that lettuce may be up-regulating these metabolites to increase tolerance against W. trilobata EO. In EO-exposed root samples, changes in fatty acid biosynthesis, elongation, degradation, phenylalanine, tyrosine and tryptophan metabolism were linked to a decrease in lyxose, palmitic acid, octadecanoic acid, aspartic acid, phenylalanine and myo-inositol. These results indicate that W. trilobata EO could cause alterations in fatty acid compositions and lead to inhibition of roots growth. Together, these findings provide insight into the metabolic responses of lettuce upon W. trilobata EO exposure, as well as potential mechanisms of action of W. trilobata EO as bio-herbicides.
Bacterial communication or quorum sensing (QS) is achieved via sensing of QS signaling molecules consisting of oligopeptides in Gram-positive bacteria and N-acyl homoserine lactones (AHL) in most Gram-negative bacteria. In this study, Enterobacteriaceae isolates from Batavia lettuce were screened for AHL production. Enterobacter asburiae, identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was found to produce short chain AHLs. High resolution triple quadrupole liquid chromatography mass spectrometry (LC/MS) analysis of the E. asburiae spent supernatant confirmed the production of N-butanoyl homoserine lactone (C4-HSL) and N-hexanoyl homoserine lactone (C6-HSL). To the best of our knowledge, this is the first report of AHL production by E. asburiae.
A crude methanol extract of Goniothalamus andersonii J. Sinclair strongly inhibited elongation of lettuce (Lactuca sativa L.) radicles. We conducted bioassay-guided purification of G. andersonii bark extract and obtained goniothalamin as the major bioactive compound. Its EC50 values against elongation of lettuce radicles and hypocotyls were 50 and 125 micromol L(-1), respectively. Among the six species tested, timothy was the most sensitive to goniothalamin. Quantification of this compound in other Goniothalamus species suggested that the plant inhibitory activity of this genus is explainable by goniothalamin, with G. calcareus as an exception.
Matched MeSH terms: Lettuce/drug effects*; Lettuce/growth & development
Weed infestation is a persistent problem for centuries and continues to be major yield reducing issue in modern agriculture. Chemical weed control through herbicides results in numerous ecological, environmental, and health-related issues. Moreover, numerous herbicides have evolved resistance against available herbicides. Plant extracts are regarded as an alternative to herbicides and a good weed management option. The use of plant extracts is environmentally safe and could solve the problem of herbicide resistance. Therefore, laboratory and wire house experiments were conducted to evaluate the phytotoxic potential of three Fabaceae species, i.e., Cassia occidentalis L. (Coffee senna), Sesbania sesban (L.) Merr. (Common sesban) and Melilotus alba Medik. (White sweetclover) against seed germination and seedling growth of some broadleaved weed species. Firstly, N-hexane and aqueous extracts of these species were assessed for their phytotoxic effect against lettuce (Lactuca sativa L.). The extracts found more potent were further tested against germination and seedling growth of four broadleaved weed species, i.e., Parthenium hysterophorus L. (Santa-Maria), Trianthema portulacastrum L. (Pigweed), Melilotus indica L (Indian sweetclover). and Rumex dentatus L. (Toothed dock) in Petri dish and pot experiments. Aqueous extracts of all species were more toxic than their N-hexane forms for seed germination and seedling growth of lettuce; therefore, aqueous extracts were assessed for their phytotoxic potential against four broadleaved weed species. Aqueous extracts of all species proved phytotoxic against T. portulacastrum, P. hysterophorus, M. indica and R. dentatus and retarder their germination by 57, 90, 100 and 58%, respectively. Nevertheless, foliar spray of C. occidentalis extract was the most effective against T. portulacastrum as it reduced its dry biomass by 72%, while M. alba was effective against P. hysterophorus, R. dentatus and M. indica and reduced their dry biomass by 55, 68 and 81%, respectively. It is concluded that aqueous extracts of M. alba, S. sesban and C. occidentalis could be used to retard seed germination of T. portulacastrum, P. hysterophorus, M. indica and R. dentatus. Similarly, aqueous extracts of C. occidentalis can be used to suppress dry biomass of T. portulacastrum, and those of M. alba against P. hysterophorus, R. dentatus. However, use of these extracts needs their thorough testing under field conditions.
Klebsiella pneumoniae (K. pneumoniae) is one of the most important members of Klebsiella genus in Enterobacteriacae family, which is responsible for pneumonia (the destructive lung inflammation disease). Vegetables are known as source of contamination with K. pneumonia. Raw vegetables are usually consumed in salads and other dishes. The aim of this study was to investigate the occurrence of K. pneumoniae in raw vegetables marketed in Malaysia. Two hundred commonly used salad vegetables (lettuces, parsley, cucumber, tomato and carrot) from hypermarkets and wet markets were investigated for presence of K. pneumoniae using Most Probable Number-Polymerase Chain Reaction (MPN-PCR). K. pneumoniae was found to be significantly more frequent (100%) and (82.5%) in lettuce and cucumbers, respectively. K. pneumoniae contamination was lowest in carrot samples (30%). All samples were contaminated with K. pneumoniae ranging from
While there is good epidemiological evidence for foods as vehicles for norovirus transmission, the precise means of spread and its control remain unknown. The feline calicivirus was used as a surrogate for noroviruses to study infectious virus transfer between hands and selected types of foods and environmental surfaces. Assessment of the potential of selected topicals in interrupting such virus transfer was also made. Ten microliters of inoculum of feline calicivirus deposited onto each fingerpad of adult subjects was allowed to air dry and the contaminated area on individual fingerpads was pressed (10 s at a pressure of 0.2 to 0.4 kg/cm2) onto 1-cm-diameter disks of ham, lettuce, or brushed stainless steel. The virus remaining on the donor and that transferred to the recipient surfaces was eluted and plaque assayed. Virus transfer to clean hands from experimentally contaminated disks of ham, lettuce, and stainless steel was also tested. Nearly 46 +/- 20.3, 18 +/- 5.7, and 13 +/- 3.6% of infectious virus was transferred from contaminated fingerpads to ham, lettuce, and metal disks, respectively. In contrast, approximately 6 +/- 1.8, 14 +/- 3.5, and 7 +/- 1.9% virus transfer occurred, respectively, from ham, lettuce, and metal disks to hands. One-way analysis of variance test showed that pretreatment (washing) of the fingerpads either with water or with both topical agent and water significantly (P < 0.05) reduced virus transfer to < or = 0.9%, as compared with < or = 2.3 and < or = 3.4% transfer following treatments with either 75% (vol/vol) ethanol or a commercial hand gel containing 62% ethanol, respectively. Despite wide variations in virus transfer among the targeted items used, intervention agents tested reduced virus transfer significantly (P < 0.05) when compared with that without such treatments (71 +/- 8.9%). These findings should help in a better assessment of the potential for cross-contamination of foods during handling and also assist in developing more effective approaches to foodborne spread of norovirus infections.
A soil cooling system that prepares soil for temperate soil temperatures for the growth of temperate crops under a tropical climate is described herein. Temperate agriculture has been threatened by the negative impact of temperature increases caused by climate change. Soil temperature closely correlates with the growth of temperate crops, and affects plant processes and soil microbial diversity. The present study focuses on the effects of soil temperatures on lettuce growth and soil microbial diversity that maintains the growth of lettuce at low soil temperatures. A model temperate crop, loose leaf lettuce, was grown on eutrophic soil under soil cooling and a number of parameters, such as fresh weight, height, the number of leaves, and root length, were evaluated upon harvest. Under soil cooling, significant differences were observed in the average fresh weight (P<0.05) and positive development of the roots, shoots, and leaves of lettuce. Janthinobacterium (8.142%), Rhodoplanes (1.991%), Arthrospira (1.138%), Flavobacterium (0.857%), Sphingomonas (0.790%), Mycoplana (0.726%), and Pseudomonas (0.688%) were the dominant bacterial genera present in cooled soil. Key soil fungal communities, including Pseudaleuria (18.307%), Phoma (9.968%), Eocronartium (3.527%), Trichosporon (1.791%), and Pyrenochaeta (0.171%), were also recovered from cooled soil. The present results demonstrate that the growth of temperate crops is dependent on soil temperature, which subsequently affects the abundance and diversity of soil microbial communities that maintain the growth of temperate crops at low soil temperatures.
To explore new approaches of phage-based bio-process of specifically pathogenic Escherichia coli bacteria in food products within a short period. One hundred and forty highly lytic designed coliphages were used. Escherichia coli naturally contaminated and Enterohemorrhagic Escherichia coli experimentally inoculated samples of lettuce, cabbage, meat, and egg were used. In addition, experimentally produced biofilms of E. coli were tested. A phage concentration of 10(3) PFU/ml was used for food products immersion, and for spraying of food products, 10(5) PFU/ml of a phage cocktail was used by applying a 20-s optimal dipping time in a phage cocktail. Food samples were cut into pieces and were either sprayed with or held in a bag immersed in lambda buffer containing a cocktail of 140 phages. Phage bio-processing was successful in eliminating completely E. coli in all processed samples after 48 h storage at 4°C. Partial elimination of E. coli was observed in earlier storage periods (7 and 18 h) at 24° and 37°C. Moreover, E. coli biofilms were reduced >3 log cycles upon using the current phage bio-processing. The use of a phage cocktail of 140 highly lytic designed phages proved highly effective in suppressing E. coli contaminating food products. Proper decontamination/prevention methods of pathogenic E. coli achieved in this study can replace the current chemically less effective decontamination methods.
In June 2011, lettuce (Lactuca sativa) plants cultivated in major lettuce growing areas in Malaysia, including the Pahang and Johor states, had extensive leaf spots. In severe cases, disease incidence was recorded more than 80%. Symptoms on 50 observed plants initially were as water soaked spots (1 to 2 mm in diameter) on leaves, and then became circular spots spreading over much of the leaves. In this research, main lettuce growing areas infected by the pathogen in the mentioned states were investigated and the pathogen was isolated onto potato dextrose agar (PDA). Colonies observed were greyish green to light brown. Single conidia were formed at the terminal end of conidiophores that were 28.8 to 40.8 μm long and 11.0 to 19.2 μm wide, and 2 to 7 transverse and 1 to 4 longitudinal septa. To produce conidia, the fungus was grown on potato carrot agar (PCA) and V8 juice agar media under 8-h/16-h light/dark photoperiod. Fourteen isolates were identified Stemphylium solani based on morphological criteria described by Kim et al. (1). To confirm morphological characterization, DNA of the fungus was extracted from mycelium and PCR was done using universal primers ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'), which amplified the internal transcribed spacer (ITS) region of rDNA (2). The sequencing result was subjected to BLAST analysis which was 99% identical to the other published sequences in the GenBank database (GenBank Accession Nos. AF203451 and HQ840713). The nucleotide sequence was deposited in GenBank under Accession No. JQ736022. Pathogenicity testing of representative isolate was done using 20 μl of conidial suspension with a concentration of 1 × 105/ml in droplets (three drops on each leaf) on four detached 45-day-old lettuce leaves cv. BBS012 (3). Fully expended leaves were placed on moist filter paper in petri dishes and were incubated in humid chambers at 25°C. The leaves inoculated with sterile water served as control. After 7 days, disease symptoms were observed, which were similar to those symptoms collected in infected fields and the fungus was reisolated and confirmed as S. solani based on morphological criteria (1) and molecular characterization (2). Control leaves remained healthy. Pathogenicity testing was completed twice. To our knowledge, this is the first report of S. solani on lettuce in Malaysia and it may become a serious problem because of its broad host range, variability in pathogenic isolates, and prolonged active phase of the disease cycle. Previous research has shown that S. solani is a causal agent of gray leaf spot on lettuce in China (4). References: (1) B. S. Kim et al. Plant Pathol. J. 20:85, 2004. (2) Y. R. Mehta et al. Current Microbiol. 44:323, 2002. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) F. L. Tai. Sylloge Fungorum Sinicorum, Sci. Press, Acad. Sin., Peking, 1979.
The use of allelopathy concept in weed management has received attentions to minimize extensively the reliance on herbicide applications on the agriculture industry in Malaysia. A laboratory study was conducted to evaluate the allelopathic potential of 15 Malaysian common weed species of different morphological characteristics (broadleaves, sedges and grasses). They were screened using the Sandwich method (from leaf litter leachate) and the Dish pack method (for testing the presence and content of volatile compounds in weeds). Among the 15 weed species tested, the leaf litter leachate of Centrosema pubescens was observed to be the most sensitive plant material inhibiting the growth of lettuce radicle (84%) and hypocotyl (55%) in the Sandwich bioassay compared to the control. This was followed by Asystasia gangentica (81%) and Cynodon dactylon (80%) inhibiting the lettuce radicle growth. In the Dish pack bioassay, Rhynchelytrum repens demonstrated maximum inhibition on the radicle and hypocotyl elongations by 44% and 29%, respectively, (over control) at 41 mm distance from the source well. Meanwhile, at the same distance, Cynodon dactylon was observed to have the least inhibitory effect on lettuce radicle growth by 12%. The results presented can be utilized as benchmark information for further research on the identification and isolation of allelochemicals for weed control strategies.
Since the inception of global industrialization, the growth of steroid estrogens becomes a matter of emerging serious concern for the rapid population. Steroidal estrogens are potent endocrine-upsetting chemicals that are excreted naturally by vertebrates (e.g., humans and fish) and can enter natural waters through the discharge of treated and raw sewage. Steroidal estrogens in plants may enter the food web and become a serious threat to human health. We evaluated the uptake and accumulation of ethinylestradiol (EE2) and 17β-estradiol (17β-E2) in lettuce plants (Lactuca sativa) grown under controlled environmental condition over 21 days growth period. An effective analytical method based on ultrasonic liquid extraction (ULE) for solid samples and solid phase extraction (SPE) for liquid samples with gas chromatography-mass spectrometry (GC/MS) has been developed to determine the steroid estrogens in lettuce plants. The extent of uptake and accumulation was observed in a dose-dependent manner and roots were major organs for estrogen deposition. Unlike the 17β-E2, EE2 was less accumulated and translocated from root to leaves. For 17β-E2, the distribution in lettuce was primarily to roots after the second week (13%), whereas in leaves it was (10%) over the entire study period. The distribution of EE2 at 2000 μg L-1 in roots and leaves was very low (3.07% and 0.54%) during the first week and then was highest (12% in roots and 8% in leaves) in last week. Bioaccumulation factor values of 17β-E2 and EE2 in roots were 0.33 and 0.29 at 50 μg L-1 concentration as maximum values were found at 50 μg L-1 rather than 500 and 2000 in all observed plant tissues. Similar trend was noticed in roots than leaves for bioconcentration factor as the highest bioconcentration values were observed at 50 μg L-1 concentration instead of 500 and 2000 μg L-1 spiked concentration. These findings mainly indicate the potential for uptake and bioaccumulation of estrogens in lettuce plants. Overall, the estrogen contents in lettuce were compared to the FAO/WHO recommended toxic level and were found to be higher than the toxic level which is of serious concern to the public health. This analytical procedure may aid in future studies on risks associated with uptake of endocrine-disrupting chemicals in lettuce plants.
Kitchen mishandling practices contribute to a large number of foodborne illnesses. In this study, the transfer and cross-contamination potential of Vibrio parahaemolyticus from bloody clams to ready-to-eat food (lettuce) was assessed. Three scenarios were investigated: 1) direct cross-contamination, the transfer of V. parahaemolyticus from bloody clams to non-food contact surfaces (hands and kitchen utensils) to lettuce (via slicing), was evaluated; 2) perfunctory decontamination, the efficacy of two superficial cleaning treatments: a) rinsing in a pail of water, and b) wiping with a kitchen towel, were determined; and 3) secondary cross-contamination, the microbial transfer from cleaning residuals (wash water or stained kitchen towel) to lettuce was assessed. The mean of percent transfer rates through direct contact was 3.6%, and an average of 3.5% of total V. parahaemolyticus was recovered from sliced lettuce. The attempted treatments reduced the transferred population by 99.0% (rinsing) and 94.5% (wiping), and the relative amount of V. parahaemolyticus on sliced lettuce was reduced to 0.008%. V. parahaemolyticus exposure via secondary cross-contamination was marginal. The relative amount of V. parahaemolyticus recovered from washed lettuce was 0.07%, and the transfers from stained kitchen towel to lettuce were insubstantial. Our study highlights that V. parahaemolyticus was readily spread in the kitchen, potentially through sharing of non-food contact surfaces. Results from this study can be used to better understand and potentially raising the awareness of proper handling practices to avert the spread of foodborne pathogens.
Hepatitis A virus (HAV) is an important pathogen which has been responsible for many food-borne outbreaks. HAV-excreting food handlers, especially those with poor hygienic practices, can contaminate the foods which they handle. Consumption of such foods without further processing has been known to result in cases of infectious hepatitis. Since quantitative data on virus transfer during contact of hands with foods is not available, we investigated the transfer of HAV from artificially contaminated fingerpads of adult volunteers to pieces of fresh lettuce. Touching the lettuce with artificially contaminated fingerpads for 10 s at a pressure of 0.2 to 0.4 kg/cm(2) resulted in transfer of 9.2% +/- 0.9% of the infectious virus. The pretreatments tested to interrupt virus transfer from contaminated fingerpads included (i) hard-water rinsing and towel drying, (ii) application of a domestic or commercial topical agent followed by water rinsing and towel drying, and (iii) exposure to a hand gel containing 62% ethanol or 75% liquid ethanol without water rinsing or towel drying. When the fingerpads were treated with the topical agents or alcohol before the lettuce was touched, the amount of infectious virus transferred to lettuce was reduced from 9.2% to between 0.3 and 0.6% (depending on the topical agent used), which was a reduction in virus transfer of up to 30-fold. Surprisingly, no virus transfer to lettuce was detected when the fingerpads were rinsed with water alone before the lettuce was touched. However, additional experiments with water rinsing in which smaller volumes of water were used (1 ml instead of 15 ml) showed that the rate of virus transfer to lettuce was 0.3% +/- 0.1%. The variability in virus transfer rates following water rinsing may indicate that the volume of water at least in part influences virus removal from the fingerpads differently, a possibility which should be investigated further. This study provided novel information concerning the rate of virus transfer to foods and a model for investigating the transfer of viral and other food-borne pathogens from contaminated hands to foods, as well as techniques for interrupting such transfer to improve food safety.
Immunomagnetic beads-PCR (IM-PCR), positively-charged virosorb filters (F), or a combination of both methods (F-IM-PCR) were used to capture, concentrate and rapidly detect hepatitis A virus (HAV) in samples of lettuce and strawberries experimentally contaminated. Direct reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of the collected HAV-beads complex showed a detection limit of 0.5 plaque forming units (PFU) of the virus present in 1-ml of wash solution from the produce, which was several hundred-fold more sensitive than that demonstrated by RT-PCR. In separate trials, virus-containing wash solutions from the produce were passed through the filters and the captured virus was eluted with 10 ml volumes of 1% beef extract. Of the 62% filter-captured HAV, an average of 34.8% was eluted by the 1% beef extract. PCR amplification of 2 microl from this eluate failed to produce a clear positive band signal. As little as 10 PFU, present on each piece of the lettuce or strawberry, was detectable by the F-IM-PCR, which was almost 20 times less sensitive than the detection limit of 0.5 PFU by the IM-PCR. However, considering the large volumes (< or =50 ml) used in the F-IM-PCR, the sensitivity of detection could be much greater than that of the IM-PCR, which was restricted to < or =20 ml volumes. These data indicate that the F-IM-PCR method provides the potential for a greater sensitivity of detection than the IM-PCR, since low levels of virus could be detected from large volumes of sample than possible by the IM-PCR method. Although positively-charged filters captured a greater amount of virus than both the IM-PCR and F-IM-PCR methods, direct PCR amplification from beef extract eluates was not successful in detecting HAV from produce.
This study investigated the removal of bacterial surface structures, particularly flagella, using sonication, and examined its effect on the attachment of Salmonella Typhimurium ATCC 14028 cells to plant cell walls. S. Typhimurium ATCC 14028 cells were subjected to sonication at 20 kHz to remove surface structures without affecting cell viability. Effective removal of flagella was determined by staining flagella of sonicated cells with Ryu's stain and enumerating the flagella remaining by direct microscopic counting. The attachment of sonicated S. Typhimurium cells to bacterial cellulose-based plant cell wall models and cut plant material (potato, apple, lettuce) was then evaluated. Varying concentrations of pectin and/or xyloglucan were used to produce a range of bacterial cellulose-based plant cell wall models. As compared to the non-sonicated controls, sonicated S. Typhimurium cells attached in significantly lower numbers (between 0.5 and 1.0 log CFU/cm2) to all surfaces except to the bacterial cellulose-only composite without pectin and xyloglucan. Since attachment of S. Typhimurium to the bacterial cellulose-only composite was not affected by sonication, this suggests that bacterial surface structures, particularly flagella, could have specific interactions with pectin and xyloglucan. This study indicates that sonication may have potential applications for reducing Salmonella attachment during the processing of fresh produce.
In gram-negative bacteria, bacterial communication or quorum sensing (QS) is achieved using common signaling molecules known as N-acyl homoserine lactones (AHL). We have previously reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. In silico analysis of the strain L1 genome revealed the presence of a pair of luxI/R genes responsible for AHL-type QS, designated as easIR. In this work, the 639 bp luxI homolog, encoding 212 amino acids, have been cloned and overexpressed in Escherichia coli BL21 (DE3)pLysS. The purified protein (~25 kDa) shares high similarity to several members of the LuxI family among different E asburiae strains. Our findings showed that the heterologously expressed EasI protein has activated violacein production by AHL biosensor Chromobacterium violaceum CV026 as the wild-type E. asburiae. The mass spectrometry analysis showed the production of N-butanoyl homoserine lactone and N-hexanoyl homoserine lactone from induced E. coli harboring the recombinant EasI, suggesting that EasI is a functional AHL synthase. E. asburiae strain L1 was also shown to possess biofilm-forming characteristic activity using crystal violet binding assay. This is the first report on cloning and characterization of the luxI homolog from E. asburiae.
As consumer interest in organically grown vegetables is increasing in Malaysia, there is a need to answer whether the vegetables are more nutritious than those conventionally grown. This study investigates commercially available vegetables grown organically and conventionally, purchased from retailers to analyse â-carotene, vitamin C and riboflavin contents. Five types of green vegetables were selected, namely Chinese mustard (sawi) (Brassica juncea), Chinese kale (kailan) (Brassica alboglabra), lettuce (daun salad) (Lactuca sativa), spinach (bayam putih) (Amaranthus viridis) and swamp cabbage (kangkung) (Ipomoea aquatica). For vitamin analysis, a reverse-phase high performance liquid chromatography was used to identify and quantify β-carotene, vitamin C and riboflavin. The findings showed that not all of the organically grown vegetables were higher in vitamins than that conventionally grown. This study found that only swamp cabbage grown organically was highest in β-carotene , vitamin C and riboflavin contents among the entire samples studied. The various nutrients in organically grown vegetables need to be analysed for the generation of a database on nutritional value which is important for future research.
This study aims to determine the frequency and density of potentially pathogenic Vibrio parahaemolyticus, defined as those possessing thermostable-direct hemolysin (tdh) and/or tdh-related hemolysin (trh) genes, in raw salad vegetables at retail level in Selangor, Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was applied to detect the presence of tdh and/or trh gene-possessing V. parahaemolyticus and to enumerate their density in the samples. A total of 276 samples of vegetables commonly eaten raw in Malaysia (Cabbage = 30; Carrot = 31; Cucumber = 28; Four winged bean = 26; Indian pennywort = 17; Japanese parsley = 21; Lettuce = 16; Long bean = 32; Sweet potato = 29; Tomato = 38; Wild cosmos = 8) were analyzed. The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). With the MPN-PCR technique, about 12.0% of the samples were positive for the presence of V. parahaemolyticus tdh-positive, with maximum densities of up to 39 MPN/g. The total frequency of V. parahaemolyticus trh-positive in the samples was 10.1%, with maximum concentration 15 MPN/g. V. parahaemolyticus tdh-positive was most prevalent in samples from Wet Market C (20.78%) and also in vegetable type Oenanthe stolonifera (Japanese parsley) with 19.0%, while V. parahaemolyticus trhpositive was predominant in samples from Wet Market D (16.7%) and was most frequent in both Oenanthe stolonifera (Japanese parsley) and Cucumis sativus (Cucumber) with 14.3% prevalence for each type. The results highlighted the fact that raw vegetables could be contaminated with virulent V. parahaemolyticus and could act as a transmission route, thus poses risk to consumers from the consumption of raw vegetables. To the author’s knowledge, this is the first assessment of V. parahaemolyticus carrying tdh and trh genes in raw
vegetables from retail outlets in Malaysia.