Edible bird's nest (EBN) which is solidified swiftlet's saliva contains high nutritional value. It is widely consumed in countries like Malaysia, Indonesia, and Thailand. However, previous proximate analysis of Malaysia EBN was not representative of all the regions in Malaysia. In recent years, safety issues such as high nitrate and nitrite contents, presence of heavy metal, adulteration, fungal infection, and cancer cell stimulation were associated with EBN. Hence, this study aimed to determine the proximate analysis, safety profile during normal weather and hazy periods, and its effect on cancer cells stimulation in Malaysia-farmed EBN. Seven raw cleaned EBN samples were sourced from 6 different regions in Malaysia. Proximate analysis and safety profile were performed using official AOCA methods and Malaysian Standard. High protein (53.03-56.37%) and carbohydrate content (27.97-31.68%) with an acceptable level of moisture (10.8-14.04%) and ash (2.22-3.38%) were reported. A good safety profile was obtained with low nitrite and nitrate levels, with undetectable heavy metals and no significant growth of pathogenic microorganism except mould. Epidermal growth factor was detected but below the quantification level with the chicken EGF ELISA kit. The microculture tetrazolium (MTT) assay was performed for growth stimulation assessment comparing human EGF and EBN. There was no significant cell growth observed in cancer cells after EBN treatment. In conclusion, EBN Malaysia has a good nutritional profile, free of heavy metals, and an acceptable level of nitrate, nitrite, and microorganism profile except for mould contents. Furthermore, the in vitro study indicated that EBN was not associated with cancer cell growth.
The delivery of a full plasmid, encoding the green fluorescent protein gene into African monkey kidney (Vero3) cells, was successfully achieved using nanobiocomposites based on layered double hydroxides. This demonstrated the potential of using the system as an alternative DNA delivery vector. Intercalation of the circular plasmid DNA, pEGFP-N2, into Mg/Al-NO(3)(-) layered double hydroxides (LDH) was accomplished through anion exchange routes to form the nanobiocomposite material. The host was previously synthesized at the Mg(2+) to Al(3+) molar ratio R(i) = 2 and subsequently intercalated with plasmid DNA. Size expansion of the interlamellae host from 8.8 A in LDH to 42 A was observed in the resulting nanobiocomposite, indicating stable hybridization of the plasmid DNA. The powder x-ray diffraction (PXRD) results, supplemented with Fourier-transform infrared (FTIR) spectroscopy, compositional and electrophoresis studies confirmed the encapsulation episode of the biomaterial. In order to elucidate the use of this resulting nanobiocomposite as a delivery vector, an MTT assay was performed to determine any cytotoxic effects of the host towards cells. The intercalated pEGFP-N2 anion was later successfully recovered through acidification with HNO(3) after treatment with DNA-degrading enzymes, thus also showing the ability of the LDH host to protect the intercalated biomaterial from degradation. Cell transfection studies on Vero3 cells were then performed, where cells transfected with the nanobiocomposite exhibited fluorescence as early as 12 h post-treatment compared to naked delivery of the plasmid itself.
The influence of dietary cadmium on the accumulation and effects of dietary lead, examined in chicken. This experiment was conducted to investigate the toxic effects of dietary Cd and Pb on chick's body weight and organ, content of the tissues of these two metals was also detected. One day age chicks of Gallus gallus domesticus fed diet supplemented with 25, 50, 100 ppm of Cd, second group exposure to 300, 500, 1000 ppm of Pb in feed daily during 4 weeks. The control groups were fed without supplementation of metals. The concentrations of Cd and Pb resulted in increased of Cd and Pb content in liver, gizzard and muscle. While Cd 100 ppm and Pb 1000 ppm were increased metals content in feather. Body weight of chicks was not influenced by Cd treatment. In contrary Pb treatment was significantly (p < 0.05) decreased body weight of chicks after dietary treatment. On the other hand, Liver weigh in chicks was significantly (p < 0.05) decreased after Cd and Pb treatments.
Nitrate concentration in groundwater is influenced by complex and interrelated variables, leading to great difficulty during the modeling process. The objectives of this study are (1) to evaluate the performance of two artificial intelligence (AI) techniques, namely artificial neural networks and support vector machine, in modeling groundwater nitrate concentration using scant input data, as well as (2) to assess the effect of data clustering as a pre-modeling technique on the developed models' performance. The AI models were developed using data from 22 municipal wells of the Gaza coastal aquifer in Palestine from 2000 to 2010. Results indicated high simulation performance, with the correlation coefficient and the mean average percentage error of the best model reaching 0.996 and 7 %, respectively. The variables that strongly influenced groundwater nitrate concentration were previous nitrate concentration, groundwater recharge, and on-ground nitrogen load of each land use land cover category in the well's vicinity. The results also demonstrated the merit of performing clustering of input data prior to the application of AI models. With their high performance and simplicity, the developed AI models can be effectively utilized to assess the effects of future management scenarios on groundwater nitrate concentration, leading to more reasonable groundwater resources management and decision-making.
The study is to identify the extraction of intracellular inulinase (exo- and endoinulinase) and invertase as well as optimization medium composition for maximum productions of intra- and extracellular enzymes from Aspergillus niger ATCC 20611. From two different methods for extraction of intracellular enzymes, ultrasonic method was found more effective. Response surface methodology (RSM) with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. The effect of five main reaction parameters including sucrose, yeast extract, NaNO₃, Zn⁺², and Triton X-100 on the production of enzymes was analyzed. A modified quadratic model was fitted to the data with a coefficient of determination (R²) more than 0.90 for all responses. The intra-extracellular inulinase and invertase productions increased in the range from 16 to 8.4 times in the optimized medium (10% (w/v) sucrose, 2.5% (w/v) yeast extract, 2% (w/v) NaNO₃, 1.5 mM (v/v) Zn⁺², and 1% (v/v) Triton X-100) by RSM and from around 1.2 to 1.3 times greater than in the medium optimized by one-factor-at-a-time, respectively. The results of bioprocesses optimization can be useful in the scale-up fermentation and food industry.
Biofouling is a crucial factor in membrane bioreactor (MBR) applications, particularly for high organic loading operations. This paper reports a study on biofouling in an MBR to establish a relationship between critical flux, Jc, mixed liquor suspended solids (MLSS) (ranging from 5 to 20 g L-1) and volumetric loading rate (6.3 kg COD m-3 h-1) of palm oil mill effluent (POME). A lab-scale 100 L hybrid MBR consisting of anaerobic, anoxic, and aerobic reactors was used with flat sheet microfiltration (MF) submerged in the aerobic compartment. The food-to-microorganism (F/M) ratio was maintained at 0.18 kg COD kg-1 MLSSd-1. The biofouling tendency of the membrane was obtained based on the flux against the transmembrane pressure (TMP) behaviour. The critical flux is sensitive to the MLSS. At the MLSS 20 g L-1 the critical flux is about four times lower than that for the MLSS concentration of 5 g L-1. The results showed high removal efficiency of denitrification and nitrification up to 97% at the MLSS concentration 20 g L-1. The results show that the operation has to compromise between a high and a low MLSS concentration. The former will favour a higher removal rate, while the latter will favour a higher critical flux.
A modified hydrophilic penta-bismuth hepta-oxide nitrate (Bi5O7NO3) surface was synthesized via a precipitation method using TiO2 and Ag as modified agents. The synthesized product was characterized by different analytical techniques. The removal efficiency was evaluated using mono- and di-sulphonated azo dyes as model pollutants. Different kinetic, isotherm and diffusion models were chosen to describe the adsorption process. X-ray photoelectron spectroscopy (XPS) results revealed no noticeable differences in the chemical states of modified adsorbent when compared to pure Bi5O7NO3; however, the presence of hydrophilic centres such as TiO2 and Ag developed positively charged surface groups and improved its adsorption performance to a wide range of azo dyes. Dyes removal was found to be a function of adsorbent dosage, initial dye concentration, solution pH and temperature. The reduction of Langmuir 1,2-mixed order kinetics to the second or first-order kinetics could be successfully used to describe the adsorption of dyes onto the modified adsorbent. Mass transfer can be described by intra-particle diffusion at a certain stage, but it was not the rate limiting step that controlled the adsorption process. Homogenous behavior of adsorbent surface can be explored by applying Langmuir isotherm to fit the adsorption data.
This paper presents the findings of a study on solidification/stabilization (S/S) of lead-contaminated soil using ordinary Portland cement (OPC) and rice husk ash (RHA). The effects of varying lead concentrations (in the form of nitrates) in soil samples on the physical properties of their stabilized forms, namely unconfined compressive strength (UCS), setting times of early mixtures and changes in crystalline phases as well as chemical properties such as leachability of lead, pH and alkalinity of leachates are studied. Results have indicated that usage of OPC with RHA as an overall binder system for S/S of lead-contaminated soils is more favorable in reducing the leachability of lead from the treated samples than a binder system with standalone OPC. On the other hand, partial replacement of OPC with RHA in the binder system has reduced the UCS of solidified samples.
In this study, zinc oxide (ZnO) nanorod arrays were synthesized using a simple hydrothermal reaction on ZnO seeds/n-silicon substrate. Several parameters were studied, including the heat-treatment temperature to produce ZnO seeds, zinc nitrate concentration, pH of hydrothermal reaction solution, and hydrothermal reaction time. The optimum heat-treatment temperature to produce uniform nanosized ZnO seeds was 400°C. The nanorod dimensions depended on the hydrothermal reaction parameters. The optimum hydrothermal reaction parameters to produce blunt tip-like nanorods (770 nm long and 80 nm in top diameter) were 0.1 M zinc nitrate, pH 7, and 4 h of growth duration. Phase analysis studies showed that all ZnO nanorods exhibited a strong (002) peak. Thus, the ZnO nanorods grew in a c-axis preferred orientation. A strong ultraviolet (UV) emission peak was observed for ZnO nanorods grown under optimized parameters with a low, deep-level emission peak, which indicated high optical property and crystallinity of the nanorods. The produced ZnO nanorods were also tested for their UV-sensing properties. All samples responded to UV light but with different sensing characteristics. Such different responses could be attributed to the high surface-to-volume ratio of the nanorods that correlated with the final ZnO nanorods morphology formed at different synthesis parameters. The sample grown using optimum synthesis parameters showed the highest responsivity of 0.024 A/W for UV light at 375 nm under a 3 V bias.
In this study, the treatment of septage (originating from septic tanks) was carried out in a pilot-scale, two-staged, vertical-flow engineered wetland (VFEW). Palm kernel shells (PKS) were incorporated as part of the VFEW's substrate (B-PKS), to compare its organic matter (OM) and nitrogen (N) removal efficiency against wetlands with only sand substrates (B-SD). The results revealed satisfactory OM removal with >90% reduction efficiencies at both wetlands B-PKS and B-SD. No increment of chemical oxygen demand (COD) concentration was observed in the effluent of B-PKS. Ammonia load removal efficiencies were comparable (>91% and 95% in wetland B-PKS and B-SD, respectively). However, nitrate accumulation was observed in the effluent of B-SD where PKS was absent. This was due to the limited denitrification in B-SD, as sand is free of carbon. A lower nitrate concentration was associated with higher COD concentration in the effluent at B-PKS. This study has shown that the use of PKS was effective in improving the N removal efficiency in engineered wetlands.
Improper use of urea may cause environmental pollution through NH3 volatilization and NO3 (-) leaching from urea. Clinoptilolite zeolite and compost could be used to control N loss from urea by controlling NH4 (+) and NO3 (-) release from urea. Soil incubation and leaching experiments were conducted to determine the effects of clinoptilolite zeolite and compost on controlling NH4 (+) and NO3 (-) losses from urea. Bekenu Series soil (Typic Paleudults) was incubated for 30, 60, and 90 days. A soil leaching experiment was conducted for 30 days. Urea amended with clinoptilolite zeolite and compost significantly reduced NH4 (+) and NO3 (-) release from urea (soil incubation study) compared with urea alone, thus reducing leaching of these ions. Ammonium and NO3 (-) leaching losses during the 30 days of the leaching experiment were highest in urea alone compared with urea with clinoptilolite zeolite and compost treatments. At 30 days of the leaching experiment, NH4 (+) retention in soil with urea amended with clinoptilolite zeolite and compost was better than that with urea alone. These observations were because of the high pH, CEC, and other chemical properties of clinoptilolite zeolite and compost. Urea can be amended with clinoptilolite zeolite and compost to improve NH4 (+) and NO3 (-) release from urea.
The description of comammox Nitrospira spp., performing complete ammonia-to-nitrate oxidation, and their co-occurrence with canonical β-proteobacterial ammonia oxidizing bacteria (β-AOB) in the environment, calls into question the metabolic potential of comammox Nitrospira and the evolutionary history of their ammonia oxidation pathway. We report four new comammox Nitrospira genomes, constituting two novel species, and the first comparative genomic analysis on comammox Nitrospira. Unlike canonical Nitrospira, comammox Nitrospira genomes lack genes for assimilatory nitrite reduction, suggesting that they have lost the potential to use external nitrite nitrogen sources. By contrast, compared to canonical Nitrospira, comammox Nitrospira harbor a higher diversity of urea transporters and copper homeostasis genes and lack cyanate hydratase genes. Additionally, the two comammox clades differ in their ammonium uptake systems. Contrary to β-AOB, comammox Nitrospira genomes have single copies of the two central ammonia oxidation pathway operons. Similar to ammonia oxidizing archaea and some oligotrophic AOB strains, they lack genes involved in nitric oxide reduction. Furthermore, comammox Nitrospira genomes encode genes that might allow efficient growth at low oxygen concentrations. Regarding the evolutionary history of comammox Nitrospira, our analyses indicate that several genes belonging to the ammonia oxidation pathway could have been laterally transferred from β-AOB to comammox Nitrospira. We postulate that the absence of comammox genes in other sublineage II Nitrospira genomes is the result of subsequent loss.
Nitrogen (N) transport from land to water is a dominant contributor of N in estuarine waters leading to eutrophication, harmful algal blooms, and hypoxia. Our objectives were to (1) investigate the composition of inorganic and organic N forms, (2) distinguish the sources and biogeochemical mechanisms of nitrate-N (NO3-N) transport using stable isotopes of NO3- and Bayesian mixing model, and (3) determine the dissolved organic N (DON) bioavailability using bioassays in a longitudinal gradient from freshwater to estuarine ecosystem located in the Tampa Bay, Florida, United States. We found that DON was the most dominant N form (mean: 64%, range: 46-83%) followed by particulate organic N (PON, mean: 22%, range: 14-37%), whereas inorganic N forms (NOx-N: 7%, NH4-N: 7%) were 14% of total N in freshwater and estuarine waters. Stable isotope data of NO3- revealed that nitrification was the main contributor (36.4%), followed by soil and organic N sources (25.5%), NO3- fertilizers (22.4%), and NH4+ fertilizers (15.7%). Bioassays showed that 14 to 65% of DON concentrations decreased after 5-days of incubation indicating utilization of DON by microbes in freshwater and estuarine waters. These results suggest that despite low proportion of inorganic N forms, the higher concentrations and bioavailability of DON can be a potential source of N for algae and bacteria leading to water quality degradation in the estuarine waters.
Saxitoxins (STXs) constitute a family of potent sodium channel blocking toxins, causative agents of paralytic shellfish poisoning (PSP), and are produced by several species of marine dinoflagellates and cyanobacteria. Two STX-core genes, sxtA and sxtG, have been well elucidated in Alexandrium but the expression of these genes under various nutritional modes in tropical species remains unclear. This study investigates the physiological responses of a tropical Pacific strain of Alexandrium minutum growing with nitrate or ammonium, and with various nitrogen to phosphorus (N:P) supply ratios. The transcriptional responses of the sxt genes were observed. Likewise, a putative sxtI encoding O-carbamoyltransferase (herein designated as AmsxtI) was recovered from the transcriptomic data, and its expression was investigated. The results revealed that the cellular toxin quota (Qt) was higher in P-depleted, nitrate-grown cultures. With cultures at similar N:P (<16), cells grown with excess ammonium showed a higher Qt than those grown with nitrate. sxtA1 was not expressed under any culture conditions, suggesting that this gene might not be involved in STX biosynthesis by this strain. Conversely, sxtA4 and sxtG showed positive correlations with Qt, and were up-regulated in P-depleted, nitrate-grown cultures and with excess ambient ammonium. On the other hand, AmsxtI was expressed only when induced by P-depletion, suggesting that this gene may play an important role in P-recycling metabolism, while simultaneously enhancing toxin production.
Groundwater hazard assessments involve many activities dealing with the impacts of pollution on groundwater, such as human health studies and environment modelling. Nitrate contamination is considered a hazard to human health, environment and ecosystem. In groundwater management, the hazard should be assessed before any action can be taken, particularly for groundwater pollution and water quality. Thus, pollution due to the presence of nitrate poses considerable hazard to drinking water, and excessive nutrient loads deteriorate the ecosystem. The parametric IPNOA model is one of the well-known methods used for evaluating nitrate content. However, it cannot predict the effect of soil and land use/land cover (LULC) types on calculations relying on parametric well samples. Therefore, in this study, the parametric model was trained and integrated with the multivariate data-driven model with different levels of information to assess groundwater nitrate contamination in Saladin, Iraq. The IPNOA model was developed with 185 different well samples and contributing parameters. Then, the IPNOA model was integrated with the logistic regression (LR) model to predict the nitrate contamination levels. Geographic information system techniques were also used to assess the spatial prediction of nitrate contamination. High-resolution SPOT-5 satellite images with 5 m spatial resolution were processed by object-based image analysis and support vector machine algorithm to extract LULC. Mapping of potential areas of nitrate contamination was examined using receiver operating characteristic assessment. Results indicated that the optimised LR-IPNOA model was more accurate in determining and analysing the nitrate hazard concentration than the standalone IPNOA model. This method can be easily replicated in other areas that have similar climatic condition. Therefore, stakeholders in planning and environmental decision makers could benefit immensely from the proposed method of this research, which can be potentially used for a sustainable management of urban, industrialised and agricultural sectors.
Ammonia emissions is an important issue during composting because it can cause secondary pollution and a significant of nitrogen loss. Based on research adding Bacillus stearothermophilus can reduce ammonia emissions during composting because it can use sugar in organic matter fermentation to produce organic acids over 50 °C. This study conducted the batch experiments by adding different concentrations of Bacillus stearothermophilus to reduce the ammonia emissions and find out its characteristic during layer manure composting by using an aerobic composting reactor with sawdust as a bulking agent. The results show that the application of Bacillus stearothermophilus can accelerate the rate of temperature and significantly decrease pH, the warming period was 2 days in the treatment with Bacillus stearothermophilus, while it was 4 days in the treatment without Bacillus stearothermophilus. Ammonia emissions were mainly occurred in warming and high temperature period during composting. The ammonia emissions in the treatment with 8.00 g/kg initial Bacillus stearothermophilus were significantly lower than the other lower Bacillus stearothermophilus treatment and control during composting (p 0.05). MiSeq System Sequencing results find that the addition of Bacillus stearothermophilus changed the bacterial community structure under warming and high-temperature periods during composting, increased the relative abundance of lactic acid bacillus and nitrification bacteria. Therefore, the reason for the low ammonia emission in 8.00 g/kg initial Bacillus stearothermophilus treatments might be not only due to the Bacillus stearothermophilus itself, but also Bacillus stearothermophilus can change the indigenous microorganism community, including increase the relative content of lactic acid Bacillus and nitrification bacteria, thus reducing the pH and promoting nitrification, and reducing ammonia emissions.
Stormwater runoff is recognized as a cause of water quality degradation because it may carry nitrogen (N) and other pollutants to aquatic ecosystems. Stormwater ponds are a stormwater control measure often used to manage stormwater runoff by holding a permanent pool of water, which reduces the peak flow, magnitude of runoff volume, and concentrations of nutrients and pollutants. We instrumented the outlet of a stormwater pond in an urban residential neighbourhood in Florida, United States to (1) investigate the concentration and composition of N forms during the summer rainy season (May to September 2016), and (2) determine the bioavailability of organic N in the stormwater pond with a bioassay experiment. A total of 144 outflow water samples over 13 storm events were collected at the outlet of the stormwater pond that collects runoff from the residential catchment. Samples were analysed for various inorganic N [ammonium (NH4-N), nitrate (NO3-N)], and organic N forms [dissolved organic nitrogen (DON), and particulate organic nitrogen (PON)]. Flow-weighted mean concentration of total N (TN) in pond outflow for all collected storm events was 1.3±1.42 mg L-1, with DON as the dominant form (78%), followed by PON and NO3-N (each at 8%), and NH4-N (6%). In the bioassay experiment, organic N (DON+PON) was significantly decreased by 25-28% after 5 days of incubation, suggesting that a portion of the DON carried from the pond outflow to receiving water bodies may be bioavailable. These results suggest that efforts to mitigate stormwater N outflows from urban ponds should incorporate both inorganic and organic N in management plans.
The marine dinoflagellate Alexandrium minutum is known to produce saxitoxins that cause paralytic shellfish poisoning in human worldwide through consumption of the contaminated shellfish mollusks. Despite numerous studies on the growth physiology and saxitoxin production of this species, the knowledge on the molecular basis of nutrient uptakes in relation to toxin production in this species is limited. In this study, relative expressions of the high-affinity transporter genes of nitrate, ammonium, and phosphate (AmNrt2, AmAmt1 and AmPiPT1) and the assimilation genes, nitrate reductase (AmNas), glutamine synthase (AmGSIII) and carbamoyl phosphate synthase (AmCPSII) from A. minutum were studied in batch clonal culture condition with two nitrogen sources (nitrate: NO3- or ammonium: NH4+) under different N:P ratios (high-P: N:P of 14 and 16, and low-P: N:P of 155). The expression of AmAmt1 was suppressed in excess NH4+-grown condition but was not observed in AmNrt2 and AmNas. Expressions of AmAmt1, AmNrt2, AmNas, AmGSIII, AmCPSII, and AmPiPT1 were high in P-deficient condition, showing that A. minutum is likely to take up nutrients for growth under P-stress condition. Conversely, relative expression of AmCPSII was incongruent with cell growth, but was well correlated with toxin quota, suggesting that the gene might involve in arginine metabolism and related toxin production pathway. The expression of AmGSIII is found coincided with higher toxin production and is believed to involve in mechanism to detoxify the cells from excess ammonium stress. The gene regulation observed in this study has provided better insights into the ecophysiology of A. minutum in relation to its adaptive strategies in unfavorable environments.
Musty odor production by actinomycetes is usually related to the presence of geosmin and 2-methylisoborneol (2-MIB), which are synthesized by enzymes encoded by the geoA and tpc genes, respectively. Streptomyces spp. strain S10, which was isolated from a water reservoir in Malaysia, has the ability to produce geosmin when cultivated in a basal salt (BS) solid medium, but no 2-MIB production occurred during growth in BS medium. Strain S10 could produce higher levels of geosmin when the phosphate concentration was limited to 0.05 mg/L, with a yield of 17.53 ± 3.12 ✕ 105 ng/L, compared with growth in BS medium. Interestingly, 2-MIB production was suddenly detected when the nitrate concentration was limited to 1.0 mg/L, with a yield of 1.4 ± 0.11 ✕ 105 ng/L. Therefore, it was concluded that phosphate- and nitrate-limiting conditions could induce the initial production of geosmin and 2-MIB by strain S10. Furthermore, a positive amplicon of geoA was detected in strain S10, but no tpc amplicon was detected by PCR analysis. Draft genome sequence analysis showed that one open reading frame (ORF) contained a conserved motif of geosmin synthase with 95% identity with geoA in Streptomyces coelicolor A3 (2). In the case of the tpc genes, it was found that one ORF showed 23% identity to the known tpc gene in S. coelicolor A3(2), but strain S10 lacked one motif in the N-terminus.