Cannabis Grow

Hemp is an annual herb plant which is used for production fibre for such as rope, sail and clothing as well as the seed crushed for oil, food and feed. Due to the outer portion of the hemp stem contains strong and long bast, fibre provides a strength and high quality textile, while inner of the stem so called hurd also used for paper and building material. After Thai royal project interested in developing industrial hemp as an economic crop to support the hill tribes, the quality seed are needed to propagate and disperse for the farmer. In addition, the cultivation of hemp is being permitted in Thailand especially in the north, but under strict control and monitoring by office of the narcotics control board of Thai. Hemp is determined a dioecious crop and seed production is required a wider space from plant to plant than producing as fibre. In addition as an annual crop, cultivated seed are usually sown in early March and late of May, and harvested in the end of November under preferable natural temperature 14 – 27 ºC. Therefore, they are attractive to produce exceed seed supply and then stored for one or several years for cultivation. Storage of seed is an important process of plant production to avoid unfavourable environmental conditions and the acceleration of the deteriorations, which is started after harvest. Storage conditions play an important role to maintain high seed quality, which directly related to environmental conditions. However, the sensitivity of seed to high temperature is depended on the water content that the higher moisture content, the looser viability. Seed deterioration is associated with the genotype, seed history and their physiological and chemical compositions. As in hemp, seed is determined as an orthodox seed that can be dried to low moisture content and stored at low temperature for several years. Nonetheless, the seed contain 20 – 25% protein, 20 – 30% carbohydrates, 10 – 15% insoluble fibre and especially 25 -35% oil which was considered to be the major contributors to seed deterioration as previous demonstrations of soybean , peanut and sunflower. For hemp seed, it was suggested that seed should be dried until the moisture content lower than 12%. Consequently,vertical farming for further storage, either high or low relative humidity affected the moisture equilibrium of seed due to hygroscopic property. Then, the type of container was important for protecting moisture immigration. 

As in closely fibre species such as kenaf , seed stored at 8% mc remains viable for up to 5.5 years when stored at 8 ºC and stored above 10 ºC and 12% mc showed significant decrease in seed viability. In parallel, Meints and Smith, also found that germination of kenaf under ideal conditions remained high in seed stored up to 4 years at 10 ºC and did not show appreciable differences in field emergence or performance through the growing season. Determination of hemp seed qualities which were derived from segregation of the local land race would assist the Royal project and further seed supplier in reliable on fibre hemp production. The objectives of this study were to determine the impact of storage conditions and hemp seed quality losses during year storage. These experiments were conducted to compare storage condition affecting different hemp seed cultivars by observing the physiological of seed qualities. The moisture contents at which these changes are observed among cultivars, and continuously involve in the change of stored seed quality. The seed cv. RPF2 contained higher moisture content than the others and also showing lower initial germination illustrated significantly decreased with storage time when packed in both seal aluminium foil and plastic bag at ambient temperature. McDonald stated that the effect of high temperatures on seeds quality is highly dependent on their water content and the loss of viability was more rapid when the seed contain high moisture. However, the quality of seed packed in both materials was not different during 5 month, but after that aluminium foil had an adverse effect on reduction of seed viability and vigour than plastic bag during 5 – 8 month. This suggested that high moisture content seeds might be closely oppressed to dry air and then water remove to the surface because of hygroscopic of which higher moisture content was observed in seed packed in aluminium foil. The change of moisture content was not different when seed stored in aluminium foil at 15, 4 and –4 ºC, while the seed germination and vigour was also remained. The result revealed that aluminium foil has better water and oxygen proved properties than plastic bag made from polypropylene which the exchange were 0.0914/m2 /hr and 0.2472/m2 /hr for oxygen; 65 g-water/m2/hr and 1378 g-water/m2/hr for water, respectively. This suggested that initial moisture content and germination are the main factor for providing seed longevity. Hemp seed containing high moisture content stored at room temperature maintained the germination and vigour only up to 5 months of storage, after that the conditions and aging was deleterious to seed viability. Seed deterioration is associated with various biochemicals, metabolic and physiological alteration including loss of membrane integrity and cell compartmentalization, decreasing in ATP production and impairment of RNA and protein biosynthesis. 

Moreover, lipid peroxidation and free radicals were considered to be the major contributors to seed deterioration , especially, at high temperature, it was mainly related to membrane damage and alteration of energy metabolisms as demonstrated in sunflower. In the other hand, seed germination and vigour of low seed moisture content as cv. RPF3 contained in aluminium foil at room temperature remained viable. This previous study mentioned that the dehydration of seed could prevent thermal denaturation. Marijuana use dates back to 2727 BC to Chinese Emperor Shen Nung. After spreading through the Greek and Roman empires and into the Islamic empire of North Africa and the Middle East, it was brought to the Western hemisphere by the Spanish. Originally lauded for its utility as fiber, it was not until its migration into North America that it began to be used in a similar fashion as it is today. Used in the form of hemp, it was seen throughout society as rope, clothing and even paper. While marijuana has been used by Americans recreationally for years, it is a subject that is becoming more commonplace in our modern society. According to the National Institutes of Health, marijuana use in 2015e2016 rose from 4.1% to 9.5% of the U.S. adult population. With more states eliminating the legal ramifications of its use and a growing debate about its federal legality, this is a subject that routinely makes local and national headlines. With a diverse array of commercial products becoming available from chocolate squares to oral sprays, marijuana is also no longer restricted to a rolled cigarette. The medical community has also joined the debate. The most obvious correlation between medicine and marijuana is medicinal marijuana. Marijuana’s effects have been well documented, allowing the push for its use as medicine within multiple specialities. Proponents of its use point to its effects on the endocannabinoid system. Studies show that through its impact on different pathways it may be used as an analgesic, immunosuppressant, muscle relaxant, anti-inflammatory agent, appetite modulator, antidepressant, antiemetic, bronchodilator, neuroleptic, antineoplastic and antiallergen. Medical marijuana also differs in chemical composition, containing a higher tetrahydrocannabinol concentration and less cannabinol than the recreational version. This is even before taking into consideration the various extraneous agents that may be found within the available recreational drug.

There is however, very little, if any research evaluating marijuana’s use in surgery. This paucity of literature presents a problem. While many surgeons may ask about recreation drug use including marijuana, many other drugs have established evidence based outcomes that allow variation in surgical planning as needed. However, when it comes to marijuana, surgeons are left to determine what to do with this information on their own.Despite the multiple studies on the physiologic effects of marijuana use, clinical studies, if any, are not cited in the medical literature. This study reviews the literature available on marijuana’s effects and discusses potential complications that may result within the surgical setting. With a reported estimation of 10%e20% of patients between the ages of 18 and 25 years regularly using marijuana, this review seeks to become an initial step for further exploration of the subject and to reveal why there is a need for more in-depth research.Due to limited research and reviews on this subject, information was utilized from articles on surgery in various fields, such as orthopedic, dental and bariatric with anesthesia considerations and general topics related to marijuana also examined. The following information was gathered: marijuana’s prevalence in the United States, marijuana’s effects on the cardiovascular system and pulmonary system, potential coagulopathies, marijuana’s effects in relation to anesthesiology, evidence based screening methods for recreational drug use, potential surgical complications that may result from marijuana use,flood tray and recommendations on marijuana use and surgery.When marijuana is smoked, THC and other cannabinoids are absorbed rapidly through the lungs with effects peaking in 15 minutes. These effects can persist for up to a dose-dependent 4 hours in the acute setting. When ingested orally however, onset of effects is slower but has a longer duration of action , due to continued absorption in the gut. This is despite a lower bioavailability due to first-pass metabolism by the liver which results in a blood concentration 25% of what is obtained if smoked. The cognitive/psychomotor effects can be present for up to 24 hours regardless of administration route. Cannabinoids are highly lipid soluble. This leads to a slow release into the bloodstream with a single dose not fully eliminated for up to 30 days. The cardiovascular effects of marijuana use range from benign to worrisome based on the timeline of use and dosage.

In a series single blind study comparing the effects of high and low doses of THC in healthy young men, tachycardia was induced beginning within the time of inhalation, and persisting at least 90 minutes, with the maximum heart rate reached at an average of 30 minutes. The study also found a significant elevation in systolic and diastolic blood pressures as well as the presence of premature ventricular contractions in subjects who received the higher doses. These experiments showed a correlation between the dose and the tachycardic and cardiovascular changes. In addition, Malit et al.’s study on the effects of intravenous THC found the majority of patients to exceed the 100 beats per minute mark but also experience intermittent spikes in heart rate with a possible etiology of psychological distress. Beaconsfield et al. postulated a mechanism of beta adrenergic stimulation for the tachycardia as he was able to block the tachycardia with the use of propranolol. Pharmacology lays credence to this. At lower or moderate doses, marijuana increases sympathetic activity reducing parasympathetics and producing an elevation in heart rate, cardiac output and blood pressure. However, the opposite is true as the dosage increases. At high doses, the parasympathetic system takes over, leading to bradycardia and hypotension with animal studies postulating that the sympathetic inhibition occurs due to the bioactive constituent of cannabis’s effects on the CB1 receptors. In addition to sinus tachycardia, marijuana use has been linked to multiple electrocardiogram changes in various case reports. Daccarrett et al. found Brugadalike changes in a 19 year old male with a known history of cannabis use and no anatomical/functional abnormalities. A case was also reported in which cannabis use was linked to the development of atrial flutter and atrial fibrillation, while other studies have reported the presence of sinus bradycardia and AV block.Aronow et al., found that while comparing marijuana to placebo, cannabis causes an increase in carboxyhemoglobin, a resultant increase in myocardial oxygen demand, decrease in oxygen supply as well as an induction of platelet aggregation. One case report showed a 21 year old male who presented with a ST elevation myocardial infarction due to plaque rupture as a complication of marijuana use. In Mittleman et al.’s analysis of over 3,800 cases of myocardial infarction, 124 patients reported use within the last year of which 37 reported use within 24 hours, with 9 reporting use within an hour of the event. The study found a statistically significant 4.8 fold increase in myocardial infarction within the first hour of marijuana use. In fact, as THC content of marijuana increases, there are a growing number of clinical studies demonstrating the association between cannabis use and adverse cardiovascular events. One such study followed 1913 adults prospectively and demonstrated that in patients with prior myocardial infarction, marijuana use up to once per week increased risk of death 2.5 fold while more frequent use yielded a fourfold risk of dying.

Tashkin et al measured FEV1 serially in 255 habitual male and female smokers of marijuana and/or tobacco and nonsmokers of either substance on up to seven occasions at intervals of $ 1 year over a total period of up to 8 years. Random effects models were used in men and women separately to estimate mean rates of decline in FEV1 and to compare these rates between smokers of marijuana or tobacco alone, smokers of both substances, and nonsmokers of either substance. Smoking marijuana was not found to be associated with greater declines in FEV1 than nonsmoking nor was a relationship seen between the daily number of marijuana joints smoked and rate of decline in FEV1. In contrast, smoking tobacco had a significant effect on FEV1 decline in men in a dose-responsive manner. Findings are illustrated in Figure 3. Hancox et al performed serial lung function measurements in 779 members of a birth cohort in Dunedin, New Zealand, at 18, 21, 26, and 32 years of age. They assessed changes in FEV1, FVC, and FEV1/FVC associated with tobacco and marijuana smoking using regression analyses and estimates of both joint-years and pack-years as predictors, adjusting for sex and height at 32 years of age, changes in height between 15 and 32 years of age, and concomitant tobacco or marijuana use. The results are shown in Table 3. Among marijuana smokers, FVC increased significantly over time, whereasFEV1 showed a non significant increase and FEV1/FVC exhibited a non significant decline. In contrast, among tobacco smokers, FEV1 and FEV1/FVC both showed significant declines, whereas FVC showed no change.The smoke contents of marijuana cigarettes contain the same procarcinogenic components, including the very potent human carcinogen, benzpyrene, found in tobacco smoke.Bronchial biopsies obtained from heavy, habitual marijuana smokers have shown the same widespread histopathologic epithelial abnormalities noted in tobacco smokers, including squamous metaplasia and cellular disorganization,that are widely thought to be precancerous.

Immunohistologic examination of bronchial biopsies from marijuana smokers has shown higher levels of the protein products of oncogenes, including Ki67 and EGFR,whereas laryngeal cancer specimens from marijuana smokers have been shown to exhibit increased expression of oncoproteins compared both with tobacco smokers and nonsmokers.A few older epidemiologic studies from North Africa have shown a positive association with lung cancer; however,greenhouse rolling racks the common practice of admixing marijuana and tobacco within the same cigarette in these societies precludes disentangling the effects of marijuana from those of tobacco. A population-based cohort study of men 18 to 20 years of age conscripted into the Swedish military in 1969 to 1970 tracked these men until 2009 for incident lung cancer cases using linked nationwide medical registries.Cox regression was used to assess the relationship between lifetime use of marijuana self-reported at the time of conscription and risk of subsequent lung cancer diagnosis over the ensuring 40 years, adjusting for tobacco use also up to only 18 to 20 years of age.The hazard ratio for lung cancer was significantly positive in relation to lifetime marijuana use of > 50 times. However, because neither marijuana nor tobacco use was determined after the baseline assessment, the authors were unable to adjust for true lifetime use of tobacco, a crucially important residual confounder.Several investigators have demonstrated a tumor suppressive effect of THC and other cannabinoids on a variety of malignancies, including lung, in both cell culture systems and animal models, as previously reviewed by Bifulco et al and Velasco et al.These findings appear to reflect antiproliferative, proapoptotic, and antiangiogenic properties of THC that might counteract the tumor-initiating or tumorpromoting effects of the carcinogens contained with the smoke of marijuana. A large cohort study of health plan participants in Northern California failed to show an increased risk of tobacco-related cancers in association with self-reported marijuana use.A pooled analysis of six well-designed case-control studies of the association between habitual marijuana smoking and lung cancer that totaled 2,159 cases and 2,985 control subjects did not find any increased risk of lung cancer in association with marijuana use.Only one of the six studies included in the pooled analysis showed a significantly positive association between lung cancer and marijuana use, mainly in the heaviest marijuana use tertile, but the latter tertile included only four matched control subjects, making the estimates of risk imprecise.

Marijuana smoking might predispose to lower respiratory tract infection in at least three ways. First, the destruction of ciliated epithelium in the large central airways and the associated hyperplasia of mucus-secreting surface epithelial cells demonstrated in bronchial biopsies of habitual marijuana smokers may lead to increased production of mucus in the face of a diminished capacity to cleanse the lung of the excess mucus because of ciliary loss, thereby providing a substrate for potentially pathogenic microbial organisms colonizing the lower respiratory tract.Second, the immunosuppressive effect of THC leading to impairment of the bactericidal and fungicidal activity of AMs,further compromises the lung’s defense against microbial infection. Finally, marijuana has been shown to be frequently contaminated with Aspergillus fumigatus and potentially pathogenic gram-negative bacteria.Therefore, introduction of these microorganisms into the lung in the face of marijuana-related impairment in the lung’s host defense provides another possible mechanism for increasing the risk of pneumonia. The possible association of marijuana with an increased risk of pneumonia is supported by older case reports of Aspergillus pneumonia in smokers of marijuana immunocompromised by AIDS,chronic granulomatous disease,bone marrow transplantation,renal transplantation, or lung cancer treated with chemotherapy.In addition, a cluster of five patients with cavitary TB who used a marijuana water pipe was reported in Australia in 2003, followed by another report from Australia 10 years later of three additional cases of open cavitary TB in marijuana bong users.In these cases, it is not clear whether the spread of TB infection was because of close contact with patients with cavitary TB who might have shared a water pipe to smoke marijuana and/or to marijuana-related impairment in the lung’s defense against infection. Although a few older epidemiologic studies have suggested that marijuana use might be a significant independent risk factor for opportunistic infection in individuals who are HIV positive,an early analysis of data from the Multi-center AIDS Cohort Study failed to find evidence that marijuana was a risk factor for progression of individuals who are HIV positive to full-blown AIDS.Further preliminary analysis of the possible association of marijuana use with pneumonia risk using the Multi-center AIDS Cohort Study data set updated to 2013 has not shown a significant association of marijuana use with increased risk of either community-acquired or opportunistic pneumonia in either the HIV-negative or HIV-positive members of the cohort, after adjustment for tobacco, age, and among the individuals who are HIV positive, cluster of differentiation 4 cell counts and viral load.

A long-standing observation in clinical and epidemiological research into substance use has been that users of one drug typically do not limit their use to a single substance. For example, alcohol and tobacco are commonly used by the same person and often in the same setting, as are tobacco and marijuana. Though the synergistic effects of these particular drugs have been suggested as a potential explanation , another interesting possibility is that individuals have an underlying liability to drug use within and across different pharmacological classes. Support for this notion has been shown for both licit and illicit drugs in a variety of populations and drug use phenotypes. As poly-substance use is associated with problematic use and reduced treatment efficacy , identifying informative precursors to the onset of abuse and dependence remains a priority. Among the variety of factors that have been examined as an early indicator of later, more problematic use patterns, how someone experiences a drug , is one of the most interesting. Subjective experiences are thought to reflect individual differences in the pharmacological effects of a drug. Factor analytic studies of these experiences frequently yield two main factors: pleasant or positive and unpleasant or negative. Positive subjective experiences often include euphoria, relaxation, and feeling less inhibition. Negative subjective experiences include nausea,vertical grow difficulty inhaling, dizziness and sadness. Though weakly correlated , users of a drug sometimes report both positive and negative experiences. Alcohol, tobacco, and marijuana are the most commonly used licit and illicit drugs. Studies examining the subjective experiences to these drugs have generally found that how a person responds to a particular drug is predictive of more problematic use of the same drug. For example, dependent cigarette users more frequently endorse positive experiences than regular smokers and moderate-to-heavy drinkers report experiencing greater stimulant-like effects to alcohol than lighter drinkers. A similar relationship has been demonstrated for marijuana use. Although results are mixed, negative experiences to tobacco and marijuana have also been positively associated with problematic use. For alcohol consumption, low levels of response, primarily measured using negative subjective effects, have been associated with an increased risk of an alcohol use disorder as lower thresholds to the sedative effects of alcohol protected against developing abuse later in life.In particular, subjective experiences to a variety of drugs are correlated and can predict levels of involvement for substances in other pharmacological classes. This observation has been shown for pleasurable experiences of alcohol and tobacco where both drugs were predictive of current alcohol use in a college aged sample. Further, marijuana use has been shown to increase a sense of “liking” among non-smokers whereas alcohol has no effect on the subjective experiences of cigarettes. Lastly, greater rates of alcohol dependence and illicit drug use have been observed among high marijuana users as defined by greater rates of sensitivity to positive and negative subjective experiences. Though additional studies are needed, these cross-drug results indicate that how a person responds to a drug is predictive of how they will respond to other drugs. In this report we detail findings from a study of subjective experiences to alcohol, tobacco, and marijuana in a sample of young adults participating in the Colorado Center for Antisocial Drug Dependence. Subjective experiences were collected from both clinical and community participants using a questionnaire developed by Lyons et al.. Our analyses were designed to address three questions. First, how do positive and negative subjective experiences across alcohol, tobacco, and marijuana compare? Second, to what extent do subjective experiences to alcohol,tobacco, and marijuana overlap? Lastly, to what degree do subjective experiences to one drug associate with more problematic use behaviors for a different drug?Participants were drawn from the Colorado Center on Antisocial Drug Dependence [CADD] and consisted of 3853 participants between the ages of 11 and 30 years old and included both community and clinical participants. Our community-based sample was drawn from those participating in the Colorado Twin Registry , Colorado Adoption Project , with clinical controls drawn from the Colorado Adolescent Substance Abuse Family Study. Our clinical sample was drawn from adolescents in treatment for substance abuse and delinquency as a part of the ASA study. Additional clinical participants were drawn from an adjudicated sample from the Denver metropolitan area. Siblings of the clinical subjects were also included. All participants in the current study met one or more of the following criteria: they had consumed at least six drinks in their lifetime, had used tobacco daily for at least one month, or had used marijuana six or more time in their lifetime.Patterns of alcohol, tobacco and marijuana use, abuse and dependence symptomatology were collected using the Composite International Diagnostic Interview-Substance Abuse Module. Abuse and dependence status as defined by the Diagnostic and Statistical Manual of Mental Disorders was determined using scoring algorithms based on whole life substance related problems. Retrospective subjective experiences were collected using a 23- item questionnaire developed by Lyons et al.. The original Lyons questionnaire was comprised of 23 items. As discussed in Zeiger et al. , due to the CADD interview length the original Lyons questionnaire was shortened after wave 1; a factor analysis was conducted on the Lyons questionnaire and 10 items with lower or mixed loadings were dropped. Subsequently, most subjects received the shortened 13-item questionnaire, thus these analyses were conducted on the 13-item response set from all subjects.

The variations in the metabolic composition of the aqueous extracts of leaves and stems were investigated by performing an OPLS-DA.A clear separation between the two classes of samples along the first two components was obtained and the analysis of the Splot allowed for the detection of the most significant x-variables and,thus,the metabolites contributing to such clustering.Specifically,the buckets having values of VIPpredictive >1 and values of p1 >|0.5| were analyzed more deeply.The spectral regions containing signals related to glucose,fructose,and galactose moieties contributed significantly to the clustering of the stems.Also,the buckets containing the signals assigned to the organic acids were characterized by significant values of VIP and p1.An analogous trend was observed for the buckets related to the signals assigned to γ-aminobutyric acid at 1.89–1.93 ppm and 3.01–3.05 ppm,and glutamine at 2.13–2.17 ppm.Interestingly,the clustering of the leaves was affected by some regions of the spectra containing very broad signals at 0.88,1.29,2.23,and 5.31 ppm ,presumably attributable to the lipids of the cell membranes and/or walls.The comparison of the 1D 1 H NOESY spectra of the leaves collected from control plants ,vigorous plants ,weak plants indicated that such broad signals were predominant in vigorous plants.Such broad signals were not observed in the aqueous extracts of stems.Also,the buckets containing the signals of ethanol at 1.17–1.25 ppm contributed relevantly to the grouping of the leaves towards the stems.These data could be explained by the fact that the leaves and,particularly the leaves sampled from vigorous green plants,have active photosynthetic processes and intact cell structures,grow racks system whereas stems of weakest plants begin to enrich with lignin and have less active photosynthesis.

The spectral regions containing these broad signals correlated positively with the calculated amounts of residual copper in the leaves.Under physiological conditions,Cu is found in two common forms,Cu preferentially binding sulfur-containing compounds having a thiol or a thioether group,and Cu ,that coordinates mainly with oxygen or imidazole nitrogen groups.The main functions of Cu are the transport of electrons in mitochondria and chloroplasts,the control of the cellular redox state ,and the remodeling of the cell wall,which is one of the major Cuaccumulation sites in hyper accumulating plants.Moreover,Cu leaf content is inversely associated with the intensity of the buckets relating to glucose and fructose and ethanol.In weakest plants,sucrose,the main soluble component of the phloem sap and translatable product of photosynthesis might be hydrolyzed into the two constituting monomers and fermented by yeast and/or bacteria producing ethanol.In a second task,the effect of the treatment with spirulina during the cultivation of hemp towards the remediation of contaminated soil was investigated.Thus,a deeper look was taken at the higher components of the OPLS model described in paragraph 3.1.Indeed,a good clustering of samples according to the treatment was visible in the t[2] vs t[5] scores plot.The second component and the fifth one explained together 11.75% of the x-variance predictive of the y-variance ,suggesting that the treatment with spirulina affected slightly the metabolic composition of the plant.Such evidence is relevant to make possible a re-utilization of the hemp,which does not change its metabolome during the phytoremediation stage.Specifically,the samples collected from plants grown in contaminated soil and the samples obtained from plants treated with a higher concentration of spirulina tended to cluster at values of t[2] >0.Conversely,the samples derived from plants treated with a lower concentration of the blue-green alga and the control ones were distributed preferentially at values of t[2] <0.A further piece of evidence was that samples C and the samples CS1 tended to separate into two clear groups along t[5].Analysis of the loading plot suggested that the uptake of all the metals,except for lead,contributed relevantly to the clustering of the plants treated with a higher dose of spirulina.Also,the average plant biomass was generally higher in contaminated plants compared to the uncontaminated ones.In addition,the plants that grew in contaminated soil and were irrigated with a higher dose of spirulina presented a higher value of biomass,confirming the activity of these cyanobacteria to promote plant growth.

The average plant contents of Ni ,Zn ,and Cd were greater in contaminated plants grown in soils added with spirulina than those grown without it.To get deeper insights into the effects deriving from the dose of spirulina,OPLS was applied to CS1 and CS0.5 samples.The observations,namely CS1 and CS0.5 ,separated along the second predictive component P2,explaining about 9.5% of the predictive x-variance.The distribution was strongly affected by the part of the plant the samples derived from,leaves or stems,with a noticeable clustering along the first predictive component P1.The corresponding loading plot showed important dose-dependent variations.Indeed,nickel,zinc,copper,and chromium were more abundant in samples treated with a higher dose of spirulina.Generally,as the concentration of spirulina increased,the residual metal content increased both in the leaves and in the stems ,whereas the Cr content increased in leaves and decreased in stems.While no dose-effect was observed for Cd,noticeable variations were exerted by increased amounts of spirulina on the quantities of Pb incorporated into the plant.Specifically,the average content of Pb in the plant was higher in CS0.5 than in CS1,suggesting that a higher amount of the blue-green alga contained in the soil might compete with hemp to sequester this metal.A piece of further evidence was that the average soil residual of all heavy metals resulted higher when it was irrigated with a higher concentration of spirulina ,suggesting that the cyanobacteria stationing in the soil may exert a strong action of metal chelation,avoiding leaching of them during the watering stage and acting as a reservoir for hemp to absorb the chelated metals.Such results find fundamentals in the reported ability of Arthrospira platensis to adsorb and accumulate all the six metals here tested through various mechanisms.Very small effects were observed on the metabolic profile when samples CS1 were compared with CS0.5.Interestingly,according to the loading plot ,the buckets at 2.66 and 2.74 ppm,containing the signals assigned to citric acid,contributed importantly to the observed grouping between the samples treated with the two different doses of spirulina.Specifically,the signals of citric acid were more intense in samples treated with a higher dose of spirulina.Considering that citric acid is one of the main chelating agents in plants,this evidence may support the hypothesis that,at a higher concentration,the cyanobacteria can chelate the heavy metals sequestering them into the soil.

As a result,less quantity of heavy metal is available for the plant to uptake,and,thus,more chelating agents inside the plant are in the free form not coordinating any metal.Industrial hemp is an annual crop grown for its seed and fibers.The three main global producers of hemp are China,Europe,and North America.Hemp is found in countless daily products: foods ,beverages ,cosmetics,and personal care products ,textiles ,papers and cardboard ,jewelry and fashion,leisure products ,animal feed,animal litter,mulching and horticulture,detergents,bio-fuel,building materials,insulation,composites ,etc.Hemp is also the object of numerous fundamental studies for innovative potential applications in bio-pesticides,biotechnology,pharmacy and medicine,paints,energy production,and wastewater treatment.Hemp shives,as valuable co-products produced during the process of extracting fiber from hemp straw,are constituted from the xylem tissue of the stem.Long considered as a by-product of the industry,used for plant mulch or animal bedding ,hemp shives are now used for house insulation,energy,and fuel production.There is also an interest in using these co-products as adsorbent materials to complex environmental pollutants since this resource is abundant,cheap,and easily burned.The last one is interesting in the context of recovery of adsorbed metals and their revalorization for example.Moreover,these materials have a highly reactive lignocellulosic macromolecular structure capable of interacting with other substances.Finally,the industry also needs to find new outlets because the volume of shives produced annually is constantly increasing.Recently,hemp fibers have been studied as adsorbent for metal removal and for the preparation of activated carbons for similar applications.However,work on hemp in shives/hurds form for environmental applications is rare.Furthermore,the identification of interactions involved in liquid-solid adsorption phenomena is often carried out using empirical mathematical models that allow the modeling of experimental results and the drawing of conclusions from the assumptions of these models.Another approach is to couple different microscopic and spectroscopic techniques to obtain information on these mechanisms.This study is part of the European research project FINEAU which brings together academics and industrialists focused on the potential use of hemp shives to recover metals from industrial effluents aiming their valorization.Two types of materials were targeted in this study by comparing shives washed with water with those chemically modified using polycarboxylic agent ,and with shives washed with water and activated with sodium carbonate.

The last one was the most efficient biosorbent of copper in our previous study among investigated raw shives and those treated with H2O,H3PO4,or KOH.The chemical treatment with Na2CO3 was carried out on a laboratory scale by copying the process already used by one of the industrialists of the project on other cellulosic materials.The objective of these two modifications is to purify and increase the capacity of raw shives towards metals,rolling flood tables while presenting high selectivity and performances independently of the pH and ionic strength representing two important issues for wastewater from the surface treatment industry.Recently,Mongioví et al reported that in the context of copper recovery by SHI-C and SHI-BTCA samples,these two materials had similar high adsorption capacities and fast adsorption kinetics,while their adsorption behaviors were very different.For example,the pH of the solution after copper adsorption onto the SHI-BTCA sample increased significantly,while that of the SHI-C sample varied much less,suggesting different interactions between these shives samples and the adsorbed copper.In addition,the amounts of sodium released into the solutions after an exchange with metal ions increased strongly in the case of the SHI-BTCA sample due to the presence of COONa groups in this material,also confirming different adsorption mechanisms.This study has two objectives: the first is to characterize the surface state of the materials before and after copper adsorption and the second is to assess the interactions between the materials and the metal ions involved in the adsorption process using microscopic and spectroscopic tools,namely computed nano-tomography ,energy-disperse X-ray spectroscopy,X-ray photoelectron spectroscopy ,Fourier-transform infrared spectroscopy,Raman spectroscopy,and X-ray absorption near-edge structure spectroscopy.Three different treatments were applied to SHI-R samples.A water washing ,an activation using sodium carbonate ,and a grafting reaction using 1,2,3,4-butanetetracarboxylic acid were carried out to study their role in the performance of samples which were further used as adsorbents for copper presents in aqueous solution.To prepare SHI-W,SHI-R samples were simply washed with water for 2 days at room temperature.After that,they were filtered and dried in an oven at 80◦C until a constant mass was obtained.

The SHI-C shives were prepared by treating SHI-W samples in 1 M Na2CO3 for 4 h at 40◦C.Thereafter,the samples were washed extensively with water until a neutral pH was obtained and dried in an oven at 80◦C until a constant mass was obtained.To obtain SHI-BTCA,SHI-W samples were pre-treated with 1 M NaOH for 3 days at ambient temperature under mechanical stirring.The shives suspension was then filtered on a filtering funnel and thoroughly washed with distilled water until the water was colorless.After drying at 60◦C in a ventilated oven for 15 h,the samples were immersed in an aqueous solution containing 100 g/L of BTCA and 30 g/L of NaPO2H2 for 15 h at ambient temperature under mechanical stirring.After draining,shives were spread on the bottom of a glass crystallizer put in a ventilated oven at 160◦C for 90 min.They were suspended in distilled water overnight under stirring,filtered,thoroughly washed with distilled water,and dried in a ventilated oven at 60◦C for 15 h.The shives were finally activated by converting the acidic form of carboxylic groups into their basic carboxylate form by immersing materials in a NaHCO3 solution under mechanical stirring for 24 h.SHI-BTCA samples were filtered,washed with distilled water until neutral pH was reached,and dried in a ventilated oven at 60◦C for 15 h.The interactions between copper and hemp-based samples were assessed by using different microscopic and spectroscopic techniques such as computed nano-tomography ,energy-disperse X-ray spectroscopy,Fourier-transform infrared spectroscopy,Raman spectroscopy,X-ray photoelectron spectroscopy ,and X-ray absorption near-edge structure spectroscopy.Nano-CT images given in Fig.3a show the typical cross-section of raw hemp shives.Namely,different tissues such as the woody part constituting the main part of the shives and some remaining pith on the right side can be observed.The woody part is made of different cell types: i) isolated or grouped in groups of two or three vessels which have a quite thin cell wall and a diameter of approximately 50-150 µm; ii) the fibers with a thick cell wall,an irregular polygonal cross-section and a diameter of only a few µm and iii) the rays which are oriented in the radial direction and are generally composed of one cell in width.

A similar experiment was performed with THC,this time only evaluating the FBBB stability at refrigerated temperatures.The characteristic red color indicative of the presence of THC did form; however,like the hemp,the color was duller than when performing the usual procedure.Finally,FBBB was preloaded onto PSPME substrates and left in refrigerated temperatures for a week and were then used to evaluate 1000 ppm THC and 1000 ppm CBD solutions.Red and orange colors did form,respectively,but they were duller like all the other chromophores formed using FBBB as a preloaded salt.There is a growing interest in natural bioactive compounds both by producers and consumers in the food and pharmaceutical industries.Particularly,consumers are looking for foods without artificial and harmful preservatives that can promote their health.Essential oils as natural food preservatives are,generally,extracted from medicinal plants and herbs for their noticeable biological activities.EOs are complex combination of secondary metabolites,including terpenoid hydrocarbons,phenol derivatives and oxygenated terpenoids ; however,due to having volatile components,their usage is limited; the volatile compounds can be degraded easily by adverse external conditions such as light,oxygen,temperature,pressure,and pH.In addition,controlled release of EOs is needed when used for specific purposes like food additives.In recent years,the isolation and utilization of EOs from agroindustrial wastes and by-products have gained much research interest.Industrial hemp’s inflorescences are usually discharged during the conventional hemp processing,resulting in an underused biomass for future uses.They are a rich source of EOs and contain mainly monoterpenes and sesquiterpene hydrocarbons like -caryophyllene,α-pinene,myrcene and α-humulene,which exhibit important biological activities.Valorization of hemp by-products is a matter of interest for producers,allowing them to increase the market value of hemp cultivation.Some studies have recently been carried out on the functional and pharmacological applications/properties of industrial hemp essential oil due to their antioxidant,trimming trays for weed antimicrobial,and anti-inflammatory activities.

There is much interest in developing biodegradable nanoparticles as an effective delivery system for delivering lipophilic food bioactives.Proteins are attractive options for designing polymeric NPs as suitable wall materials thanks to their amphiphilic nature compatible with many active substances,as well as excellent functional properties.Whey protein,as a typical cheese processing by-product,is commonly used to foods due to its great nutritional value and techno-functional properties such as gelling,foaming and emulsifying.Protein fibrillation is used nowadays for fabrication of protein fibrils having novel functionalities and improved structure stability.Heating the protein above 80 ◦C for 5–24 h at low ionic strength and acidic conditions can result in production of WPI fibrils ,which are more suitable as delivery carriers comparing to native proteins,due to having multiple functional groups,and thus,promoting different interactions with numerous and drugs and nutrients.Protein complexation has recently been proposed as a promising and efficient way for improving the bio-availability,chemical stability and dispersion of bio-active compounds in an aqueous environment.Mung bean protein is a significant plant protein that demonstrates high potential as sustainable protein source for its availability,nutritional value,hypoallergenic,and desirable foaming,emulsifying,gelling,and film-forming capabilities.The rainbow trout as an extremely perishable food belongs to the Salmonidae family.Recently,the demand for rainbow trout has increased remarkably,and this could be due to its desirable characteristics.Due to the demand for fresh refrigerated fish with extended shelf life,notable study has been directed toward prolonging the shelf life of this fish.Although,there are scarce studies on the characterization and fabrication of IHEO encapsulated in complexation of WP nanofibrils and MBP NPs.The present study is to manufacture and characterize WPNFMBP NPs as a novel wall material for loading IHEO.Furthermore,application of IHEO-reinforced WPNF-MBP NPs as a potential natural additive for increasing the shelf-life of Rainbow trout fillets during refrigerated storage is going to be evaluated.The chemical changes of Rainbow trout fillets were monitored through the measurement of TVB-N,TBA and PV values of the fish oil during storage.Hydroperoxides are the main products of lipid oxidation; thus,measurement of peroxide values is helpful for indicating oxidative rancidity.As depicted in Fig.3a,PV in the control samples increases from 1.33 to 8 meq O2/kg at day 10 and declines after this day,maybe due to the reaction of hydroperoxide with protein as well as the collapse of primary oxidation products into secondary oxidation products.PV level tends to increase toward the end of the storage period.

A similar pattern of hydroperoxide content has also been reported in Rainbow trout fillets during storage.The samples coated with WPNF-MBP and IHEO-reinforced WPNF-MBP NPs showed significantly lower PV levels than the control during the storage period.The literature considers a PV value of 20 meq O2/kg oil is as the maximum limit for fish.Our results indicated that the IHEO-reinforced WPNF-MBP NPs coating is significantly reduced the amount of primary lipid oxidation in fish during storage; this is in agreement with the results of other similar investigations.TBA index is a common indicator for evaluating the lipid oxidation level by measuring such oxidation products as aldehydes like malondialdehyde.The recommended perceivable level of TBA in food as objectionable odour is about 1–2 mg MDA/kg ; however,Raeisi et al.proposed the maximal acceptable level 5 mg MDA eq/kg in Rainbow trout with no negative effects on its safety and quality.As indicated in Fig.3b,the TBA level remained below the maximal acceptable level during the 14-day storage.However,in the uncoated samples,the TBA value was much higher than in the coated samples ; however,the initial value of TBA was in the range of 0.09–0.13 mg MDA/kg,consistent with reports of other researchers for fresh Rainbow trout.The slightly lower oxidation rate in the IHEO-reinforced WPNFMBP NPs coatings can be because of the oxygen barrier and antioxidant activity features of IHEO and WPNF.The antioxidant mechanism of IHEO can be due to its polyphenols,which show scavenging activity against free radicals through providing hydrogen atoms to free radicals,preventing radical chain initiation,and thus,preventing the formation of metal catalyzed free radicals.On the other hand,the WP coatings provided a great protection against oxidation.IHEO addition improved the WPNF-MBP NPs coatings’ antioxidant properties.WPs exhibit antioxidant activity through different ways: 1) formation of a coating,which is a good barrier for O2 permeability coated samples during storage,2) enjoying a free radical scavenging capacity by some amino acids and metal chelation by bovine serum albumin and lactoferrin,3) having sulfhydryl groups partially responsible for their antioxidant properties,and 4) containing β-lactoglobulins and α-lactalbumin with good antioxidant activity for having amino acid residues.Furthermore,particle size reduction of IHEO and WPNF-MBP coatings after nanoencapsulation can increase these ingredients’ specific surface; thereby achieving an efficient amount of free radical absorption would be achieved.

TVB-N is produced from degradation of proteins and non-protein nitrogenous compounds,mainly as a result of microbial and enzymatic activities,as an indicator of meat and fish spoilage.The TVB-N level in the control samples was initially 6.68 mg N/100 g,and there was no significant difference among the different samples at the first day of storage.By increasing the bacterial counts,the TVB-N value increased gradually in all groups; however,in the samples coated with WPNF-MBP and IHEO-reinforced WPNF-MBP NPs,the TVBN value was significantly lower than in the controls.Considering the maximal acceptable level of 25 mg N/100 g in fish flesh ,the TVB-N limit was achieved at day 8 of storage in the controls,while in the WPNF-MBP coated samples,this limit was reached by the 10th days,and in the IHEO-reinforced WPNF-MBP NPs coated samples,it was lower than this limit during the whole period of storage.At the last day of storage,the TVB-N value for the IHEO-reinforced WPNF-MBP NPs coated samples was 12.1 and 9 mg N/100 g lower than the control and WPNF-MBP coated samples,respectively.We previously mentioned that TVB-N is produced mainly because of the bacterial degradation of the nitrogenous compounds of proteins and non-protein products,its low value in the coated samples can be due to the microbial inhibitory effects of treatments that decrease the formation of TVB-N.Similarly,Shokri et al.found that coating of Rainbow trout fillets with chitosan-Ferulago angulate EO nano-emulsion retarded the increasing rate of TVB-N index during storage at 4 ◦C.Ozogul et al.found that nano-emulsions based on plant EOs significantly inhibited the TVBN formation in Rainbow trout fillets during ice storage.To the best of our knowledge,there are no studies on the effect of IHEO-reinforced WPNFMBP NPs coating on the formation of TVB-N in Rainbow trout fillets; however,trimming tray with screen the lower TVB-N in the samples coated with IHEO-reinforced WPNF-MBP NPs in the present work could result from the antibacterial efficiency of WPNF-MBP coating facilitated by nano-encapsulation of IHEO.Fig.4 illustrates the overall acceptable scores of the control and coated Rainbow trout fillets with WPNF-MBP and IHEO-reinforced WPNF-MBP NPs.In the present study,appearance,color,odor,and texture were taken into consideration in the overall acceptance scoring.The overall acceptance scores were in the range of 1–5.High preference levels represent high element scores.All samples exhibited high sensory and quality scores at the first day of analysis.An overall acceptance below 3 of fish is considered to be unacceptable for human consumption.The overall acceptance scores of the coated and control fish samples showed a decreasing trend up to the end of storage time.The samples coated with IHEO-reinforced WPNF-MBP NPs exhibited a higher score comparing to the other samples during the refrigerated storage.The control samples’ sensory properties were ‘unacceptable’ by the 10th day.Also,at the day 12,the samples coated with WPNF-MBP did not achieve acceptable scores due to their unpleasant appearance and sticky surface; however,the incorporation of IHEO reduced these defects.Hence,the samples coated with IHEO-reinforced WPNF-MBP NPs had better sensory scores than the others for control the lipid oxidation and bacterial population.

These results are consistent with the findings of O˘ guzhan Yıldız and Yangılar and Farsanipour et al.for Rainbow trout WP-based coated samples incorporated with EOs stored at refrigerator condition.Industrial hemp,Cannabis sativa L.is a multifaceted crop that is increasingly being considered by farmers looking to diversify and reduce their ecological footprint.Industrial hemp is grown for both fiber and food,and is also used in cosmetics and for medicinal and nutraceutical/ pharmaceutical purposes.A major factor contributing to the sustainability of hemp is that most parts of the plant can be used,and innovation is contributing to additional uses of hemp for biofuels,high-performance textiles and natural insecticides.Hemp has long been used as a food source and in traditional medicine ,and this has stimulated research into its use as a functional food.In addition to its healthy oil and protein profiles,several bioactive peptides have been found in hemp seed,as well as polyphenols and other effective antioxidants.Hemp seed should be relatively easy to get to food markets as seed processing can use existing production facilities with some adjustments.Hemp seed is actually an achene or nut,with a hard outer shell ,a papery testa,and an inner seed.Industrial hemp is classified as Cannabis plants with low levels of Δ9- tetrahydrocannabinol ,depending on the country and state.It was recently estimated that the global production of hemp will double in the four years from 2016 to 2020,and some states in Australia have already more than doubled their production.This resurgence of interest in hemp has led to an increasing number of states in the USA growing low THC hemp,and as of 2017 all states in Australia have legalized cultivation of industrial hemp for food and fiber applications.In comparison to other crops hemp has been described as sustainable because it is relatively resistant to biotic and abiotic stresses thereby reducing agronomic inputs.In temperate Australia industrial hemp is grown as a summer crop and requires supplemental irrigation.Hemp has a relatively high water requirement compared to dryland cereals,although much less than cotton.A major advantage of hemp from an agronomic point of view is the rapid growth of seedlings such that it effectively outcompetes weeds.This and the fact that it has relatively few pests and diseases makes hemp better suited for organic production than many other row crops.In Australia,some pests and diseases have been observed,but in most cases there has been no significant impact on productivity.Industrial hemp is suited to a wide variety of soil types as long as they are fertile with good drainage and water holding capacity.When C.sativa is grown for fiber,tall varieties are used because of their increased biomass.Shorter varieties are being bred with higher harvest index that are specifically targeted to food markets.Characterization of FINOLA seed showed that it contains approximately 30% oil and 25% protein.Analysis of lipids of FINOLA seed revealed that over 80% are polyunsaturated including two essential fatty acids,linoleic acid and α-linolenic acid.Hemp seed contains a ratio of omega-6 to omega-3 in a desirable range between 2:1 and 3:1.Current western diets are generally deficient in omega-3 fatty acids,with a high ratio of omega-6:omega-3,due to the increased intake of vegetable oils including sunflower and corn oils,which have ratios of >50:1 omega-6:omega-3.