Microorganisms in Biostimulants

In March 2016, the EU Commission presented a proposal for new regulations on fertilising material. The regulation includes product rules for a wide range of organic and inorganic products. Microbial biostimulants is one of the categories of products that are included. Biostimulants, in the draft EU regulation, are defined as fertilising materials that affect nutrient processes independently of the product's own nutrient content and with the purpose of improving nutrient utilisation, tolerance for abiotic stress or quality of the crop. Positive list in which species of these bacterial genera are listed: Azotobacter spp, Rhizobium spp., Azospirillum spp and Mycorrhizal fungi are a part of the regulation. Since the import and use of these organisms are the responsibility of both the Norwegian Food Safety Authority and the Norwegian Environment Agency, they asked VKM to submit a joint report on effects on health (humans, plants and animals), biodiversity and dispersal, quality of agricultural land and on soil environment. Conclusions: Health risks: Based upon our literature review, we have found no indication of any specific diseases in plants, animals or humans induced by the discussed microorganisms. A few reported cases of human disease are caused through wound infections or injections in immunocompromised patients. These represent a situation where any microorganism may induce infections and is not specific for the agents discussed in this report. In summary, the risk of any disease caused by the discussed microorganisms is considered negligible. Environmental risks: In soil the biodiversity, competition, adaptation and functional redundancy of microorganisms are extremely high. This means that introduced microorganisms have a very small chance for establishing, and even less so for affecting biodiversity and soil functioning. Introduction of nitrogen fixing species or fungi that can transport P to plants (mycorrhiza) will lead to an increase in the primary production. However, even a large increased activity for these processes will not outcompete naturally occurring symbiotic N-fixation or growth of inherently non-mycorrhizal plant species. Thus, the risks associated with introduced non-pathogenic microorganisms are very low.

Risk Assessment of "Other Substances" – Inositol

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating the addition of “other substances” to food supplements and other foods. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals that have a nutritional and/ or physiological effect. It is added mainly to food supplements, but also to energy drinks and other foods. In this series of risk assessments of "other substances", VKM has not evaluated any potential beneficial effects from these substances, only possible adverse effects. The present risk assessment is based on previous risk assessments of inositol and articles retrieved from a literature search. According to information from NFSA, inositol is an ingredient in energy drinks sold in Norway. NFSA has requested a risk assessment of 10 mg/100 ml inositol in energy drinks. Drinking patterns reflecting a high acute intake, a mean chronic intake and a high chronic intake were assessed. Inositol (CAS no. 6917-35-7) is a sugar alcohol. Among the nine possible stereoisomers, myo inositol (CAS no. 87-89-8) is the most abundant. The name inositol is frequently used as a synonym for myo -inositol. Inositol occurs naturally in all organisms including humans, and is an important component in all human cells. Inositol-containing lipids and phosphates are required for various structural and functional processes, including membrane formation, signalling, membrane trafficking and osmoregulation. Endogenous production of inositol in humans amounts to about 4 g/day (about 57 mg/kg bw per day in a 70 kg adult) (EFSA, 2014). The total dietary intake of inositol in adults is estimated to range between 500 to 1000 mg/day (about 7-14 mg/kg bw per day). Inositol added to energy drinks in Norway denotes the compound myo -inositol, according to information from NFSA. M yo -inositol is a water-soluble compound naturally occurring in the cells of all living organisms including humans, animals, plants and microorganisms. Certain plant (fruits and vegetables) and foods from animals contain inositol, and seeds of cereals and legumes show high levels of the inositol storage form, phytic acid (inositol hexaphosphate). With regard to hazard identification and characterisation of inositol, most of the adverse effects observed in several human studies were related to gastrointestinal symptoms such as nausea, flatulence, loose stools and diarrhoea. Drinking patterns reflecting a high acute intake, a mean chronic intake and a high chronic intake were assessed for energy drinks containing 10 mg inositol per 100 ml, for the age groups children (3 to <10 years and 10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years). For the high acute drinking pattern, the intake was estimated to be 1000, 1500, 2000 and 2000 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (>18 years), respectively. For the mean chronic drinking pattern, the intake was estimated to be 58, 65, 64 and 71 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. For the high chronic drinking pattern, the intake was estimated to be 163, 180, 210 and 320 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. The data on toxicity of inositol was very limited. The human study with the longest exposure at highest doses (3 months treatment at maximum tolerated dose) that was available for risk assessment was a clinical study of 40-74 year old smokers with bronchial dysplasia, from which a NOAEL of 18 g/day of myo -inositol was established (Lam et al. 2006). VKM estimated the margins of exposure (MOE) based on the NOAEL established in this study. The MOE is the ratio of the NOAEL value to the exposure. An acceptable MOE value for a NOAEL-based assessment of inositol based on a human study is ≥10, taking into account a factor 10 for the interindividual variation between humans in toxicokinetics and toxicodynamics. Due to the uncertainty regarding the relevance of the study by Lam et al. (2006) for the general healthy population, an additional safety factor of 3 was used. Therefore, an acceptable MOE value was 30. For all age groups, the MOE values were in the range of 857 to 2570 for mean chronic intake and in the range of 367 to 857 for high chronic intake of energy drinks, respectively, i.e. far above the acceptable MOE value of 30. Since neither the sub-optimal human study by Lam et al. (2006) or the animal studies in rodent models of chronic diseases available were on healthy subjects, as a supplement to the MOE values calculated from the human study, comparisons with endogenous production and amounts in food of inositol were also performed. No studies specifically on children (3 to <10 years and 10 to <14 years) and adolescents (14 to <18 years) were identified. Based on the included literature there was no evidence indicating that age affects tolerance or endogenous production of inositol. Therefore, in this risk characterisation a tolerance and an endogenous production of inositol as for adults, based on body weight, was assumed for these age groups. For the high acute drinking pattern, and for the mean chronic and the high chronic drinking patterns all estimated intakes of inositol from energy drinks containing 10 mg/100 ml were far below the endogenous production (57 mg/kg bw per day), and also below the dietary intake (7-14 mg/kg bw per day). VKM concludes that it is unlikely that the exposure to inositol from the high acute, the mean chronic or the high chronic drinking patterns causes adverse health effects in children (3 to <10 years and 10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years).

Risk Assessment of "Other Substances" – Taurine

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses in food supplements and concentrations in energy drinks given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating the addition of “other substances” to food supplements and other foods. "Other substances" are described in the food supplement directive 2002/46/EC as s ubstances other than vitamins or minerals that have a nutritional and/or physiological effect . It is added mainly to food supplements, but also to energy drinks and other foods. VKM has not in this series of risk assessments of "other substances" evaluated any claimed beneficial effects from these substances, only possible adverse effects. The present report is a risk assessment of taurine, and it is based on previous risk assessments and articles retrieved from a literature search. According to information from NFSA, taurine is an ingredient in food supplements and energy drinks sold in Norway. NFSA has requested a risk assessment of 750, 800, 900, 1000 and 2000 mg/day of taurine in food supplements, and of 300, 350 and 400 mg/100 ml of taurine in energy drinks. Drinking patterns reflecting a high acute intake, a mean chronic intake and a high chronic intake were assessed. For food supplements, the intake of taurine was estimated for the age groups children (10 to <14 years), adolescents (14 to <18 years) and adults (>18 years), whereas for energy drinks the age group children (3 to <10 years) was also included. Other sources of taurine, such as foods and cosmetics, have not been included in the present risk assessment. Taurine (CAS No. 107-35-7) is synthesised endogenously (average 50-125 mg per day), and participates in the formation of bile salts and is involved in a number of crucial physiological processes, including modulation of calcium flux and neuronal excitability, osmoregulation and membrane stabilisation. Taurine occurs naturally in food, especially in meat and seafood. The mean daily intake of taurine from the diet has been estimated to vary between 40 and 400 mg/day. There are indications that taurine may have cardiovascular and neurological effects in humans. However, based on the human studies, an intake of approximately 21 mg/kg bw per day is considered unlikely to cause adverse health effects. Based on a 13-week neurotoxicity study in rats, a no observed adverse effect level (NOAEL) of 1000 mg/kg bw per day for pathological changes was set in 2009 by the European Food Safety Authority (EFSA). In the present risk assessment, VKM has used this NOAEL of 1000 mg/kg bw per day from rats. The human studies available were not of sufficient quality (due to low number of participants, non-healthy populations, short duration) to be used as the sole basis for the risk characterisation. The risk characterisation is based on the margin of exposure (MOE) approach; the ratio of the NOAEL to the exposure. An acceptable MOE value for a NOAELbased assessment of taurine based on an animal study is ≥100, which includes a factor 10 for extrapolation from animals to humans and a factor 10 for interindividual human variation. However, since the NOAEL set by EFSA was based on the highest tested dose and there is a possibility that the actual NOAEL is higher than 1000 mg/kg bw per day, the intake that was considered unlikely to cause adverse health effects based on human studies (21 mg/kg bw per day) was also taken into consideration in the risk characterisation. Food supplements: For children (10 to <14 years), the estimated daily intakes of taurine were 17.3, 18.4, 20.7, 23.0 and 46.1 mg/kg bw per day from daily doses of 750, 800, 900, 1000 and 2000 mg taurine, respectively, from food supplements. The margin of exposure (MOE) values was in the range of 22-58 for the various taurine doses, i.e. all below 100. However, from a daily intake of 750, 800 or 900 mg taurine from food supplements, the estimated intakes were below 21 mg/kg bw per day (the intake considered unlikely to cause adverse health effects based on human studies). VKM therefore concludes that it is unlikely that a daily intake of 750, 800 or 900 mg taurine from food supplements causes adverse health effects in children (10 to <14 years). The estimated exposure from a daily intake of 1000 or 2000 mg taurine was above 21 mg/kg bw per day. Thus, VKM concludes that a daily intake of 1000 or 2000 mg taurine from food supplements may represent a health risk in children (10 to <14 years). For adolescents (14 to <18 years), the estimated daily intakes were 12.2, 13.1, 14.7, 16.3 and 32.6 mg/kg bw per day from daily doses of 750, 800, 900, 1000 and 2000 mg taurine, respectively, from food supplements. For adults (≥18 years), the estimated intakes were 10.7, 11.4, 12.9, 14.3 and 28.6 mg/kg bw per day from a daily intake of 750, 800, 900, 1000 and 2000 mg taurine, respectively, from food supplements. For adolescents (14 to <18 years) and adults (≥18 years), the MOE values were in the range of 31-82 and 35-93, respectively, i.e. all below 100. However, from a daily intake of 750, 800, 900 or 1000 mg taurine from food supplements the estimated intakes were below 21 mg/kg bw per day (the intake considered unlikely to cause adverse health effects based on human studies) for both age groups. Thus, VKM concludes that it is unlikely that a daily intake of 750, 800, 900 or 1000 mg of taurine causes adverse health effects in adolescents (14 to <18 years) and adults (≥18 years). For adolescents (14 to <18 years) and adults (≥18 years) the estimated MOE values were 31 and 35, respectively, i.e. below 100, after a daily intake of 2000 mg taurine from food supplements. In addition, the estimated intakes were above the intake level of 21 mg/kg bw per day (the intake considered unlikely to cause adverse health effects based on human studies) for both age groups. Thus, VKM concludes that a daily intake of 2000 mg of taurine may represent a risk of adverse health effects in adolescents (14 to <18 years) and adults (≥18 years). Energy drinks: High acute drinking pattern, all age groups: For the high acute drinking pattern, the estimated consumption of energy drinks was 1000, 1500, 2000 and 2000 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. For the concentrations of 300, 350 and 400 mg taurine/100 ml energy drink, the intake levels of taurine after a high acute consumption of energy drinks (in mg/kg bw per day) were 130, 152 and 173; 104, 121 and 138; 97.9, 114 and 131; and 85.7, 100 and 114, for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. Due to lack of an acute reference dose or other data for acute toxicity of taurine, it was not possible to characterise the risk related to an acute intake of taurine for any of the age groups. Mean chronic drinking pattern, all age groups: For the mean chronic drinking pattern, the estimated consumption of energy drinks was 58, 65, 64 and 71 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. For the concentrations of 300, 350 and 400 mg taurine/100 ml energy drink, the intake levels of taurine after a mean chronic drinking pattern (in mg/kg bw per day) were 7.5, 8.8 and 10.0; 4.5, 5.2 and 6.0; 3.1, 3.7 and 4.2; and 3.0, 3.6 and 4.1, for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. In all age groups, the estimated MOE values were 100-333, i.e. 100 or above, for all three taurine concentrations. In addition, the estimated intakes were all below 21 mg/kg bw per day (the intake considered unlikely to cause adverse health effects based on human studies) for all age groups. Thus, VKM concludes that it is unlikely that the mean chronic intake of all three concentrations of taurine causes adverse health effects in children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years). High chronic drinking pattern, all age groups: For the high chronic drinking pattern, the estimated consumption of energy drinks was 163, 180, 211 and 320 ml/day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. For the concentrations of 300, 350 and 400 mg taurine/100 ml energy drink, the intake levels of taurine after a high chronic drinking pattern (in mg/kg bw per day) were 21.2, 24.7 and 28.2; 12.4, 14.5 and 16.6; 10.3, 12.0 and 13.8; and 13.7, 16.0 and 18.3 mg/kg bw per day for children (3 to <10 years), children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), respectively. For children (3 to <10 years), the estimated MOE values were 47, 40 and 35, for the three taurine concentrations of 300, 350 and 400 mg/ml, respectively, i.e. all below 100. In addition, the estimated intakes were all above 21 mg/kg bw per day (the intake considered unlikely to cause adverse health effects based on human studies) for all three taurine concentrations. Thus, VKM concludes that a high chronic intake of all three concentrations of taurine from energy drinks may represent a health risk in children (3 to <10 years). For children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years), the estimated MOE values were in the range of 55-97, i.e. all below 100 for all three taurine concentrations. However, the estimated intakes were all below the intake level of 21 mg/kg bw

Risk Assessment of "Other Substances" – Coenzyme Q10

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating the addition of “other substances” to food supplements and other foods. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals t hat have a nutritional and/or physiological effect. It is added mainly to food supplements, but also to energy drinks and other foods. In this series of risk assessments of "other substances", VKM has not evaluated any potential beneficial effects from these substances, only possible adverse effects. The present risk assessment of coenzyme Q10 (CoQ10) is based on previous risk assessments and articles retrieved from a literature search. According to information from NFSA, CoQ10 is an ingredient in food supplements sold in Norway. NFSA has requested a risk assessment of intake of 100 mg/day of CoQ10 in food supplements. CoQ10 (CAS no. 303-98-0) is a naturally-occurring, lipid-soluble compound present in all tissues in humans. Ubiquinone is the totally oxidized form (CoQ10), whereas ubiquinol (CoQ10H2) is the totally reduced form. Meat and fish are the food sources richest in CoQ10. CoQ10 intake from the diet ranges between 3 and 6 mg/day in developed countries. The total body pool of CoQ10 is estimated to be approximately 0.5–1.5 g in an adult. Several studies of CoQ10 (both oxidized and reduced form) have been performed in healthy humans (adults) and animals, showing fairly similar results. The adverse effects reported in a small number of human subjects were generally limited to mild gastrointestinal symptoms such as nausea and stomach upset. In humans, orally ingested CoQ10 was well tolerated at doses up to 900 mg/day (corresponding to 12.9 mg/kg bw per day in a 70 kg adult) over periods up to one month. With regard to animal studies, the lack of adverse effects of CoQ10 doses up to 1200 mg/kg per day in long-term toxicity studies supported and extended the results from the human studies. No studies on children (10 to <14 years) and adolescents (14 to <18 years) were identified. Based on the included literature there was no evidence indicating that age affects tolerance for CoQ10. Therefore, in this risk characterisation the same tolerance as for adults was assumed for these age groups (adjusted for body weight). From a daily dose of 100 mg CoQ10, the daily exposure is 2.3 mg/kg bw for children (10 to <14 years), 1.6 mg/kg bw for adolescents (14 to <18 years), and 1.4 mg/kg bw for adults (≥18 years). For the risk characterization, the values used for comparison with the estimated exposure are 900 mg/day (corresponding to 12.9 mg/kg bw per day in a 70 kg adult) based on human studies (4 weeks) and the no observed adverse effect level (NOAEL) of 1200 mg/kg bw per day based on a long-term toxicity study in rats (52 weeks). The margin of exposure (MOE) approach is used for the rat study; that is the ratio of the NOAEL to the exposure. An acceptable MOE value for a NOAEL-based assessment of CoQ10 based on an animal study is ≥100, which includes a factor 10 for extrapolation from animals to humans, and a factor 10 for interindividual human variation. Comparing the NOAEL from a long-term toxicity study in rats with the estimated exposure for the different age groups, it is unlikely that a daily dose of 100 mg/day of CoQ10 causes adverse health effects in children above 10 years, adolescents and adults. Comparing the dose reported to be well tolerated for healthy adults directly with the estimated exposure, it is unlikely that a daily dose of 100 mg/day of CoQ10 causes adverse health effects in children above 10 years, adolescents and adults. VKM concludes that it is unlikely that a daily dose of 100 mg of CoQ10 from food supplements causes adverse health effects in children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years).

Risk Assessment of "Other Substances" – Lycopene

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses in food supplements and concentrations in energy drinks given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating the addition of “other substances” to food supplements and other foods. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals that have a nutritional and/or ph ysiological effect. It is added mainly to food supplements, but also to energy drinks and other foods. VKM has not in this series of risk assessments of "other substances" evaluated any claimed beneficial effects from these substances, only possible adverse effects. The present report is a risk assessment of lycopene, and it is based on previous risk assessments and articles retrieved from a literature search. According to information from NFSA, lycopene is an ingredient in food supplements sold in Norway. NFSA has requested a risk assessment of 10 mg/day of lycopene in food supplements. The intake of lycopene was estimated for the age groups children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years). Other sources of lycopene, such as foods and cosmetics, have not been included in the present risk assessment. Lycopene belongs to a large group of naturally-occurring pigments known as carotenoids, and is known to have antioxidant properties. Lycopene is a natural constituent of red fruits and vegetables and of certain algae and fungi. The major sources of natural lycopene in the human diet are tomatoes and tomato-based products. Fruits like pink grapefruit, water melon, rosehip, papaya and guava are also sources of lycopene. Lycopene can be obtained by solvent extraction of the natural strains of red tomatoes (Lycopersicon esculentum L.) with subsequent removal of the solvent. Synthetic lycopene can be produced by the Wittig condensation of synthetic intermediates commonly used in the production of other carotenoids used in food. Lycopene biosynthesis by the fungus B. trispora follows the same pathway as the synthesis of lycopene in tomatoes. There are case reports of yellow-orange skin discoloration and/or gastrointestinal discomfort after prolonged high intakes of lycopene-rich food and supplements, those effects being reversible upon cessation of lycopene ingestion. The results from one study indicated that lycopene increased the incidence of the preterm labor and low birthweight babies. However, due to weaknesses in the reporting, VKM cannot use the results from this study in the risk characterisation. An ADI of 0.5 mg/kg bw per day was established by EFSA in 2008. The ADI was derived from the NOAEL of 50 mg/kg bw per day from a 52-week toxicity study in rats, based on a partly reversible increased level of the liver enzyme alanine transaminase (ALT). An ADI is set to cover the general population, including children. This ADI-value was used for comparison with the estimated exposure in the risk characterization. From a daily dose of 10 mg lycopene, the daily exposure is 0.23 mg/kg bw for children (10 to <14 years), 0.16 mg/kg bw for adolescents (14 to <18 years), and 0.14 mg/kg bw for adults (Table 3.1-1). Thus, the intakes are below the ADI of 0.5 mg/bw per day for all age groups. VKM concludes that it is unlikely that a daily dose of 10 mg lycopene from food supplements causes adverse health effects in children (10 to <14 years), adolescents (14 to <18 years) and adults (≥18 years).