The Risk of Development of Antimicrobial Resistance with the Use of Coccidiostats in Poultry Diets

Background: Antimicrobials revolutionized human as well as animal medicine in the 20th century by providing effective treatment of diseases caused by pathogenic microorganisms. However, microorganisms have the ability to develop antimicrobial resistant strains. This occurs when microorganisms mutate or when resistance genes are exchanged between them. The use of antimicrobial drugs accelerates the emergence of drug-resistant strains. A priority is to safeguard the efficacy of antimicrobial drugs we depend on for treatment of infectious diseases in humans. Use of antimicrobials in food animals can create a source of antimicrobial resistant bacteria that can spread to humans both by direct contact and through the food supply. Coccidiosis is an intestinal disease in animals caused by unicellular parasites called coccidia. As most of the damage of this infection is done by the time signs of the disease are widespread, preventive measures are preferred. Coccidiostats are animal feed additives used to prevent coccidiosis by inhibiting or killing coccidia. There are two major groups of coccidiostats; ionophores and non-ionophores, the latter also referred to as “non-ionophore coccidiostats” (but also called chemicals). One main difference between these groups is that ionophores also inhibit or kill some bacterial species, whereas non-ionophore coccidiostats do not. Consequently, some bacterial infections may also be controlled by ionophore coccidiostats, e.g. the poultry disease necrotic enteritis caused by the bacterium Clostridium perfringens (C. perfringens). Eleven different coccidiostats have been authorised for use in the EU, both ionophores and non-ionophore coccidiostats. Norway has been exempted from the EEA Agreement in this field and has approved only five; all ionophores. The two ionophore coccidiostats currently used in Norway are narasin for broilers and monensin for turkeys. Resistance to coccidiostats in coccidia and bacteria: Development of resistance in coccidia to all eleven coccidiostats has been described in the scientific literature, but the prevalence of resistance is unknown. Cross-resistance between various ionophore coccidiostats has also been shown, i.e. development of resistance to one ionophore may also render the coccidia resistant to another ionophore. Various rotation and shuttle programmes with exchange between ionophores and non-ionophore coccidiostats are believed to prevent or delay development of resistance in coccidia. In Norway, such programmes will have little effect as long as only ionophores and not non-ionophore coccidiostats are approved for use. Development of resistance against ionophores has also been observed in bacteria. In the Norwegian surveillance programme NORM-VET during the years 2002 - 2013, between 50 - 80% of the tested flocks had narasin resistant faecal enterococci, which are bacteria that are part of the normal intestinal microbiota. However, the pathogenic bacterium C. perfringens has not been shown to be resistant against any ionophore. Cross-resistance in bacteria to more than one ionophore has been observed. In addition, a limited amount of data may indicate an association between narasin and resistance to the antibacterials bacitracin and vancomycin. As these are antibacterials used for treatment in humans, more research should be performed to validate these results. Non-ionophore coccidiostats, which do not have antibacterial effect, are not approved in Norway. If such coccidiostats were approved in Norway, coccidiostats with negligible probability of inducing resistance in bacteria would be available. Human exposure to resistant bacteria and coccidiostats: Humans may theoretically be exposed to coccidiostat resistant bacteria from poultry in a number of ways, e.g. by handling live animals and their manure, through slaughtering and processing, and by preparation and consumption of poultry meat. Furthermore, bacteria of the human normal microbiota, which cover all skin and mucosal surfaces, might develop resistance if they are exposed to coccidiostats. In this assessment, the probabilities of exposure are classified as: Negligible (extremely low), Low (possible, but not likely), Medium (likely), High (almost certain) and Not assessable. The Panel has estimated the following probabilities of human exposure: Handling manure from coccidiostat fed poultry without sufficient risk-reducing measures entails a high probability of exposure to both resistant bacteria and coccidiostats. Without proper protection, the probability of exposure to coccidiostats is also high when handling coccidiostat premixes and feeds containing coccidiostats without proper protection measures. Various treatments, e.g. composting, of the manure may reduce the probability. The probability of exposure to resistant bacteria is medium for workers handling carcasses and raw meat on a daily basis if risk-reducing measures are not applied, whereas the probability of exposure to coccidiostats is negligible. For consumers, the probability of exposure to coccidiostats is negligible. The probability for exposure to resistant bacteria is also negligible in heat treated food since heat treatment kills the bacteria. The probability of exposure to coccidiostat resistant bacteria is low to medium if handling raw meat without proper hygienic procedures, because raw meat may harbour resistant bacteria. Risk-reducing measures will lower the probabilities. However, little is known concerning the consequences of human exposure to coccidiostat resistant bacteria or to to coccidiostats. There is little information in scientific literature indicating whether such bacteria in fact will colonize the human body, either transitionally or permanently. Furthermore, there is no information on the probability of exchange of resistance genes from transferred bacteria to bacteria of the human natural microbiota or to pathogens. Likewise, the Panel has no information on the level of exposure, e.g. the amount of coccidiostats and their metabolites, or the time period, necessary for the various bacteria to give rise to resistant variants. As coccidiostats are not used to treat infectious diseases in humans, concern of resistance is related to possible cross- or co-resistance with antibacterials considered important in human medicine. Such resistance has so far not been confirmed. Use of therapeutic antibacterials for poultry: If the ionophore coccidiostats used in Norway are replaced by one or more non-ionophore coccidiostat with no antibacterial effect and no other changes are done, the coccidiostats used will no longer inhibit the bacterium Clostridium perfringens, which is the cause of necrotic enteritis. Over time this will likely to lead to a need for intermittent or continuous use of higher levels of therapeutic antibacterials due to increased incidence of this desease in poultry production. The magnitude of the increase is difficult to predict. Alternatives to in-feed antimicrobials: Eradication from the birds’ environment of coccidia causing coccidiosis is difficult to achieve because the coccidia form oocysts that survive outside the host and resist commonly used disinfectants. Vaccination with non-pathogenic vaccines is now used increasingly in commercial Norwegian broiler farms, instead of in-feed coccidiostats. So far coccidiosis has not been reported as a problem in this transition process to broiler rearing without in-feed coccidiostats in Norway. Non-antimicrobial feed additives with purported health-promoting benefits, i.e. acid-based products, probiotics, prebiotics, synbiotics, yeast-based products, plant-derived products, combinations of these, and other products have been developed and used in feed. These products have been tested for efficacy against coccidia with conflicting, non-consistent or non-convincing results. The majority of these products appear to target the bacterial microbiota rather than coccidia. The Panel has not assessed possible effects of other types of management changes.

The Link between Antimicrobial Resistance and the Content of Potentially Toxic Metals in Soil and Fertilising Products

Potentially toxic metals (PTM), along with PTM-resistant bacteria and PTM-resistance genes may be introduced to soil and water through sewage systems, direct excretion, land application of biosolids (organic matter recycled from sewage, especially for use in agriculture) or animal manures as fertilisers, and irrigation with wastewater or treated effluents. The Norwegian Food Safety Authority (NFSA) asked the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an assessment of the link between antimicrobial resistance (AMR) and potentially toxic metals (PTM) in soil and fertilising products. The NFSA would like VKM to give an opinion on the following question related to the influence of potentially toxic metals on antimicrobial resistance: Can the content of arsenic (As), cadmium (Cd), chromium (CrIII + CrVI), copper (Cu), lead (Pb), mercury (Hg), nickel (Ni), and zinc (Zn) in soil and fertilising products that are relevant for Norway play a role in the development, spreading, and persistence of bacterial resistance to these elements, as well as cross or co-resistance to antimicrobial agents? VKM appointed a working group, consisting of two members of the Panel on Biological Hazards, to prepare a draft Opinion document and answer the questions. The Panel on Biological Hazards has reviewed and revised the draft prepared by the working group and approved the Opinion document “The link between antimicrobial resistance and the content of potentially toxic metals in soil and fertilising products”. In this report we assess the following PTM: arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), mercury (Hg), lead (Pb), and zinc (Zn), because of their possible presence in fertilisers and their potential to induce AMR in bacteria. This assessment is based on internationally published data. There is no systematic monitoring for toxic metals in soils in Norway, and the levels are expected to be highly variable depending on the input sources, previous and current agricultural practices, and the characteristics of the soil. Data on PTM in fertilising products added to soil are also fragmented and limited. Fertilising materials, in the form of sewage sludge or livestock manure, will add toxic metals to the existing levels in soil, and in areas of intensive agriculture, the levels will be expected to be highest. The additive effect of toxic metals in fertilising materials must be assessed from a long-term perspective, as these metals accumulate in the environment. Development of AMR can be partly regarded as a dose- and time-dependant response to exposure to different drivers for resistance. There is an indication that the presence of potentially toxic metals is a driver for development of AMR in exposed bacteria, but the dose and time exposure most likely to cause this effect is not known. Investigation of PTM-driven co-selection of AMR in environments impacted by agriculture and aquaculture should focus especially on Cu and Zn, which are added to animal feed, and on Cd because of its high concentration, in comparison with other PTM, in inorganic fertilising products. The naturally occurring background resistance in environmental bacteria complicates the estimation of the effect of PTM exposure on development of resistance. In addition, it is difficult to distinguish between the natural resistome and an elevated abundance of AMR in environmental samples. Spreading of resistance towards the PTM evaluated in this assessment involves cross- and co-resistance to antimicrobial agents used in prophylaxis and therapy in animals and people. Most important are those cases where toxic metal resistance is coupled to resistance towards highly important and critically important antibiotics. This has been described in some of the published articles included in this assessment. We do not fully understand the mechanisms behind persistence of AMR, and removing drivers for development and spread of resistance may result in a decrease in the levels of resistance, but not necessarily full disappearance. There is lack of knowledge regarding links between the level and concentration of PTM in fertilising products and soil and development of resistance in bacteria. Data regarding the routes and frequencies of transmission of AMR from bacteria of environmental origin to bacteria of animal and human origin were lacking in the published articles reviewed here. Due to the lack of such data, it is difficult to estimate the probability of development, transmission, and persistence of PTM resistance in the Norwegian environment. More research is needed to explain the relationship between development of resistance against potential toxic metals and resistance toward antimicrobial agents in bacteria.

Risk Assessment of Specific Strains of Lactobacillus rhamnosus Used as "Other Substances"

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 sold in Norway. These risk assessments will provide NFSA with the scientific basis for regulation of 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 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 Lactobacillus rhamnosus Rosell-11 ND, Lactobacillus rhamnosus W71, Lactobacillus rhamnosus GG and Lactobacillus rhamnosus Lr-329 based on previous risk assessments and also publications retrieved from literature search. The risk of the Lactobacillus strains listed above was assessed for the general population. However, in previous assessments of probiotics published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups with mature gastro-intestinal tract. VKM has also assessed the risk of L. rhamnosus Rosell-11 ND, L. rhamnosus W71, L. rhamnosus GG and L. rhamnosus Lr-329 in food supplements and other foods independent of the dose and have assessed exposure in general terms. VKM concludes that it is unlikely that L. rhamnosus Rosell-11 ND, L. rhamnosus W71, L. rhamnosus GG and L. rhamnosus Lr-329 would cause adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Health Risk Assessment of a Food Supplement Containing Lactobacillus reuteri Protectis®

The Norwegian Scientific Commitee for Food Safety (VKM) appointed a working group of experts to answer a request from the Norwegian Food Safety Authority regarding health risk assessment of Lactobacillus reuteri Protectis® in a food supplement intended for use by infants and young children. The mandate of this health risk assessment was not to evaluate the health claims related to the products as such health claims are assessed by EFSA. The specific strain DSM 17938 is a “daughter strain” of the strain ATCC 55730 which was originally isolated from normal human milk. ATCC 55730 harbours two plasmids carrying transferable resistance genes against tetracycline and lincosamides respectively. The “daughter strain” DSM 17938 was established in 2008 by curing the ATCC 55730 for these plasmids, but is in all other respects claimed to be identical to ATCC 55730 and bioequivalence of the two strains has been suggested. The strain DSM 17938 was still resistant to tetracycline (although at a considerably lower level than ATCC 55730) and a number of other antibiotics, but these resistances were all considered being intrinsic by FBO. The absence of possible transferable/mobile genes has, to our knowledge, not been confirmed in later studies. We are not aware of any data indicating that L. reuteri has been the cause of serious human diseases – and none of the studies examined has reported any adverse or undesirable short time effects. It has also been used in preterm infants with dosage corresponding to the actual recommended doses - without reporting any adverse, short term reaction. There is therefore no evidence leading to consider the strain DSM 17938 at the dosage recommended as unsafe. However, more long-term data are still lacking and the long-term safety for the age groups considered in this assessment cannot be established. As evidence is accruing that the early microbial composition of the infant gut is important for the development of the gut flora and the immune system of the growing child, it is not possible to exclude that a daily supply of a particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, albeit still unknown, adverse effects on it’s development. As the long-term data are lacking it is not possible to answer whether the amount of the food supplement or the age of the infant or young child is of importance. However, if later long-term data should reveal any adverse reaction, it is reasonable to assume that the actual age group will be the most vulnerable. As the safety was not entirely established, the question of whether there are any vulnerable groups (i.e. premature, infants or children with diseases) where there are health risks associated with the intake of Lactobacillus reuteri Protectis®, as a food supplement was not considered.

CWD in Norway

The Norwegian Food Safety Authority (NFSA) and Norwegian Environmental Authority (NEA) asked the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion on factors associated with the introduction of Chronic Wasting Disease (CWD) to Norway. VKM appointed a working group consisting of two members of the Panel on Biological Hazards, one member of Panel on Animal Health and Welfare, and two external experts to prepare the answer to the questions. The Panel on Biological Hazards has reviewed and revised the draft prepared by the working group and approved the opinion. CWD was diagnosed in March 2016 in a wild reindeer (Rangifer tarandus) from the Nordfjella mountain area in Norway and in May and June in two mooses (Alces alces) in Selbu in South Trøndelag County, approximately 300 km north from the first case. There is currently no information to determine the origin(s) of CWD agents in Norway. However, the sporadic or genetic (somatic mutation) occurrence of prion disease in cervids cannot be excluded, nor can introduction from North America or other countries. Furthermore, there is no evidence that it has not been circulating at low levels in the Norwegian cervid populations for years, but has not previously been identified. In this scientific opinion, information on prion diseases in general, and CWD in particular, is presented in the light of experiences with this disease in North America. Prions are among the most resilient pathogens known and dissemination of prions into ecosystems is likely to result in long-term problems. Prions bind strongly to soil and remain infectious. In CWD, prions are present in most peripheral organs and also shed into the environment via saliva, faeces, and urine, as well as with the placenta. CWD transmits easily among cervids, either through direct contact, or indirectly via the environment. Migration of animals is relevant for the spread between areas. Strain diversification might occur in CWD and may influence transmission properties of the agents. Clinical signs of CWD are non-specific and do not alone enable confirmation of the diagnosis. Analysis of tissue from the brainstem at the level of the obex by approved methods is necessary for diagnosis of CWD. Prion infectivity is assessed by bioassays, often involving transgenic mice. In vitro conversion assays, like protein misfolding cyclic amplification (PMCA), provide sensitive quantification of converting activity, which is a good approximation of infectivity. Genetic variation (polymorphisms) in the gene that encodes PrP (PRNP) can modulate sensitivity towards CWD. The level of such genetic variation in Norwegian wild and semi-domesticated cervids is currently unknown. Cattle and sheep are at very low risk of developing CWD and it is highly unlikely that prion diseases in sheep or cattle are the origin of CWD. Although transmission of CWD to humans has never been known to occur, and animals other than cervids have not been found to be infected, indicating a species barrier, this possibility cannot be excluded. Thus, measures for reduction of human exposure are recommended. Taking into account uncertainties regarding the plasticity of the CWD agents and the lack of transmission data from the Norwegian isolates, this scientific opinion considers the zoonotic risk of CWD to be very low.

Risk Assessment of Specific Strains of Lactobacillus acidophilus Used as "Other Substances"

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 sold in Norway. These risk assessments will provide NFSA with the scientific basis for regulation of 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 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 Lactobacillus acidophilus W37, Lactobacillus acidophilus DDS-1, Lactobacillus acidophilus La-5 and Lactobacillus acidophilus La-14 based on previous risk assessments and also publications retrieved from literature search. The risk of the Lactobacillus strains listed above was assessed for the general population. However, in previous assessments of probiotics published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups with mature gastro-intestinal tract. VKM has also assessed the risk of L. acidophilus W37, L. acidophilus DDS-1, L. acidophilus La-5 and L. acidophilus La-14 in food supplements and other foods independent of the dose and have assessed exposure in general terms. VKM concludes that it is unlikely that L. acidophilus W37, L. acidophilus DDS-1, L. acidophilus La-5 and L. acidophilus La-14 would cause adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Specific Strains of Bifidobacterium spp. Used as "Other Substances"

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 sold in Norway. These risk assessments will provide NFSA with the scientific basis for regulation of 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 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 Bifidobacterium lactis Bi-07, Bifidobacterium bifidum W23, Bifidobacterium longum Rosell-175, Bifidobacterium breve Rosell-70, and Bifidobacterium animalis sub. lactis Bb12 based on previous risk assessments and also publications retrieved from literature search. The risk of the Bifidobacterium strains listed above was assessed for the general population. However, in previous assessments of probiotics published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups with mature gastro-intestinal tract. VKM has also assessed the risk of Bifidobacterium spp. in food supplements and other foods independent of the dose and have assessed exposure in general terms. VKM concludes that it is unlikely that B. lactis Bi-07, B. bifidum W23, B. longum Rosell-175, B. breve Rosell-70, and B. animalis sub. lactis Bb12 would cause adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Specific Strains of Lactobacillus plantarum Used as "Other Substances"

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 sold in Norway. These risk assessments will provide NFSA with the scientific basis for regulation of 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 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 Lactobacillus plantarum W62, Lactobacillus plantarum 299v and Lactobacillus plantarum HEAL9 based on previous risk assessments and also publications retrieved from literature search. The risk of the Lactobacillus strains listed above was assessed for the general population. However, in previous assessments of probiotics published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups with mature gastro-intestinal tract. VKM has also assessed the risk of L. plantarum W62, L. plantarum 299v and L. plantarum HEAL9 in food supplements and other foods independent of the dose and have assessed exposure in general terms. VKM concludes that it is unlikely that L. plantarum W62, L. plantarum 299v and L. plantarum HEAL9 would cause adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Streptococcus thermophilus Used as "Other Substances"

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 sold in Norway. 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 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 Streptococcus thermopilus, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of S. thermophilus was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal flora (age group 0-36 months), population with mature gastro-intestinal flora (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of S. thermophilus in food supplements independent of the dose and have assessed exposure in general terms. Other sources of S. thermophilus, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that S. thermophilus causes adverse health effects in the general healthy population with mature gastro-intestinal tract. Acquired resistance genes have been detected in this species and the assessment of susceptibility to antibiotics for each single strain is required. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut flora and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactobacillus helveticus Rosell-52 ND Used as "Other Substances"

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 sold in Norway. 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 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 Lactobacillus helveticus Rosell-52 ND, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. helveticus Rosell-52 ND was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. helveticus Rosell-52 ND in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. helveticus Rosell-52 ND, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. helveticus Rosell-52 ND causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactobacillus delbrueckii subsp. bulgaricus Used as "Other Substances"

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 sold in Norway. 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 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 Lactobacillus delbrueckii subsp. bulgaricus, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. delbrueckii subsp. bulgaricus was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. delbrueckii subsp. bulgaricus in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. delbrueckii subsp. bulgaricus, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. delbrueckii subsp. bulgaricus causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactobacillus casei W56 Used as "Other Substances"

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 sold in Norway. 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 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 Lactobacillus casei W56, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. casei W56 was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. casei W56 in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. casei W56, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. casei W56 causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactococcus lactis W58 Used as "Other Substances"

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 sold in Norway. 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 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 Lactococcus lactis W58, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. lactis W58 was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. lactis W58 in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. lactis W58, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. lactis W58 causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactobacillus salivarius W24 Used as "Other Substances"

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 sold in Norway. 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 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 Lactobacillus salivarius W24, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. salivarius W24 was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. salivarius W24 in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. salivarius W24, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. salivarius W24 causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Lactobacillus paracasei 8700:2 Used as "Other Substances"

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 sold in Norway. 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 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 Lactobacillus paracasei 8700:2, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. paracasei 8700:2 was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal flora (age group 0-36 months), population with mature gastro-intestinal flora (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. paracasei 8700:2 in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. paracasei 8700:2, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. paracasei 8700:2 causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut flora and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Risk Assessment of Bacillus coagulans Used as "Other Substances"

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 sold in Norway. 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 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 Bacillus coagulans, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of B. coagulans was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of B. coagulans in food supplements independent of the dose and have assessed exposure in general terms. Other sources of B. coagulans, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that B. coagulans causes adverse health effects in the general healthy population with mature gastro-intestinal tract. Acquired resistance genes have been detected in this species and the assessment of susceptibility to antibiotics for each single strain is required. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Assessment of Antimicrobial Resistance in the Food Chains in Norway

The Norwegian Food Safety Authority (NFSA) asked the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an assessment of antimicrobial resistance (AMR) in the food chains in Norway, with focus on each of the following food chains: pigs and pork products; poultry, eggs and poultry products; cattle and bovine products; aquaculture and aquaculture products; fresh produce (fruit, berries, and vegetables); and drinking water. AMR in imported food has not been assessed in this report. AMR in Norwegian food chains has been assessed in terms of probability of exposure to humans. Due to data constraints, it has not been possible to assess the consequences of this exposure for human health. VKM appointed a working group consisting of three members of the Panel on Biological Hazards, one member of Panel on Animal Health and Welfare, and four external experts to prepare a draft Opinion document and the answer the questions. The Panel on Biological Hazards has reviewed and revised the draft prepared by the working group and approved the Opinion document «Assessment of antimicrobial resistance in the food chains in Norway”. AMR can be described as the ability of a bacterium to withstand the effects of an antimicrobial. The clinical antimicrobial resistance crisis has focused attention on all uses of antimicrobial agents, including their use in human medicine, veterinary medicine, and in agriculture and aquaculture. AMR is considered the greatest challenge to face health care in 21st century, and there is increasing concern and debate about which roles the food production chains play as reservoirs and disseminators of AMR. This assessment addresses several food chains. The report does not characterise all forms of AMR that may occur in these chains, but puts emphasis on the resistant bacteria and resistance determinants that have emerged at the animal-human interface in recent decades. VKM’s choice is based on zoonotic potential and the limited alternatives available for treatment of infections. In order for a comprehensive and detailed assessment to be conducted, these particular resistance forms need to be characterised and assessed separately. At an overall level, the hazard regarding exposure of humans to antimicrobial resistant bacteria from cattle, milk/milk products, fish/fish products/seafood, fresh produce, water, and food processing in Norway is considered by VKM to be negligible. Current data regarding possible pathways for transmission of LA-MRSA via contaminated food/meat to the broader human population fail to implicate LA-MRSA from pigs as a foodborne pathogen. Compared with other animal products, poultry and poultry products are regarded as the most important reservoirs of ESBL/AmpC-producing Enterobacteriaceae, quinolone-resistant E. coli (QREC), and their corresponding resistance determinants. The probability of human exposure of ESBL/AmpC-producing Enterobacteriaceae and QREC via poultry is assessed as being non-negligible. Probability of AMR Transfer Associated with Food and Uncertainties: In this assessment, the probability of transmission of AMR from food chains to humans has been either categorized as negligible or non-negligible according to the following definitions: Negligible – the probability of transfer of AMR is extremely low. Negligible probability should be considered insignificant. Non-negligible – the probability of transfer of AMR is greater than negligible. Non-negligible probability should be considered significant, but the available data are currently insufficient to enable discrimination between the different levels. Lack of data has made it difficult to reach any firm conclusions regarding the probability of AMR transmission from food to humans in Norway. Similarly, ranking the probabilities with regard to relative importance is largely not possible with the data available. The probability of transfer of AMR from cattle, milk/milk products, fish, seafood, and drinking water has been assessed to be negligible. The probability of transfer of LA-MRSA from live pigs to humans is considered to be non-negligible, while the probability of transfer from pork to humans has been assessed to be negligible. The probability of transfer of ESBL/AmpC-producing Enterobacteriaceae, quinolone-resistant E. coli, and their respective corresponding genes from live poultry and poultry meat is considered as non-negligible. Processing of food, such as cooking or preservation, can reduce the number of bacteria in the products and thus decrease the transmission of antimicrobial resistant bacteria from food to humans. It should be noted that both categories of probabilities (negligible and non-negligible) in this assessment are associated with a number of uncertainties. Bacteria are living organisms that are under continuous evolution, and are able to adapt rapidly to changing living conditions. This report is an assessment of the current situation with regards to development and dissemination of antibiotic resistant bacteria and their resistance genes in the food chain. This situation may change as the bacteria continue to adapt to the selection pressures exerted by the worldwide use of antimicrobials. Such bacterial changes, sometimes occurring VKM Report 2015:29 9 in “quantum leaps” due to horizontal gene transfer (HGT), may also rapidly change the probability of transfer of resistance to specific antimicrobials. Data Gaps: There is a lack of knowledge regarding the vast reservoir of AMR in the environmental, animal, and food reservoirs. Furthermore, there is lack of data regarding the routes and frequencies of transmission of AMR from live, food-producing animals and foodstuffs of different origins to humans and vice versa.

Risk Assessment of Insect-resistant and Herbicide Tolerant Genetically Modified Maize Bt11 for Cultivation, Import, Processing, Food and Feed Uses under Directive 2001/18/EC and Regulation (EC) Opinion

The Norwegian Environment Agency (NEA) and the Norwegian Food Safety Authority (NFSA) requested the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion of potential risks to biodiversity and agriculture in Norway associated with import of seeds for sowing and cultivation of insect-resistant and herbicide tolerant genetically modified maize Bt11 under Directive 2001/18/EC (Notification C/F/96.05.10). The notification is still pending for authorisation in the European Union. VKM is also requested to assess the applicant´s post-market environmental monitoring plan, and the management measures suggested in the draft implementing decision of the European Commission. As the scope of the notification does not cover food and feed uses of maize Bt11, VKM was not asked for a health risk assessment of maize Bt11. However, VKM has decided to update a previous safety evaluation of the food and feed uses of maize Bt11 and derived products (VKM, 2014). VKM appointed a working group consisting of members from the Panel on Genetically Modified Organisms, the Panel on Alien Organisms and trade in Endangered Species (CITES) and the VKM staff to answer the requests. The Panel on Genetically Modified Organisms assessed and approved the final report. The genetically modified maize Bt11 was developed to provide protection against certain lepidopteran target pests, such as the European corn borer (ECB, Ostrinia nubilalis), and some species belonging to the genus Sesamia . The insect resistence is achieved by the expression of a truncated form of a Cry1Ab protein encoded by a modified cry1Ab gene derived from the soil microorganism Bacillus thuringiensis subsp kurstaki HD-1. Maize Bt11 also expresses the phosphinothricin - N - ace tyltransferase (pat) gene, derived from the soil microorganism Streptomyces viridochromogenes strain Tu494, which encodes the enzyme: phosphinothricin acetyl transferase (PAT). PAT protein confers tolerance to the herbicidal active substance glufosinate-ammonium. The PAT protein expressed in Bt11 was used as a selectable marker to facilitate the selection process of transformed plant cells and is not intended for weed management purposes. Since the scope of the notification C/F/96.05.10 does not cover the use of glufosinate-ammonium-containing herbicides on maize Bt11, potential effects due to the use of such herbicides on maize Bt11 are not considered by VKM. In delivering its scientific opinion, VKM considered relevant peer-reviewed scientific publications and information provided by the applicant in the notification C/F/96.05.10, the renewal application EFSA/GMO/RX/Bt11, and scientific opinions and comments from EFSA and other EU-member states. VKM has evaluated maize Bt11 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010a), selection of comparators for the risk assessment of GM plants (EFSA, 2011b) and for the post-market environmental monitoring of GM plants (EFSA, 2011c). The scientific risk assessment of maize Bt11 includes molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity. An evaluation of unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes, the post-market environmental monitoring plan and coexistence measures at the farm level has also been undertaken. It is emphasised that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM. Molecular Characterization: Appropriate analyses of the integration site, inserted DNA sequence, flanking regions, and bioinformatics have been performed. The molecular characterisation reported by the applicant shows that the DNA-fragment containing the cry1Ab and pat genes, is integrated as a single copy at a single locus in the nuclear genome of maize Bt11 and that it is stably inherited as a dominant trait. VKM considers the molecular characterisation of maize Bt11 satisfactory. Comparative Assessment: Comparative analyses of data from field trials located at representative sites and environments in North America and Europe indicates that maize Bt11 is compositionally equivalent to its conventional counterpart, with the exception of the herbicide tolerance and insect resistance traits, conferred by the expression of the PAT and Cry1Ab proteins. However, data on the amino acid tryptophan, is only given in one out of six studies. Based on current knowledge, VKM concludes that maize Bt11 is compositionally equivalent to conventional maize. The data provided by the applicant are not sufficient to show that Bt11 maize is phenotypically and agronomically equivalent to conventional near-isogenic maize lines. The agronomic assessment data are provided from one growing season in the North America and one growing season in France. This is not considered to be sufficient for representative testing of agricultural environments. Food and Feed Risk Assessment: Whole food feeding studies have not indicated any adverse health effects of maize Bt11. These studies further support that maize Bt11 is nutritionally equivalent to conventional maize. The Cry1Ab and PAT proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. Some studies have however indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM concludes that maize Bt11 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry1Ab and PAT proteins will introduce a toxic or allergenic potential in food or feed based on maize Bt11 compared to conventional maize. Environmental Risk Assessment: Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivated maize with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. Since maize Bt11 has no altered agronomic and phenotypic characteristics, except for the specific target insect resistance and herbicide tolerance, the likelihood of unintended environmental effects as a consequence of spread of genes from maize Bt11 is considered to be extremely low. There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. Published scientific studies showed that the likelihood of negative effects of Cry1Ab protein on non-target arthropods that live on or in the vicinity of maize plants is low. In Norway, the maize cultivation is marginal. The total crop area of forage maize is estimated to 2000-2800 decares, equivalent to less than 0.1% of the areas with cereal crops. The area of individual fields is limited by the topography such that the quantity of maize pollen produced under flowering is also limited. The potential exposure of Cry1Ab-containing maize pollen on non-target lepidopteran species in Norway is therefore negligible. Cultivation of maize Bt11 is not considered to represent a threat to the prevalence of red-listed species in Norway. Exposure of nontarget organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Cry proteins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. VKM concludes that, although the data on the fate of the Cry1Ab protein and its potential interactions in soil are limited, the relevant scientific publications analysing the Cry1Ab protein, together with the relatively broad knowledge about the environmental fate of other Cry1 proteins, do not indicate significant direct effects on the soil environment. Despite limited number of studies, most studies conclude that effects on soil microorganisms and microbial communities are transient and minor compared to effects caused by agronomic and environmental factors. However, data are only available from short-term experiments and predictions of potential long-term effects are difficult to deduce. Coexistence: VKM concludes that separation distances of 200 meters most likely will ensure coexistence between genetically modified maize and conventional and organic maize varieties in Norway. Overall Conclusion: Based on current knowledge, VKM concludes that maize Bt11 is nutritionally equivalent t

Risk Assessment of Insect-resistant and Herbicide Tolerant Genetically Modified Maize 1507 for Cultivation, Import, Processing, Food and Feed Uses under Directive 2001/18/EC and Regulation (EC) No 1829/2003

The Norwegian Environment Agency (NEA) and the Norwegian Food Safety Authority (NFSA) requested the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion of potential risks to biodiversity and agriculture in Norway associated with import of seeds for sowing, and cultivation of insect-resistant and herbicide-tolerant genetically modified maize 1507 under Directive 2001/18/EC (Notification C/ES/01/01). The notification is still pending for authorisation in the European Union. VKM is also requested to assess the applicant´s post-market environmental monitoring plan, and the management measures suggested in the draft implementing decision of the European Commission. As VKM delivered a scientific opinion on this application including cultivation in 2014 (VKM, 2014), VKM is asked to assess whether the previous risk assessment is still valid concerning cultivation, and to update the opinion after current knowledge. The assessment shall specifically consider Norwegian conditions. Furthermore, as the notification does not cover food and feed uses of maize 1507, VKM was not asked for a health risk assessment of maize 1507. However, VKM has decided to update the previous safety evaluation of the food and feed uses of maize 1507 and derived products from 2014. VKM appointed a working group consisting of members from the Panel on Genetically Modified Organisms, the Panel on Alien Organisms and trade in Endangered Species (CITES) and the VKM staff to answer the requests. The Panel on Genetically Modified Organisms has assessed and approved the final report. The genetically modified maize 1507 was developed to provide protection against certain lepidopteran target pests, such as the European corn borer (ECB, Ostrinia nubilalis), and some species belonging to the genus Sesamia. The insect resistence is achieved by the expression of a synthetic version of the truncated c ry1F gene derived from Bacillus thuringiensis subsp. aizawai, a common soil bacterium. Maize 1507 also expresses the phosphinothricin - N - acetyltransferase (pat) gene, derived from the soil bacterium Streptomyces viridochromogenes. PAT protein confers tolerance to the herbicidal active substance glufosinate-ammonium. The PAT protein expressed in maize 1507 was used as a selectable marker to facilitate the selection process of transformed plant cells and is not intended for weed management purposes. Since the scope of the notification C/ES/01/01 does not cover the use of glufosinate-ammonium-containing herbicides on maize 1507, potential effects due to the use of such herbicides on maize 1507 are not considered by VKM. In delivering its scientific opinion, VKM considered relevant peer-reviewed scientific publications and information provided by the applicant in the notifications C/ES/01/01, C/NL/00/10, the applications EFSA/GMO/NL/2004/02 and EFSA/GMO/RX/1507, and scientific opinions and comments from EFSA and other EU member states. VKM has evaluated maize 1507 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. VKM has also decided to take into account, the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA, 2011a), the environmental risk assessment of GM plants (EFSA, 2010a), selection of comparators for the risk assessment of GM plants (EFSA, 2011b) and for the post-market environmental monitoring of GM plants (EFSA, 2011c). The scientific opinion of maize 1507 include molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity. An evaluation of unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes, the postmarket environmental monitoring plan and coexistence measures at the farm level has also been undertaken. It is emphasised that VKM’s mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by VKM. Molecular Characterization: Appropriate analyses of the transgenic DNA insert, its integration site, number of inserts and flanking sequences in the maize genome, have been performed. The results show that one copy only of the insert is present in maize 1507. Homology searches with databases of known toxins and allergens have not indicated any potential production of harmful proteins or polypeptides caused by the genetic modification in maize 1507. Southern blot analyses and segregation studies show that the introduced genes cry1F and pat are stably inherited and expressed over several generations along with the phenotypic characteristics of maize 1507. VKM considers the molecular characterisation of maize 1507 satisfactory. Comparative Assessment: Comparative analyses of maize 1507 to its non-GM conventional counterpart have been performed during multiple field trials located at representative sites and environments in Chile (1998/99), USA (1999) and in Europe (1999, 2000 and 2002). With the exception of small intermittent variations, no biologically significant differences were found between maize 1507 and the conventional maize. Based on the assessment of available data, VKM concludes that maize 1507 is compositionally, agronomically and phenotypically equivalent to its conventional counterpart, except for the introduced characteristics, and that its composition fell within the normal ranges of variation observed among non-GM varieties. The field evaluations support a conclusion of no phenotypic changes indicative of increased plant weed/pest potential of 1507 compared to conventional maize. Food and Feed Safety Assessment: Whole food feeding studies on rats, broilers, pullets, pigs and cattle have not indicated any adverse health effects of maize 1507. These studies further indicate that maize 1507 is nutritionally equivalent to conventional maize. The PAT and Cry1F proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. Some studies have however, indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, VKM concludes that maize 1507 is nutritionally equivalent to conventional maize varieties. It is unlikely that the PAT and Cry1F proteins will introduce a toxic or allergenic potential in food or feed based on maize 1507 compared to conventional maize. Environmental Risk Assessment: Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivated maize with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. Since maize 1507 has no altered agronomic and phenotypic characteristics, except for the specific target insect resistance and herbicide tolerance, the likelihood of unintended environmental effects as a consequence of spread of genes from maize 1507 is considered to be extremely low. There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. There are a limited number of published scientific studies on environmental effects of Cry1F protein. Published scientific studies show that the likelihood of negative effects of Cry1F protein on non-target arthropods that live on or in the vicinity of maize plants is low. In Norway, the maize cultivation is marginal. The total crop area of forage maize is estimated to 2000-2800 decares, equivalent to less than 0.1 % of the areas with cereal crops. The area of individual fields is limited by the topography such that the quantity of maize pollen produced under flowering is also limited. The potential exposure of Cry1F-containing maize pollen on non-target lepidopteran species in Norway is therefore negligible. Cultivation of maize 1507 is not considered to represent a threat to the prevalence of red-listed species in Norway. Exposure of non-target organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Cry proteins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. VKM concludes that, although the data on the fate of the Cry1F protein and its potential interactions in soil are limited, the relevant scientific publications analysing the Cry1F protein, together with the relatively broad knowledge about the environmental fate of other Cry1 proteins, do not indicate significant direct effects on the soil environment. Despite limited number of studies, most studies conclude that effects on soil microorganisms and microbial communities are transient and minor c

Assessment of Probiotics in Infant Formula and Cereal Based Baby Foods Containing Bifidobacterium lactis Bb12– Update 2014.

“Assessment of benefits and risks of probiotics in processed cereal-based baby foods supplemented Bifidobacteriumn lactis Bb12” from 2010 answered a request from the Norwegian Food Safety Authority focusing on the age groups 4-6 months, 6-12 months and 1-3 years. However, the use of infant formula intended for newborns, supplemented with this probiotic, was neither asked by the NFSA nor assessed by VKM. The notifier of the baby foods intended for infants and small children has provided information on three different cereal-based products intended for age-groups over 4 months and one infant formula intended for newborns, all supplemented with B. lactis. In its letter the company concludes that their products supplemented with B. lactis do not pose any health and safety risk. Regarding health effect, we have already mentioned in our assessment (Halvorsen et al. 2010) that: “It is not the mandate of this report to evaluate the health claims related to the products as these health claims are assessed by EFSA.” Our main conclusions regarding safety were as follows: “No serious adverse events are reported, but neither has the effect of long-term intake of a single bacterial strain been studied. Furthermore, cereals supplemented with B. lactis Bb12 intended for infants and toddlers have not been studied regarding safety. We are not aware of any in vivo studies explicitly concerning the ability of B. lactis Bb12 to influence gene expression of epithelial cells”. Furthermore, we were concerned regarding presence of antibiotic resistance gene against tetracycline (tetW) in the B. lactis Bb12. In the answer to the question from NFSA regarding antibiotic resistance gene in L. lactis Bb12, we concluded that: “Consumption of probiotic microororganism B. lactis Bb12 that harbour gene encoding resistance against tetracycline (tetW) may increase the risk of the transfer of such genes to the resident microbiota and pathogenic bacteria and hence increase development of bacterial resistance. High similarity has been observed between tetW gene in bacteria of human and environment origin and B. lactis Bb12. This suggests the spread of tetracycline resistance gene (tetW) between bacteria of various origins. However, the transfer of tetracycline resistance gene (tetW) to other bacteria as a consequence of consumption of Bb12 has not been studied.” As we have already mentioned in our assessment (Halvorsen et al. 2010). “It is important to note that the infant’s diet comprises a restricted variety of foods, which often are taken several times a day during a period of life when a stable intestinal flora is not yet established. The establishment of a normal intestinal microbiota takes at least two years and thus the intake of large numbers of probiotic bacteria in monoculture during the first years of life may greatly influence this process.” According to the “Guideline for evaluation of probiotics in food” (FAO/WHO 2002): ‘‘….the onus is on the producer to prove that any given probiotic strain is not a significant risk with regard to transferable antibiotic resistance or other opportunistic virulence properties.” The tet(W) gene in Bifidobacterium seems to be integrated in the chromosome and its surrounding regions vary depending on the strain, but very often the gene is flanked by transposase target sequences or genes coding for transposase, an enzyme that catalyzes the movement of DNA fragments between different locations by recognizing specific target sequences, suggesting that, under adequate conditions, the gene could be transferred (Gueimonde et al. 2013). The presence of a tetracycline resistance gene, tet(W), flanked by a putative transposase gene in B. animalis subsp. lactis was also confirmed in other strains of Bifidobacterium than Bb12 (Stahl & Barrangou 2012). Among the data provided by the notifier, we could not identify any new studies regarding the above mentioned concerns. As already mentioned, our assessment from 2010 did not include probiotic-supplemented infant formula intended for use by newborns. It seems likely that the same concerns as for the cerealbased products will be valid in this age group and possibly of even greater importance. Among the literature provided by the notifier was also the position paper from 2011 of the ESPGHAN Committee on Nutrition (ESPGHAN 2011). Among their general conclusions are: • (Conclusion 1): “For healthy infants, the available scientific data suggest that the administration of currently evaluated probiotic-supplemented formula to healthy infants does not raise safety concerns with regard to growth and adverse effects”. But none the less: • (Conclusion 5): “In general, there is a lack of data on the long-term effects of the administration of formula supplemented with probiotics. Such data would be of particular importance if the effects persisted after the administration of the probiotics has ceased.” And concludes lastly; • (Conclusion 6): “Considering the above, the Committee does not recommend the routine use of probiotic-supplemented formula in infants.” Our view is in accordance with these conclusions.