Prevention and amelioration of lead accumulation in a human

Abstract

The present invention relates to a Lactiplantibacillus strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human.

Description

[0001] Ref 16302-WO-PCT

[0002] PREVENTION AND AMELIORATION OF LEAD ACCUMULATION IN A HUMAN

[0003] Reference to a deposit of biological material

[0004] This application contains a reference to a deposit of biological material, which deposit is incorporated herein by reference. For complete information see last paragraph of the description.

[0005] FIELD OF THE INVENTION

[0006] The present invention relates to a Lactiplantibacillus strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human.

[0007] BACKGROUND OF THE INVENTION

[0008] Heavy metals like e.g. lead, cadmium, and arsenic pose significant health risks by accumulating in the body and affecting nearly all human organs. Lead exposure is particularly detrimental to children, causing irreversible damage. Despite improvements, heavy metal poisoning remains prevalent in some countries.

[0009] Lead is a neurotoxic pollutant that can cause serious damage to the kidney, liver, central nervous, and hematologic systems. The Centers for Disease Control and Prevention (US CDC) and the World Health Organization (WHO) both report that even low doses of lead exposure are harmful, and WHO notes that any exposure can hinder children's cognitive development and intelligence quotient (IQ). Up to 80% of daily lead intake in some developing countries comes from food.

[0010] Global lead exposure is especially concerning in low- and middle-income countries. Cognitive deficits in children are among the most substantiated effects of lead exposure. In China, despite declining blood lead levels in children since 2000, the average blood lead levels remain higher than in the USA and Europe. Even at blood lead levels below 5 pg / day, children under the age of 5 are most vulnerable to adverse effects from lead, including behavioral and learning problems, lower IQ, hyperactivity, hearing problems, and impaired growth. Iron, zinc, calcium and vitamin C level can influence the gastrointestinal absorption and distribution of lead (Zhai Q. et al., J Agric Food Chem. Apr 10 2019; 67(14): 3956-3965; Dawson E.B. et al., J Am Coll Nutr. Apr 1999; 18(2): 166-70). Also, diets low in calcium have been reported to enhance lead accumulation (Goyer R.A., Am J Clin Nutr. Mar 1995; 61 (3 Suppl): 646S-650S) while iron-deficient children have higher gastrointestinal absorption of lead (Barton J.C. et al., J Lab Clin Med. Oct 1978; 92(4): 536-47)).

[0011] Probiotics have shown effect against the uptake of heavy metals in the human body. Lactiplantibacillus plantarum strain CCFM8661 has been shown to bind cadmium (Cd) and affect it’s uptake in the intestine, kidney and liver while higher levels of cadmium in feces was observed (Zhai et al. (July 2016, ASM Journals, Appl. Environ. Microbiol., Vol. 82(14), p. 4429-4440). Cd exposure has been shown to cause alterations of calcium (Ca), zinc (Zn), iron (Fe), and magnesium (Mg) Ref 16302-WO-PCT levels in liver and kidney tissues. Most of such alterations were reversed by Lactiplantibacillus plantarum strain CCFM8661 while Lactiplantibacillus plantarum CCFM8661 treatment by itself did not result in significant differences in the levels of these essential elements from those in the control groups (Zhai et al, 2014, ASM Journals, Appl. Environ. Microbiol., July 2014, Vol 80(13), p. 4063-4071).

[0012] Studies have shown mixed results with probiotics against lead uptake in the body. One study found no improvement in blood lead levels (BLLs) with Lactobacillus rhamnosus GR-1 , while another reported benefits with Lactiplantibacillus plantarum CCFM8661 in lead-exposed adults (Bisanz JE, et al. mBio, Oct 072014;5(5):e01580-14. doi:10.1128 / mBio.01580-14; Chen F. et al., Int J Biol Macromol. Mar 2024;261 (Pt 2): 129815).

[0013] There is still a need for alternative methods for preventing or ameliorating lead accumulation in a human.

[0014] SUMMARY OF THE INVENTION

[0015] The invention provides a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use in preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0016] The strain or a dietary supplement comprising such may be for use in preventing or ameliorating lead accumulation in the body of a child.

[0017] The strain or a dietary supplement comprising such may upon administration to a human result in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, prevention of lead absorption as measured in the urine lead level, increase in excretion of lead from the body, decrease in the lead level in other organs or body parts, reduction of the gastro-intestinal discomfort, reduction in abdominal pain, and reduction in behavioral discomfort.

[0018] Also described herein is a method of administering a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0019] Further described herein is a dietary supplement composition comprising a Lactiplantibacillus plantarum strain and a multivitamin composition, wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition. Ref 16302-WO-PCT

[0020] DEFINITIONS

[0021] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. For the sake of brevity and / or clarity, well-known functions or constructions may not be described in detail.

[0022] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0023] Throughout this disclosure, unless the context requires otherwise, the words "comprise," "comprises," and "comprising" will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

[0024] The term "consisting of" means including, and limited to, whatever follows the phrase "consisting of." Thus, the phrase "consisting of' indicates that the listed elements are required or mandatory, and that no other elements may be present. The term "consisting essentially of" means including any elements listed after the phrase and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they materially affect the activity or action of the listed elements.

[0025] The terms “administer”, “administration” or “administering” is used to mean the action of providing or applying substance such as e.g. a dietary supplement to a human, i.e. the action used by a human to achieve an intake of a substance. Administer is herein used interchangeably with ingest, consume, provide, apply, absorb, drink, eat, swallow, take into the body, give, and dispense. Administration is herein used interchangeably with ingestion, consumption, provision, application, absorption, drinking, eating, swallowing, taking into the body, giving, and dispension. Administering is herein used interchangeably with ingesting, consuming, providing, applying, absorbing, drinking, eating, swallowing, taking into the body, giving, and dispensing.

[0026] The term “dissolve” is used to describe the process by which a solute is incorporated into a solvent, resulting in a homogeneous mixture or a dispersed phase where the solute is uniformly distributed throughout the dispersing medium without visible lumps.

[0027] As used herein, “essential minerals” are nutrients that are essential for maintaining a healthy body. They are inorganic molecules that play a multitude of functional roles in human cells both Ref 16302-WO-PCT physiologically and biochemically and are known to the person skilled in the art. Non-limiting examples of “essential minerals” include, but is not limited to, sodium, potassium, phosphorus, magnesium and calcium.

[0028] As used herein, “heavy metal” refers to a metallic chemical element that has a relatively high density and is toxic or poisonous at low concentrations and include without limitations lead, cadmium, arsenic and mercury.

[0029] The term “improve lead excretion in fecal samples” is used to describe increase of lead in fecal samples from a human, e.g. after treatment with a lead intervention agent such as Lactiplanti- bacillus plantarum compared to fecal samples from a human before the treatment. Alternative terms for “lead excretion” include, but are not limited to, lead detoxification, lead detoxifying, decreasing lead, lead depollution, and lead decontamination.

[0030] The term “Lactiplantibacillus plantarum" is herein used for species previously named Lactobacillus plantarum and is thus the same species.

[0031] As used herein, “lead binding product” refers to a product that binds to lead ions e.g. in the gastrointestinal (Gl) tract of the human body. Lead binding in the Gl tract may e.g. be measured in vivo as the reduction of lead in a blood sample obtained from a person after consumption of a lead binding product compared to a blood sample from the same person without consumption of the lead binding product, or by measuring lead ions excreted in the human faeces of a person before and after receiving the lead binding product.

[0032] As used herein, the term ’’multivitamin composition” is used for any composition containing vitamins and / or minerals. Included is e.g. dietary compositions, sachets, capsules, tablets, lozenges, chewable tablets, effervescent tablets, oils, liquids, syrups, gummies, gel capsules, jelly pops, powders, chewing gums, multivitamin injections, food, food enriched with vitamins, medical type foods, foods for special medical purposes, foods naturally rich in vitamins, infant formula, and topical patches.

[0033] The term “preventing or ameliorating lead accumulation” is herein used for the support of the natural function of the gut microbiota by preventing or ameliorating lead absorption in the body and thereby preventing or ameliorating lead poisoning, also called lead accumulation. Alternative terms for lead accumulation include, but are not limited to, lead poisoning, build-up of lead, lead build-up in the body, increasing lead, lead intoxication, saturnism, and lead pollution.

[0034] While certain aspects of the present disclosure will hereinafter be described with reference to embodiments thereof, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the claims. Ref 16302-WO-PCT

[0035] DETAILED DESCRIPTION OF THE INVENTION

[0036] The present invention relates to a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human. The inventors of the present invention have surprisingly found that said Lactiplantibacillus plantarum strain decreases the lead binding in a human when the strain is administered to said human less than 12 hours before or after intake of a multivitamin composition.

[0037] In one aspect, the Lactiplantibacillus plantarum or the dietary supplement comprising such prevents intestinal absorption of lead and thereby prevents lead induced tissue damage when administered less than 12 hours before or after intake of a multivitamin composition. The prevention of intestinal absorption of lead may e.g. be measured by measuring the lead excretion in fecal samples. Thus, in one aspect the effect of administering the Lactiplantibacillus plantarum or the dietary supplement comprising such results upon administration in increase in excretion of lead from the body such as increase in excretion of lead via the intestinal tract and / or excretion of lead via the feces.

[0038] In one aspect, the Lactiplantibacillus plantarum or the dietary supplement comprising such is for use in supporting the natural function of the gut microbiota to prevent lead absorption in the body. In another or further aspect, the Lactiplantibacillus plantarum or the dietary supplement comprising such is for use in improving lead excretion in fecal samples.

[0039] In one aspect, the human is a child, such as a child that is below 18 years old, e.g. between 0 to 16 years old, between 1 to 14 years old or between 3 to 12 years old. In another or further aspect, the human is an infant that is below 1 year old. In yet another aspect the human is a prenatal infant, where the Lactiplantibacillus plantarum strain and the multivitamin composition are administered to the prenatal infant via the mother.

[0040] The multivitamin composition to be taken less than 12 hours before or after administration of the Lactiplantibacillus plantarum or the dietary supplement comprising such may be any composition comprising one or more vitamins and / or one or more minerals such as e.g. zinc, iron and / or calcium. In one aspect, the multivitamin composition is selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a chewing gum, a food, a food enriched with vitamins, a food naturally rich in vitamins, and infant formula.

[0041] The Lactiplantibacillus plantarum strain for use or administration herein may in one aspect be Lactiplantibacillus plantarum strain deposited as DSM 33464.

[0042] In one aspect, the Lactiplantibacillus plantarum or the dietary supplement comprising such results upon administration in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, decrease in the lead level in the gut luminal content, prevention of lead absorption as measured in the urine lead level, increase in excretion of lead from the body such as increase in excretion of lead via the intestinal tract and / or Ref 16302-WO-PCT excretion of lead via the feces, decrease in the lead level in other organs such as the liver, kidney end / or brain, decrease in the lead level in other body parts such as the hair and / or nails, reduction of the gastro-intestinal discomfort such as abdominal pain, reflux, indigestion, diarrhea and / or constipation, and reduction in behavioral discomfort such as psychosomatic feeling, learning difficulties, conduct problems, anxiety, impulsive behavior and / or hyperactive behavior.

[0043] In one aspect, the Lactiplantibacillus plantarum or the dietary supplement comprising such prevents or ameliorates lead accumulation in a human by removing lead from the gut luminal content. The removal of lead from the gut luminal content may be obtained via one or more mechanisms including, but not limited to, sequestration of lead inside the Lactiplantibacillus plantarum bacterial cells and / or binding of lead to the cells wall.

[0044] Administration

[0045] The Lactiplantibacillus plantarum strain or dietary supplement comprising such for use or administration herein should not be mixed directly with the multivitamin composition at consumption but may e.g. be administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition such as a layered, e.g. a multilayered composition.

[0046] In one aspect, the Lactiplantibacillus plantarum strain or dietary supplement comprising such is provided as a powder and is dissolved before administration. The time for dissolving the strain or dietary supplement before administration may take any time such as up to 15 minutes, 10 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute or may be instantaneous.

[0047] In one aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 12 hours before or after intake of a multivitamin composition. In one aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 12 hours and at least 1 hour before or after intake of a multivitamin composition. In another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 12 hours and at least 2 hours, 3, hours, 4 hours, 5, hours, 6 hours, 7 hours or 8 hours before or after intake of a multivitamin composition. In yet another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 10 hours and at least 1 hour, 2 hours, 3, hours, 4 hours, 5, hours, 6 hours, 7 hours or 8 hours before or after intake of a multivitamin composition. In still another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 8 hours and at least 1 hour, 2 hours, 3, hours, 4 hours, 5, hours, 6 hours or 7 hours before or after intake of a multivitamin composition. In yet another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human less than 6 hours and at least 1 hour, 2 hours, 3, hours, 4 hours or 5, hours before or after intake of a multivitamin composition. Ref 16302-WO-PCT

[0048] In one aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 1 hour and less than 10 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours or 2 hours before or after intake of a multivitamin composition. In another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 2 hours and less than 12 hours, 10 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours or 3 hours before or after intake of a multivitamin composition. In yet another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 3 hours and less than 12 hours, 10 hours, 8 hours, 7 hours, 6 hours, 5 hours or 4 hours before or after intake of a multivitamin composition. In still another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 4 hours and less than 12 hours, 10 hours, 8 hours, 7 hours, 6 hours or 5 hours before or after intake of a multivitamin composition. In yet another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 6 hours and less than 12 hours, 10 hours, 8 hours or 7 hours before or after intake of a multivitamin composition. In yet another aspect, the Lactiplantibacillus plantarum strain or a supplement comprising such is administered to a human at least 8 hours and less than 12 hours or 10 hours before or after intake of a multivitamin composition.

[0049] In one aspect, the Lactiplantibacillus plantarum strain for use or administration is deposited as DSM 33464.

[0050] Composition

[0051] In one aspect, the dietary supplement composition comprising the Lactiplantibacillus plantarum strain is in a form, wherein the Lactiplantibacillus plantarum strain is present in a first set of microcapsules and the multivitamin composition is present in a second set of microcapsules.

[0052] The dietary supplement composition may additionally or alternatively be in a form wherein the Lactiplantibacillus plantarum strain and the multivitamin composition are present in a layered composition, where the Lactiplantibacillus plantarum strain is present in one layer and the multivitamin composition is present in a separate layer, the Lactiplantibacillus plantarum strain is present in the core of the composition and the multivitamin composition is present in a separate layer around the core, or the multivitamin composition is present in the core of the composition and the Lactiplantibacillus plantarum strain is present a separate layer around the core.

[0053] An example of a layered composition is e.g., where the Lactiplantibacillus plantarum strain is coated as a layer around a pre-formed inert core particle to form a first layer and the multivitamin composition is coated as a layer around the Lactiplantibacillus plantarum strain to form a second layer. Another example is e.g. where the multivitamin composition is coated as a layer around a pre-formed inert core particle to form a first layer and the Lactiplantibacillus plantarum strain is coated as a layer around the multivitamin composition to form a second layer. Ref 16302-WO-PCT

[0054] In one aspect, the layer or layers are in the form of substantially continuous layer(s). A substantially continuous layer is herein to be understood as a coating having few or none holes, so that the core unit it is encapsulating has few or none uncoated areas. The layer or coating should preferably be homogenous in thickness.

[0055] Each layer in a layered composition may e.g. be applied by adding an atomized solution to the core respectively the first layer. This is typically performed in a fluid bed apparatus wherein the pre-formed core particles are fluidized, and the layer solution adheres to the core particles respectively the first layer and dries up to leave a layer of dry active compound on the surface of the core particle / first layer. The layered composition may comprise further ingredients e.g. in one, two or more additional coating layers on the inside or outside surface of the Lactiplantibacillus plantarum strain layer and / or the multivitamin composition layer.

[0056] In one aspect, the multivitamin composition is further divided into two layers where one layer comprises the vitamins of the multivitamin composition and another layer comprises the minerals of the multivitamin composition. In another aspect, the multivitamin composition is further divided into two layers where one layer comprises the hydrophilic vitamins and minerals of the multivitamin composition and another layer comprises the hydrophobic vitamins and minerals of the multivitamin composition.

[0057] The dietary supplement composition may e.g. be solid or liquid. When the dietary supplement composition is a solid it may e.g. be selected from the group consisting of capsules, tablets, enteric-coated tablets, chewing gum, sachets, a lozenge, a chewable tablet, an effervescent tablet, a powder and a chewing gum.

[0058] When the dietary supplement composition is a liquid, it may be formed from a powder before administration, such as e.g. 1 , 2, 3, 4, 5, 10 or 15 minutes before administration. In one aspect, the liquid is formed by dissolving the powder in water or another drink such as juice, syrup, shake, tea e.g. cold tea, milk, plant drink, plant milk, smoothie or the like.

[0059] PREFERRED EMBODIMENTS

[0060] The invention is also described by the following numbered embodiments:

[0061] 1. A Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use in preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0062] 2. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to embodiment 1 , wherein the human is a child. Ref 16302-WO-PCT

[0063] 3. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the human is a child that is below 18 years old, between 0 to 16 years old, between 1 to 14 years old or between 3 to 12 years old.

[0064] 4. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the human is an infant that is below 1 year old, or a prenatal infant, where when the human is a prenatal infant the Lactiplantibacillus plantarum strain and the multivitamin composition are administered to the prenatal infant via the mother.

[0065] 5. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the multivitamin composition is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch.

[0066] 6. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the Lactiplantibacillus plantarum strain is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch.

[0067] 7. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the Lactiplantibacillus plantarum strain or dietary supplement comprising such is provided as a powder and is dissolved before administration.

[0068] 8. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the multivitamin composition comprises at least one or more minerals and / or vitamins selected from the group consisting of: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin B12, Niacin (Vitamin B3), Folic acid (Vitamin B9), Pantothenic acid (Vitamin B5), Biotin (Vitamin B7), Vitamin A, Vitamin D, Zinc, Iron and Calcium.

[0069] 9. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the multivitamin composition comprises calcium, zinc and / or iron.

[0070] 10. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the multivitamin composition further comprises one or more minerals and / or vitamins selected from the group consisting of Ref 16302-WO-PCT vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, vitamin K1 , vitamin K2, vitamin K3, vitamin B, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B6 (pyridoxine), vitamin B12 (cyanocobalamin), vitamin B5 (Pantothenic acid), vitamin B7 (Biotin), vitamin B9 (Folate or folic acid), magnesium and iodine..

[0071] 11. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the Lactiplantibacillus plantarum strain is deposited as DSM 33464.

[0072] 12. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the strain is administered less than 10 hours, such as less than 8 hours, 7 hours, 6 hours or 5 hours before or after intake of the multivitamin composition.

[0073] 13. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the strain is administered at least

[0074] 1 hour before or after intake of the multivitamin composition.

[0075] 14. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the strain is administered at least

[0076] 2 hours, such as at least 3 hours, 4 hours, 5 hours, 6 hours, 7 hours or 8 hours before or after intake of the multivitamin composition.

[0077] 15. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the strain is administered simultaneously with the intake of the multivitamin composition.

[0078] 16. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the strain is administered together with a meal.

[0079] 17. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the composition is administered at least once per day, up to 5 times per day.

[0080] 18. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, wherein the dietary supplement is given with a meal.

[0081] 19. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding embodiments, which upon administration to a human results in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, prevention of lead absorption as measured in the urine lead level, increase in excretion of lead from the body, decrease in the lead level in other organs or body parts, reduction of the gastro-intestinal discomfort, reduction in abdominal pain, and reduction in behavioral discomfort. Ref 16302-WO-PCT

[0082] 20. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to embodiment 19, which decreases the lead level in the body and / or decreases the blood lead level upon administration compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such.

[0083] 21. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to embodiment 19, which decreases the lead level in the body and / or decreases the blood lead level upon administration compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such, wherein the lead level is measured after at least 2 weeks of administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such.

[0084] 22. A method of administering a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0085] 23. The method according to embodiment 22, wherein the human is a child.

[0086] 24. The method according to any one of embodiments 22 to 23, wherein the human is a child that is below 18 years old, between 0 to 16 years old, between 1 to 14 years old or between 3 to 12 years old.

[0087] 25. The method according to any one of embodiments 22 to 24, wherein the human is an infant that is below 1 year old, or a prenatal infant, where when the human is a prenatal infant the Lactiplantibacillus plantarum strain and the multivitamin composition are administered to the prenatal infant via the mother.

[0088] 26. The method according to any one of embodiments 22 to 25, wherein the multivitamin composition is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch.

[0089] 27. The method according to any one of embodiments 22 to 26, wherein the Lactiplantibacillus plantarum strain is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch. Ref 16302-WO-PCT

[0090] 28. The method according to any one of embodiments 22 to 27, wherein the Lactiplantibacil- lus plantarum strain or dietary supplement comprising such is provided as a powder and is dissolved before administration.

[0091] 29. The method according to any one of embodiments 22 to 38, wherein the multivitamin composition comprises at least one or more minerals and / or vitamins selected from the group consisting of: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin B12, Niacin (Vitamin B3), Folic acid (Vitamin B9), Pantothenic acid (Vitamin B5), Biotin (Vitamin B7), Vitamin A, Vitamin D, Zinc, Iron and Calcium.

[0092] 30. The method according to any one of embodiments 22 to 29, wherein the multivitamin composition comprises calcium, zinc and / or iron.

[0093] 31. The method according to any one of embodiments 22 to 30, wherein the multivitamin composition further comprises one or more minerals and / or vitamins selected from the group consisting of vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, vitamin K1, vitamin K2, vitamin K3, vitamin B, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B6 (pyridoxine), vitamin B12 (cyanocobalamin), vitamin B5 (Pantothenic acid), vitamin B7 (Biotin), vitamin B9 (Folate or folic acid), magnesium and iodine..

[0094] 32. The method according to any one of embodiments 22 to 31 , wherein the Lactiplantibacil- lus plantarum strain is deposited as DSM 33464.

[0095] 33. The method according to any one of embodiments 22 to 32, wherein the strain is administered less than 10 hours, such as less than 8 hours, 7 hours, 6 hours or 5 hours before or after intake of the multivitamin composition.

[0096] 34. The method according to any one of embodiments 22 to 33, wherein the strain is administered at least 1 hour before or after intake of the multivitamin composition.

[0097] 35. The method according to any one of embodiments 22 to 34, wherein the strain is administered at least 2 hours, such as at least 3 hours, 4 hours, 5 hours, 6 hours, 7 hours or 8 hours before or after intake of the multivitamin composition.

[0098] 36. The method according to any one of embodiments 22 to 35, wherein the strain is administered simultaneously with the intake of the multivitamin composition.

[0099] 37. The method according to any one of embodiments 22 to 36, wherein the strain is administered together with a meal.

[0100] 38. The method according to any one of embodiments 22 to 37, wherein the composition is administered at least once per day, up to 5 times per day.

[0101] 39. The method according to any one of embodiments 22 to 38, wherein the dietary supplement is given with a meal.

[0102] 40. The method according to any one of embodiments 22 to 39, which upon administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such to a human results in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, prevention of lead absorption as measured Ref 16302-WO-PCT in the urine lead level, increase in excretion of lead from the body, decrease in the lead level in other organs or body parts, reduction of the gastro-intestinal discomfort, reduction in abdominal pain, and reduction in behavioral discomfort.

[0103] 41. The method according to embodiment 40, which decreases the lead level in the body and / or decreases the blood lead level upon administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such.

[0104] 42. The method according to embodiment 40, which decreases the lead level in the body and / or decreases the blood lead level upon administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such, wherein the lead level is measured after at least 2 weeks of administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such.

[0105] 43. Use of a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0106] 44. The use according to embodiment 43, wherein the human is a child.

[0107] 45. The use according to any one of embodiments 43 to 44, wherein the human is a child that is below 18 years old, between 0 to 16 years old, between 1 to 14 years old or between 3 to 12 years old.

[0108] 46. The use according to any one of embodiments 43 to 45, wherein the human is an infant that is below 1 year old, or a prenatal infant, where when the human is a prenatal infant the Lactiplantibacillus plantarum strain and the multivitamin composition are administered to the prenatal infant via the mother.

[0109] 47. The use according to any one of embodiments 43 to 46, wherein the multivitamin composition is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch.

[0110] 48. The use according to any one of embodiments 43 to 47, wherein the Lactiplantibacillus plantarum strain is provided in one or more formulations selected from the group consisting of: a dietary composition, a sachet, a capsule, a tablet, a lozenge, a chewable tablet, an effervescent tablet, an oil, a liquid, a syrup, gummy, a gel capsule, a jelly pop, a powder, a chewing gum, a Ref 16302-WO-PCT multivitamin injection, a food, a food enriched with vitamins, a medical type food, a food for special medical purposes, a food naturally rich in vitamins, infant formula, and a topical patch.

[0111] 49. The use according to any one of embodiments 43 to 48, wherein the Lactiplantibacillus plantarum strain or dietary supplement comprising such is provided as a powder and is dissolved before administration.

[0112] 50. The use according to any one of embodiments 43 to 49, wherein the multivitamin composition comprises at least one or more minerals and / or vitamins selected from the group consisting of: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin B12, Niacin (Vitamin B3), Folic acid (Vitamin B9), Pantothenic acid (Vitamin B5), Biotin (Vitamin B7), Vitamin A, Vitamin D, Zinc, Iron and Calcium.

[0113] 51. The use according to any one of embodiments 43 to 50, wherein the multivitamin composition comprises calcium, zinc and / or iron.

[0114] 52. The use according to any one of embodiments 43 to 51 , wherein the multivitamin composition further comprises one or more minerals and / or vitamins selected from the group consisting of vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, vitamin K1 , vitamin K2, vitamin K3, vitamin B, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B6 (pyridoxine), vitamin B12 (cyanocobalamin), vitamin B5 (Pantothenic acid), vitamin B7 (Biotin), vitamin B9 (Folate or folic acid), magnesium and iodine..

[0115] 53. The use according to any one of embodiments 43 to 52, wherein the Lactiplantibacillus plantarum strain is deposited as DSM 33464.

[0116] 54. The use according to any one of embodiments 43 to 53, wherein the strain is administered less than 10 hours, such as less than 8 hours, 7 hours, 6 hours or 5 hours before or after intake of the multivitamin composition.

[0117] 55. The use according to any one of embodiments 43 to 54, wherein the strain is administered at least 1 hour before or after intake of the multivitamin composition.

[0118] 56. The use according to any one of embodiments 43 to 55, wherein the strain is administered at least 2 hours, such as at least 3 hours, 4 hours, 5 hours, 6 hours, 7 hours or 8 hours before or after intake of the multivitamin composition.

[0119] 57. The use according to any one of embodiments 43 to 56, wherein the strain is administered simultaneously with the intake of the multivitamin composition.

[0120] 58. The use according to any one of embodiments 43 to 57, wherein the strain is administered together with a meal.

[0121] 59. The use according to any one of embodiments 43 to 58, wherein the composition is administered at least once per day, up to 5 times per day.

[0122] 60. The use according to any one of embodiments 43 to 59, wherein the dietary supplement is given with a meal.

[0123] 61 . The use according to any one of embodiments 43 to 60, which upon administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such to a human Ref 16302-WO-PCT results in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, prevention of lead absorption as measured in the urine lead level, increase in excretion of lead from the body, decrease in the lead level in other organs or body parts, reduction of the gastro-intestinal discomfort, reduction in abdominal pain, and reduction in behavioral discomfort.

[0124] 62. The use according to embodiment 61 , which decreases the lead level in the body and / or decreases the blood lead level upon administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such.

[0125] 63. The use according to embodiment 61 , which decreases the lead level in the body and / or decreases the blood lead level upon administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or the dietary supplement comprising such, wherein the lead level is measured after at least 2 weeks of administration of the Lactiplantibacillus plantarum strain.

[0126] 64. A dietary supplement composition comprising a Lactiplantibacillus plantarum strain and a multivitamin composition, wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

[0127] 65. The dietary supplement composition according to embodiment 64, wherein the Lactiplantibacillus plantarum strain is deposited as DSM 33464.

[0128] 66. The dietary supplement composition according to any one of embodiments 64 to 65, wherein the Lactiplantibacillus plantarum strain is present in a first microcapsule and the multivitamin composition is present in a second microcapsule.

[0129] 67. The dietary supplement composition according to any one of embodiments 64 to 66, wherein the Lactiplantibacillus plantarum strain and the multivitamin composition are present in a layered composition, where the Lactiplantibacillus plantarum strain is present in one layer and the multivitamin composition is present in a separate layer, the Lactiplantibacillus plantarum strain is present in the core of the composition and the multivitamin composition is present in a separate layer around the core, or the multivitamin composition is present in the core of the composition and the Lactiplantibacillus plantarum strain is present a separate layer around the core, where the multivitamin composition is optionally further divided into two layers.

[0130] 68. The dietary supplement composition according to embodiment 67, where the multivitamin composition is further divided into two layers, wherein one layer comprises the vitamins of the multivitamin composition and another layer comprises the minerals of the multivitamin composition, or wherein one layer comprises the hydrophilic vitamins and minerals of the multivitamin composition and another layer comprises the hydrophobic vitamins and minerals of the multivitamin composition. Ref 16302-WO-PCT

[0131] 69. The dietary supplement composition according to any one of embodiments 64 to 68, where the composition is solid or liquid.

[0132] 70. The dietary supplement composition according to any one of embodiments 64 to 69, where the dietary supplement composition is solid.

[0133] 71. The dietary supplement composition according to embodiment 70, where the dietary supplement composition is selected from the group consisting of capsules, tablets, enteric- coated tablets, chewing gum, sachets, a lozenge, a chewable tablet, an effervescent tablet, a powder and a chewing gum.

[0134] 72. The dietary supplement composition according to any one of embodiments 64 to 71 , where the dietary supplement composition is a liquid which is formed from a powder before administration.

[0135] 73. The dietary supplement composition according to any one of embodiments 64 to 72 for use in a method of any one of embodiments 22 to 42.

[0136] 74. Use of the dietary supplement composition according to any one of embodiments 64 to 72 in a method of any one of embodiments 22 to 42.

[0137] EXAMPLES

[0138] Deposit of Biological Material

[0139] The following biological material has been deposited under the terms of the Budapest Treaty with the Leibniz-lnstitut DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ), Inhoffenstr. 7 B, D-38124 Braunschweig, Germany, and given the following accession number:

[0140] Deposit Accession Number Date of Deposit Country of origin

[0141] SmartGuard DSM 33464 05 March 2020 China

[0142] The strain has been deposited under conditions that assure that access to the culture will be available during the pendency of this patent application to one determined by foreign patent laws to be entitled thereto. The deposit represents a substantially pure culture of the deposited strain. The deposit is available as required by foreign patent laws in countries wherein counterparts of the subject application, or its progeny are filed. However, it should be understood that the availability of a deposit does not constitute a license to practice the subject invention in derogation of patent rights granted by governmental action. Ref 16302-WO-PCT

[0143] Example 1 - Survival of L. plantarum DSM 33464 in simulated gastrointestinal conditions in the absence or presence of multivitamin & mineral

[0144] The survival of L. plantarum DSM 33464 was assessed in an in vitro model simulating the oral, gastric and small intestinal conditions encountered in the human gastrointestinal tract, as described by Minekus et al (Minekus et al, 2014, Food Fund, 2014, 5, 1113-1124). These experiments were conducted in a simulated “fasted” or “fed” state in the presence or absence of 100 mg multivitamin and mineral supplement diluted in 0.75 mL of simulated salivary fluid. The composition of the multivitamin and mineral supplement had the respective amounts of vitamins and minerals as follows: Vitamin B1 1.5 mg; vitamin B2 0.8 mg; vitamin B3 5 mg; vitamin B5 1 mg; vitamin B6 0.5 mg; vitamin B7 10 g; vitamin B9 80 pg; vitamin B12 1 pg; vitamin A 600 III; vitamin D 200 IU; iron pyrophosphate 5 mg; calcium 180 mg; zinc gluconate 4 mg; maltodextrin 1.4 g. In brief, freeze-dried L. plantarum DSM 33464 strain powder (100 mg) was incubated first for 10 min in 0.75 mL of a simulated salivary fluid as described by Minekus et al. Then this preparation was further incubated for 110 min in a simulated gastric intestinal fluid (as described in Minekus et al). Finally, the preparation was incubated in a small intestinal fluid (as described in Minekus et al) for 120 min. All experiments were carried out at 37°C.

[0145] The viability of the preparation (Mean log 10 CFU=10.72) was assessed in the absence of any additional ingredients (“fasted” state), or in the presence of 1 :1 milk containing 3,8% fat (“fed” state). The viability of the cells was evaluated by plate counts on MRS agar (37C, 48h, anaerobic incubation) at time T=0h, 10 min (oral phase), 120 min +(gastric phase), 240 min (+small intestinal phase). Log 10 loss of the bacterial cells was calculated as referred to TO.

[0146] Results:

[0147] Results indicated that the viability of the strain was well maintained for the strain in the oral and gastric phase, with a maximum of a -0.5 log after 120 min co-incubation in all tested conditions (Table 1). In fed-state, without vitamin and mineral additions, log 10 loss were small (-1.60) after 240 min co-incubation, after small intestinal conditions. In fasted state, log 10 loss were higher in fasted state compared to fed state after 240 min co-incubation after small intestinal conditions compared, reaching -2.61 log 10 loss. In the presence of multivitamin and mineral, loss after 240 min of co-incubation were also the highest in fasted state compared to the fed state. In conclusion, the presence of vitamins and minerals decreased the viability of the cells compared to the same conditions without vitamin and minerals, when the vitamins and minerals provided simultaneously with the strain.

[0148] Incubation time Multivitamin and Fed or mean LOG 10 LOG 10 (min) mineral added fasted CFU loss

[0149] Start CFU 0 1 / 10,72

[0150] SSF 10 No Fasted 10,33 0,40

[0151] SSF+SGF 120 No Fasted 10,11 0,32 Ref 16302-WO-PCT

[0152] Table 1. Viable cells number of freeze-dried L. plantarum DSM 33464 strain at TO (0 min), after incubation in SSF (simulated salivary fluid) (10 min), after further incubation in SGF (simulated gastric fluid) (120 min), and after further incubation in SIF (small intestinal fluid) (240 min).

[0153] The strain was tested in different simulated fasted and fed conditions.

[0154] Example 2 - Lead binding measurement in simulated gastrointestinal intestinal conditions From the same experiment described in example 1 , samples from the fed state after 240 min of co-incubation were analyzed for lead binding capacity, using a lead binding assay. In short, the samples were co-incubated 60 min with a 26.2-ppm lead acetate solution, and centrifuged. Lead analysis was conducted as described previously using Inductively Coupled Plasma-Mass Spectrometer (ICP-MS) method, after treatment of the samples with nitric acid.

[0155] Results:

[0156] In absence of the strain, the concentration of lead in the small intestinal fluid hardly changed after 60 min of co-incubation (Table 2). The absence or presence of multivitamins and minerals did not change this measurement either (25.5 and 25.3 ppm lead respectively).

[0157] In contrast, in presence of the L. plantarum DSM 33464 strain, the concentration of lead present in the small intestinal fluid after 60 min of co-incubation with a lead acetate solution strongly decreased from 26.2 to 5.36 ppm. This result suggest that less lead would be available in vivo in the gut for absorption when the strain is present. An effect was also seen in the fluid where multivitamins and minerals were added together with the strain, however the effect was smaller than when the strain was added alone. In this set up, the lead concentration decreased from 26.2 to 8.56 ppm. This smaller decrease compared to the condition where the strain was added alone is could likely be attributed mostly to the fact that the survival of the strain was lower in presence of these vitamins and minerals (see table 1), and therefore was slightly less able to bind as much lead. In conclusion, the presence of vitamins and minerals, when given simultaneously with the strain, decreased the lead binding capacity of the strain by 16% under simulated gastrointestinal intestinal conditions compared to the same conditions without vitamin and minerals. Ref 16302-WO-PCT

[0158] Co-incubation ..1X. .x. , ,

[0159] . . „ „ Multivitamin Lead time with lead . . Strain DSM .

[0160] . . , .. composition . concentration acetate solution , , , 33464 present

[0161] (min)added (ppm>

[0162] Solution before

[0163] 0 / / 26.2 co-incubation SSF+SGF+SIF 60 No NO 25.5 SSF+SGF+SIF 60 YES NO 25.3 SSF+SGF+SIF 60 No YES 5.36 SSF+SGF+SIF 60 YES YES 8.56

[0164] Table 2 concentral ion of lead in small intestinal fluid before and after co-incubation. The strain was tested in different simulated fed conditions.

[0165] Example 3 - L . plantarum DSM 33464 effects in children with elevated blood lead levels

[0166] A randomized, double-blind, placebo-controlled, parallel-group, multi-center clinical trial was conducted. The trial included five clinic visits: visit 1 screening (day -14-day -2), visit 2 randomization (day -1), visit 3 follow-up (day 28±3; week 4), visit 4 follow-up (day 56±3; week 8), visit 5 (day 84±3; week 12), and visit 6 follow-up phone call four weeks post-intervention (day 112±3; week 16). Children aged 3-12 years having a blood lead level (BLL) between 3.5-24.9 pg / dL were included in the trial. The children were randomized into an experimental group receiving L . plantarum DSM 33464 as the intervention and a control group receiving placebo as the intervention, both groups furthermore received health education and daily nutrition multivitamin / mineral supplement throughout the 12-week intervention period. Fifty-four children received at least one intervention (probiotic; n=30 and control; n= 24). The majority (94.4%) of children had a slightly elevated blood lead level at baseline (median 4.2(1.3) pg / dL). Data collected showed that those randomized to each group were similar on all demographic and physiological measures and a questionnaire on lead exposure sources showed no major differences in both groups.

[0167] The experimental group intervention consisted of L. plantarum DSM 33464 (probiotic) given at a daily dose of 1 x 109colony forming units (CFUs) and dextrin in one sachet (2 g) , and a daily multivitamin / mineral supplement sachet containing vitamin B1 1.5 mg; vitamin B2 0.8 mg; vitamin B3 5 mg; vitamin B5 1 mg; vitamin B6 0.5 mg; vitamin B7 10 pg; vitamin B9 80 pg; vitamin B12 1 pg; vitamin A 600 III; vitamin D 200 IU; iron pyrophosphate 5 mg; calcium 180 mg; zinc gluconate 4 mg; maltodextrin 1.4 g. The intervention took place for 12 weeks.

[0168] The control group intervention consisted s of one daily sachet of placebo (dextrin only, 2 g) and the same multivitamin / mineral supplement sachet as in the experimental group intervention (containing vitamin B1 1.5 mg; vitamin B2 0.8 mg; vitamin B3 5 mg; vitamin B5 1 mg; vitamin B6 0.5 mg; vitamin B7 10 pg; vitamin B9 80 pg; vitamin B12 1 pg; vitamin A 600 IU; vitamin D 200 IU; Ref 16302-WO-PCT iron pyrophosphate 5 mg; calcium 180 mg; zinc gluconate 4 mg; maltodextrin 1.4 g). The intervention took place for 12 weeks.

[0169] All children were instructed to take one intervention sachet (i.e. dependent of intervention group a sachet of probiotic or placebo) orally 0.5 hours before having a meal. The intervention sachet was taken after dissolving the sachet into moderate amount of lukewarm water, once daily. Additionally, one sachet of the multivitamin / mineral supplement was taken orally by dissolving it into moderate amount of warm water or food with a dosing interval of at least 4 hours before or after the intake of either probiotic or placebo. The probiotic and placebo sachets had the same visual appearance to avoid compromising the study blinding.

[0170] Results:

[0171] Blood lead levels reduced from baseline to the end of supplementation in both groups by about 40% from 5.3 pg / dL to 3.1 pg / dL in the probiotic and from 4.8 pg / dL to 2.9 pg / dL in the placebo group, respectively . Urine lead levels were also reduced from baseline in both groups throughout the study. At week 8, the urine lead levels (Lean Squares Means) for probiotic was -30.0 while control was -13.2, and the trend persisted in week 12. The blood levels of all essential trace elements (calcium, zinc, copper, magnesium and iron) at week 4, 8 and 12, stayed within normal physiological ranges, without any deficiency reported. At end of intervention, there was no statistically significant difference between groups for all the essential elements. (Table 3) Ref 16302-WO-PCT

[0172] Table 3 Blood Lead Levels, Urine Lead Levels and Essential Trace Elements Levels in Blood. N = number of subjects.

[0173] GSRS (Gastrointestinal Symptoms Rating Scale) total score improved for both groups from base- line to week 12, with abdominal pain score reduced significantly in probiotic group compared to control at week 12 (Table 4).

[0174] Probiotic Control

[0175] N Mean N Mean Ref 16302-WO-PCT

[0176] Table 4 Gastrointestinal Symptom Rating Scale (GSRS) Questionnaire Scores. Safety Set Population. N = number of subjects.

[0177] CPRS (Conners’ Parent Rating Scale) total score and all subscores improved for both groups from baseline to week 12; with improvement in psychosomatic feelings and learning problems in probiotic group more pronounced than control at week 12 (Table 5).

[0178] Probiotic Control

[0179] N Mean N Mean

[0180] Table 5 48-item Conners’ Parent Rating Scale (CPRS). Safety Set Population. N = number of subjects.

Claims

Ref 16302-WO-PCTCLAIMS1 . A Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use in preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

2. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to claim 1 , wherein the human is a child.

3. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to claim 1 or claim 2, wherein the multivitamin composition comprises at least one or more minerals and / or vitamins selected from the group consisting of: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin B12, Niacin (Vitamin B3), Folic acid (Vitamin B9), Pantothenic acid (Vitamin B5), Biotin (Vitamin B7), Vitamin A, Vitamin D, Zinc, Iron and Calcium.

4. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding claims, wherein the multivitamin composition comprises calcium, zinc and / or iron.

5. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding claims, wherein the Lactiplantibacillus plantarum strain is deposited as DSM 33464.

6. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding claims, wherein the strain is administered at least 1 hour before or after intake of the multivitamin composition.

7. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to any one of the preceding claims, which upon administration to a human results in one or more effects selected from the group consisting of: Decrease in the lead level in the body, decrease in the lead level in the blood, prevention of lead absorption as measured in the urine lead level, increase in excretion of lead from the body, decrease in the lead level in other organs or body parts, reduction of the gastro-intestinal discomfort, reduction in abdominal pain, and reduction in behavioral discomfort.

8. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to claim 7, which decreases the lead level in the body and / or decreases the blood lead level upon administration compared to the lead level before the administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such.

9. The Lactiplantibacillus plantarum strain or a dietary supplement comprising such for use according to claim 7, which decreases the lead level in the body and / or decreases the blood lead level upon administration compared to the lead level before the administration of theRef 16302-WO-PCTLactiplantibacillus plantarum strain or the dietary supplement comprising such, wherein the lead level is measured after at least 2 weeks of administration of the Lactiplantibacillus plantarum strain or dietary supplement comprising such.

10. A method of administering a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.11 . Use of a Lactiplantibacillus plantarum strain or a dietary supplement comprising such for preventing or ameliorating lead accumulation in a human, wherein the strain is administered to a human less than 12 hours before or after intake of a multivitamin composition, and wherein the strain is administered at a different time point than the multivitamin composition and / or in a composition wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

12. A dietary supplement composition comprising a Lactiplantibacillus plantarum strain and a multivitamin composition, wherein the Lactiplantibacillus plantarum strain is physically separated from the multivitamin composition.

13. The dietary supplement composition according to claim 12, wherein the Lactiplantibacillus plantarum strain is present in a first microcapsule and the multivitamin composition is present in a second microcapsule.

14. The dietary supplement composition according to claim 12 or 13, wherein the Lactiplantibacillus plantarum strain and the multivitamin composition are present in a layered composition, wherein the Lactiplantibacillus plantarum strain is present in one layer and the multivitamin composition is present in a separate layer, the Lactiplantibacillus plantarum strain is present in the core of the composition and the multivitamin composition is present in a separate layer around the core, or the multivitamin composition is present in the core of the composition and the Lactiplantibacillus plantarum strain is present a separate layer around the core, wherein the multivitamin composition is optionally further divided into two layers.

15. The dietary supplement composition according to any one of claims 12 to 14, wherein the composition is solid or liquid.

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