Safety Assessment of Bacillus subtilis MB40 for Use in Foods and Dietary Supplements

Abstract

With the growing popularity of probiotics in dietary supplements, foods, and beverages, it is important to substantiate not only the health benefits and efficacy of unique strains but also safety. In the interest of consumer safety and product transparency, strain identification should include whole-genome sequencing and safety assessment should include genotypic and phenotypic studies. Bacillus subtilis MB40, a unique strain marketed for use in dietary supplements, and food and beverage, was assessed for safety and tolerability across in silico, in vitro, and in vivo studies. MB40 was assessed for the absence of undesirable genetic elements encoding toxins and mobile antibiotic resistance. Tolerability was assessed in both rats and healthy human volunteers. In silico and in vitro testing confirmed the absence of enterotoxin and mobile antibiotic resistance genes of safety concern to humans. In rats, the no-observed-adverse-effect level (NOAEL) for MB40 after repeated oral administration for 14 days was determined to be 2000 mg/kg bw/day (equivalent to 3.7 × 1011 CFU/kg bw/day). In a 28 day human tolerability trial, 10 × 109 CFU/day of MB40 was well tolerated. Based on genome sequencing, strain characterization, screening for undesirable attributes and evidence of safety by appropriately designed safety evaluation studies in rats and humans, Bacillus subtilis MB40 does not pose any human health concerns under the conditions tested.

RETRACTED: Expression of SARS-CoV-2 Spike Protein Receptor Binding Domain on Recombinant B. subtilis on Spore Surface: A Potential COVID-19 Oral Vaccine Candidate

“…B. subtilis has been used as an adjuvant in chickens treated with inactivated avian influenza virus H9N2 [10]. The use of recombinant B. subtilis has become an economical and practical solution, as it can be mass-produced, due to its easy cultivation [11] and safety [12]. It has been applied in industry [13], agriculture [14], medicine [15], health and food [16] and other fields [17,18].…”

Preclinical Safety Assessment of Bacillus subtilis BS50 for Probiotic and Food Applications

“…In addition to work utilizing in silico and in vitro studies, several animal toxicity studies have demonstrated the safety of B. subtilis for human use [ 15 , 16 , 17 , 18 , 19 , 20 ]. Clinical trials of B. subtilis and W. coagulans (formally Bacillus coagulans ) strain supplementations have also shown safety and tolerance in humans, as well as digestive and GI health benefits in subjects with inflammatory bowel syndrome [ 21 , 22 , 23 , 24 , 25 ], dyspepsia [ 26 , 27 ], as well as individuals with or without mild symptoms of GI distress [ 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 ].…”

“…Clinical trials of B. subtilis and W. coagulans (formally Bacillus coagulans ) strain supplementations have also shown safety and tolerance in humans, as well as digestive and GI health benefits in subjects with inflammatory bowel syndrome [ 21 , 22 , 23 , 24 , 25 ], dyspepsia [ 26 , 27 ], as well as individuals with or without mild symptoms of GI distress [ 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 ]. For example, the B. subtilis strain MB40 has been shown to be safe and support GI health in a randomized, double-blind, placebo-controlled trial of 100 healthy adults [ 17 , 35 ]. Additionally, B. inaquosorum DE111 supplementation has been shown to be safe in both adult and pediatric human subjects [ 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ].…”

Lysinibacillus macrolides 38328 Isolated from Agricultural Soils as a Promising Probiotic Candidate for Intestinal Health

“…The mice in the experimental groups exhibited normal growth compared to the control group, and there were no pathological changes in various organs. According to the oral toxicity study of rats by Spears et al [25], the safety of B. subtilis was determined. Kim et al [43] have veri ed the safety of Bacillus through observation of pulmonary conditions.…”

“…According to the method of Spears et al [25] with some modi cations, a challenge test was conducted to evaluate the pathogenicity of the selected three strains. Six-week-old female BALB/c mice weighing 17-21 g (Liaoning Changsheng Biotech Co., Ltd., China) was randomly divided into four groups consisting of ve mice per group.…”