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Food Microbiology 28 (2011) 214e220 Contents lists available at ScienceDirect Food Microbiology Bacillus probiotics Simon M. Cutting* School of Biological Sciences, Royal Holloway University of London, Egham, Surrey TW20 0EX, UK Bacterial spore formers are being used as probiotic supplements for use in animal feeds, for human Available online 24 March 2010 dietary supplements as well as in registered medicines. Their heat stability and ability to survive thegastric barrier makes them attractive as food additives and this use is now being taken forward. While often considered soil organisms this conception is misplaced and Bacilli should be considered as gut commensals. This review summarises the current use of Bacillus species as probiotics, their safety, mode of action as well as their commercial applications.
Ó 2010 Elsevier Ltd. All rights reserved.
1. Bacterial spores 2. The use of Bacillus as probiotics Bacterial spores are produced in nature as a means to survive Probiotics are live microbes, which when administered in extreme environmental conditions enabling long-term survival in adequate amounts confer a health benefit to the host (Araya et al., conditions that could otherwise kill vegetative bacteria (Nicholson 2002). Bacillus species have been used as probiotics for at least 50 et al., 2000). The decision to sporulate is very much dependant years with the Italian product known as EnterogerminaÒ registered upon the decline in nutrients in the immediate vicinity of the live 1958 in Italy as an OTC medicinal supplement. The scientific cell. Sensing this, the bacterium enters an irreversible program of interest in Bacillus species as probiotics though, has only occurred development that results in the production of a spore some 8 h later in the last 15 years and three principal reviews have covered the (Fig. 1) (Errington, 2003). Intrinsic to survival is the structure of the field (Hong et al., 2005; Mazza, 1994; Sanders et al., 2003). Of the bacterial endospore, that contains, at its core, a condensed and species that have been most extensively examined these are inactive chromosome. Additional layers surround the spore, Bacillus subtilis, Bacillus clausii, Bacillus cereus, Bacillus coagulans including a peptidoglycan-rich cortex and one or more layers of and Bacillus licheniformis. Spores being heat-stable have a number proteinaceous material referred to as the spore coat (Henriques and of advantages over other non-spore formers such as Lactobacillus Moran, 2007). Together these protect the spore from UV radiation, spp., namely, that the product can be stored at room temperature in extremes of heat (typically up to 80e85
C in most species), a desiccated form without any deleterious effect on viability. A exposure to solvents, hydrogen peroxide and enzymes such as second advantage is that the spore is capable of surviving the low lysozyme (Nicholson et al., 2000). The spore itself, is dehydrated pH of the gastric barrier (Barbosa et al., 2005; Spinosa et al., 2000) and if exposed to appropriate nutrients will germinate, a process which is not the case for all species of Lactobacillus (Tuohy et al., taking just a few minutes, allowing water to enter the spore, 2007) so in principle a specified dose of spores can be stored breakage and removal of the spore coats, and outgrowth and indefinitely without refrigeration and the entire dose of ingested resumption of vegetative cell growth (Fig. 1) (Moir, 2006).
bacteria will reach the small intestine intact.
Depending on species spores are spherical or ellipsoidal in shape, Spore probiotics are being used extensively in humans as die- between 0.8 and 1.4 mm in length, have a negative surface charge tary supplements (Table 1), in animals as growth promoters and competitive exclusion agents (Table 2) and lastly in aquaculture for commonly fall under two genera, Bacillus and the strictly anaerobic enhancing the growth and disease-resistance of cultured shrimps, Clostridia although a surprisingly large number of other, lesser- most notably the Black Tiger shrimp (Penaeus monodon) (Table 3).
known, genera include spore formers.
This review will focus primarily on the use of spore products forhuman use. Interestingly, a number of Bacillus products are licensedas medicinal supplements. Rather than describing specific productsa short summary of the major Bacillus species used in commercial * Tel.: þ44 (0) 1784 443760; fax: þ44 (0) 1784 414224.
E-mail address: [email protected] products will be summarised.
0740-0020/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fm.2010.03.007 Author's personal copy
S.M. Cutting / Food Microbiology 28 (2011) 214e220 Vegetative Cell Growth e3 vials are taken each day with the aim of pre- venting infantile diarrhoea (Figs. 2e4). The suspension of spores inwater is thought to enhance delivery of spores to the mucosa anddemonstrates the versatility of spore formulations. The product carries four antibiotic resistant strains of B. clausii that are recom- mended for use with antibiotics (Coppi et al., 1985; Green et al.,1999; Senesi et al., 2001). The four strains are each derived from ATCC 9799, a penicillin-resistant strain originally designated asB. subtilis. Through a multi-step process strains resistant to novo- biocin þ rifampin (strain N/R), chloramphenicol (strain O/C), streptomycin þ neomycin (strain SIN) and tetracycline (strain T)have been obtained (Ciffo, 1984; Mazza, 1994). Interestingly, theseB. clausii strains also carry resistance to a number of other antibi-otics including erythromycin, cephalosproins and cycloserine, Fig. 1. The sporulation life cycle. A schematic showing the opposed life cycles of kanamycin, tobramycin, and amikacin (Mazza et al., 1992). It has bacterial spore formers. Under conditions of nutrient starvation the growing, vegeta-tive cell (VC) will undergo a series of morphological changes that create a forespore (F) now been demonstrated that the resistance genes within these within the mother cell (MC) of the sporangium. After approximately 8 h the spore (S) is B. clausii strains are stable and are unable to transfer (Bozdogan released by lysis of the MC.
et al., 2004; Mazza, 1983; Mazza et al., 1992).
Although the initial scientific studies used to register this product in 1958 are obscure clinical trials have subsequently been performed demonstrating efficacy, although a number of thesetrials lack completeness in terms of controls. Of note are clinical B. clausii spores are used in the product EnterogerminaÒ which studies assessing the effect of Enterogermina modulating the is registered as an OTC medicinal supplement. Unlike most pro- immune responses in allergic children with recurrent respiratory biotic formulations that are supplied in tablet or capsule form the infections (Ciprandi et al., 2004, 2005a,b). After administration of Enterogermina product carries, spores (2  109) suspended in 5 ml the probiotic nasal symptoms and eosinophil counts in allergic Table 1Bacillus probiotics for human use.a Produced by Marion Merrell (Levallois-Perret, France) Capsule carrying 1  109 spores of Bacillus cereus strain IP5832b but also by Hoechst and then Aventis Pharma following (ATCC 14893) [n.b., originally deposited as B. subtilis.
merger acquisitions. Also cited as being produced byCasella-Med, Cologne, Germany Protexin Health Care B. subtilis is one component of 14 strains carried in this UK probiotic (1) Biofarm, Dniepropetrovsk, Ukraine BiosporinÒ is a mixture of two strains of living antagonistic bacteriaB. subtilis 2335 (sometimes referred to as B. subtilis 3) and B. licheniformis2336 (ratio is 3:1). Originally isolated from animal fodder.
(2) Garars, Russia There are a number of versions of this products produced in differentcountries including a recombinant form, Subalin.
Geyer Medicamentos S.A. Porto Alegre, RS, Brazil B. cereus strain GM Suspension of 106 spores ml1.
Binex Co. Ltd., Busan, S. Korea Tablet carrying spores (1.7  107) of B. polyfermenticus SCD.c BioProgress SpA, Anagni, Italy Vial carrying 1  109 spores of Bacillus clausii in suspension, labelled as carrying B. subtilis. No longer marketed.
Sanofi Winthrop SpA, Milan, Italy Vial (5 ml) carrying 1  106 spores of B. clausii in suspension. At least four different strains of B. clausii present and product originally labelled ascarrying B. subtilis.
Flora-Balance, Montana, USA Capsules labelled as carrying Bacillus laterosporus BODc but containing Brevobacillus laterosporus BOD.
Ganeden Biotech Inc., Ohio, USA B. coagulans GanedenBC30 This is a patented strain that has GRAS approval in the USA.
Istituto Biochimico Italiano SpA, Milan, Italy Capsule carrying spores of B. subtilis labelled as carrying 2  109 spores ofLactobacillus sporogenes.c Sabinsa Corp., Piscataway, NJ, USA Labelled as Lactobacillus sporogenesc but contains B. coagulans 6e15  109 g1.
Hanmi Pharmaceutical Co. Ltd., Beijing, China B. subtilis strain RO179 (at 108 g1) in combination with Enterococcus faecium.
Nature's First Food Nature's First Law, San Diego, CA, USA 42 species listed as probiotics including: B. subtilis, B. polymyxa,c B. pumilus and B. laterosporus.c Newpharma S.r.l., Milan, Italy Mixture of lactic acid bacteria inc. L. acidophilus, B. bifidum and L. sporogenes.cL. sporogenes at 3.3  105 CFU g1 whose valid name is B. coagulans and ismislabelled as a strain of B. subtilis.
B. subtilis.
Palm Beach, Florida, USA.
www.gardenoflife.com/ a This list is likely incomplete and excludes Vietnamese products that are shown in Table 4.
b Contains the same strain used in the now discontinued animal feed product Paciflor.
c Not recognised as a Bacillus species (www.bacterio.cict.fr).
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S.M. Cutting / Food Microbiology 28 (2011) 214e220 Table 2Bacillus probiotics for veterinary use.a Alpharma Inc., Melbourne, Australia B. licheniformis (NCTC 13123) at 1091010 spores kg1. This is a non-bacitracin producing strain. Not licensed in the EU.
Poultry, calves and swine Provita Eurotech Ltd., Omagh, Listed as containing spores of B. licheniformis (1.6  109 CFU g1) and Northern Ireland, UK B. subtilis (1.6  109 CFU g1).
Piglets,a Chickens, Christian Hansen Hoersholm, Mixture (1/1) of B. licheniformis (DSM 5749) and B. subtilis (DSM 5750) turkeys for fatteningc at 1.6  109 CFU g1 of each bacterium. EU approved.a Esporafeed PlusÒ Norel, S.A. Madrid, Spain 1  109 B. cereus (CECT 953). Not licensed in the EU.
Poultry, calves and swine Pharmed Medicare, Bangalore, India Labelled as Lactobacillus sporogenesb but contains B. coagulans.
Poultry, calves and swine Sanofi Sante Nutrition Animale, France 2 strains of B. clausii (CNCM MA23/3V and CNCM MA66/4M). Not licensedin the EU.
Calves, poultry, rabbits and Asahi Vet S.A., Tokyo (Head Off.), Japan B. cereus var. toyoi (NCIMB-40112/CNCM-1012) at a minimum swine. Possible use also for concentration of 1  1010 CFU g1 mixed with maize flour (4% by weight) and calcium carbonate (90% by weight). Licensed in the EU.a a Authorised for unlimited use by the EU.
b Not recognised as a Bacillus species (www.bacterio.cict.fr).
children were significantly reduced. In these studies a Th1 it is being used in a number of products such as Sustenex and is also (T-helper 1) bias was observed showing that ingestion of Enter- being incorporated into foods where spores can survive the mild ogermina could enhance the cellular immunity in allergic children heat-treatments used to sterilise foods. A recently published who normally carry a Th2 bias. These studies have been supported randomized, double-blind, placebo-controlled, parallel-design, has by later studies by Marseglia et al. (2007) who have examined the shown significant effects of B. coagulans as an adjunct therapy for duration and rate of respiratory infections in 40 children (mean age relieving symptoms of rheumatoid arthritis (Mandel et al., 2010).
4.3  1.5 years). After administration of Enterogermina for 90 days Other than this the value of B. coagulans as a probiotic has, however, they observed a decrease in the duration of respiratory infection, recently been questioned (Drago and De Vecchi, 2009) and but not the frequency of infection. Other clinical trials have undoubtedly, further scientific evidence supporting the efficacy of examined the positive effect of Enterogermina on the side effects of this species is required.
antibiotic-based Helicobacter pylori therapy (Nista et al., 2004), andon urinary tract infections (Fiorini et al., 1985).
2.3. B. subtilis and B. licheniformis The product was originally labelled as carrying spores of B. subtilis but subsequent studies have identified the species as B. subtilis has been extensively studied at a genetic and physi- B. clausii (Green et al., 1999; Senesi et al., 2001). This product is not ological level. Numerous probiotic products are labelled as carrying specifically referred to as a probiotic but claims to enhance the B. subtilis and in part, this probably results historically from body's immune system following germination of the spores in the a carelessness in assuming that most aerobic spore formers are small intestine.
B. subtilis. Accordingly, numerous products claiming to carryB. subtilis have been shown to carry other species (see Table 1 and 2.2. B. coagulans Table 4). However, B. subtilis var. Natto is worthy of comment. Thisbacterium is used in the fermentation of soybeans that is used to This species is often labelled, incorrectly, as Lactobacillus spor- prepare the Japanese staple known as Natto. Natto carries as many ogenes which is an unrecognised species name. The origin of this as 108 viable spores per gram of product and for decades health species for use in probiotics stems from India where a number of benefits have been associated with consumption of Natto including manufacturers produce B. coagulans as a food ingredient for export stimulation of the immune system (Hosoi and Kiuchi, 2004). A and relabelling in Europe and the US. B. coagulans secretes serine protease known as Nattokinase is secreted from vegetative a bacteriocin, Coagulin, which has activity against a broad spectrum cells of B. subtilis var. Natto and has been shown to reduce blood of enteric microbes (Hyronimus et al., 1998). Recently one strain, clotting by fibrinolysis (Sumi et al., 1987, 1995). There are several labelled as GanedenBC30 has been granted self-affirmed GRAS important points here, firstly, the serine protease that is named status by the FDA in the US. Marketed by Ganeden, as GanedenBC30 Nattokinase is in fact produced by all strains of B. subtilis but in the Table 3Bacillus probiotics for aquaculture.a Sino-Aqua Corp., Kaohsiung, Taiwan B. subtilis strains Wu-S and Wu-T at 108 CFU g1, product also contains Lactobacillus and Saccharomyces spp.
Microbial Solutions, Johannesburg, South Africa and Mixture of: B. megaterium, B. licheniformis, Paenibacillus polymyxa Advanced Microbial Systems, Shakopee, MN, USA and two strains of B. subtilis.
Cargill, Animal Nutrition Division Undefined Bacillus species.
Sino-Aqua company Kaohsiung, Taiwan Carries four strains of B. subtilis.
Sanocare Sanolife Sanoguard INVE Technologies nv Dendermonde, Belgium Various Bacillus species.
a This shows just a selection of registered products from international companies. In shrimp-producing countries the number of ‘local' products is substantial, for example, in Vietnam over 30 different products are sold.

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S.M. Cutting / Food Microbiology 28 (2011) 214e220 Fig. 2. EnterogerminaÒ. This is a licensed OTC product containing 2  109 of GMP-produced spores of B. clausii in 5 ml of water. 2e3 vials are consumed per day to helpprevent gastroenteritis in infants and children.
Natto strain it is produced at high levels. Second, it cannot be ruledout that health benefits ascribed to Natto require consumption ofboth soybeans and bacteria, rather than just the bacterium. In anyevent, Nattokinase has GRAS status as an enzyme produced froma bacterium in the US and is purified and sold as a health supple-ment worldwide. In poultry studies controlled trials have shownthat oral administration of B. subtilis spores reduce infection bySalmonella enterica serotype Enteritidis, Clostridium perfringens andEscherichia coli O78:K80 (La Ragione et al., 2001; La Ragione andWoodward, 2003).
Fig. 4. Natto. Natto is normally consumed as a fermented soybean product either hot B. subtilis and B. licheniformis are used together in two products, or cold. In this example it is sold as a snack with dried soybeans coated with a fine Biosporin and BioPlusÒ 2B. BioPlusÒ 2B is used in animal feed while white powder of B. subtilis var. Natto, the active ingredient required for the taste andtexture of Natto.
Biosporin is licensed as a medicine in the Ukraine and Russia.
Biosporin is sold in glass vials that must be reconstituted in waterbefore consumption. The two Bacillus strains, B. subtilis 2335 and H. pylori (Pinchuk et al., 2001). In the case of BioPlusÒ 2B this animal B. licheniformis 2336 are well characterised and a number of clinical feed product has also been extensively studied with numerous studies have been used to demonstrate probiotic effects although efficacy studies focused on the suppression of gastrointestinal none been performed with the rigour of a full clinical trial (Bilev, pathogens completed resulting in the registration of this product as 2002; Osipova et al., 2003, 2005; Sorokulova, 1997; Sorokulova a feed supplement in Europe (SCAN, 2000b). It remains unclear et al., 1997). Interestingly, B. subtilis 2335 has been shown to whether there is any added benefit in the combined use of the two produce the antibiotic Amicoumacin with in vitro activity against B. cereus is a known human pathogen that is the cause of mild food poisoning due to the production of up to three enterotoxinsand one emetic toxin (Stenfors Arnesen et al., 2008). Not all strainsof B. cereus carry enterotoxin genes yet a number of B. cereus pro-biotics have been shown to carry the enterotoxin genes (Hoa et al.,2000) and one product, Paciflor, used in animal feed has beenwithdrawn from use in the EU (SCAN, 2001a). Despite this B. cereusproducts are still being used for example, ToyerocinÒ, an animalfeed product is registered for use in Europe (SCAN, 2001b) andBactisubtilÒ as a registered medicinal supplement for human use.
Interestingly, the strain of B. cereus used in BactisubtilÒ known asIP5832 is the same as that in the withdrawn animal productPaciflorÒ.
3. How do spore probiotics work? Fig. 3. Biosubtyl and Biosubtyl DL. Typical Vietnamese products, in this case, Biosubtyl Bacillus species are often considered soil organisms since spores that carries spores of B. cereus IP5832 and Biosubtyl DL carrying a mixture of B. cereus they can readily be retrieved from soil. However, attempting to IP5832 and Lactobacillus acidophilus. Neither product is labelled properly nor carriesthe stated dose.
isolate vegetative bacteria from soil is more problematic and it now Author's personal copy
S.M. Cutting / Food Microbiology 28 (2011) 214e220 Table 4Vietnamese Bacillus OTC products licensed for human use.
Viet-Duc Pharmaceutical Co. Ltd., Hanoi Labelled as containing B. subtilis, L. acidophilus, S. faecalis but B. subtilis isB. cereus at 107 g1.
Tediphar Corporation (TEDIPHARCO), Ho Chi Minh City, Vietnam Sachet (1 g) carrying 107108 spores of B. subtilis.
Bidiphar. Binh Dinh Pharmaceutical and Medical Equipment Labelled sachets carrying 1  106 spores of B. cereus but mislabelled as Company, 498 Nguyen Thai Hoc, Qui Nhon, Vietnam B. subtilis.
Biophar Company, Da lat, Vietnam Sachet (1 g) carrying 106107 of B. cereus spores mixed with tapioca.
Product labelled as B. subtilis. The strain is closely related by 16S rRNAanalysis to IP 5832 used in BactisubtilÒ.
IVAC, 18 Le Hong Phong, Da Lat, Vietnam Sachets (1 g) carrying 107108 CFU of B. subtilis andLactobacillus acidophilus.
Biosubtyl I and II Biophar Company, Nha Trang, Vietnam Sachet (1 g) carrying 106107 of B. pumilus spores mixed with tapioca.
Product labelled as B. subtilis.
Pasteur Institute of Ho Chi Minh City, Vietnam Sachets (1 g) carrying 108 spores of B. subtilis.
Mekophar, Pharmaceutical Factory No. 24, Ho Chi Minh City, Capsule carrying 106107 spores of a B. cereus species termed B. cereus var. vietnami. Product labelled as carrying B. subtilis.
ILdong Pharm Co., Ltd., 60-1, SinKeonji-Dong, Ansung-Si, Each gram of granules contains: Kyong Ki-Do, Korea Lactobacillus sporogenes 5.0  107 cfuClostridium butyricum 1.0  107 cfuBacillus subtilis 3.0  106Thiamine Nitrate 0.3 mgRiboflavin 0.2 mgAscorbic Acid 5.0 mgNicotinamide 0.1 mgDibasic calcium photphate 20.0 mgDried yeast 50.0 mg ILdong Pharm Co., Ltd., 60-1, SinKeonji-Dong, Ansung-Si, Each gram of granules contains: Kyong Ki-Do, Korea Lactobacillus sporogenes 5.0  107 cfuClostridium butyricum 1.0  107 cfuBacillus subtilis 3.0  106Thiamine Nitrate 0.3 mgRiboflavin 0.2 mgAscorbic Acid 5.0 mgNicotinamide 0.1 mgDibasic calcium phosphate 20.0 mgDried yeast 50.0 mg seems likely that spores are designed to survive transit across the B. subtilis spores were shown to suppress infection with pathogenic gastric barrier of animals that ingest them. This view originates S. enterica (La Ragione and Woodward, 2003), C. perfringens from studies that show that a percentage (>10%) of an inoculum of (La Ragione and Woodward, 2003) and E. coli (La Ragione et al., B. subtilis spores can germinate in the small intestine, grow and 2001). A mouse model has been used to show suppression of Cit- proliferate and then re-sporulate (Hoa et al., 2001; Tam et al., 2006).
robacter rodentium (a model for the traveller's diarrhoea pathogen, Peristalsis ensures that spores are shed in faeces resulting in their ETEC) by administration of B. subtilis spores (D'Arienzo et al., 2006).
accumulation in the soil. An intestinal habitat of spore formershelps explain why spores can be found in the gut of insects, animals and humans (Barbosa et al., 2005; Fakhry et al., 2008; Hong et al.,2009a). Recent work has shown that Bacilli can readily be Two spore formers, Bacillus anthracis and B. cereus are known as obtained from the human GI-tract using analysis of both biopsies human pathogens. The former requires no elaboration while the and faeces (Fakhry et al., 2008; Hong et al., 2009a). In the latter, use of B. cereus appears to be a cause for concern on a case-by-case Bacillus spores can be found at levels of approximately 104 spores/g basis. The safety of Bacillus species has been extensively reviewed of faeces which is several logs higher than can reasonably be pre- elsewhere (de Boer and Diderichsen, 1991; Ishibashi and Yamazaki, dicted from food intake alone (Hong et al., 2009b).
2001; Logan, 2004; Osipova et al., 1998; Sanders et al., 2003; SCAN, Numerous studies have shown that germinating spores can 2000a) and most incidences of illness associated with Bacillus elicit potent immune responses in the GI-tract of mouse models appear to result for opportunistic infections or miss-diagnosis.
and this immune stimulation may be the underlying reason why Extensive animal studies including acute and sub-chronic toxicity spores exert a probiotic effect (Hong et al., submitted for testing as well as in vitro studies have now been performed on publication). One of the most informative, yet least recognised a number of species, including B. subtilis var. Natto (Hong et al., studies was one examining the effect of orally administered 2008), Bacillus indicus (Hong et al., 2008), B. coagulans (Endres bacteria on the development of the gut-associated lymphoid tissue et al., 2009) and B. subtilis 2335 (Sorokulova et al., 2008) and (GALT) in infant rabbits (Rhee et al., 2004). In these studies B. licheniformis 2336 (Sorokulova et al., 2008). All appear to show no B. subtilis was shown to be of greater importance than other indicators of adverse effects.
commensal bacteria in GALT development. Of course, other prop-erties such as the secretion of antimicrobials such as Coagulin, 5. Approved products in Europe and the USA Amicoumacin and Subtilisin may also further provide a probioticeffect by suppressing growth of competing microbes as well as Bacillus products that have been formally approved in the West enteric pathogens. Studies showing efficacy are less easy to distil are few. Numerous authors routinely cite B. subtilis as having GRAS yet a few convincing examples are as follows. In a poultry model (Generally Regarded as Safe) status but this is incorrect. Nattokinase, Author's personal copy
S.M. Cutting / Food Microbiology 28 (2011) 214e220 the proteolytic enzyme that is purified from B. subtilis var. Natto does spores could be added to beverages and foods yet retain their carry GRAS status as a microbially produced enzyme but not the probiotic properties. Indeed, such probiotic foods have already bacterium. In 2008 B. coagulans strain GanedenBC30 was the first entered the market with "Activate Muffins" containing Gane- Bacillus strain to be given self-affirmed GRAS approval. In Europe, for denBC30 launched by Isabella's Health Bakery in the USA in 2008.
approval, for use as a supplement a case must be made based on prioruse. The application is first made by authorities in the host country and then assessed by an EU committee. To date B. subtilis has beenapproved for use as a supplement in Italy and the UK. B. clausii that is The use of Bacillus species as probiotic dietary supplements is used in the medicinal OTC product EnterogerminaÒ and B. cereus expanding rapidly with increasing number of studies demon- IP5832 (BactisubtilÒ) are registered as medicines with specific claims strating immune stimulation, antimicrobial activities and compet- regarding the prevention of childhood diarrhoea and, as a medicine, itive exclusion. The single and most important advantage of these are not marketed under the probiotic label.
products is that they can be produced easily and the stability of thefinished product can be assured, further they can be incorporated 6. The Vietnamese market into everyday foods. Studies are showing that these bacteria areable to grow within the intestinal tract and possibly be considered In SE Asia, notably, Vietnam, where no concept of dietary temporary residents. This is important because it shows that these supplements exists, Bacillus products are licensed with the Ministry bacteria are not foreigners but rather may exert a unique symbiotic of Health as medicinal supplements (Table 4) with claims ranging relationship with their host.
from prevention of rotavirus infection (infant diarrhoea) and foodpoisoning to immune stimulation. It is unclear whether theirapproval requires formal clinical trials but in any event these products are easily obtained and often used as the first line ofdefence against enteric infections both prophylactically but more This article was based in part on a publication in Nutrafoods often therapeutically. The use of Bacillus probiotics in Vietnam is (2009 Vol. 8:7e14). Research in the laboratory of SMC and TCD is more developed than in any other country and the reason for this is supported by an EU 7th FP grant, KBBE-2007-207948.
unclear. There is also intense interest in using heat-stable Bacillusspores in aquaculture and it is not uncommon for shrimp farms to use products produced for human use.
Agnew, M.D., Koval, S.F., Jarrell, K.F., 1995. Isolation and characterisation of novel 7. Recent innovations: functional foods alkaliphiles from bauxite-processing waste and description of Bacillus vedderisp. nov., a new obligate alkaliphile. Syst. Appl. Microbiol. 18, 221e230.
Araya, M., Morelli, L., Reid, G., Sanders, M.E., Stanton, C., 2002. Joint FAO/WHO In recent work pigmented Bacillus species have been charac- Working Group Report on Guidelines for the Evaluation of Probiotics in Food terised and the pigment has been shown to be due one or more carotenoids (Duc et al., 2006; Khaneja et al., 2009). These carotenoids Barbosa, T.M., Serra, C.R., La Ragione, R.M., Woodward, M.J., Henriques, A.O., 2005.
Screening for Bacillus isolates in the broiler gastrointestinal tract. Appl. Environ.
have been shown to carry anti-oxidant activity in vitro and thus could Microbiol. 71, 968e978.
be of nutritional value (SM Cutting; unpublished data). Yellow, Bilev, A.E., 2002. Comparative evaluation of probiotic activity in respect to in vitro orange, red and pink Bacillus species can be easily obtained from soil, pneumotropic bacteria and pharmacodynamics of biosporin-strain producers inpatients with chronic obstructive pulmonary diseases. Voen. Med. Zh. 323, 54e57.
river and pond sediments as well as from the intestinal tracts of Bozdogan, B., Galopin, S., Leclereq, R., 2004. Characterization of a new erm-related animals (Hong et al., 2009a; Yoon et al., 2001, 2005). This includes macrolide resistance gene present in probiotic strains of Bacillus clausii. Appl.
a red pigmented Bacillus megaterium (Mitchell et al., 1986) a pink Environ. Microbiol. 70, 280e284.
pigment found in some isolates of Bacillus firmus (Pane et al., 1996) Ciffo, F., 1984. Determination of the spectrum of antibiotic resistance of the Bacillus subtilis strains of Enterogermina. Chemioterapia 3, 45e52.
and red pigment found in Bacillus atrophaeus (Fritze and Pukall, 2001; Ciprandi, G., Tosca, M.A., Milanese, M., Caligo, G., Ricca, V., 2004. Cytokines evalu- Nakamura, 1989). A variable yellow-orange pigmentation has ation in nasal lavage of allergic children after Bacillus clausii administration: been found in a number of species including, B. indicus (Suresh et al., a pilot study. Pediatr. Allergy Immunol. 15, 148e151.
Ciprandi, G., Vizzaccaro, A., Cirillo, I., Tosca, M.A., 2005a. Bacillus clausii effects in 2004), Bacillus cibi (Yoon et al., 2005), Bacillus vedderi (Agnew et al., children with allergic rhinitis. Allergy 60, 702e703.
1995), Bacillus jeogali (Yoon et al., 2001), Bacillus okuhidensis (Li Ciprandi, G., Vizzaccaro, A., Cirillo, I., Tosca, M.A., 2005b. Bacillus clausii exerts et al., 2002), Bacillus clarkii (Nielsen et al., 1995), Bacillus pseudo- immuno-modulatory activity in allergic subjects: a pilot study. Eur. Ann. Allergy Clin. Immunol. 37, 129e134.
rmus (Nielsen et al., 1995) and B. firmus (Ruger and Koploy, 1980).
Coppi, F., Ruoppolo, M., Mandressi, A., Bellorofonte, C., Gonnella, G., Trinchieri, A., 1985.
The carotenoids are found in the vegetative cell as well as in the spore Results of treatment with Bacillus subtilis spores (Enterogermina) after antibiotic and they help protect spores from UV radiation (Khaneja et al., 2009).
therapy in 95 patients with infection calculosis. Chemioterapia 4, 467e470.
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