Lactobacillus gasseri

Description and significance

Like the other members of the Lactobacillus genus, Lactobacillus gasseri is an anaerobic, grampositive bacterium that falls into the category of lactic acid bacteria. It is also a rod shaped and of the non-spore-forming type. It is typically found in the gastrointestinal tracts of humans and animals due to its largely fermentative function (1,2,3). Although mainly in the GI tract, it can also be found in many other places as well (see Ecology). Isolation of this bacterium was achieved by taking a sample from the gastrointestinal tract and was discovered to be part of the lactobacillus acidophilus complex (4). This bacterium is very important to many applications in daily life. One of its roles, as described above, is fermentation in the GI tract. Recently, its function as a probiotic has been the area of most interest (1).

Genome structure
The complete genome has been sequenced by a combination of efforts from Joint Genome Institute, Fidelity Systems Inc., and North Carolina State University. The final draft was accepted on October 13, 2006. Lactobacillus gasseri ATCC 33323 was the strain they used to come up with the sequence (5). (ATCC stands for American Type Culture Collection) The genome is circular and contains 1.894362 Mb (5). The GC content of the sequence was found to be 35.3% which makes the AT content to be 64.7%. There were also 1755 different proteins identified as well as 98 different RNAs present (6). The mapping of the sequence gives great information about the role and function of this bacterium. Dr. Todd Klaenhammer states L. gasseri is substantially more amenable to DNA introduction and manipulation which has lead to the development of more genetic tools that will be useful in the functional genomic analysis of this species (5).

Cell structure and metabolism

Lactobacillus gasseri is rod shaped and a gram-positive bacterium. This implies that it has a very thick outer cell wall. It is also classified as a non-spore-forming type. One feature that is still being researched is how this adheres to intestinal cells. It was determined that mannose-specific proteinaceous adhesion is responsible for L. gasseris ability to attach to the intestinal linings when the mucosal layer is encountered (7). Another interesting feature is the use of Aggregation-

promoting factor (APF) to create and support the rod shape. The APF protein was examined and the researchers concluded that it truly did alter the cell shape based on its amount present in the cell (8). L. gasseri is an anaerobe so it lives mostly in regions where no oxygen is present. It also participates in fermentative actions which produce lactic acid as well as the energy required for growth. One other interesting thing that is made by this bacterium is hydrogen peroxide which is found in the vaginal tract. Research was done with cattle to examine the effectiveness of this hydrogen peroxide producing bacteria as a probiotic (9). (See Current Research)

Ecology
Even though this bacterium is most commonly found in the gastrointestinal tract, it can also be found in various other places. One very important location is in the vaginal tract of reproductive, normal women. Its role is to protect the vagina from infections (10). This correlates with the hydrogen peroxide produced, as described in the Cell Structure and Metabolism section. It is can also be found in the fecal waste products of adult humans as well as in the mucosa layer of the rectum and oral cavity. With L. gasseri heavily found in humans and animals, a significant amount of research has been conducted to try and find out exactly what this bacterium and its fellow genus members actually do as well as how it can operate as a probiotic (10).

Pathology
In 2004, Lactobacillus gasseri was identified as a cause of Fournier's gangrene (11). Fournier's gangrene is a bacterial infection of the skin that affects the genitals and perineum. It is most typically due to a wound that is infected on the skin between the genitals and the anus. Some typical symptoms include soft, spongy skin, discolored skin, pus, genital pain, and an odor (12).

Application to Biotechnology
Lactobacillus gasseri as well as the rest of the lactobacillus genus are known for their role in fermentation of many different food products. One example is the use of lactobacillus gasseri in fermenting meat. Many of the lactic acid bacteria are useful for this type of process, but L. gasseri was found to be the most dominant in the human gut. With this being said, it will provide less complications in the digestive system which makes it the best choice to ferment the meat. This bacterium will not only decrease the ability of pathogens like Staphylococcus aureus to grow in the meat, but would also function as a probiotic for the intestinal flora of the human GI tract. This would produce a much healthier meat product for all to consume (13).

Current Research

Inhibition of Staphylococcus aureus by H202-producing Lactobacillus gasseri isolated from vaginal tract of cattle (9) The use of antibiotics in dairy farms to keep the cattle healthy from reproductive problems has left residues in the food products obtained. Also an increased resistance to these drugs has been observed as well as increased costs of supplying these drugs. Lactobacillus gasseri is found in the vaginal tract. Research was conducted on the amount of hydrogen peroxide two strains (CRL1421 and CRL141) generated in different conditions. Also the interaction with Staphylococcus aureus was looked at in the cultures. It was observed that the lactic acid and the hydrogen peroxide had a very strong effect on the pathogen. S. aureus had a different type of morphological damage from the lactic acid and hydrogen peroxide. This concluded that lactobacillus gasseri could be used as a probiotic to help combat the infections seen in cattle. Lactobacillus gasseri GasserAM63T degrades oxalate in a multistage continuous culture simulator of the human colonic microbiota (14) Oxalate can cause many problems in the human body including excess urinary oxalate and cause kidney stones to form. This compound is introduced to the body by plants that are eaten. Lactobacillus gasseri has formyl coenzyme A transferase (frc) and oxalyl coenzyme A decarboxylase (oxc) on its genome which may have a role in oxalate degradation. Research found that L. gasseri does in fact break down oxalate in vitro. To test this finding in vivo the bacteria was cultured in human fecal matter to simulate colon conditions. It was found that oxalate degradation occurred in the section which corresponded to the proximal colon. These findings give some insight on using L. gasseri as a possible treatment for those with kidney stones. Preliminary human study for possible alteration of serum immunoglobin E production in perennial allergic rhinitis with fermented milk prepared with Lactobacillus gasseri TMC0356 (15) Allergic disease due to enhanced immunoglobulin E (IgE) responses to common stimuli has become more common in the population. The internal microbiota has been identified to play a part in regulating the immune function in humans. Due to this observation, an experiment was setup with 15 people that showed the enhanced IgE responses. These subjects drank milk fermented with Lactobacillus gasseri TMC0356 and were observed. The results were that the levels of the IgE were decreased. This data shows that IgE was reduced, but no data was found on whether allergic responses were reduced as well. A promising future is ahead of this type of fermented milk, but further research on the effects on the person must be conducted to see if this is a safe way to go.

References
1. Alatossova, T., Munro, K., Ng, J., Tannock, G. W., & Tilsala-Timisjarvi, A. (1999) Identification of Lactobacillus Isolates from the Gastrointestinal Tract, Silage, and Yoghurt by 16S-23S rRNA Gene Intergenic Spacer Region Sequence Comparisons. Applied and Enviromental Microbiology. 65(9). 4364-4267

2. Falsen, E, Pascual, C, Sjoden, B, Ohlen, M, & Collins, MD. (1999) Phenotypic and phylogenetic characterization of a novel Lactobacillus species from human sources: description of Lactobacillus iners sp. Nov. Int J Syst Bacteriol. 49. 217-221 3. Mitsuoka, T. (1992) The human gastrointestinal tract. In, The Lactic Acid Bacteria: Volume 1, The Lactic Acid Bacteria in Health and Disease. B.J.B. Wood (ed), pp69-114.Elsevier Science Publishers, Ltd. Essex, England. 4. Kullen, M.J., R.B. Sanozky_Dawes, D.C. Crowell and T.R. Klaenhammer. (2000) Use of DNA sequence of variable regions of the 16SrRNA gene for rapid and accurate identification of bacteria in the Lactobacillus acidophilus complex. J. Appl. Microbiol. 89:511-518. 5. DOE Joint Genome Institute http://genome.jgipsf.org/finished_microbes/lacga/lacga.home.html Accessed 8/23/07 6. NCBI Genome Project http://www.ncbi.nlm.nih.gov/sites/entrez?db=genomeprj&orig_db=&term=Lactobacillus%20gas seri%20&cmd Accessed 8/23/07 7. Horie, M, & Horie. (2005). Anaerobic induction of adherence to laminin in Lactobacillus gasseri strains by contact with solid surface. Current microbiology, 51(4), 275-282. 8. Jankovic, I., M. Ventura, V. Meylan, M. Rouvet, M. Elli, and R. Zink. (2003) Contribution of aggregation-promoting factor to maintenance of cell shape in Lactobacillus gasseri 4B2. J. Bacteriol. 185:3288-3296 9. Otero, M. C., and Nader-Macas M. E. (2006) Inhibition of Staphylococcus aureus by H202producing Lactobacillus gasseri isolated from vaginal tract of cattle. Animal Reproductive Science. 96(1-2). 35-46 10. Tailliez, P. (2004). Lactobacilli: properties, habitats, physiological role and importance in human health. Antibiotiques, 6(1), 35-41. 11. Tleyjeh, I. M., Routh, J., Qutub, M. O., Lischer, G., Liang, K. V., & Baddour, L. M. (2004). Lactobacillus gasseri causing Fournier's gangrene. Scandinavian Journal for infectious Diseases. 36(6-7). 501-503 12. Urological Emergencies: Fournier's gangrene, Causes, Symptoms, Diagnosis, Treatment. http://www.urologychannel.com/emergencies/fgangrene.shtml Accessed 8/25/07 13. Arihara, K., Ota, H., Itoh, M., Kondo, Y., Sameshima, T., Yamanaka, H., Akimoto, M., Kanai, S., & Miki, T. (1998) Lactobacillus acidophilus Group Lactic Acid Bacteria Applied to Meat Fermentation. Journal of Food Science. 63(3). 544547.

14. Lewanika, TR, & Lewanika. (2007). Lactobacillus gasseri Gasser AM63(T) degrades oxalate in a multistage continuous culture simulator of the human colonic microbiota. FEMS microbiology ecology, 61(1), 110-120 15. Morita, H., He, F., Kawase, M., Kubota, A., Hiramatsu, M., Kurisaki, J., & Salminen, S. (2006). Preliminary human study for possible alteration of serum immunoglobin E production in perennial allergic rhinitis with fermented milk prepared with Lactobacillus gasseri TMC0356. Microbiology and Immunology. 50(9). 701-6.

Lactobacillus plantarum

Description and significance

Lactobacillus plantarum is a Gram-positive, bacteria which ferments plants and is used industrially to make yogurt, cheese, and pickled vegetables. Apart from plants, L. plantarum is found in the human GI tract and acts as a probiotic bacteria. Its ability to survive in the GI tract has made this bacteria a target for further research in using it as a vehicle to deliver recombinant vaccines and treatments for diseases.[1]

Genome structure
In 2003, Michiel Kleerebezem mapped out the complete genome of Lactobacillus plantarum WCFS1. Lactobacillus plantarum strain WCFS1 is known to have 3,308,274 base pairs and contains 3,052 protein-encoding genes as well as three plasmids (1,917-bp, 2,365-bp, and 36,069-bp,) which accounts for its high adaptability. The G+C content of the chromosome is 44.5%, but the plasmids contain slightly less G+C content. [1]

Cell structure and metabolism

Lactobacillus plantarum is a Gram-positive, rod-shaped, organotrophic, aerotolerant, bacteria often called lactic acid bacteria because it gets most of its energy from converting glucose to lactate via homolactic and heterolactic fermentation. Homolactic fermentation uses the EMP pathway and heterolactic fermentation uses the phosphoketolase pathway. Therefore, L. plantarum is a facultative heterofermentative lactobacilli. [2] [1]

Ecology
Lactobacillus plantarum is a highly adaptive bacteria that can survive at temperatures between 1 to 60 Celsius and also a great range of atmospheric pressures. The name "plantarum" indicates that this bacteria is a "species of the plants." L. plantarum is found particularly in fermented plants, such as kimchee. However, it is also found in other fermented foods such as yogurt and cheese and it is also found in animal GI tracts, human saliva and vagina. [3] [4] [1]

Cheese Yogurt

Kimchee Human GI Tract

Pathology
Lactobacillus plantarum 299v is actually considered to be a probiotic. Due to its ability to survive in the human GI tract for >6 days, L. plantarum 299v can inhibit the growth of other harmful bacteria such as E. coli in the host after contaminated meat consumption. L. plantarum 299v is found in various fermented foods such as yogurt, cheese, kimchee, and is recommended to be consumed regularly to support the immune system. In addition, the ability of L. plantarum 299v to produce lactic acid helps maintain healthy, low pH levels of the GI tract. [1] [3] [4]

Applications to Biotechnology
In 2004, Jean-Marie Perrier-Cornet discovered a way to control food fermentation by using Lactobacillus plantarum, a common food bacteria with resistance to a good range of temperature, pressure, and pH level. A cold pasteurization method without freezing the food and thus altering the composition of the food is a new technology of interest to the food industry. Perrier-Cornet found optimization of the procedure when pasteurizing L. plantarum for 10 min at temperatures between 20 and 25 C and pressure between 100 and 350 MPa. [5]

Current Research
In 2006, Michael Schultz studied the effects of treatment with Lactobacillus plantarum 299v on immunodeficient mice prone to the development of colitis. The study found that L. plantarum 299v administered orally with water to interleukin-10 deficient mice produced significant results of reduced immune-mediated colitis. This suggests that L. plantarum 299v has the potential to be used against clinical inflammatory bowel disease. [6]

In 2008, Beatriz del Rio studied the use of live recombinant Lactobacillus plantarum as an oral delivery vehicle for a Lyme disease vaccine to invoke a protective immune response in mice. Because L. plantarum can survive in the GI tract, it has been long considered for this type of research. L. plantarum successfully expressed vaccine antigens inducing immunity in the mice infected with Lyme disease. [7] In 2009, Alistair W. G. Waugh studied the effects of treatment with Lactobacillus plantarum 299v on healthy mice induced with irritable bowel syndrome via rectal administration of 1% allyl isothiocyanate in 30% ethanol. The mice were then force-fed L. plantarum 299v for up to 28 days. The results showed that L. plantarum 299v reduced inflammation and normalized the bowels.[8]

References
1. 1.0 1.1 1.2 1.3 1.4 Kleerebezem, Michiel, Jos Boekhorst, Richard van Kranenburg, Douwe Molenaar, Oscar P. Kuipers, Rob Leer, Renato Tarchini, Sander A. Peters, Hans M. Sandbrink, Mark W. E. J. Fiers, Willem Stiekema, Ren M. Klein Lankhorst, Peter A. Bron, Sally M. Hoffer, Masja N. Nierop Groot, Robert Kerkhoven, Maaike de Vries, Bjrn Ursing, Willem M. de Vos*, and Roland J. Siezen. (2003). [1] Complete genome sequence of Lactobacillus plantarum WCFS1. Proceedings of the National Academy of Sciences, 100 (4), 1990-1995. 2. "Lactobacillus." MicrobeWiki. 8 April 2009 Kenyon College. 21 April 2009. [2] 3. 3.0 3.1 "Bacteria Genomes - LACTOBACILLUS PLANTARUM" European Bioinformatics Institute. European Molecular Biology Laboratory. 21 April 2009. [3] 4. 4.0 4.1 Hwang, Hamin. "KimChee and Fermentation." 19 April 2005. 21 April 2009. [4] 5. Perrier-Cornet, Jean-Marie, Sandra Tapin, Serenella Gaeta and Patrick Gervais. (2004). [5]High-pressure inactivation of Saccharomyces cerevisiae and Lactobacillus plantarum at subzero temperatures. Journal of Biotechnology, 115 (4), 405-412. 6. Schultz, Michael, Claudia Veltkamp , Levinus A. Dieleman , Wetonia B. Grenther , Priscilla B. Wyrick , Susan L. Tonkonogy , R. Balfour Sartor, M.D. (2006). [6]Lactobacillus plantarum 299V in the treatment and prevention of spontaneous colitis in interleukin-10-deficient mice. Wiley Interscience, 8 (2), 71-80. 7. Beatriz del Rio, Raymond J. Dattwyler, Miguel Aroso, Vera Neves, Luciana Meirelles, Jos F. M. L. Seegers, and Maria Gomes-Solecki1. (2008) [7]Oral Immunization with Recombinant Lactobacillus plantarum Induces a Protective Immune Response in Mice with Lyme Disease. Clinical and Vaccine Immunology, 15. (9), 1429-1435 8. Waugh, Alistair W. G., Rae Foshaug, Sarah Macfarlane, Jason SG Doyle, Thomas A. Churchill, Beate C. Sydora, Richard N. Fedorak. (2009). [8]Effect of Lactobacillus plantarum 299v treatment in an animal model of irritable bowel syndrome. Microbial Ecology in Health and Disease, 21. (1), 33-37. Superscript text

Lactobacillus casei

Description and significance

Lactobacillus casei is a rod-shaped Gram-positive bacteria. It is non-sporing, non-motile, and anaerobic. Consistent with other lactic acid bacteria, this species is acid tolerant. Lactobacillus casei dwells in environments such as the intestinal tracts of animals and fermented dairy products. It can be found naturally in both the human intestine and the mouth. They have a wide temperature range as well as a wide pH range. The organism is mesophilic, which means it has an optimum temperature range around 30C to 40C. The optimum pH is at approximately 5.5. Scientists have found it to possess beneficial properties that support human health. It is able to improve and promote digestion. Some strains of the bacteria help control diarrhea, while other strains have an anti-inflammatory effect on the gut. Other advantageous effects include reducing lactose intolerance, alleviating constipation, and even modulation of the immune system. Numerous strains have been proven to be probiotics, that is according to the World Health Organization, are "live microorganisms which when administered in adequate amounts confer a health benefit on the host." Because it is lactic acid producer, it has several applications in biotechnology and in the food industry.[1] [2]

Genome structure
The genome of Lactobacillus casei ATCC 334 comprise of a circular chromosome with 2,895,264 base pairs and a circular plasmid 1 with 29,061 base pairs. According to US DOE Joint Genome Institute the strain contains a total of 2,924,325 nucleotides, 2,771 protein genes, and 75 RNA genes. L. casei have a 45-47% G+C content.[3]

Cell structure and metabolism

The cell structure of Lactobacillus casei is typically straight, rod-shaped, and arranged in chains. The cell size tends to be around 0.7-1.1 x 2.0-4.0 micrometer. It is also a facultative anaerobe. This means it is an organism that is able to grow under both aerobic and anaerobic environments but develops better and more rapidly in the presence of oxygen. The microbe is an organotroph and its metabolism is a homofermentative one. Unlike heterofermentative lactobacteria which can produce either alcohol or lactic acid from carbohydrates, L. casei participates in a homolactic fermentation process that can only result in one single major end product. It obtains most of its energy by converting glucose into lactic acid. Some varieties of Lactobacillus casei can produce lactic acid utilizing galactose, fructose, or even mannose. Stress, poor diet, and antibiotics can lead to a deficient growth of the bacteria.[4]

Ecology

L.casei inhabits the oral and gastrointestinal tracts of animals. The bacterium bears a resistance to both gastric acid and bile enabling it to endure the harsh conditions throughout the gastrointestinal tract. Studies have shown that this bacterium generates health-promoting effect on the host. Therefore they are best known as probiotics. Their presence helps sustain a stabilized distribution of microflora in the intestine. It fulfills this role through antimicrobial activity. The mechanism involves creating an acidic habitat that restricts the growth of other bacteria that may be detrimental or cause infections. The production of bacteriocins (a toxin) by L.casei inhibits the growth of similar or closely related bacterial strains preventing overpopulation. Another mechanism is by competition inhibition and exclusion. L. casei Shirota strain is able to directly compete with pathogens that resides in the gastrointestinal tract for adhesion sites. This reduces pathogenic bacteria from adhering to the intestinal wall. Their occupation within the intestine is significant in maintaining the homeostasis of the gut immune system as well. It is also known to produce DL-lactic acid and amylase that complements the growth of Lactobacillus acidophilus, another probiotic lactic acid bacterium. Similar to many probiotics it does not permanently stay in the host, instead it usually lives in the intestine for a duration of around 10 days after being taken.[5][6]

Pathology
Lactobacillus casei does not cause any diseases. Not only is it generally considered to be harmless, it is well recognized as a beneficial microorganism and a nonpathogenic.

Application to Biotechnology
L. casei strains are of industrial significance since they can be applied in a range of fermentation processes, whereas other strains are utilized for their probiotic properties. Some are used in the production of cheese, yogurt, fermented milks, fermented Sicilian green olives, and other products. The natural end product of fermentation for this microbe is lactic acid, which inhibits the development of other organisms as well as decreasing the pH level in the food or beverage product. It is also known to be used in the development of flavor for selected cheeses. Numerous food industries use the probiotic bacteria in their products to promote health. Yogourmet a probiotic yogurt starter includes Lactobacillus Casei, Bifidus, and Acidophilus. Other name brand that also uses L. casei includes Lifeway Yogurt and Stonyfield Farm Yogurt.

Yakult (Lactobacillus casei Shirota strain) by Casey Yancey at Flickr.com. Add image caption here. DANNON the worldwide leader in fresh dairy products, began producing a probiotic drink called DanActive. The product is contains their own particular strain of the bacteria called L. casei Immunitas, L. bulgaricus, S. thermophilus, and milk as the main ingredient. L. casei Immunitas is also known as L. casei DN-114001. It claims that "daily consumption of DanActive with L. casei Immunitas helps strengthen your body's defenses." [7][8] Since 1955, the Lactobacillus casei Shirota strain has been used by Yakult Honsha a Japanese food company to produce a probiotic milk-like product called Yakult. It is generated from the fermentation of the bacteria with skimmed milk. It contains 6.5 billion cfu of the bacteria per 65 ml bottle.[9]

Current Research
"Lactobacillus casei DN-114 001 Inhibits the Ability of Adherent-Invasive Escherichia coli Isolated from Crohn's Disease Patients To Adhere to and To Invade Intestinal Epithelial Cells" by Isabelle Ingrassia, Antony Leplingard, and Arlette Darfeuille-Michaud Lactobacillus casei have been found to be helpful to people with a chronic inflammatory bowel disease (IBD) called Crohn's disease. This research purpose was to demonstrate the in vitro inhibitory effects of Lactobacillus casei DN-114 001 on adherent-invasive E. coli. The adherentinvasive E. coli was extracted from Crohn's disease patients. The experiment included the preincubation of cultured intestinal epithelial cells with L. casei DN-114 001 before infecting it with adherent-invasive E. coli LF82. Intestine-407 and Caco-2 cells are intestinal epithelial cells. The probiotic was able to inhibit the adherent-invasive ability of LF82 to undifferentiated Intestine407 cells by 62%. It also was able to inhibit E.coli LF82 from adhering to undifferentiated Caco2 cells by 47% and differentiated Caco-2 cells by 43%. The inhibitory effect on LF82 invasion was only found for undifferentiated cells, but the effect was slightly higher than the effect on adhesion. When the adherent-invasive E. coli and L. casei DN-114 001 were added together with intestinal epithelial cells in co-incubation, reduction in adhesions and invasions were observed as well. These results illustrates that the strain can be effectively used to inhibit the interaction of

those pathogenic bacteria with intestinal epithelial cells. This suggests a new possible option for the treating Crohn's disease.[10] "Daily ingestion of fermented milk containing Lactobacillus casei DN114001 improves innate-defense capacity in healthy middle-aged people" by Parra MD, Martnez de Morentin BE, Cobo JM, Mateos A, Martnez JA at Department of Physiology and Nutrition, University of Navarra, Spain. The goal of this experiment was to examine the effects of Lactobacillus casei DN114001 intake from fermented milk on the immune response capacity in middle-age participants. This case consisted of 45 healthy subjects between the ages of 51 to 58: 24 women and 21 men. The two groups were formed randomly. One group received three cups per day of the fermented milk with the bacteria and the other group received a placebo for a period of eight weeks. The results showed no significant changes in immune cell proportions. However there was an increased in oxidative burst capacity of monocytes and an increase in tumoricidal activity of NK cells seen in the group treated with DN114001 strain. They concluded that "that daily intake of fermented milk containing Lactobacillus casei DN114001 could have a positive effect in modulating the innate immune defense in healthy-middle-age people".[11] "Modulation of natural killer cell activity by supplementation of fermented milk containing Lactobacillus casei in habitual smokers" by Morimoto K, Takeshita T, Nanno M, Tokudome S, Nakayama K at Department of Social and Environmental Medicine, Osaka University Graduate School of Medicine, Osaka, Japan. This research involves testing L. casei's ability in restoring natural killer cell activity in habitual smokers which tend to have a significant reduction in natural killer cell activity. The 99 habitually smoking volunteers were randomly placed in to two groups. One group received the fermented milk containing the probiotic daily and the other group received a placebo daily. This experiment was carried out for a duration of three weeks. The researchers recorded the NK cell activity in the peripheral blood mononuclear cells before and after the intake. According to the journal the natural killer cell activity in participants had an inverse relationship with the number of cigarettes smoked. The results revealed that there was no significant difference in the proportion of natural killer cells between the two groups. The averaged natural killer cell activity adjusted by the numbers of cigarettes was significantly higher for the individuals in the L. casei group than those in the placebo group. They concluded that the consumption of fermented milk with lactic acid bacteria, which in this case is Lactobacillus casei, can be effective in restoring the natural killer cell activity in habitual smokers.[12]

References
1. "Lactobacillus casei." Wikipedia, The Free Encyclopedia. 22 Mar 2009, 22:52 UTC. 31 Mar 2009 <http://en.wikipedia.org/w/index.php?title=Lactobacillus_casei&oldid=279033109>. 2. http://genome.jgi-psf.org/finished_microbes/lacca/lacca.home.html genome.jgi-psf.org 3. http://www.kegg.com/kegg-bin/show_organism?org=lca kegg.com

4. "Lactobacillus casei." Microbewiki, <http://microbewiki.kenyon.edu/index.php/Lactobacillus_casei> 5. http://www.probiotics-lovethatbug.com/lactobacillus-casei.html www.probioticslovethatbug.com 6. http://jmm.sgmjournals.org/cgi/content/full/52/10/925 jmm.sgmjournals.org 7. "Actimel." Wikipedia, The Free Encyclopedia. 2 Apr 2009, 08:50 UTC. 4 Apr 2009 <http://en.wikipedia.org/w/index.php?title=Actimel&oldid=281256710>. 8. http://www.danactive.com/danactive_whatIs_casei.html 9. "Yakult." Wikipedia, The Free Encyclopedia. 14 Mar 2009, 07:19 UTC. 4 Apr 2009 <http://en.wikipedia.org/w/index.php?title=Yakult&oldid=277140860>. 10. http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1151832 pubmedcentral.nih.gov 11. http://www.ncbi.nlm.nih.gov/pubmed/15457926?ordinalpos=1&itool=EntrezSystem2.PE ntrez.Pubmed.Pubmed_ResultsPanel. Pubmed_DiscoveryPanel.Pubmed_Discovery_RA&linkpos=2&log$=relatedarticles&log dbfrom=pubmed www.ncbi.nlm.nih.gov 12. http://www.ncbi.nlm.nih.gov/pubmed/15749143?ordinalpos=1&itool=EntrezSystem2.PE ntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DiscoveryPanel. Pubmed_Discovery_RA&linkpos=3&log$=relatedarticles&logdbfrom=pubmed www.ncbi.nlm.nih.gov/pubmed