See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/327700552

Quantitative and qualitative evaluation of three commercial probiotic

products from India

Article  in  Research Journal of Biotechnology · December 2017

CITATIONS READS

0 588

3 authors:

Kavya Sudha Ramya Mohandass

SRM Institute of Science and Technology SRM Institute of Science and Technology
4 PUBLICATIONS   10 CITATIONS    62 PUBLICATIONS   785 CITATIONS   

SEE PROFILE SEE PROFILE

Parani Madasamy
SRM Institute of Science and Technology
93 PUBLICATIONS   2,240 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Development of Natural Anti-Mycotics for Ringworm Infection View project

Isolation and Characterization of mesenchymal stem cells derived from human dental pulp View project

All content following this page was uploaded by Kavya Sudha on 29 December 2018.

The user has requested enhancement of the downloaded file.

Research Journal of Biotechnology
Quantitative and qualitative evaluation of three
commercial probiotic products from India
Chavali Kavyasudha, Mohandass Ramya and Madasamy Parani*
Department of Genetic Engineering, SRM University, Chennai 603 203, Tamil Nadu, INDIA
*parani.m@ktr.srmuniv.ac.in
Abstract
Studies from other countries have shown that many
commercial probiotics did not contain the organisms
that were listed in the label or they contained species
that were not listed – some of them were even
pathogenic. There are no regulatory guidelines
governing the production and sale of probiotics in
India and thus it is very important to bring the
regulatory guidelines. Therefore, a pilot study was
conducted to evaluate the microbial contents of the
probiotics in terms of quality and quantity.
Commercial probiotics were purchased from local
drug stores. As per the label, all the three probiotics
should contain Clostridium butyricum, Streptococcus
faecalis and Bacillus mesentricus. In addition,
BIFILAC® and ViBact® should contain Lactobacillus
sporogenes while Pre-Pro® should contain
Lactobacillus acidophilus.
In analysis, S. faecalis and C. butyricum were present
in all the three probiotics but in case of Pre-Pro®,
CFU/sachet was several fold less than what was
claimed in the label. Both BIFILAC® and ViBact®
contained L. sporogenes in expected quantity but
according to latest nomenclature it should be
renamed as B. coagulans. Bacillus mesentricus was
absent in all the three products and L. acidophilus
was absent in Pre-Pro®. All the three products
contained B. subtilis which was not claimed in the
label. Fortunately, we did not find any pathogenic
bacteria in any of the probiotic products analyzed.
Our study clearly demonstrates mislabeling of the
organisms, absence of claimed organisms, presence of
unclaimed organisms and significantly less number of
certain claimed organisms in the probiotics tested.
Keywords: Commercial probiotics, Clostridium
butyricum, Streptococcus faecalis, Bacillus subtilis,
Bacillus coagulans, 16S rDNA sequencing.
Introduction
According to the Food and Agricultural Organization of
World Health Organization, probiotics are the live
microorganisms which when administered in adequate
amounts confer a health benefit on the host. Probiotic
activity is strain specific and it is important to link a strain
to a specific health effect. Though probiotics are commonly
used for treating acute diarrhea, lactose intolerance and to
69
Vol. 12 (12) December (2017)
Res. J. Biotech
offset side effects of antibiotics on gut microflora, recently
they are also used for treating or preventing specific
disorders such as irritable bowel syndrome, eczema,
allergies, Helicobacter pylori infection and
sequelae.1,15,16,18,19,20,24. Probiotic therapy is becoming
increasingly common in veterinary and human medicine
and numerous probiotic products are now available
commercially.27. Commercial probiotics are available in the
form of conventional foods, food supplements and dietary
supplements containing probiotic bacteria.8,22,23
According to the 'Probiotics Market’ report (2009-2014),
the global probiotics market is expected to increase 31.1bn
USD by 2015 with the Europe and Asia accounting for
nearly 42 and 30% of the total revenues respectively. The
probiotics industry in India was estimated to worth Rs. 20.6
million in 2009 and its annual growth rate was predicted to
increase about 22.5 percent until 2015.
Only in 2002, WHO recommended that the information on
genus, species, strain designation and minimum viable
number of each probiotic strain at the end of the shelf life
should be described on the label of probiotic products
FAO/WHO, 2002.7 There were instances of unsubstantiated
claims on contents as well as their beneficial effects.
Studies from developed and developing countries like
USA, UK, Japan, South Africa etc. have reported low
viable counts resulting in a loss of probiotic
effects,3-6,12,13,27 absence of specified organisms,14 and more
dangerously, the presence of pathogenic organisms.2 We
conducted a pilot study wherein the contents of three
commercial probiotics were evaluated against the labeled
claims by using morphological, biochemical and molecular
methods.
Material and Methods
Materials and Media used: Commercial probiotics
BIFILAC® (Tablets India Limited, Puducherry), ViBact®
(Allianz Biosciences Pvt. Ltd, Puducherry], Pre-Pro®
(Swiss Garnier Life Sciences, Himachal Pradesh) were
purchased from local drug stores. As per the label, all the
three products were manufactured in December 2010 and
due to expire by May 2012. This study was conducted four
months after the date of manufacturing and 12 months
before the date of expiry. The following media were used
for growing the probiotics.
Clostridium butyricum medium - CB medium: Meat
extract 3.0 g l-1, Liver extract 5.0 g l-1, Yeast extract 5.0 g l-
1, Peptone 10.0 g l-1, Tryptone 10.0 g l-1, Soy peptone 3.0 g
l-1, Soluble starch 5.0 g l-1, Glucose 10.0 g l-1, NaH2PO4 0.5
Research Journal of Biotechnology
g l-1, Na2HPO4 0.5 g l-1, MgSO4 0.2 g l-1, MnSO4 0.062 g l-
1
, NaCl 0.1 g l-1, FeSO4 0.1 g l-1, Tween 80 1.0 ml l-1, L-
cysteine-HCl 0.5 g l-1, Agar 15.0 g l-1, pH adjusted to 7.2.
Streptococcus faecalis medium - SF medium: Yeast
extract 5.5 g l-1, Peptone 12.5 g l-1, Dextrose 11.0 g l-1,
CaCO3 5.0 g l-1, NaH2PO4 0.25 g l-1, Na2HPO4 0.25 g l-1,
MgSO4 0.1 g l-1, MnSO4 0.05 g l-1, FeSO4 0.05 g l-1,
CH3COONa 0.2 g l-1, Agar 15.0 g l-1, pH adjusted to 6.8.
Bacillus mesentericus medium - BM medium: Meat
extract 10.0 g l-1, Peptone 10.0 g l-1, NaCl 5.0 g l-1,
Na2HPO4 5.0 g l-1, Soluble starch 10.0 g l-1, Agar 15.0 g l-1,
pH adjusted to 7.0.
MRS medium for Lactobacillus: Protease peptone 10.0 g
l-1, Beef extract 10.0 g l-1, Yeast extract 5.0 g l-1, Dextrose
20.0 g l-1, Polysorbate 80 1.0 g l-1, Ammonium citrate 2.0 g
l-1, CH3COONa 5.0 g l-1, MgSO4 0.10 g l-1, MnSO4 0.05 g
l-1, Na2HPO4 2.0 g l-1, Agar 12.0 g l-1, pH adjusted to 6.5.
Screening for probiotic strains: The lyophilized probiotic
samples in the sachet were weighed and thoroughly
dissolved in 100 ml of diluent solution (0.2% NaCl) and
used for serial dilution. Serial dilutions were done up to
10-7 by mixing 9 ml of diluent solution and 1ml of sample
solution. Appropriate dilutions were used for pour plating
in which 1.0 ml of the diluted sample was taken in sterile
Petri dish, 20 ml of the medium (with 1.5% agar) was
poured over and incubated as required. For Clostridium
butyricum, the sample from 10-6 dilution was plated in CB
medium and incubated in anaerobic condition at 37°C for
12 h.
For Streptococcus faecalis, the sample from 10-7 dilution
was plated in SF medium and incubated at 37°C for 72 h.
For Bacillus mesentericus, the sample from 10-5 dilution
was plated in BM medium and incubated at 37°C for 24 h.
For Lactobacillus sporogenes and Lactobacillus
acidophilus, the sample from 10-6 dilution was plated in
MRS medium and incubated at 37°C for 48 h. Plating was
done in triplicate for each sample. At the end of the
incubation period, the number of colonies were counted
manually and average number of colonies for each sample
was found. This count was used to calculate CFU/sachet of
500 mg probiotic sample.
Species identification by 16S rDNA sequencing: For
each species, twelve colonies were randomly selected for
sequencing of 16S rDNA. 16S rDNA sequence (1500bp)
was amplified by colony PCR using universal forward
primer 8f (5'-GAGTTTGATCATGGCTCAG-3') and
reverse primer 1495r (5'-CTACGGCTACCTTGTTACG-
3').10 The PCR program was one cycle of initial
denaturation at 95°C for 5 min, 30 cycles of denaturation at
95°C for 30 s, annealing at 45 °C for 30 s and extension at
72°C for 60 s and a final extension cycle at 72°C for 5
min.2, 17 The PCR product was purified and sequenced with
70
Vol. 12 (12) December (2017)
Res. J. Biotech
primer 8f by automated DNA sequencing using Genetic
Analyzer 3130xl (Applied Biosystems, USA). The 16S
rDNA sequences obtained from the samples were analyzed
by using BLASTN tool of GenBank database
(www.blast.ncbi.nlm.nih.gov) for species identification.
Biochemical characterization of the strains: The colonies
selected for 16S rDNA sequencing were subjected to
biochemical characterization as per the Bergey’s Manual of
Systematic Bacteriology28 for verification purpose. The
isolates were subjected to series of biochemical tests which
included gram staining, endospore forming test, catalase
test, aerobic growth, Voges–Proskauer (VP) test, utilization
of glucose, glycogen, mannose, starch hydrolysis test,
casein hydrolysis test, citrate utilization test, growth at
different NaCl concentrations (2%, 5%, 6.5% NaCl) and
temperatures (20°C, 30°C, 40°C and 65°C).
Results and Discussion
Probiotic supplements provide beneficial effects in the host
by improving the intestinal microbial balance.11 There are
several probiotics in the market which are prescribed for
children and adults. Previous studies conducted in other
countries have shown that the commercial probiotics do not
often contain the organisms that are claimed to be present.
On the other hand, they may also contain organisms that
are not claimed to be present and some of which may be
even pathogenic.27 Therefore, we have taken up the current
study to evaluate three commercial probiotics that are sold
in India namely BIFILAC®, ViBact® and Pre-Pro®.
According to the label, BIFILAC® and ViBact® are a
mixture of four bacteria namely Clostridium butyricum,
Streptococcus faecalis, Bacillus mesentericus and
Lactobacillus sporogenes while Pre-Pro® contains L.
acidophilus in place of L. sporogenes. The main objective
of this study was to evaluate the contents of these
commercial probiotic products for the presence of the right
organism in right quantity as claimed in the label.
When the contents of BIFILAC® and ViBact® were grown
in plates containing respective medium, bacteria were
present in higher amount than what was claimed on the
label (table 1). Since there was no difference in colony
morphology, 12 colonies were randomly selected from each
selective medium and tested by 16S rDNA sequencing. The
colonies from the plates with CB and SF medium showed
100% nucleotide identity with C. butyricum and S. faecalis
as claimed in the label (table 2). However, the colonies
from the plates with BM medium showed only 96%
nucleotide identity with B. pumilus (Accession No.
JF732754) which is a synonym of B. mesentericus but
100% nucleotide identity with B. subtilis which was not
claimed to be present.
Biochemical analyses were done to verify if these colonies
represent B. subtilis. The colonies were gram positive, rod
shaped and positive for endospore
Research Journal of Biotechnology Vol. 12 (12) December (2017)
Res. J. Biotech

formation, catalase test, aerobic growth, Voges-Proskauer
(VP) test, starch hydrolysis, casein hydrolysis and citrate
utilization. They were able to grow at 2%, 5% and 6.5%
NaCl concentrations and at the temperatures of 20°C, 30°C
and 40°C. They were able to utilize glucose, glycogen and
mannose as carbon source (table 3). These biochemical
features confirmed the colonies from the plates with BM
medium as B. subtilis. B. pumilus (= B. mesentericus) and
B. subtilis are phylogenetically related in same group but
are not identical.26
Though a previous study has reported unclaimed
pathogenic organisms in probiotic products,27 B. subtilis is
not pathogenic. It is in fact a probiotic organism indicated
for oral bacteriotherapy and bacterioprophyl axis of
gastrointestinal disorders9 and is used in commercial
probiotic products such as Enterogermina and Biosubtyl.
However, this does mean that it can be added in any
probiotic product without testing it for indicated health
benefits and reflecting the same in the label.
The colonies from the plates with MRS medium that were
used for growing L. sporogenes of BIFILAC® and
ViBact® showed 100% nucleotide identity with B.
coagulans. Biochemical analyses were done to verify if
these colonies represent B. coagulans. The colonies were
gram positive, rod shaped, positive for endospore staining,
catalase test and starch hydrolysis test. They were negative
for casein hydrolysis test, citrate utilization test (table 2).
They were able to utilize glucose and mannose but not
glycogen as carbon source. They were not able to grow at
5% NaCl, 6.5% NaCl and at 65°C. These biochemical
features confirmed the colonies from the plates with MRS
medium as B. coagulans.
Neither 16S rDNA sequences in the GenBank database
(www.ncbi.nlm.nih.gov) nor biochemical data in the
Table 1
Details of the bacterial strains present in the three probiotic products (BIFILAC®, ViBact®, Pre-Pro®)
S. N. Product Organism mentioned in the
label
Clostridium butyricum
1 BIFILAC® Streptococcus faecalis
Bacillus mesentericus
Lactobacillus sporogenes
Clostridium butyricum
2 ViBact® Streptococcus faecalis
Bacillus mesentericus
Lactobacillus sporogenes
Clostridium butyricum
3 Pre-Pro® Streptococcus faecalis
Bacillus mesentericus
Lactobacillus acidophilus
71
Bergeys Manual of Systematic Bacteriology28 exists for L.
sporogenes. It shows characteristics of both Lactobacillus
and Bacillus and its taxonomic position between the
families Lactobacillaceae and Bacillaceae has often been
discussed.
A detailed study on spore forming probiotics21 has reported
that in some cases commercial products containing B.
coagulans use the invalid name L. sporogenes. Studies3,4
have also reported that L. sporogenes should be correctly
classified as Bacillus coagulans. Thus, from the above
evidence, L. sporogenes in BIFILAC® and ViBact®
should be renamed as B. coagulans.
When the contents of Pre-Pro® were grown in plates
containing respective medium, two species were found to
be present in much lower amount when compared to the
labeled claim and two species were completely absent
(table 1). 16S rDNA sequencing of the colonies from the
plates with CB and SF medium showed 100% nucleotide
identity with C. butyricum and S. faecalis as claimed in the
label. But their counts were only 0.4 million and 2.0
million as against 15 million CFU/sachet claimed in the
label which is 37.5 and 7.5 - fold less for C. butyricum and
S. faecalis respectively. This may be due to inadequate
quality control at the time of manufacturing and/or death
during storage.
Similar to BIFILAC® and ViBact®, both 16S rDNA
sequencing and biochemical characterization showed the
presence of B. subtilis instead of B. mesentericus in the
plates with BM medium. However, unlike BIFILAC® and
ViBact®, its colony count in Pre-Pro® was only 0.04
million CFU/sachet. Pre-Pro® did not contain L.
acidophilus as claimed in the label.
CFU/Sachet of 500g
Claimed in the label Present in the sample
(million) (million ± SD)
2 74.8 ± 0.6
30 124 ± 1.7
1 8.1 ± 0.3
50 66 ± 1.5
2 41.08 ± 0.8
30 416 ± 3.4
1 5.6 ± 0.3
50 51.73 ± 0.4
15 0.4 ± 0.3
15 2.0 ± 2.3
15 0.04 ± 0.03
15 0
Research Journal of Biotechnology Vol. 12 (12) December (2017)
Res. J. Biotech

Table 2
Details of the bacterial strains present in the three probiotic products (BIFILAC®, ViBact®, Pre-Pro®)
showing the nucleotide identity
S.N. Product Organism mentioned in the Organism detected in the sample using % nucleotide
label 16S rDNA sequencing identity

Clostridium butyricum Clostridium butyricum 100

Streptococcus faecalis Streptococcus faecalis 100
1 BIFILAC® Bacillus mesentericus Bacillus subtilis 100
Lactobacillus sporogenes Bacillus coagulans 100
Clostridium butyricum Clostridium butyricum 100
Streptococcus faecalis Streptococcus faecalis 100
2 ViBact® Bacillus mesentericus Bacillus subtilis 100
Lactobacillus sporogenes Bacillus coagulans 100
Clostridium butyricum Clostridium butyricum 100
Streptococcus faecalis Streptococcus faecalis 100
3 Pre-Pro® Bacillus mesentericus Bacillus subtilis 100
Lactobacillus acidophilus None -

Table 3
Biochemical characterization of the colonies that were identified to be Bacillus subtilis and Bacillus coagulans
based on 16S rDNA sequencing
S. N. Biochemical test Bacillus subtilis Bacillus coagulans
1 Endosperm forming test + +
2 Catalase test + +
3 Aerobic test + +
4 Voges-proskauer (VP) + -
5 Glucose utilization test + +
6 Glycogen utilization test + -
7 Mannose utilization test + +
8 Starch hydrolysis test + +
9 Casein hydrolysis test + -
10 Citrate utilization test + -
Growth at different NaCl
concentration:
11 2% + +
5% + -
6.5% + -
Growth at different temperature
ranges:
12 20°C + -
30°C + +
40°C + +
65°C - -

In summary, none of the probiotics that were analyzed in
the current study conformed to the labeled claims in terms
of species as well the number of live organisms to be
present. BIFILAC® and ViBact® contained only three of
72
the four bacteria of which one should be renamed correctly.
Pre-Pro® contained only two of the four bacteria that too in
significantly less quantity than what was claimed in the
label. All the three probiotics contained B. subtilis which
Research Journal of Biotechnology
was not claimed in the label. Presence of an unclaimed
organism cannot be taken as a substitution for the missing
organism because probiotic effects are specific even at
strain level25.
While the current study has evaluated the contents only at
the species level, efforts should be taken to evaluate the
contents at strain level as well. Strain identification is much
more challenging than species identification. Probably, the
manufacturers of the probiotics should provide the markers
for identification of their microorganisms at the strain level.
This would not only benefit the consumers and quality
control agencies but also the manufacturers to establish the
identity of their microorganisms in case of intellectual
property right violations.
Considering the data presented here and the fact that
awareness about and the use of probiotics is rapidly
increasing among all age groups, it is important to take
immediate steps to bring the products containing probiotic
organisms under proper legally binding regulations and
monitoring similar to that of drugs.
Conclusion
Our study clearly demonstrates mislabeling of the
organisms, absence of claimed organisms, presence of
unclaimed organisms and significantly less number of
certain claimed organisms in the probiotic products tested.
Thus, serious measures have to be taken to evaluate the
probiotic products commercially available in the market
using microbiological and molecular methods.
Acknowledgement
The authors wish to thank SRM University for constantly
providing financial support throughout the project.
References
1. Ambalam P., Raman M., Purama R.K. and Doble M.,
Probiotics, prebiotics and colorectal cancer prevention, Best
Pract. Res. Cl. Ga., 30, 119-131 (2016)
2. Cerritos R., Vinuesa P., Eguiarte L.E., Herrera-Estrella L.,
Alcaraz- Peraza L.D., Arvizu-Gimez J.L., Olmedo G., Ramirez
E., Siefert J.L. and Souza V., Bacillus coahuilensis sp. nov a
moderately halophilic species from a desiccation lagoon in the
Cuatro Cienegas Valley in Coahuila, Mexico, Int. J. Syst. Evol.
Microbiol., 58(4), 919–923 (2008)
3. De Vecchi E. and Drago L., Lactobacillus Sporogenes or
Bacillus Coagulans: misidentification or mislabelling?, Int. J.
Probiot. Prebiot., 1, 3–10 (2006)
4. Drago L., De Vecchi E., Nicola L., Colombo A. and Gismondo
M.R., Microbiologica evaluation of commercial probiotic
products available in Italy, J. Chemotherapy, 16, 436-467 (2004)
5. Duc H.L., Hong H.A., Barbosa T.M., Henriques A.O. and
Cutting S.M., Characterisation of Bacillus probiotics available for
Human Use, Appl. Environ. Microbiol., 70, 216-217 (2004)
73
Vol. 12 (12) December (2017)
Res. J. Biotech
6. Elliott E. and Teversham K., An evaluation of nine probiotics
available in South Africa, August 2003, S. Afr. Med., 94, 2–29
(2004)
7. FAO/WHO, Report of a joint FAO/WHO expert consultation
on guidelines for the evaluation of probiotics in food. London,
Ontario, Canada, World Health Organization and Food
Agriculture Organization of the United Nations (2002)
8. Fasoli S., Marzotto M., Rizzotti L., Rossi F., Dellaglio F. and
Torriani S., Bacteria composition of commercial probiotic
products as evaluated by PCR DGGE analysis, Int. J. Food
Microbiol., 82, 59–70 (2003)
9. Green D.H., Wakeley P.R., Page A., Barnes A., Baccigalupi L.,
Ricca E. and Cutting S.M., Characterization of two bacillus
probiotics, Appl. Environ. Microbiol., 65, 4288–4291 (1999)
10. Grifoni A., Bazzicalupo M., DiSerio C., Fancelli S. and Fani
R., Identification of Azospirillum strains by restriction fragment
length polymorphism of the 16S rDNA and of the histidine
operon, FEMS Microbiol. Lett., 127, 85–91 (1995)
11. Guarner F. and Schaafsma G.J., Probiotics, Int. J. Food
Microbiol., 39, 237–238 (1998)
12. Higuchi W., Muramatsu M., Dohmae S., Takano T., Isobe H.,
Yabe S., Da S., Baranovich T. and Yamamoto T., Identification
of probiotic lactobacilli used for animal feeds on the basis of 16S
ribosomal RNA gene sequence, Microbiol. Immunol., 52, 559–
563 (2008)
13. Jose N.M., Bunt C.R. and Hussain M.A., Comparison of
Microbiological and Probiotic Characteristics of Lactobacilli
Isolates from Dairy Food Products and Animal Rumen Contents,
Microorganisms, 3, 198–212 (2015)
14. Lata J., Jurankova J., Doubek J., Pribramska V., Fric P., Dite
P., Kolar M., Scheer P. and Kosakova D., Labelling and content
evaluation of commercial veterinary probiotics, Acta Vet. Brno.,
75, 139–144 (2006)
15. Lesbros-Pantoflickova D., Helicobacter pylori and probiotics,
J. Nutr., 137, 812–818 (2007)
16. McFarland L.V. and Dublin S., Meta-analysis of probiotics
for the treatment of irritable bowel syndrome, World J.
Gastroenterol., 14, 2650–2661 (2008)
17. Mohkam M., Nezafat N., Berenjian A., Mobasher M.A. and
Ghasemi Y., Identification of Bacillus Probiotics isolated from
soil rhizosphere using 16S rRNA, recA, rpoB Gene sequencing
and RAPD-PCR, Probiotics & Antimicro. Prot., 8, 8-18 (2016)
18. Ouwehand A.C., Antiallergic effects of probiotics, J. Nutr.,
137, 794–797 (2007)
19. Quigley E.M., Probiotics in irritable bowel syndrome: an
immunomodulatory strategy, J. Am. Coll. Nutr., 26, 684–690
(2007)
20. Rastall R.A., Gibson G.R., Gill H.S., Guarner F.,
Klaenhammer T.R., Pot B., Reid G., Rowland I.R. and Sanders
M.E., Modulation of the microbial ecology of the human colon by
Research Journal of Biotechnology
probiotics, prebiotics and synbiotics to enhance human health: an
overview of enabling science and potential applications, FEMS
Microbiol. Ecol., 52, 145– 152 (2005)
21. Sanders M.E., Morelli L. and Tompkins T.A., Sporeformers
as Human probiotics: Bacillus, Sporolactobacillus and
Brevibacillus, Compr. Rev. Food Sci. Food Saf., 2, 101–110
(2003)
22. Simmering R. and Blaut M., Pro- and prebiotics—the tasty
guardian angels, Appl. Microbiol. Biotechnol., 55, 19–28 (2001)
23. Singh T.P., Kaur G., Malik R.K., Schillinger U., Guigas C.
and Kapila S., Characterization of intestinal Lactobacillus reuteri
strains as potential probiotics, Probiotics & Antimicrob. Prot., 4,
47-58 (2012)
24. Spiller R., Review article: probiotics and prebiotics in irritable
bowel syndrome, Alim. Pharmacol, Ther., 28, 385–396 (2008)
74
View publication stats
Vol. 12 (12) December (2017)
Res. J. Biotech
25. Tambekar D.H. and Bhutada S.A., An evaluation of probiotic
potential of Lactobacillus sp. from milk of domestic animals and
commercial available probiotic preparations in prevention of
enteric bacterial infections, Recent Res. Sci. Technol., 2(10), 82–
88 (2010)
26. Wang W. and Sun M., Phylogenetic relationships between
Bacillus species and related genera inferred from 16S rDNA
sequences, Braz. J. Microbiol., 40, 505–521 (2009)
27. Weese J.S., Microbiologic evaluation of commercial
probiotics, J. Am. Vet. Med. Assoc., 220, 794–797 (2002)
28. Whitman W.B, Goodfellow M., Kämpfer P., Busse H.J.,
Trujillo M.E., Ludwig W. and Suzuki K.I., Bergey’s Manual of
Systematic Bacteriology, 2nd ed., vol. 5, parts A and B, Springer-
Verlag, New York, NY (2012).
(Received 30th June 2017, accepted 24th August 2017)