The outcome of present study enumerates that the skin microbiota is definitely important to the health and wellness of human being. The toluidin blue agar medium,where enterobacter species(Hs6) shown elevated level pathonogenesity due to the presence of exotoxin. The study also reveals that presence of haemolysin and coagulase enzymatic properties. Once they reach opportunities pathogenesity under stress condition. Additional study depects, how skin resedint microbes influence immune response as a primary defence mechanism against invading foreign bodies even though their are to many beneficiary from host, skin microbiota but One have to proper hygiene life style to maintain the balance of communsle microflora that prevents the colonization of pathogrnic bacteria the observation shown out of present study suggest not only the phenotyping of human skin microbiota but also effects skin microbiota on immune system they have wide rangeing systematic sequalae that are ripe for exploration in near future. Future work will need to investigate the context-dependent behaviour of resident microbes—how microbe–microbe interactions, host–microbe interactions, and strain-specific differences may govern a microbe’s tendency towards cooperation or aggression.

1. Presentation
on ;
PHENOTYPING
OF HUMAN EPIDERMAL
MICROBIOTA
Presented
By ;
Shivakumar V B
PS182934
Kuvempu university
Department Of Biotechnology,Shanakaragatta

2. ‘PHENOTYPING OF HUMAN
EPIDERMAL MICROBIOTA
Presented by ;
Shivakumar V B
PS182934
Dept. of biotechnology
Jnana sahyadri,kuvempu university,
shanakaragatta
Project guide;
Dr.Prabha R
Associate professor
Dept. of dairy science microbiology,
Dairy science college ,
Bengalore
Co-guide;
Dr.Riaz mahmood
Professor
Dept. of biotechnology,
Kuvempu university,
shankaragatta

3. INTRODUCTION
The skin is the human body’s
largest organ.
The primary role of the skin is to
serve as a physical barrier, protecting
our bodies from potential assault by
foreign organisms or toxic
substances.
colonized by a diverse milieu of
microorganisms, most of which are
harmless or even beneficial to their
host.

4. Skin microbiota
 The human body, which contains
about 1013 cells, routinely
harbours about 1014 bacteria.
 A diverse microbial flora is
associated with the skin and
mucous membranes of every
human being from shortly after
birth until death.
 skin is also an interface with the
outside environment and, as
such, is colonized by a diverse
collection of microorganisms
including bacteria, fungi and
viruses.
Skin Microbiome

5. Objectives ;
1
• To enumerate & isolate the microflora
of human epidermis (skin).
2
• To characterize the bacterial isolates
obtained from human epidermis.
3
• To screen the selected bacterial
isolates for special characteristics.

6. Methods Adopted ;
1.Collection of
human skin swab
samples
2.Serial dilution of
swab sample
3.Pouring the
medium to plates
incubate at 50°C
4.Isolation and
maintenance of
isolates of skin
swab
5.Identification of
the bacterial
isolates of skin
swab
6.Preliminary tests
7.Specific tests
8.Direct
Microscopic
Count(DMC)
9.bacterial isolates
for special
characteristics

7. 1) To enumerate & isolate the microflora of human epidermis (skin)
The hand swab samples (SWI, SWII & SWIII) collected
subjected for serial dilution pour plating using
A) TBC on SPCA B) Staph On MSA C) Coliforms on VRBA
Enumeration of total bacteria, coliform and staphylococci from epidermal swab samples
Broth culture of bacterial isolates obtained from swab of epidermi
Growth appeared in the broth tubes in the form of turbidity as pure isolates

8. 2) To characterize the bacterial isolates obtained from human epidermis
Phenotypic identification of staphylococcus isolates
• Simple staining
• Gram staining
• Catalase test
• Oxidative test
A)
Preliminary
Tests
• Aerobic growth
• Anaerobic growth
B)
SPECIFICT
(OF ) Test
CATALASE TEST
SIMPLE STAINING GRAM STAINING
OXIDASE TEST
OXIDATION &FERME(TION
(OF Test)

9. Table 01; Result of preliminary test and specific test
Tests
Bacterial isolate code
Hs1 Hs2 Hs3 Hs4 Hs5 Hs6
Preliminary tests
Simple staining Cocci, Cocci Cocci Cocci Cocci Rod
Gram staining + + + + + -
Catalase test + + + + + +
Oxidative test - - - - - -
Specific tests
OF test
Aerobic
growth
O+ O+ O+ O+ O+ O+
Anaerobic
growth
F+ F+ F+ F+ F+ F+
TSI H2S Test
Acid slant,
acid
butt
Acid slant,
acid
butt
Acid
slant
acid
butt
Acid
slant
acid
but
Acid slant
acid butt
Acid slant
acid
butt

10. 3) To screen the selected bacterial isolates for special
characteristics.
selected bacterial isolates
Blood serum media Toluidine blue agar media
Pour Pour
Enterobacter sp.Hs6 showed
presence of three exotoxins
indicating their pathogenicity
Staphylococcus sp. Hs1 &
Hs2 revealed the presence
of haemolysin & coagulase
investigated the role of skin bacterial isolates and few isolates showed production of
haemolysisn, coagulase and thermonuclease.

11. Table 02: Screening of bacterial isolates for special activity
Bacterial
isolate code
Haemolysin production on
blood agar
Coagulase activity
In blood serum
Thermonuclease
activity on TDNAse
agar
Hs6 clear zone indicating
complete haemolysis
Coagulation of serum once
culture is mixed with blood
serum
Pink colouration on
agar due to break
down of DNA.
Hs1 clear zone indicating
complete haemolysis
Coagulation of serum once
culture is mixed with blood
serum
No pink colourization
indicating no
Thermonuclease
activity.
Hs2 clear zone indicating
complete haemolysis
Coagulation of serum once
culture is mixed with blood
serum
No pink colourization
indicating no
Thermonuclease
activity.
i.e All The Three Isolates Showed Presence Of Three Exotoxins Indicating Their Pathogenicity

12. Conclusion ;
The outcome of present study enumerates that the skin
microbiota is definitely important to the health and wellness of human
being. The toluidin blue agar medium, where enterobacter species(Hs6)
shown elevated level pathonogenesity due to the presence of exotoxin.
The study also reveals that presence of haemolysin and coagulase
enzymatic properties. Once they reach opportunities pathogenesity
under stress condition.

13. REFERENCE ;
Byrad A.L., Belkaid L., and SegreJ.L., 2018. The human skin microbiome.
Nature Reviews Microbiology 16(3):157-165
Delhaes L, Monchy S, Frealle E, Hubans C, Salleron J, Leroy S, et al. 2012.The
airway microbiota in cystic fibrosis: a complex fungal and bacterial
community—implications.
Gao, Z.,Guillermo I. Perez-Perez, Yu Chen, Martin J. Blaser, 2010. Quantitation
of Major Human Cutaneous Bacterial and Fungal
Populations.teriology,J.Clinical Microbiology, DOI: 10.1128/JCM.00597-10
Grice E.A. and Segre, J.A. 2011. The skin microbiome. Nat. Rev. Microbiol.
9(4): 244–253