BIOACTI
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Cochin partial fulf
UNDER
RTMENT OF COCHIN
CAN AND ACTERIZA
Thes University fillment of
DOCTOR
MIC R THE FAC
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MARINE BIO SCHOOL N UNIVERSITY
KOCH De
MELANI ATION AN
sis submitt y of Scienc f the require
R OF PHIL in CROBIOLO ULTY OF M
by LSY WILS Reg. No: 418
OLOGY, MICRO OF MARINE S Y OF SCIENC HI- 682016, I ecember 20
N FROM ND EVALU
ted to ce and Tech
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OGY MARINE SCI
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OBIOLOGY AN SCIENCES E AND TECH INDIA
16
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To the Lord Almighty...
Professor Cochin University of Science and Technology Fine Arts Avenue, Kochi-16
Certificate
This is to certify that the thesis entitled “Bioactive Glucan and Melanin from Marine Yeasts: Characterization and Evaluation” is an authentic record of the research work carried out by Ms. Wilsy Wilson under my supervision and guidance in the Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Microbiology under the Faculty of Marine Sciences of Cochin University of Science and Technology, and no part thereof has been presented for the award of any other degree, diploma or associateship in any university. All the relevant corrections and modifications suggested by the audience during the pre-synopsis seminar and recommended by the Doctoral committee have been incorporated in the thesis.
Dr. Rosamma Philip (Supervising Guide) Kochi-16
December 2016
Declaration
I hereby declare that the thesis entitled “Bioactive Glucan and Melanin from Marine Yeasts: Characterization and Evaluation” is a genuine record
of research work done by me under the supervision and guidance of Dr. Rosamma Philip in the Department of Marine Biology, Microbiology and
Biochemistry, Cochin University of Science and Technology. The work presented in this thesis has not been submitted for any other degree or diploma earlier.
Wilsy Wilson Kochi-16
December 2016
It is generous su completion. M
I was Professor and express my de sincere gratit enough kind accessibility, appreciable.
student, rese During my c driving force been possibl
I gra of Marine Bi providing me
I wis coordinator, especially for express my h needed.
I am encourageme Dr. Aneykutty valuable advi
I than the departme of Marine Scie I tak Technology a and Senior R
my great pr upport durin My cherished s immensely b d Head of the eep sense of g tude and love dness, affecti
, coupled wi Mam, you ex earcher and s course of res e. Without yo
e. I am enoug atefully ackno
ology, Microb e all the help, sh to express National Cen r the shrimp heartfelt grati
very much th ent. I thank y Joseph, Dr. S ice, suggestion
nkfully ackno ent for their h ences, CUSAT ke this oppo
and the Univ Research Fello
ivilege to exp ng the cours
d ambition…
blessed with d e Department gratefulness to e towards you
on, motivatio th meticulou xceptionally i so on. You we search, at the
our support gh fortunate t
owledge Dr.
biology and B , support and s my deepest ntre for Aqua bioassays, th itude to the g
hankful to all Dr. Babu Ph S. Bijoy Nanda n and constan owledge all th help and assis T, for extendin ortunity to versity Grant
owships.
press sincere se of the pr
divine mercy w t as my resear o you mam. On u mam, for y on and bless us way of ap inspired a lot ere a real mod e dawn of ea and encoura to be called a C.K. Radhakr Biochemistry d encouragem gratitude and tic Animal he hroughout my genuine effor
the teachers hilip, Dr. A.V.
an, Dr. Sajeeva nt support.
he office staff, stance. I would ng the necessa
acknowledge Commission
e gratitude to resent thesis
who directed m rch guide. Th n this occasion your valuable
sings. Your p pproach to so t and enriche del to me and ach day you agement, this as your studen
rishnan, Profe y, for the eno ment.
d sincere than ealth for prov y work perio rt and timely
of the depart Saramma, D an T.P and Dr.
technical sta d like to than ary facilities.
e Cochin Un n for the fello
o each one o work for i
me to Dr. Ros e words are n n, I would like
guidance, en pleasant natu
olve a probl ed my overal d I learnt a lot were my ins s work woul
nt.
essor (Retd).
ough kindnes
nks to Dr. I.S.
viding all the od. Sir, I have help provide
tment for thei Dr. A. A. Moh
. Valsamma Jo
ff and non-tea k the library
niversity of owship awar
of you for the its successfu
samma Philip not enough to to express my ndless support ure, patience em is deeply ll growth as a t from you….
spiration and d never have
., Departmen ss offered and
. Bright Singh e lab facilities e no words to ed whenever
ir support and hamed Hatha oseph for thei
aching staff o staff of Schoo
Science and ded as Junio
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p, o y t, e, y a ..
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from Arabian Sea and Bay of Bengal. The marine yeast isolates used for my study were isolated by Dr. Reema.
Words are not enough to express my sincere gratitude to Dr. Swapna P. Antony, for teaching me the basics of molecular biology and taking care of me as a student of you.
Your valuable suggestions, encouragement, timeless help throughout my research and thorough revision of manuscript is deeply appreciable. Thanks a lot for your invaluable help and friendship….
I am much obliged to Dr. Jayesh Puthumana for his heartfelt support and timeless help all the way through my research and always being a helping hand whenever I needed.
My immense thanks to Ms. Sruthy K.S, for the invaluable help received for the molecular works and also for her concern, encouragement and support during various stages of research.
I am much thankful to Ms. Divya T. Babu for the help and assistance received in setting the aquarium tanks and the help rendered on all occasions.
My whole hearted thanks to Ms. Aiswarya Nair, Ms. Manomi S and Mr. Jayanath G for their timely help and support.
I sincerely acknowledge the assistance received from Dr. Douglas Lawman for NMR analysis of glucan and melanin samples for the accomplishment of research.
Heartfelt thanks to Dr.Sudhir N S for the timely help in WSSV inoculum preparation for bioassays.
I would like to express my sincere thanks to my seniors, Ms. Chaithanya E.R, Ms. Jimly C. Jacob, Ms. Ramya K.D, Dr. Deepthi Augustine, Dr. Naveen Satyan, Dr. Afsal V.V and Mr. Anil Kumar P.R. for their help and support.
I cherish the friendship of my fellow colleague Ms. Neema Job, and a cheerful group of juniors, Ms. Archana K., Ms. Bhavya Kachiprath, Ms. Dhanya Kesavan, Mr. Solly Solomon, Ms. Sephy Rose and Ms. Anju M V., for your timely help, support and joyful time.
My special thanks to all fellow colleagues and research fellows of Marine Biology, especially, Ms. Aneesa, Ms. Emilda Rosmine, Ms. Jesmi Youssaf, Ms. Deborah Alexander, Ms Reshma S, Ms. Susan, Dr. Nifty John, Dr. Divya P.S, Dr. Neethu C.S, Dr. Mujeeb Rahman, Ms. Ally, Ms. Bini, Ms. Thasnim, Ms. Melji, Ms. Soumya Krishnankutty, Mr. Rithin, Mr. Jabir T, Mr. Ajin and too many…. for your friendliness, help and support. I also would like to thank the research scholars of NCAAH for their help during my research.
blessing that helped me in fulfilling this effort. I humbly offer you with prayers, my thankfulness and admiration.
I am so much grateful to my parents-in-law, Mr. C.V Jose and Mrs. Rosily Jose for their concern, prayers and encouragement. I express my sincere thanks and deep sense of gratitude to you for the genuine interest, initiative and effort taken to continue my studies. My special thanks to my sister-in-law, brother-in-law and their family for the concern and help whenever I needed.
I would like to thank my uncles, aunts, cousins and family friends for their encouragement, prayers, help and support.
The abundant prayers and immense blessings of my parents Mr. T.D Wilson and Mrs. Alphonsa Wilson was my strength throughout my life. Your invaluable sacrifice and hard work is behind all the achievements of my life. I take this opportunity to express my deepest gratitude and immense thanks to them. I also thank my sisters for their love, prayers and support.
Finally, I must tribute to my husband Mr. Munna Chirayath, who has always been my pillar of support, my joy, my strength. Thank you so much for your love, care and constant encouragement. My research endeavor would not have fulfilled without your support and sacrifice.... I deeply appreciate the immense affection and sacrifice of my son Akhil and my daughter Ann mariya who have adjusted well in my strain.
I thank all the experimental animals (Penaeus monodon) who sacrificed their lives for my research work.
The Lord is my strength and my shield (Psalm 28:7). I bow my head and surrender my life to the God Almighty for granting me the strength, health and wisdom to overcome the hardships of research and empowering me for the fulfillment of this endeavor. I do believe that with God all things are possible.
Thank you so much one and all…….
Wilsy Wilson
Chapter 1 General introduction --- 1
Chapter 2 Screening of marine yeast glucans as immunostimulant in P. monodon post larvae --- 17-78 2.1 Introduction --- 17
2.1.1 Shrimp aquaculture --- 18
2.1.2 Diseases in shrimp aquaculture --- 19
2.1.3 Crustacean immune system --- 21
2.1.3.1 Haemocytes --- 23
2.1.3.2 Cellular immune responses --- 23
2.1.3.2.1 Phagocytosis --- 23
2.1.3.2.2 Nodule formation --- 24
2.1.3.2.3 Encapsulation --- 24
2.1.3.3 Humoral immune responses --- 25
2.1.3.3.1 The clotting system --- 25
2.1.3.3.2 Prophenoloxidase system (proPO system) --- 26
2.1.3.3.3 Antimicrobial peptides --- 27
2.1.3.3.4 Reactive oxygen compounds --- 28
2.1.3.3.5 Pattern recognition proteins (PRPs) --- 28
2.1.3.3.6 Lectins/Agglutinins --- 29
2.1.3.3.7 Peroxinectin --- 30
2.1.4 Disease control strategies in shrimp aquaculture --- 30
2.1.4.1 Use of chemicals and antibiotics --- 31
2.1.4.2 Vaccination --- 31
2.1.4.3 Probiotics --- 31
2.1.4.4 Use of immunostimulants --- 32
2.1.4.5 Bacterial cell wall components as immunostimulant --- 33
2.1.4.5.1 Peptidoglycan --- 33
2.1.4.5.2 Lipopolysaccharides --- 33
2.1.4.6 Glucan as an immunostimulant in aquaculture --- 34
2.2 Materials and Methods --- 37
2.2.1 Microorganisms used for glucan extraction --- 37
2.2.2 Glucan extraction --- 38
2.2.3 Structural characterization of the cell wall glucans --- 40
2.2.4 Determination of biocompatibility --- 40
2.2.5 Preparation of glucan incorporated diet --- 41
2.2.6 Experimental animals and rearing conditions --- 42
2.2.7 Experimental design and WSSV challenge --- 42
2.2.8 Gene expression analysis --- 43
2.2.8.1 Total RNA extraction --- 43
2.2.8.2 Reverse transcription --- 44
2.2.8.3 Semi-quantitative RT-PCR analysis --- 44
2.2.8.4 Agarose gel electrophoresis --- 44
2.2.8.5 Expression analysis of target genes --- 45
2.2.9 Statistical analysis --- 45
2.3 Results --- 47
2.3.1 NMR spectra and Glucan yield of different marine yeast isolates --- 47
2.3.2 Biocompatibility of yeast glucans --- 51
2.3.3 Estimation of the immunostimulant potential of yeast glucans --- 51
2.3.4 Expression profile of control genes in P. monodon post larvae in response to various marine yeast glucan and WSSV challenge --- 53
2.3.5 Expression profile of AMP genes in P. monodon post larvae in response to various marine yeast glucan and WSSV challenge --- 55
2.3.6 Expression profile of immune genes in P. monodon post larvae in response to various marine yeast glucans and WSSV challenge --- 59
post larvae in response to various marine yeast
glucans on challenge with WSSV --- 63
2.4 Discussion --- 68
Chapter 3 Characterization and evaluation of selected glucans as immunostimulants in adult P. monodon against WSSV challenge --- 79-116 3.1 Introduction --- 79
3.2 Materials and Methods --- 85
3.2.1 Microorganisms used as source of glucans --- 85
3.2.2 Glucan extraction --- 86
3.2.3 Structural characterization of the cell wall glucan --- 86
3.2.4 Preparation of glucan incorporated feed --- 87
3.2.5 Experimental animals and rearing conditions --- 87
3.2.6 Virus inoculum preparation --- 88
3.2.7 Determination of WSSV titer --- 89
3.2.8 Experimental design and WSSV challenge --- 89
3.2.9 Hemolymph collection --- 90
3.2.10 Gene expression analysis --- 90
3.2.10.1 Total RNA isolation and reverse transcription --- 90
3.2.10.2 Quantitative Real Time PCR analysis of gene expression --- 91
3.2.11 Statistical analysis --- 91
3.3 Results --- 92
3.3.1 Glucans from marine yeast isolates --- 92
3.3.2 Comparison of the immunostimulatory efficacy of the yeast glucans --- 97
3.3.3 Expression of immune genes in haemocytes of P. monodon in response to marine yeast glucan and WSSV challenge --- 98
3.4 Discussion --- 105
from a marine black yeast, Hortaea werneckii --- 117-165
4.1 Introduction --- 118
4.1.1 Sources of melanin --- 120
4.1.2 Fungus as a source of melanin --- 122
4.1.3 Enzymes for microbial melanin synthesis --- 124
4.1.4 Types of melanin --- 126
4.1.5 Characterization of melanin --- 126
4.1.6 Functions of melanin --- 127
4.1.6.1 Melanin as a radioprotectant --- 127
4.1.6.2 Melanin as a factor of virulence --- 129
4.1.6.3 Melanin as antioxidants --- 129
4.2 Materials and Methods --- 132
4.2.1 Microorganism used for melanin production --- 132
4.2.2 Extraction of melanin --- 132
4.2.3 Characterization of black yeast melanin --- 133
4.2.3.1 Solubility --- 133
4.2.3.2 UV-VIS spectra of black yeast melanin --- 133
4.2.3.3 Nuclear Magnetic Resonance (NMR) Spectroscopy --- 133
4.2.3.4 Fourier Transform Infrared Spectroscopy (FTIR) --- 134
4.2.3.5 Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) --- 134
4.2.3.6 Scanning Electron Microscope - Energy Dispersive Spectrometer analysis (SEM- EDS) --- 135
4.2.3.7 Elemental analysis of R23 melanin (CHNS analysis) --- 135
4.2.4 Assays for antioxidant activity --- 135
4.2.4.1 ABTS radical scavenging assay --- 136
4.2.4.2 DPPH radical scavenging assay --- 136
4.2.5 Biocompatibility and cytotoxicity of R23 melanin --- 135
4.2.5.1 2, 3-Bis [2-Methoxy -4-Nitro-5-Sulfophenyl]- 2H-Tetrazolium-5- Carboxanilide (XTT) assay --- 138
4.2.5.2 Sulforhodamine B (SRB) assay --- 139
4.2.6 Phototoxicity assay of R23 melanin --- 140
4.2.7 Statistical analysis --- 141
4.3 Results --- 141
4.3.1 Production and extraction of melanin from black yeast ---141
4.3.2 Physicochemical characterization of black yeast melanin ---142
4.3.2.1 Solubility of black yeast melanin --- 142
4.3.2.2 UV-VIS spectra of R23 melanin --- 142
4.3.2.3 Fourier Transform Infrared Spectroscopy (FTIR) --- 143
4.3.2.4 Nuclear Magnetic Resonance (NMR) Spectra of melanin --- 144
4.3.2.5 Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) --- 145
4.3.2.6 Elemental analysis of melanin (CHNS analysis) --- 146
4.3.2.7 SEM-EDS analysis of melanin --- 146
4.3.3 Antioxidant activity --- 148
4.3.3.1 ABTS radical scavenging assay --- 148
4.3.3.2 DPPH radical scavenging assay --- 149
4.3.3.3 Lipid peroxidation inhibition assay --- 150
4.3.4 Biocompatibility and cytotoxicity of R23 melanin --- 151
4.3.5 Phototoxicity assay of R23 melanin --- 152
4.4 Discussion --- 154
Chapter 5 Evaluation of marine yeast glucan and melanin as anticancer agents by in vitro gene expression analysis --- 167-206 5.1 Introduction --- 167
5.1.1 Anticancer agents from plant sources --- 169
5.1.3 Anticancer agents from marine sources --- 172
5.1.4 Genes in cancer regulation --- 175
5.2 Materials and methods --- 180
5.2.1 Microorganisms used for the study --- 180
5.2.2 Preparation of marine yeast glucan compound --- 180
5.2.3 Preparation of marine yeast melanin compound --- 181
5.2.4 Cell lines and testing anticancer activity in vitro --- 182
5.2.5 Gene expression analysis --- 182
5.2.5.1 Total RNA isolation and reverse transcription --- 182
5.2.5.2 Real-time quantitative PCR analysis of target gene expression --- 184
5.3 Results --- 186
5.3.1 Effect of marine yeast β-D-glucan on the expression of cancer-related genes in lung cancer cells ---186
5.3.2 Effect of marine yeast melanin on the expression of cancer-related genes in lung cancer cells --- 189
5.4 Discussion --- 192
Chapter 6 Summary and Conclusion --- 207-212 References --- 213-263 Appendix I - GenBank Submissions --- 265
Appendix II - List of Publications --- 266-277
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become an have been nical marine edicine. The hniques have nisms and i omano et al.
re one of the otentials. To roorganisms e secondary awat, 2005
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n n e e e it ., e o s y
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Yea either prim formation.
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sts are pred tely 1,500 sp ycetes and being char d yeast biod
solated from ey can grow nd Inacio, 20 rine yeasts, nts, are able water (Chi et
011). The p marine yeas active potent karyotic, n udding and t
yeast cells e commonl on age and re unicellula
the forma hae or false varying colo s similar to molecular an
mical, genet dominant me pecies have the basidio racterized in diversity on m different s w as symbi 006; Xu et a defined a to grow bet t al., 2010).
present stud sts, which i tial over the on-motile, transverse d s are larger y spherical d environme
ar, some sp tion of stri e hyphae, a our ranging that of high nd cell biol
tic and mole embers of th been descri omycetes ( n a regular n Earth. Ye
sources such iotic or par al., 2011; Yu
s the yeast tter on medi The first m
dy is exclu s still an un eir terrestrial unicellular division or r than bact , ovoid or e ent (Waym pecies of ye ings of con as seen in m from yellow her eukaryo logy due to ecular biolo he fungal ec ibed which (Kurtzman
manner an easts are ub
h as soil, wa rasitic in as
urkov et al., ts that are ium prepare marine yeasts
usively focu nderexploite l counterpar fungi that sexually thr eria and th elongated in man and Par
ast forms m nnected bu most molds w to black. Y otic cells. It o their ease ogical analy cosystem an
are distribu and Fell, 2 nd there is c biquitous in ater, plants, ssociation w
, 2012).
isolated fr ed using sea s were isolat
used on the ed microbia rts.
t reproduce rough spore he size vary n shape and rekh, 1990) may become udding cells s. Yeast can Yeasts have is used as a e of growth yses.
nd at presen uted between
2006). New considerable n nature and animals and with animals
from marine awater rather ted from the e al
e e y d ).
e s n e a h,
nt n w e d d s
e r e
Atlantic Oc identified a yeasts wer seawater, se grouped as marine or terrestrial h (Kohlmeyer
Diff depending o For deep Researchers of marine y medium for seawater n dextrose ag researchers 1999; Loure of the medi water. Ant commonly growth of m Bengal, dich
Gen morphology techniques advancemen
cean by Fi as Torula sp re isolated eaweeds, inv obligate ma estuarine h habitats but a r and Kohlm ferent samp
on the locat sea sample s use a num yeasts. Wick
r the isolatio nutrient aga gar medium
for the isola eiro et al., 2 ia preparatio tibiotics suc added in the moulds, var hloran and p nerally, yea
y, physiolog have been nt in molecu
ischer in 1 p. and Myco
from diffe vertebrates, arine yeasts habitat whe are able to g meyer, 1979
lers have b tions and de e collection mber of medi
kerham’s ye on of marine ar medium,
and YPD ation of mar 005; Masud ons, compon ch as chlo
e medium t rious inhibit propionate (J ast species gical and bio
applied to ular biology
894. Those oderma sp.
erent sourc , seabirds, m s are those w ereas facult
grow and sp 9).
been design epth of the n, research ia with diffe east malt me e yeasts. M
YM agar agar medium rine yeasts (W da et al., 200 nents were s ramphenico o inhibit the tors have b Jarvis, 1973 were phe ochemical an o differentia
and emerge
e were red respectively es such as marine fishe
which grow tatives are porulate in t
ned for the sea (Dorsch h submarine erent compo edium is th oreover, ma r medium,
m are some Wickerham, 08; Sarlin an suspended in ol, streptom e growth of been added 3; King et al.
enotypically nalysis. Rece ate or ident ence of new
and white y. Subseque s estuaries,
s, mammals and sporula those origi the marine e
collection hel, 2007; S es can als ositions for e most com alt extract ag modified e of the me , 1951; Naga nd Philip, 20
n natural or mycin or pe
f bacteria. T to the med ., 1979; Kutt y described ently, variou tify yeast s
methods ha
yeasts and ently marine sediments s etc. Yeasts ate solely in inated from environmen
of samples ingh, 2011) o be used the isolation mmonly used gar medium
Sabauroud’s edia used by ahama et al.
011). In mos artificial sea enicillin are o inhibit the dia viz., rose
ty, 2009).
d based on us molecular species. The ave made the d e s, s n m nt
s ).
d.
n d m, s y ., st a e e e
n ar e e
D
A
H
identificatio identificatio imparts mor strains (Rec
The specific gen genera, suc Rhodotorul black yeast 1963; Godi 2001; Gada 2010; Koni Debaryomy among this distribution
The Branco (19 Trough, of Aureobasid Adriatic co Hortaea we halotolerant et al., 200 werneckii i 2012; Kutty pigment me to coastal w
on and ch on methods re powerful cek et al., 20 previous st nus or grou ch as Cand la, Saccharo ts (Bhat and
nho et al., 1 anho and S ishi et al., 2 yces and Rh
Candida, D n.
presence of 963) from ff San Die dium pullula oast (Zalar e
erneckii wer t eukaryotic 5; Cantrell n Arabian y et al., 20 elanin. The waters of the
aracterizatio , genetic ch l means of s 002).
tudy report up, but are r dida, Crypto omyces, Tri d Kachwalla 1978; Kohlm Sampaio, 20
2010; Galk hodotorula Debaryomyc
f black yeas Pacific sub ego. Hortae ans are the b
et al., 1999 re isolated f c model org et al., 20 Sea and Ba 013a) and t
distribution e Atlantic, P
on more s haracterizati strain identif
states that represented ococcus, D ichosporon, a, 1955; Fel meyer and K 005; Kutty
iewicz et a are the mos ces and Rho
st has been r b surface w
ea werneck black yeast s 9). Moreove from the hyp
anism (Gun 06). The p ay of Benga these isolate n studies of
acific and In
systematic.
ion through fication and
marine yea by a wide Debaryomyce Torulopsis ll et al., 196 Kohlmeyer,
and Philip, al., 2012). C
st frequentl odotorula ex
reported by waters and kii, Phaeot strains isolat er, most of persaline wa nde-Cimerm presence of al have bee es were cap
marine yeas ndian Ocean
Instead of h molecular differentiat
asts do not variety of es, Pichia, s, Sporobol 60; Fell and
1979; Naga 2008; Bur Candida, Cr ly observed xhibited a co
Van Uden a deep water theca triang
ted from sal the black y aters and it man et al., 20
f black yea en reported
pable of pr sts are most ns.
f traditiona r techniques tion amongs
belong to a well-known Hansenula lomyces and d van Uden ahama et al.
rgaud et al.
ryptococcus genera and osmopolitan
and Castelo rs of Loma gularis and lt pans at the yeast strains is known as 000; Butinar ast, Hortaea (Kuriakose oducing the tly restricted al
s st
a n a, d n, ., ., s, d n
- a d e s s r a e, e d
Mar sources of Cragg, 2004 been done t are promis temperature microorgan biomolecule biotechnolo has been r substances and vitamin and chemic protection ( most of th research ha Sinc production alcohol dist (SCP) whic and its poss ability of y alkaline pro were exten Martini, 20 Chi et al., 2 application
rine organis enormous n 4; Lordan e to explore t ing. The e e, salinity, p nisms comp es. The biom ogical poten
reported tha such as am ns with pot cal industri (Chi et al., 2 e terrestrial s been exten ce ancient tim
of alcohol tillation and ch is highly sible utiliza yeasts to pr otease, acid sively studi 002; Gupta 2007; Bhadr in food,
sms have number of b et al., 2011;
the bioactive extreme env pH and man
prising ma molecules p ntial and its at marine y mino acids, g
tential appli ies as well 2009; Sarkar lly isolated nded to the m mes, yeasts
and carbon d baking ind substantial ation as feed
roduce diffe protease, ce ied (Ogrydz et al., 2003 ra et al., 20 feed, pha
considerabl biologically Chen et al.
e potential o vironmental ny other fact arine yeasts produced by
s use as cos yeasts are glucans, glu ication in th l as for m r et al., 2010
bioactive marine envi were used in
dioxide, w dustries. Yea
in aquacultu d for anima erent extrac ellulase, phy ziak, 1993;
3; Vakhlu an 008;). Most armaceutical
le importan y active com
, 2016). Ext of marine m l conditions
tors have be s to produ these organ smeceutical
able to pr utathione, to he food, ph marine cultu 0). Due to th
compounds ironment.
n many indu hich are im ast, also use ure owing t als or in aqu cellular enzy ytase, lipase,
Pandey et nd Kour, 20 of the enzy l, maricultu
nce as new mpounds (N tensive rese microbes and
s such as een fortified uce potenti nisms have g
s and nutra roduce man oxins, enzym harmaceutica ure and en
he exhauste s appear rep
ustrial proce mportant to t ed as single to the nutriti uaculture. S ymes such
, xylanase a al., 2001; B 006; Rocha ymes may fi ural and f
w promising Newman and earches have d the results variation in d the marine ially active gained grea aceuticals. I ny bioactive mes, phytase al, cosmetic nvironmenta
ed resources petitive and
esses such as the brewing
cell protein ional quality imilarly, the as amylase and inulinase
Buzzini and et al., 2006 ind potentia fermentation
g d e s n e e at It e e c al s, d
s g, n y e e, e d 6;
al n
industries. F as thiamin pyridoxine, one of the b been notice Ergosterol, important p main materi extracted fr et al., 2011) Sprenger et acts as an an most of its Pohanka, 20 Kille yeast strain Cryptococc Metschniko Zygosaccha producing k 2004; Oro, yeast strain Metschniko pathogenic expansum, vitro and in
Furthermore ne, pantothe choline, glu biomolecule ed in Debary
another be harmaceutic ial to produc om several ). Productio t al. (1999).
ntioxidant. T effect sugg 011; Rodrigu er toxins are ns which cus, Debary owia, Pichia aromyces a killer toxins 2014; Meh ns belonging owia pulch
fungi such P. digitatum n vivo wound
e, yeast cells enic acid, utathione, fo which invo yomyces (T eneficial bio cal intermed ce sterol dru yeasts (You on of melato It is known The diverse gests a clea uez-Naranjo e one of the can be us yomyces, H a, Saccharo are some o s (Young a lomakulu et g to Cryptoc
errima and h as Botryt
m and Mon d assay (Oro
s are good so riboflavin, olic acid and olves in many
ovar et al., omolecule w diate, precurs ugs such as c ung et al., 20 onin in yeast
n as a regul functions o ar neurohorm
o et al., 2012 e remarkabl sed as nat Hanseniaspo omyces, Us
of the ide and Yagiu, t al., 2014).
coccus albi d Aureoba tis sp., Asp nilia fructico
o, 2014). Ye
ources of a n vitamin B d p-amino be ny of the cell 2002; Reye with varying sor of vitam cortisone an 003; He et a t S. cerevisia lator of circ of melatonin
monal activ 2).
le compoun tural bioco ora, Hanse tilago, Toru ntified yea
1978; Staib Furthermo idus, Wicker asidium pu pergillus ca ola were al easts or its m
number of vi B6, and vi
enzoic acid.
lular functio es-Becerril e g functions min D2 and c
nd progestero al., 2007; G ae has been cadian rhyth have been r vity (Jacob e
nds produced ntrol agent enula, Kluy ulopsis, Wi ast genera b, 1999; Pfe re, antifung rhamomyces ullulans ag arbonarius, lso reported metabolites
itamins such itamin B12
Polyamines ons also have
et al., 2011) such as an cortisone, the one was also omez-Lopez n reported by hms and also reported and et al., 2002
d by severa t. Candida yveromyces illiopsis and
capable o feiffer et al.
gal effects o s anomalus gainst some
Penicillium d through in can be used h 2, s, e ).
n e o z y o d 2;
al a, s, d f ., f s, e m n d
as biocontro pre- and po
The bioactive d made up of The cell w extraction.
health indu modifiers (B of the imm infections.
antigenotox application β-Gl wall (~50-6 in animals.
The first r (Chihara et including an et al., 2005 the immun against pat inflammato Geastrum s vitro cytoto cancer cells extracted fr
ol agents an st-harvest d cell wall p due to the d
f glucans, m wall polysac
These poly ustries. The BRMs) due mune system Kogan et xic activitie in anticanc lucans are h 60 %) and a
A variety reported ma
al., 1970).
ntifungal, an ) and chole ity towards thogens tha ory, antioxid
saccatum m oxic study o s exhibited s rom various
nd it will be decay of man polysacchari
diverse func mannans, ma ccharides ca ysaccharides ese polysac e to their ab m against tu al. (2008) s of the yea cer preventio homopolysa are potent st of biologic ajor functio
Many other ntiinfection sterol reduc s various di at are alre dant and cyt mushroom ha
of β-glucan strong antic s mushroom
a valid alte ny fruits.
ides of yeas ctions/appli annoprotein an be separ
s have vario ccharides ar bility to trigg umor cells, i
reported th ast polysacc on/therapy.
accharides o timulators o cal functions
on of β-glu r biological
(Onderdon ction (Wolev
iseases (Ho eady active
totoxic activ ave also be
from Maita cancer effect ms exhibited
ernative to fu
st also have ications. In ns and mino rated from
ous applicat re known a ger a usuall inflammatio he antioxida charides and
of glucose, p of nonspecif
s of β-gluca ucans was activities h nk et al., 199 ver et al., 20 ofer and Po
in human vities of β-g en reported ake mushro t (Fullerton d a positive
fungicides in
e been dem yeast, the or compone each other tions in foo as biologic ly non-speci
ons, viral a ant, antimu d suggest th
present in th fic defense m
an have be its antitum have also be
92), radiopro 011). Yeast ospisil, 2011
ns and anim glucan-rich e d (Dore et a
oom in hum et al., 2000 impact aga
n controlling
onstrated as cell wall is ent of chitin
by alkaline od, feed and cal response ific reaction nd bacteria utagenic and
eir potentia
he yeast cel mechanisms en reported mor activity een reported
otective (Gu ts strengthen 1) and figh
mals. Anti extract from al., 2007). In man prostatic 0). β-glucans ainst a range g
s s n.
e d e n al d al
ll s d.
y d;
u n ht - m
n c s e
of cancers Due to its e can prevent screens, oil (1993) repo in vaccines In re have attract For instanc multiple b immunostim special atten threat by va increases t responses, b has been rep enhance d immunostim
Aqu various viru shellfish ag immunomo been studi immunostim 2011c). Va (THC), ph phosphatase
(Okamura e emulsion-sta t skin injurie s and gels ( orted that ye against furu ecent years,
ted much a ce, polysacc biological mulating, an
ntion in aqu arious patho the resistan
but by enha ported that disease resi
mulation.
uaculture is uses and ba gainst bacter odulator (Ch ied by pre mulant (Saje arious imm henoloxidase
e activity an
et al., 1986 abilizing eff es caused by (Mason, 200 east glucan a unculosis in , bioactive p attention in charides or activities ntioxidant uaculture sec
ogens. Subs nce of the
ancing nons polysacchar istance in
facing sev acteria. Gluc rial or viral hang et al., 2
evious rese eevan et al., munological
e (PO) ac nd acid pho
6; Kimura e ffects and an
y solar radia 01; Michiko also showed n Atlantic sa
polysacchar the field of their glyco
including etc. Simila ctor as an im stances used
host, not specific defe rides from y shrimp, a
vere threat can enhance l infection a 2003). Marin
earchers an , 2009; Suk
parameters ctivity, res osphatase ac
et al., 1994;
ntioxidant a ation and th o and Yutak d an adjuvan almon (Salm
rides isolate f biochemis conjugates
anticarcino arly, polysa
mmunostim d for immun
by enhanc fense mecha
yeasts also c a process
due to the es the defen and thus act ne yeast der nd proved kumaran et a s such as t spiratory bu ctivity have
Kodama e activity, yea herefore are ka, 2007). R nt effect wh mo salar L.).
ed from natu stry and pha were show ogenic, an accharides h mulant agains nostimulatio cing specif anisms (Sak confer prote often refe
e continuou nse potential ts as immun rived glucan d its poten al., 2010; A
total haemo urst activit
been used t
et al., 2002) ast β-glucans
used in sun Rorstad et al hen included
ural sources armacology wn to exhibi nticoagulant have gained
st the severe on therefore fic memory kai, 1999). I
ection and/or erred to as
us attack o l of fish and nostimulant ns have also ntial as an Antony et al.
ocyte coun ty, alkaline to detect the ).
s - l.
d
s y.
it t, d e e y It r s
f d t/
o n ., nt e e
immunolog aquaculture significantly serious risk S169, Cand of the mar aquaculture Subramania of attentio chemothera In ad the yeast ce important r D-mannose glucose, gal supplement immune sy polysacchar gut health ( Huu and Jo growth of b also used as have been p such as imm and binding
In th not only du
gical effect e. Earlier
y enhance t k to aquacu dida tropica rine yeasts e (Sajeevan
an and Phili on as a apeutics in th
ddition to gl ell wall. Yea role in vari , but some lactose and s and provi ystem of th rides in feed
Ponton et al ones, 2014).
beneficial ba s adjuncts fo proposed to p munomodul g of toxins an
he same wa ue to the abil
of glucan studies rep the survival ulture indus alis S186, C used for et al., 2009;
ip, 2013). T valuable a he fight aga lucans; mann ast derived m ious biologi
etimes cont xylose. Man ide a range he animal,
d helps to m l., 2001; Gri
Mannan c acteria in the or fish and a promote ani lation, oxida nd pathogen ay, natural p lity to incre
n against port that g l of shrimps try. Debary C. haemulon the study
; Sukumaran he use of im alternate to ainst infectio
nans and ma mannans/ma
ical activity tain minor nnans have b of benefici growth and maintain a h
isdale-Hella can also fun e gut. These animal feed imal growth ative status, ns (Kurtzman
pigments ha ase the mar
bacterial o glucan inco s against W yomyces ha nii S27 and of glucan n et al., 201 mmunostimu
o the use ous diseases annoprotein annan-oligos y. These are
saccharide been added ial effects. I d survival.
healthy gut f and et al., 20 nction as pre e yeast cell w ds. In genera h and health b
, interaction n et al., 201 ave also bec rketability o
or viral in orporated f WSSV attack ansenii S8, C. sake S16 and its ap 0; Antony e ulants have e of antib s in aquacult ns represent ~
saccharides e composed e componen in many of It helps to Incorporatio flora and th 008; Refstie ebiotic by fa wall polysac al, these poly
by various m ns with gut
1).
come a cho f products;
nfections in feeds could k, one of the D. hanseni 65 are some pplication in et al., 2011c
gained a lo biotics and ture sector.
~ 30-40% o (MOS) have d mainly o nts such as f animal feed improve the on of these hus promotes et al., 2010 avouring the ccharides are ysaccharides mechanisms
constituents
ice of study but also due n d e ii e n c;
ot d
f e
f s
d e e s 0;
e e s s, s
y e
to advantag Melanins a preparation for the prod industries.
Yea property; p point of vi yellow to b common pi the pigmen frequently a function ag and extrem an importa preparations 2009), free r effects (El- melanin.
Mela microorgan Pathogenic immune sys brasiliensis pathogenic produce me source of m
geous biolo are frequen ns. Earlier st
duction of v
sts are able igmented y iew. It prod bright orang igments pro nted yeasts b
associated w gainst damag e climate co ant candid s. Photoprot radical-scav -Obeid et a
anin is an isms. Melan yeasts pro stem. Crypt and Wang yeasts (da elanin and a melanin prod
ogical activ ntly used i tudies revea various pigm
e to produc yeasts are an
duces differ ge, red, pin duced by ye belong to a with the cell ges caused onditions. T date in ma tection (Par venging (Roz al., 2006) a
n ubiquitou nin producin duce melan tococcus neo giella derm
Silva et al., apart from te
ducing yeas
vities as an in medicine aled that, yea ments and it
ce varying h n interesting rent colours nk and dark easts compr ascomycota wall of yea by ultravio These protec any of th ramonov et zanowska et are some of
us pigment ng yeasts are nin as a pr
oformans, C atitidis are , 2009). Ter errestrial ha st and most
ntioxidants e, pharmac asts are also t may find p
hues of pig g group fro s of pigme k brown or
rise melanin and basidi asts, which c olet (UV) li ctive effects e pharmac al., 2002), a t al., 1999) a f the biolog
t found in e generally k rotective me Candida alb
some of t rrestrial fun abitat, marin t of them as
and antican cology, and o a promisin potential app
gments and om the biote ents ranging even black n and carote omycota. M can provide
ght, oxidan of melanin cology and antioxidatio and immuno gical effects
n plants, a known as ‘b easure again
icans, Para the melanin ngus has bee ne environm
ssociated w
ncer agents d cosmetics ng candidate plications in
due to this echnologica g from ligh k. The mos ene. Most o Melanins are
a protective nts, radiation n make them d cosmetics n (Tu et al.
omodulatory s of natura
animals and black yeasts’
nst the hos coccidioides n producing en shown to ment is also a ith high sal
s.
s e n
s al ht st f e e n m s ., y al
d
’.
st s g o a lt
concentratio triangularis halophilic f marine blac species in h Cimerman a Caro colours of f colour of th nutritional of function antioxidant have antic aquaculture their antiox stressors (C 2011). Rec AY-01 with (Yoo et al., been isolate Rhodotorula al., 2001).
capacity to biological a Cont is a relevan pathogens a
on. The asc s, Trimmat fungi that i ck yeasts, H hypersaline and Plemeni otenoid is
fruits, flowe his pigmen supplement s, especially ts. In vitro cancer prop e animal he xidant defen Chien et al., cently, caro h significant , 2016). Car ed and mo a, Rhodospo Former stud increase the activities suc
tinuous outb ant problem are various
comycetous tostroma s inhabit hyp H. wernecki waters on itas, 2006).
a pigment ers, birds, in nt make it a
ts. Apart fro y in relation cell cultur perties (Fr ealth, carot nse ability a
, 2003; Pan otenoid pigm
t antioxidant rotenoid pro ostly they b oridium, Spo dies have re e marketabi ch as antioxi breaks of dis m in aquacu viruses and
black yeast salinum an persaline wa
ii has been three contin
responsible nsects and m a colourant om this, car
n to human re experime raser and
enoids play and resistan n et al., 200 ment from t and antiba oducing yea belong to b
oridiobolus eported that lity of produ idant and ant
eases and th ulture. Main d bacteria.
ts, Hortaea nd Aureoba ater of sola identified a nents (Koge
e for most marine inve
in many o otenoids als
health and ents have s Bramley, y an impor nce to disea 03; Amar et yeast Rhod cterial activ asts from ma basidiomyce
and Sporob natural pigm ducts, but als
ticancer pro he constant u nstream dise
Bacteria su
werneckii, asidium pu ar salterns.
as the dom ej et al., 20
of the ye ertebrates. T of the foods so have exte their role a hown that 2004). Si rtant role in ases and env t al., 2004;
dotorula m vities have b arine enviro etous classe bolomyces (N
ments not on so exhibit a operties.
use of chemo ease causin uch as Vibri
Phaeotheca ullulans are Among the inant funga 004; Gunde
llow to red The pleasan
s, feeds and ensive array as biologica
carotenoids imilarly, in n enhancing vironmenta Yang et al.
mucilaginosa een reported onment have es, including Nagahama e
nly have the dvantageous
otherapeutics ng infectious io vulnificus a e e al -
d nt d y al s n g al , a d e g et e s
s s s
(Sung et al alginolyticu harveyi (Am (WSSV) (C Taura synd hematopoie myonecrosi causing age most alarm worldwide Different str have been Administrat been consid use of immu use of antib farming (Su The paved the w cancer. Eve hallmark is in increased through apo involved in cancer effec concern of Espinosa-A
l., 1994), V.
us (Wang an mparyup et Chang et al.
drome virus etic necrosi
is virus (IM ents in aquac ming shrimp shrimp cult rategies suc proposed fo tion of imm dered as a p unostimulan biotics and v ubasinghe, 1 e emergence way for the t en though m
the loss of b d cell repli optosis (Han n the apopto cts of extrac f many of t Aguirre, 2004
V. campbellii nd Chen, 20 al., 2012) a ., 2003; Saj (TSV) (So s virus (IH MNV) (Nune
culture. Amo pathogens, ture industry ch as the use
or the contr munostimula potential mo nts has gaine
vaccines in 997).
e of new bio treatment of many factors balance betw cation and nahan and W osis, an acti
cts from va the research 4; Park et
i, V. proteo 005), Aerom and viruses
jeevan et al ong et al., HHNV) (Y es, 2010) ar ong the dive , and it has y (Takahash e of probioti rol and prev ants especial ode of protec
ed lot of atte the fight ag
oactive com f many of the s contribute
ween cell pr the failure Weinberg, 20 ive form of arious natura
h study (Le al., 2006; B
lyticus (Ma monas veron such as wh l., 2009; Su
2003), infe Yeh et al.,
re some of t erse pathoge s caused sev hi et al., 199 ics, vaccines vention of d
lly glucan i ction agains ention as a v gainst infect
mpounds fro e life threate
to the deve roliferation
of damage 011). Netwo f cell suicid al sources h eone et al., Bachmeier e
arques et al nii (Lin et a ite spot syn ukumaran e ectious hypo
2009) and the problem ens, WSSV
vere devast 94; Wang e s and immun diseases in in shrimp cu st WSSV in valuable sub tious disease
om natural s ening diseas elopment of and cell dea ed cells to orks of gene de (Wong, 2 have been em
, 2003; Ab et al., 2008;
l., 2006), V al., 2011), V ndrome virus et al., 2010)
odermal and d infectious matic disease is one of the tation to the et al., 1995)
nostimulants aquaculture ultures have nfection. The bstitute to the es in shrimp
sources have ses including f cancer, one ath, resulting be removed es have been 2011). Anti merged as a bdullaev and Zikri et al.
V.
V.
s ), d s e e e ).
s e.
e e e p
e g e g d n - a d .,
2009; Leon various nat (Giovannuc Beca have been c compounds longa L., m signaling pa survival pa pathway (ca death recept pathway (JN extracts of t with signifi apoptosis in Recently, an exhibited st viz., MCF-7 and DU-145 Duri metabolites have been i Amador et become imp Rece culture extr
nardi et al., 2 tural source cci et al., 199
ause of the i considered a for their an modulates g
athways inc athway (Bc aspase-8, ca tor pathway NK, Akt, an the dark fun cant change n human pro
n in vitro stu trong antica 7, MDAMB 5 by MTT a ing the pas
with divers isolated from
al., 2003).
portant sourc ently Sekar racts of mari
2010; Kim es have also
95; Narisaw intricacy of as a tool for nticancer eff growth of tu cluding cell cl-2, Bcl-x, aspase-3, cas y (DR4, DR nd AMPK) (R ngus Ganode es in the exp ostate cancer udy of the re ancer potenti B231, T47D
assay (Athav st 20 years se biologica m various m
Subsequent ces of new b et al. (2015 ine yeast, C
et al., 2011 o been mad wa et al., 1996 this disease r the better u fects. Curcum umor cells t
proliferation cFLIP, X spase-9), tum
5), mitochon Ravindran e erma lucidu pression of g r cell lines P ed pigment e ial on a pan
and prostate vale et al., 20 s, thousand al activities r
marine sourc tly, marine- bioactive mo
) reported th Candida albi
). Similarly de an area 6; Nahum et , gene expre understandin min, major through regu
n pathway ( XIAP, c-IAP mor suppres
ndrial pathw et al., 2009).
um have exh genes associ PC3 and DU
extracted fro nel of three e cancer cell 013).
s of novel ranging from ces (Garcia- -derived mic olecules (La hat selected icans can ac
y, pigment e of anticanc t al., 2001).
ession based ng of the ne component ulation of m (cyclin D1, P1), caspas ssor pathway ways, and pr . Ethanolic a hibited antica iated with ce U145 (Kao e om Serratia
breast canc l lines viz., P
compound m antiviral t -Fernandez croorganism am, 2007).
compounds ct as an antic
extracts from cer research
d approaches ewly isolated of Curcuma multiple cel
c-myc), cel e activation y (p53, p21) rotein kinase and spore oi ancer effects ell cycle and et al., 2014) a marcescens
cer cell lines PC3, LNCaP
ds and thei to anticancer
et al., 2002 ms have also
s from crude cancer agen
m h
s d a ll ll n ), e il s d ).
s s P
r r 2;
o
e nt
by the prot subjected fo fucoxanthin pathways in several othe metastasis (
The Candida al from coast carcinoma Green Mon Kathiresan, cancer cell three yeast normal Ver extract ind cancer act environmen
Sear expanding o Henceforth provide the perspective yeasts as Saccharomy immunostim branching a
ein ligand d or anticance n exerts its ncluding the er molecules (Kumar et al
cytotoxic a lbicans, Kur
al mangrov cells (MCF nkey kidney , 2015). Al
s (MCF-7, t strains, S ro cell line dicate more
tivity. Auth nt will be pr rching for n
our ability t the marin e novel bio
, the presen a source o yces cerevi mulant. Eff as well as th
docking stud er studies in
antiprolifera e Bcl-2 prot
s that are in l., 2013a).
activity of raishia caps ve ecosyste F7), human y cell lines ll extracts w
HepG2) an S. cerevisiae es and at th
efficiency hors sugge romising fo natural prod to find nove ne environm oactive com nt study was
of bioactive isiae has al ficacy of y he number
dy. A marin various cel ative activit teins, MAPK nvolved in e
three differ sulate and S em have be
hepatocarc (VERO) u were able t nd the norm e showed he same tim
against M est that th r further de ducts in rel el, potent an ment, the l
mpounds w s undertaken e compoun lready been yeast glucan
of glucose
ne caroteno ll lines, has ty through d K, NFκB, C either cell cy
rent extract Saccharomy een studied inoma cells using MTT to inhibit th mal Vero c
more than me the IC50
MCF7 cells his yeast evelopment latively unt nd selective argely unex with immen
n to explore nds. Glucan n proved to ns depends molecules i
id pigment shown that different mo Caspases, GA ycle arrest,
ts of the m yces cerevis
against hu s (HepG2)
assay (Sen he prolifera cell viability 80% cell value of S suggesting species fr as an antica tapped sour e bioactive xplored res nse possibil the potenti n from Bak o be a good s on the fr
in the β- 1,
fucoxanthin the pigmen olecules and ADD45, and apoptosis o
arine yeasts siae isolated uman breas
and African nthilraja and ation of the y. Amongs
viability in S. cerevisiae g anti-breas
om coasta ancer drug.
rces will be compounds servoir may lity. In this al of marine ker’s yeast d source o requency o 6 branching
n nt d d r
s d t n d e t n e t al
e s.
y s e t, f f g
points. Earl property an mortality da glucans fro shrimps and in aquacultu Prev melanins. In since the in and functio undertaken Obje 9 Extr yeas 9 Test imm 9 Fun
expl The topic is gi primary scr against WS characteriza adult P. mo of melanin melanin an
lier studies nd the survi ata. The pre om various d selection o ure.
vious studi n this study nformation onal charact
to understan ectives of th raction and st isolates ting the b munostimula nctional cha loring its bi Thesis is c iven in Ch reening of g SSV infect ation and e onodon agai n from mar nd evaluatio
have show ival in shrim esent study
marine yea of the poten
ies on mel , marine bla on this me terization o nd its prope he study are d characteri
bioactive p ant in Penae aracterizatio ioactive pot comprised o
apter1. Ext glucan as an ion is pres evaluation inst WSSV rine black on of its bio
wn yeast-wis mps estima
was undert asts in term ntial strains a
lanins are ack yeast, H elanin is ver of the marin erty for poss
as follows:
ization of g
potential o eus monodo on of marin tential of six chapt
traction of n immunost sented in C
of selected infection. C
yeast, phy oactivity. Ch
se variation ated by imm
taken to und ms of immu as source of
mainly foc Hortaea wern ry much lim ne black y sible comme
glucan and
of marine on
ne yeast gl
ters. A gene glucan fro timulant in P Chapter 2.
d glucans a Chapter 4 il ysicochemic hapter 5 de
n in immuno munological
derstand the une gene ex f glucans for
cused on p neckii has b mited. Phys east melani ercial applic
melanin fr
yeast glu
lucan and m
eral introdu om marine
P. monodon Chapter 3 as immunos
llustrates th cal characte
als with the
ostimulation l assays and e efficacy o xpression in r application
plant/anima een selected sicochemica
in has been cations.
from marine
ucan as an
melanin for
uction to the yeasts and n postlarvae deals with stimulant in he extraction erization o e evaluation
n d f n n
al d al n
e
n
r
e d e h n n f n
of glucan analysis. A followed by
and melani A summary y references
in as an an and conclu s and appen
nticancer ag usion of the ndices.
********
gent by in e study is p
vitro gene presented in
e expression n Chapter 6
n 6,
2.1 Introd
Aqu plays a crit developing food and nu source of in acquired by single comm value of int shrimp prod as a result America. A prevention
Dise causing hug developmen
duction
uaculture is tical role in and develo utritional se ncome and y FAO in modity in v ternationall duction volu
of disease- As the shellf and control ease outbrea ge economi ntal stages
one of the n global foo oped countr ecurity, man livelihood i 2014 indica value terms, ly traded fis umes decre -related pro fish aquacul l increases c ak has been ic loss durin
of the anim
fastest grow od security a
ries. In addi ny millions in the aquac ates that sh , accounting shery produ
ased in 201 oblems in s lture industr concurrently a major thr ng the last mals are be
wing food and nutritio ition to its
of people a culture sect hrimp conti g for about ucts in 2012 2 and partic some count
ry grows, th y.
reat to the s few decade elieved to b
sectors in th onal needs o growing im around the w tor. Recent inues to be 15 percent 2. Globally, cularly in 2 tries in Asi he necessity
hrimp farm es. Generall be more su
he world. I of people in mportance in world find a information e the larges
t of the tota , the farmed
013, mainly a and Latin y for disease
ming industry ly, the early usceptible to It
n n a n t al d y n e
y y o
pathogens t Penaeus m developing White spot problems a reaching 10 immunostim shrimp farm and protec components immunostim Burgents et al., 2010; Su 2005); mann (Boonyaratp et al., 2000) Leung et al.
rendering hi 2.1.1 Shri
Shri aquaculture the world m more than cultured shr the top pro producing c and now re farming ind
than adults monodon is countries in syndrome v and is usua 00% within mulants as ming help to ction from
s are used a mulants of al., 2004; R ubramanian noproteins ( palin et al., 1 ) known as ., 2006) hav igher surviv mp aquac mp culture e crops worl market. Mos 50 countri rimps come oducers. In country. Shr epresents a dustry produ
(Lightner the most n Asia and i virus (WSSV ally associa n 3-10 days prophylact o control infe
drug resis as immunost
biological o Rodriguez et n and Philip, (Tizard et al 995; Zhang biological r ve been prov val rate and
ulture has grown ldwide. All st coastal co
ies practice e from Asia
the Wester rimp aquacu multi-billio uced an esti
et al., 1983 farmed cru is under per V) infection ated with h of infectio tic agents r fections and stant patho timulants in origin such t al., 2007; S , 2013;Mast ., 1989;Abu g et al., 2014) response mo ved to enhan
resistance to
n into one o kinds of sh ountries have
shrimp aq with Thaila rn hemisphe ulture expan
n dollar ind imated 1.6 m
3a; Momoy ustacean sp rsistent threa n is consider high inciden on (Liu et a rather than
therefore le ogens. Yeas n aquaculture h as β-gluca Sajeevan et
tan, 2015); c u-Elala et al
) and lipopol odifiers (BR nce immune
o infections.
of the large hrimps are h e a harvest i quaculture.
and, China, ere, Ecuado nded signific dustry. In 2
million met
yama and S pecies in m at from vira red as one o nce of larv al., 2009a).
n chemother ead to reduc
st cell and e over the y an (Chang e
al., 2009; Su chitin (Wan ., 2013); pep lysaccharide RMs) (Yan responses in .
est and mos highly desir
industry for Around 80 Indonesia a or is the m
cantly durin 2002, the gl
tric tons of
Sano, 1989) many of the
al infections f the serious al mortality The use o rapeutics in ced mortality d cell wal years. Use o et al., 2003 ukumaran e ng and Chen ptidoglycans es (Takahash
et al., 2005 n organisms
st importan rable now in r shrimp and 0 percent o
and India as major shrimp ng the 1980s lobal shrimp shrimp, and ).
e s.
s y f n y ll f
; et n, s hi 5;
s,
nt n d f s p s p d
