Effect of Use of Lactobacillus plantarum as a Probiotics to improve Cocoon

Production of Mulberry Silkworm, Bombyx mori (!"#

K.K.Singh, R.M.Chauhan, A.B.Pande, S.B.Gokhale and N.G.Hegde
Journal of Basic and Applied Sciences Vol. (1), JuneNov. pp 1-8.
$bstract

The sil!or" larvae !ere fed on "ul#err$ leaves %rea%ed !i%h a pro#io%ic #ac%eria
na"el$ Lactobacillus plantau! durin& differen% ins%ars and i%s effec% on larval #od$
!ei&h%, "a%ure #od$ !ei&h%, cocoon !ei&h%, pupa !ei&h%, shell !ei&h%, shell ra%io and
pupa%ion percen%a&e !ere s%udied. Appro'i"a%el$ all %he %rea%"en% !i%h L. plantau!
a pro#io%ics !as si&nifican%l$ superior %o %ha% of con%rol. L. plantau! %rea%ed &roups
recorded si&nifican%l$ hi&her values for larval #od$ !ei&h%, cocoon !ei&h%, shell
!ei&h%, pupa%ion percen%a&e a% all %he ins%ars na"el$ 1
s%
, (nd, )rd, *%h and +
%h
ins%ar
respec%ivel$ as co"pared %o con%rol. The s%ud$ indica%ed %ha% %he pro#io%ic #ac%eria
e'hi#i% induce"en% of cer%ain &ro!%h fac%ors leadin& %o increase in sil $ields and
pupa%ion , in sil!or" rearin& ho!ever %he "echanis" of in%erac%ion of hos% na%ive
"icro#io%a !i%h %he pro#io%ic #ac%eria influencin& ph$siolo&$ and inna%e i""uni%$ of
sil!or" re-uire fur%her inves%i&a%ion.
%eywords. /ro#io%ics, Lactobacillus plantau!, Bo!b"# !oi $L.% and 0ocoon
produc%ion
&ntroduction

/ro#io%ics are %he live "icro#ial food supple"en%s #eneficiall$ affec%in& %he hos% #$
i"provin& %he "icro#ial #alance (1uller, 1221and Aus%in et al., 122+).Several
researchers have repor%ed a#ou% #eneficial role pla$ed #$ use of pro#io%ics in 3u"ans
(0han et al., 128+4 Bri&idi et al., (555 6 (551), ru"inan%s ( ), a-uacul%ure (7ouille%
and 8an&don, 122* and 9ild#er& et al., 122:) and insec%s (7illon and 7illon,
(55*).Therefore, produc%s con%ainin& pro#io%ic #ac%eria are &ainin& populari%$,
increasin& %he i"por%ance of %heir accura%e specia%ion(;eun& et al ,(55( ) .Si"ilarl$,
in "ul#err$ sil!or" (Bo!b"# !oi ) presence of differen% %$pes of #ac%eria in %he
&u% have #een repor%ed (<o$ et al, (555 and =oda"a,(551).>os% of %he species
#elon&in& %o 9enus Steptococcus are found %o #e pa%ho&enic %o Bo!b"# !oi larvae
!hile #ac%eria fro" 9enus Pediococccus, Leuconostoc and Lactobacillus didn?%
produce an$ infec%ed sil!or". The precise "echanis" of #eneficial effec% on hos% or
in%erac%ion a"on& %he differen% #ac%erial s%rains presen% as "icro-flora is no% no!n
(S%einhaus, 12*2 and =oda"a, (551).

7ifferen% species of lac%ic acid #ac%eria have #een e'%ensivel$ s%udied
(Bruno et al .,122)4 Berne%-0a"ard et al.,122: 49i#son et al 12284 =oda"a,(551)
and found %o #e #eneficial as pro#io%ics (1uller, 12214Berne% 0a"ard,
122:4Saa"o%o et al 4 (551).Therefore i% !as planned %o s%ud$ %he effec% of a lac%ic
acid #ac%eria na"el$ Lactobacillus plantau! a no!n pro#io%ics on econo"ic
para"e%ers of Bo!b"# !oi $L..%

Materials and Met'ods

S%andard cul%ure of Lactobacillus plantau! procured fro" Na%ional 0ollec%ion of
@ndus%rial >icro-or&anis"s (N0@> (58)), Na%ional 0he"ical 8a#ora%or$, /une !as
cul%ured in ><S #ro%h "ediu". The #ro%h cul%ure of L. plantau! (15
A
cfuB"l) !as
used for %he e'peri"en%. The leaves of "ul#err$ S-1A)+ varie%$ and an indi&enous
"ul%ivol%ine sil!or" race na"el$ BA@1-1 !ere used no!n %o #e suscep%i#le %o
nuclear pol$hedrosis virus. C'peri"en% !as carried ou% under con%rolled la#ora%or$
condi%ions (%e"pera%ure (8D)
5
0 and hu"idi%$ of :5 %o 8+,).
>ul#err$ leaves !ere fed %o Bo!b"# !oi (8.) larvae as per rearin& "e%hods
su&&es%ed #$ =rishnas!a"i et. al. ., 12:8 .C'peri"en%al as !ell con%rol &roups had
) replica%ions consis%in& of 155 larvae each . >ul#err$ leaves of e-ual !ei&h% and
appro'i"a%el$ sa"e nu"#er !ere used for feedin& B. !oi larvae in con%rol and
e'peri"en%al &roups. C'peri"en%al &roups !ere fed !i%h "ul#err$ leaves %rea%ed #$
dippin& %he leaves in %he suspension of L .plantau! (15
A
cfu B "l) and par%iall$
dr$in& a% roo" %e"pera%ure, onl$ once soon af%er ha%chin& and firs% feed af%er 1
s%
, (
nd
,
)
rd
and *
%h
"oul% pass larvae. The con%rol &roup of B. !oi $L.% larvae (155 larvae in )
replica%ions) !ere fed !i%h "ul#err$ leaves !i%hou% %rea%"en% of %he L. plantau!
cul%ure. /ara"e%ers lie sin&le larva !ei&h% (A
%h
da$ of +
%h
ins%ar) and "a%ure larva
!ei&h% ( Eus% #efore "oun%in& ) , %o%al cocoon !ei&h% (&"), shell !ei&h% (&"), pupa
!ei&h% (&"), shell ra%io (,) and effec%ive pupa for"a%ion (,) !ere recorded and da%a
!as anal$sed s%a%is%icall$ %o s%ud$ %he effec%.
(esults and discussion
The resul%s on effec% of pro#io%ic #ac%eria L.plantau! on econo"ical para"e%ers of
sil!or" are presen%ed in Ta#le 1. <esul%s sho!ed %ha% %he larvae of %rea%"en% &roups
had si&nifican%l$ hi&her larval !ei&h%, cocoon !%. pupal !%. shell ra%io and pupa%ion
ra%e as co"pared %o con%rol &roup. Also overall anal$sis (Ta#le () of in%erac%ion
#e%!een differen% %rea%"en% &roups and con%rol &roup sho!ed si&nifican% hi&her
values for larval !ei&h%, cocoon, pupa, and shell !ei&h% and ra%io and pupa%ion ,.
L. plantau! %rea%ed "ul#err$ leaves fed %o larvae a% 1
s%
,(
nd
,)
rd
,*
%h
and +
%h
ins%ar
recorded "a'i"u" larval !ei&h%s ((.25, (.8A, (.25, (.8A and (.2)&" respec%ivel$ ),
cocoon !ei&h% (1.52,1.(:,1.)5,1.(2,1.(( and 1.))&"), pupal !ei&h% (1.5:,1.15,1.58
and 1.5(&") shell !%.(5.(5,5.(5,5.(1,5.(5,and 5.(1&"), shell ra%io
(1+.28,1+.)),1A.(1,1A.)( and 1+.A+,) respec%ivel$ as co"pare %o con%rol &roup. The
hi&her value of larval !ei&h%s and cocoon para"e%ers indica%ed %he &rea%er
sui%a#ili%$ Baccep%a#ili%$ of pro#io%ic L. planteu! alon& !i%h food plan%s. The a"oun%
of food consu"ed #$ a larva influences i%s &ro!%h ra%e, develop"en%, final #od$
!ei&h% and pro#a#ili%$ of survival (Slans$ and Scri#er, 128+).These resul%s sho!
%ha% a funda"en%al shif% in %he "icro#ial profile in sil!or" larval &u% is #eneficial %o
%he hos% !hich in %urn "a$ si&nifican%l$ con%ri#u%e %o increase sil produc%ion. 1ood
in%ae is also re&ula%ed #$ %he ph$sical na%ure of food and also presence of pha&o-
s%i"ulan%s in %he food (7add, 12:5). Sil!or" B. "ori (8) reared on "ul#err$ leaves
supple"en%ed !i%h "inerals, oral pro%ein supple"en%a%ion, cereal flours, "edicinal
e'%rac%s, plan% &ro!%h hor"ones ( Sunder <aE e% al.,(555 Sin&h ,122:) are repor%ed %o
have #eneficial effec%s on econo"ic para"e%ers. Suu"ar (128)) has repor%ed
enhancin& effec% on sil $ield usin& "ul#err$ ph$lloplane $eas% Spoobolo!"ces
oseus .@n presen% s%ud$ si"ilar o#serva%ion "a$ #e a%%ri#u%ed %o increased efficienc$
of di&es%ion and assi"ila%ion of food "a%erials leadin& %o increased pro%ein s$n%hesis

and su#se-uen% accu"ula%ion of s%ora&e pro%eins in %he #od$ on accoun% of ac%ivi%$ of
a pro#io%ic "icro#ial flora in &u% of hos%. 0o"para%ivel$, %he food consu"p%ion in %he
con%rol ca%e&or$ !i%h indi&enous "icro flora reflec%s (!i%h respec% %o %rea%"en%
&roups) %he lo! sil produc%ion a#ili%$ as feedin& -uali%$ leaves influences %he
s$n%hesis of %o%al <NA and %ransla%ion of fi#roin "essen&er <NA and 7NA s$n%hesis
(0havanc$ and 1lournier, 12:2). S%a%is%icall$ no si&nificance (%a#le () !as o#served
!i%h respec% %o "a%ure larva !ei&h%s in %rea%"en% and con%rol &roups !hich "a$ due
%o co""on %i"in& of picin& %he ripe larvae in %rea%ed and con%rol &roups.
Si&nifican%l$ hi&her pupa%ion , (Ta#le1 and () !as o#served in all %rea%"en% &roups
e'cep% (
nd
ins%ar %rea%"en% &roup. L.plantau! %rea%ed larvae recorded "a'i"u"
pupa%ion , in *
%h
ins%ar (8),) follo!ed #$ +
%h
ins%ar (8(,), 1
s%
ins%ar (:8,) and )
rd
(:),) respec%ivel$ as co"pare %o con%rol &roup. This "a$ #e par%l$ due %o hi&her
food di&es%i#ili%$ leadin& %o hi&her survival ra%e (Slans$ and Scri#er, 128+) and due
%o increase in ac%ivi%$ of red florescen% pro%ein (<1/) a pro%ein responsi#le for
providin& inna%e i""uni%$ #$ inac%iva%in& virus a"on& %he sil!or" ( =oda"a,
(551). @% is !ell su#s%an%ia%ed #$ 3a$ashi$a,e% al .,12:1 , %he concen%ra%ion of <1/ is
"a'i"u" durin& %he "oul%in& period (!hen %he larvae do no% feed) especiall$ durin&
*
%h
and +
%h
ins%ar !hile i% is lo! in earlier ins%ar. Since in sil!or" "ore %han 25, of
%o%al larval feed is in&es%ed and also <1/ is "a'i"u" durin& *%h and +
%h
ins%ar of
B.!oi, (=oda"a, (551) %he supple"en%a%ion of 8.plan%aru" soon af%er %he "oul%
"a$ have influenced <1/ as !ell alon& !i%h feedin&. This defini%el$ have resul%ed in
hi&her pupa%ion , as co"pare %o con%rol &roup in presen% s%ud$ durin& *
%h
ins%ar
follo!ed #$ +
%h
ins%ar )
rd
ins%ar and 1
s%
ins%ar. Thou&h (
nd
ins%ar sho!ed enhance"en%
in cocoon charac%er #u% pupa%ion , !as si"ilar %o con%rol &roup "a$ #e due %o less
influence of 8.plan%aru" on ac%ivi%$ of <1/ durin& %his s%a&e of B."ori.
Beneficial effec%s #$ "odula%ion of &u% "icro-flora and influence on "ucosal
i""uni%$ or %hrou&h al%erin& enF$"a%ic ac%ivi%ies has #een e'%ensivel$ s%udied in
hu"an ,ani"als and "an$ insec%s (;eun& et al ,(55( ) leadin& %o co""ercialiFa%ion
of produc%s as pro#io%ics. /resen% preli"inar$ s%ud$ indica%e %he #eneficial effec%s of a
pro#io%ic L. plantau! on econo"ic %rai%s of sil!or" B.!oi, ho!ever %he
"echanis" of in%erac%ion of hos% na%ive "icro#io%a !i%h %he pro#io%ic #ac%eria
influencin& direc%l$ and indirec%l$ on food assi"ila%ion , ph$siolo&$ and inna%e
i""uni%$ of sil!or" re-uire fur%her inves%i&a%ion.

)able *+ Effect of !plantarum on economic parameters of a multivoltine B.mori

Parameters
)reatment
,roup
Sin,le larva
wt! (,m"
Mature larva
wt! (,m"
Cocoon wt!
(,m"
Pupae wt!
(,m"
S'ell wt! (,m" S'ell ratio
-
Pupation
-
Control (.:)D5.5)2 (.):D5.5A5 1.52D5.5)) 5.21D5.5)5 5.1+D5.55+ 1).+1D5.*A( A(D5.+(*
*
st
instar (.25D5.5** (.+(D5.5)+ 1.(:D5.5)( 1.5:D5.5)( 5.(5D5.55+ 1+.28D.+*5 :8D1.+:(
.
nd
instar (.8AD5.5*: (.+(D5.5+A 1.)5D5.5*8 1.15D5.5*) 5.(5D5.55+ 1+.))D5.))) A)D1.*(8
/
rd
instar (.25D5.5** (.+(D5.5)+ 1.(2D5.5*+ 1.58D5.5*1 5.(1D5.55+ 1A.(1D5.*:: :)D1.15)
0
t'
instar (.8AD5.5*: (.+(D5.5+A 1.((D5.5)2 1.5(D5.5)A 5.(5D5.55A 1A.)(D5.**2 8)D5.+(*
1
t'
instar (.2)D5.5*+ (.+(D5.5+) 1.))D5.5+( 1.1(D5.5*: 5.(1D5.55A 1+.A+D5.)21 8(D1.(22
SE(m" 2 3!3*4 3!3.* 3!3*5 3!3*6 3!33/ 3!.33 3!4/1
C7 at 1- 3!340 3!340 3!353 3!368 3!3*/ 3!540 0!*64
C9 - 8!53 5!83 *1!// *8!8/ *8!36 */!00 */!*4
)able .! $nalysis of variance on effect of L. planatrum treatment on economic
parameters of a multivoltine B.mori (L.)

Source of
variation
7f Mean of s:uare value
Sin,le
larva
wt!
Matur
e larva
wt!
Cocoon
wt!
Pupae
wt!
S'ell
wt!
S'ell
ratio
Pupation
&nteractio
n between
control
and
treatment
,roups
1 3!34* 3!38/8 3!*/8* 3!*33
3
3!334
4
*4!11/
*
*1/*!533
; <S ;; ;; GG GG GG
;Si,nificant at 1-, ;; =i,'ly si,nificant at *-, <S+ <on >si,nificant
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0<SBSC<@B(55+B
To,
Su#. <e&ardin& su#"ission of a research paper en%i%led MCffec% of use of
Lactobacillus plantau! as a pro#io%ics %o i"prove cocoon produc%ion in %he
"ul#err$ sil!or" Bo!b"# !oi (8.)N
7ear Sir,

He are su#"i%%in& here!i%h a research paper en%i%led MCffec% of use of Lactobacillus
plantau! as a pro#io%ics %o i"prove cocoon produc%ion in %he "ul#err$ sil!or"
Bo!b"# !oi (8.)N
>a$ !e re-ues% $ou %o please infor" %he a#ou% accep%ance of %he sa"e.
Than $ou for $our coopera%ion.
Hi%h ind re&ards
;ours sincerel$,
(7r.S.B.9ohale)
Vice /residen% 6 3ead
0<S,BA@1,Kruli-anchan
Cnclosures 1. 3ard cop$ of %he research paper