The Presence Of Inclusion Bodies Of Helicoverpaarmigera Nuclear Polyhedrosis Virus (Hanpv) In The Midgut Tissue Of Fourth Instars Spodopteralitura Larvae.
THE PRESENCE OF INCLUSION BODIES OF Helicoverpaarmigera NUCLEAR
POLYHEDROSIS VIRUS (HANPV) IN THE MIDGUT TISSUE OF FOURTH
INSTARS Spodopteralitura LARVAE
MIA MIRANTI, MUHAMAD RULIAWANSALIM, AND
YASMIPURNAMASARIKUNTANA
Department of Biology, Faculty of Mathematics and Natural Sciences,
University of Padjadjaran.
Jl. Raya BandungSumedang KM-21 Jatinangor, Indonesia
Contact Person : miamiantariksa@yahoo.com
ABSTRACT
The research to determine the presence of inclusion bodies of HaNPV in the midgut
tissue of fourth instars Spodopteraliturae larvae has been done. The larval has been
infected by 1.13 X 105polihedra/ml concentration of viruses and the presence of
inclusion bodies has been conducted every 8 hours from the 1st until 72nd hours after
virus infection. The histological slide of midgut tissue of larvae has been stained by
Mallory-Azan staining method. The experiment shows that in 8th hours after infection,
the inclusion bodies of virus presence in columnar cells of larvae. In 24th hours after
infection, inclusion bodies of virus through disgestive tract and appeared in faeces of
larvae. In 32nd hours after infection, the inclusion boddies of virus still appeared in
regenerative columnar cells until 56th hours after infection. After 64th hours until 72nd
hours after infection, the present of polihedra damaged of midgut tissue, made all
columnar cells lysis and inclusion bodies form into granul (polyhedral granules).
Therefore HaNPV can replicated in Spodopteraliturae larval as an alternative host.
Keywords
:Helicoverpaarmigera
Nuclear
Polyhedrosis
Virus
(Ha
NPV),
Spodopteralitura, Inclusion Bodies, Histological midgut structure,
Mallory Azan Staining Method.
INTRODUCTION
Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV) isolated from
cadaver of Helicoverpaarmigera larvae has an inclusion bodies (Maramorosch and
Sherman, 1985 ;Flipsen, 1995). In the specific host, the inclusion bodies of HaNPV can
be formed. However, H. armigera as a specific host for in vivo production is
cannibalism. Otherwise, in vivo production of HaNPV need another host as an
alternative host.
HaNPV
can
infect
another
insect larval
as
Spodopteralitura,
Spodopteraexiguaandcrocidolomiapavonana (Miranti, 2008). The virus is ideal to use
as biological agent to control the population of these insects. Spodopteralitura is an
ideal host for HaNPV production because the inclusion bodies can formed perfectly
(Miranti and Wardono, 2009). In this research, the presence of inclusion bodies of
HaNPV will observed in histological slide of midgutof S. litura larvae at 1st hour until
72nd hours after larval infection.
MATERIAL AND METHOD
HaNPV collected from Laboratory of Microbiology, Department of Biology,
Faculty of Mathematics and Natural Sciences, University of Padjadjaran, BandungIndonesia is in cadaver of H. armigera larvae. The suspension of virus has been made
with Indrayani, et al., (1993) method with modification. Larval fourth instars of S. litura
collected from BalaiPenelitianSayuran, Lembang, West Java-Indonesia.
The inclusion bodies of the virus was 1 X 105 Inclusion Bodies/ml
concentration. Every 1 ml of virus suspension mixed with 30 gram of kernel sweet corn.
S. litura larval infection within orally method. The histological slide of midgut tissue of
larval body was made every 8 hours from the 1st until the 72nd hour after larval
infection, through Mallory-Azan staining method. The inclusion bodies of virus
observed with light microscope at 400X.
RESULT
The experiment shows that in ht e histological slide of midgut tissue, the
inclusion bodies of HaNPV can be observed. At 1st hour after larval infection, the
inclusion bodies of virus is disappear from the midgut. In the 1st hour after larval
infection, the polihedrin of virus is lysis and liberated the virions. The virionscan not be
presence in midgut tissue (Flipsen, 1995). They formed a budding virus in midgut cells.
In the 8th hours after infection, the inclusion bodies of virus presence in
columnar cells of larvae. In 24th hours after infection, inclusion bodies of virus through
digestive tract and appeared in faeces of larvae. In 32nd hours after infection, the
inclusion boddies of virus still appeared in regenerative columnar cells until 56th hours
after infection. Hawtin, et al., (1992), and Sudhakar and Mathavan (1999) said that the
presence of the inclusion bodies shows that virus can replicated in S. litura perfectly.
After 64th hours until 72nd hours after infection, the present of polihedra
damaged of midgut tissue, made all columnar cells lysis and inclusion bodies form into
granul (polyhedral granules). The midgut cells were lysis and the inclusion bodies
liberated to the hemocoel. The midgut cells damaged and the colour of the hemocoel is
whitegrey. The presence of the inclusion bodies shows that HaNPV can replicated in
Spodopteraliturae larval as an alternative host.
CONCLUSION
The presence of polyhedral of HaNPV in the body of S. litura larval shows that
S. litura can be used as an alternate host for HaNPV in vivo production.
ACNOWLEDGEMENT
This research was supported in part by HibahBersaing Grant 2008-2010,
DirjenDikti, The Ministry of National Education, Republic of Indonesia.
REFFERENCES
Flipsen, H. 1995. Pathogenesis Induced by (Recombinant) Baculoviruses in Insects.
Thesis.Wageningen. 13-24
Hawtin, R.E., L.A. King, and R.D. Possee. 1992. Prospects for The Development of a
Genetically Engineered Baculoviruses. Pesticides Sciences. 34. 9-15
Indrayani, I.G.A.A., Subiyakto, dan G. Kartono. 1993. Teknik Perbanyakan
Helicoverpaarmigera Nuclear
Polyhedrosis
Virus H
( aNPV).Prosiding
Simposium Patolog Serangga.12-13 Oktober 1993.UGM-Yogyakarta.163-170.
Maramorosch, K and K.E. Sherman. 1985. Viral Insecticides for Biological Control.
London : Academic Press, INC
Miranti, M. 2008. Produksi Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV)
secara in vivo pada Inang Pengganti. Disertasi.UniversitasPadjadjaran
Miranti, M, and W. Niloperbowo
. 2009. Pengaruh Konsentrasi Infeksi
Helicoverpaarmigera Nuclear Polyhedrosis Virus pada Tingkat Kematian,
Waktu
Kematiandan
roduktivitas
Produksi
olihedradalam
Larva
Spodopteralitura F. Sebagai Inang Pengganti. Jurnal Agrikultura. 20. No.1.
April 2009.
Sudhakar, S and Mathavan. 1999. Electron Microscopical Studies and Restriction
Analysis
of Helicoverpaarmigera
Nucleo
Polyhedrosis
Virus.
http://www.iisc.er.net. Diaksestanggal 17 Oktober 2009.
POLYHEDROSIS VIRUS (HANPV) IN THE MIDGUT TISSUE OF FOURTH
INSTARS Spodopteralitura LARVAE
MIA MIRANTI, MUHAMAD RULIAWANSALIM, AND
YASMIPURNAMASARIKUNTANA
Department of Biology, Faculty of Mathematics and Natural Sciences,
University of Padjadjaran.
Jl. Raya BandungSumedang KM-21 Jatinangor, Indonesia
Contact Person : miamiantariksa@yahoo.com
ABSTRACT
The research to determine the presence of inclusion bodies of HaNPV in the midgut
tissue of fourth instars Spodopteraliturae larvae has been done. The larval has been
infected by 1.13 X 105polihedra/ml concentration of viruses and the presence of
inclusion bodies has been conducted every 8 hours from the 1st until 72nd hours after
virus infection. The histological slide of midgut tissue of larvae has been stained by
Mallory-Azan staining method. The experiment shows that in 8th hours after infection,
the inclusion bodies of virus presence in columnar cells of larvae. In 24th hours after
infection, inclusion bodies of virus through disgestive tract and appeared in faeces of
larvae. In 32nd hours after infection, the inclusion boddies of virus still appeared in
regenerative columnar cells until 56th hours after infection. After 64th hours until 72nd
hours after infection, the present of polihedra damaged of midgut tissue, made all
columnar cells lysis and inclusion bodies form into granul (polyhedral granules).
Therefore HaNPV can replicated in Spodopteraliturae larval as an alternative host.
Keywords
:Helicoverpaarmigera
Nuclear
Polyhedrosis
Virus
(Ha
NPV),
Spodopteralitura, Inclusion Bodies, Histological midgut structure,
Mallory Azan Staining Method.
INTRODUCTION
Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV) isolated from
cadaver of Helicoverpaarmigera larvae has an inclusion bodies (Maramorosch and
Sherman, 1985 ;Flipsen, 1995). In the specific host, the inclusion bodies of HaNPV can
be formed. However, H. armigera as a specific host for in vivo production is
cannibalism. Otherwise, in vivo production of HaNPV need another host as an
alternative host.
HaNPV
can
infect
another
insect larval
as
Spodopteralitura,
Spodopteraexiguaandcrocidolomiapavonana (Miranti, 2008). The virus is ideal to use
as biological agent to control the population of these insects. Spodopteralitura is an
ideal host for HaNPV production because the inclusion bodies can formed perfectly
(Miranti and Wardono, 2009). In this research, the presence of inclusion bodies of
HaNPV will observed in histological slide of midgutof S. litura larvae at 1st hour until
72nd hours after larval infection.
MATERIAL AND METHOD
HaNPV collected from Laboratory of Microbiology, Department of Biology,
Faculty of Mathematics and Natural Sciences, University of Padjadjaran, BandungIndonesia is in cadaver of H. armigera larvae. The suspension of virus has been made
with Indrayani, et al., (1993) method with modification. Larval fourth instars of S. litura
collected from BalaiPenelitianSayuran, Lembang, West Java-Indonesia.
The inclusion bodies of the virus was 1 X 105 Inclusion Bodies/ml
concentration. Every 1 ml of virus suspension mixed with 30 gram of kernel sweet corn.
S. litura larval infection within orally method. The histological slide of midgut tissue of
larval body was made every 8 hours from the 1st until the 72nd hour after larval
infection, through Mallory-Azan staining method. The inclusion bodies of virus
observed with light microscope at 400X.
RESULT
The experiment shows that in ht e histological slide of midgut tissue, the
inclusion bodies of HaNPV can be observed. At 1st hour after larval infection, the
inclusion bodies of virus is disappear from the midgut. In the 1st hour after larval
infection, the polihedrin of virus is lysis and liberated the virions. The virionscan not be
presence in midgut tissue (Flipsen, 1995). They formed a budding virus in midgut cells.
In the 8th hours after infection, the inclusion bodies of virus presence in
columnar cells of larvae. In 24th hours after infection, inclusion bodies of virus through
digestive tract and appeared in faeces of larvae. In 32nd hours after infection, the
inclusion boddies of virus still appeared in regenerative columnar cells until 56th hours
after infection. Hawtin, et al., (1992), and Sudhakar and Mathavan (1999) said that the
presence of the inclusion bodies shows that virus can replicated in S. litura perfectly.
After 64th hours until 72nd hours after infection, the present of polihedra
damaged of midgut tissue, made all columnar cells lysis and inclusion bodies form into
granul (polyhedral granules). The midgut cells were lysis and the inclusion bodies
liberated to the hemocoel. The midgut cells damaged and the colour of the hemocoel is
whitegrey. The presence of the inclusion bodies shows that HaNPV can replicated in
Spodopteraliturae larval as an alternative host.
CONCLUSION
The presence of polyhedral of HaNPV in the body of S. litura larval shows that
S. litura can be used as an alternate host for HaNPV in vivo production.
ACNOWLEDGEMENT
This research was supported in part by HibahBersaing Grant 2008-2010,
DirjenDikti, The Ministry of National Education, Republic of Indonesia.
REFFERENCES
Flipsen, H. 1995. Pathogenesis Induced by (Recombinant) Baculoviruses in Insects.
Thesis.Wageningen. 13-24
Hawtin, R.E., L.A. King, and R.D. Possee. 1992. Prospects for The Development of a
Genetically Engineered Baculoviruses. Pesticides Sciences. 34. 9-15
Indrayani, I.G.A.A., Subiyakto, dan G. Kartono. 1993. Teknik Perbanyakan
Helicoverpaarmigera Nuclear
Polyhedrosis
Virus H
( aNPV).Prosiding
Simposium Patolog Serangga.12-13 Oktober 1993.UGM-Yogyakarta.163-170.
Maramorosch, K and K.E. Sherman. 1985. Viral Insecticides for Biological Control.
London : Academic Press, INC
Miranti, M. 2008. Produksi Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV)
secara in vivo pada Inang Pengganti. Disertasi.UniversitasPadjadjaran
Miranti, M, and W. Niloperbowo
. 2009. Pengaruh Konsentrasi Infeksi
Helicoverpaarmigera Nuclear Polyhedrosis Virus pada Tingkat Kematian,
Waktu
Kematiandan
roduktivitas
Produksi
olihedradalam
Larva
Spodopteralitura F. Sebagai Inang Pengganti. Jurnal Agrikultura. 20. No.1.
April 2009.
Sudhakar, S and Mathavan. 1999. Electron Microscopical Studies and Restriction
Analysis
of Helicoverpaarmigera
Nucleo
Polyhedrosis
Virus.
http://www.iisc.er.net. Diaksestanggal 17 Oktober 2009.