ICMR BULLETIN VOL. 29-No.-1-JANUARY-1999.pdf
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ICMR
BULLETIN
January, 1999
Vol. 29, No. 1
PROSPECTS OF USING BACILLUS SPHAERICUS IN THE
CONTROL OF CULEX MOSQUITOES IN RELATION TO
RESISTANCE DEVELOPMENT
Vector resistance to insecticides, high cost and
B.sphaericus in the integrated vector management (I VM)
environmental pollution associated with insecticide use
programmes will depend on how the emergence of
resistance in the field population is managed. This review
highlights the mode of action of B. sphaericus, the current
status of the resistance in Culex mosquitoes to B. sphaericus
have imposed severe constraints on vector control
programmes. Consequently, vector control strategies have
to be redesigned to minimize the use of chemical pesticides.
Bacillus thuringiensis van israelensis and B. sphaericus
are being projected as promising biocontrol agents against
the vectors of malaria, fi lariasis and B. thuringiensis against
onchocerciasis1*6. Several field trials with B. sphaericus
carried out against Culex and Anopheles mosquitoes
breeding in different habitats produced variable results7’20.
B. sphaericus has a narrow host range21, is non-toxic to
black flies, less active against most Aedes species and
and its future implications in mosquito control
programmes.
Mode of Action of Bacillus sphaericus
B. sphaericus (Neide) is an aerobic and spore forming
soil bacterium. Laboratory assays showed that >100 spores/
ml are required to kill larvae of Cx. quinquefasciatus^.
tends to be more effective against Culex followed by
Among the 48 strains, the most studied were 1593,2297
and 2362 7«29’42«4\ Pathogenicity is caused by the production
anophelines22*37. The euphoria of control of vector
mosquitoes with this agent was short lived in view of
of two kinds of toxic proteins viz. crystal and Mtx (100
reports of resistance development from various laboratory
and field studies15-18-35-38"40, which showed that high dosage
than Mtx and is produced during sporulation and
and continuous exposure to B. sphaericus would result
in extremely high or moderate levels of resistance in Culex
quinquefasciatus.
The potential of using B. sphaericus as a biocontrol
agent to control the vectors of the filarial parasites will
and 30.8 kDa). The crystal toxin is more mosquitocidal
concentrated in the parasporal inclusions. These inclusions
have a crystalline ultrastructure and are released into the
medium along with the spore after the completion'of
sporulation. The Mtx is reported to be produced only
during the vegetative phase44*46. The crystal toxin is
depend upon the risk of emergence of resistance. This
composed of two proteins, P51 and P42 (42 kDa) that
act in synergy. The P51 part is responsible for the initial
information is essential in formulating suitable strategies
binding.of the crystal toxin to the receptor on the brush
for resistance management. The future utility of
border membrane of the epithelial cells in the gastric caecae
Division of Publication & Information, ICMR, New Delhi - 110029
and posterior midgut regions and the 42 kDa component
confers toxicity47'50 But the relationship between toxicity
of B. sphaericus to larva and spore germination is not
yet understood51. The toxic process involves (i) the
ingestion of the crystal/spore/cell complex; (ii) dissolution
and may not account for the high level of resistance19.
of the toxin at the alkaline pH of the cell wall in the
for different mechanisms of resistance, may be present
in natural populations at frequencies equal to the muta
tion rate.
midgut; (iii) activation of the protoxin (42 kDa) into the
toxic form by gut proteases; (iv) binding of the binary
toxin to the specific receptors on the brush border
membrane of epithelial cells in the gastric caecae and
posterior midgut regions; (v) entry of spores containing
binary toxin into the midgut cells by forming pores on
the cell membrane;and (vi) the pathogenic effects then
involve basal separation and sloughing of the midgut
epithelial cells, swelling of the midgut, that leads to
tremors, sluggishness and eventual death of the larvae21*52.
Differential Susceptibility to Bacillus sphaericus
Susceptibility of mosquito species to B. sphaericus
varies widely53 and at least one or more of four factors
are responsible for such wide variation. They are rate of
ingestion of $pore containing toxin, dissolution of the
toxin in the mosquitoe’s gut, activation of the toxic pro
tein and binding to the specific receptor sites. In general
Culex mosquitoes are more susceptible than anophelines
and Aedes29'54'60. The lack of susceptibility oL4erfe$ mos
quitoes (especially Ae. aegypti) to B. sphaericus is not
due to rates of toxin ingestion/concentration, dissolution
of the toxin, specificity of the gut proteases and/or acti
vation of the crystal toxin by midgut proteases38,61*66 but
is due to the failure of toxins binding to the midgut epi
thelial cells59,67 since no functional receptors were found
in this species31. For Anopheles species, B. sphaericus
toxins bind to the larval midgut cells but with low speci
Heterozygotes were only 3 times less susceptible than
the parental field population. Resistance is found to be
highly specific affecting only members ofthe strain used
in the selection38. Several mutated genes, each coding
From various laboratory and field studies it was found
that the B. sphaericus exposed populations of Cx.
quinquefasciatus were 4 to 100,000 times less susc-^
tible than unselected populations (Table). This wide
variation might be due to different factors influencing
selection71. Factors such as selection pressure, the pro
portion of population exposed, the gene frequency of the
resistance alleles in the population prior to selection,
existence of untreated refugees, reduced ingestion rate,
low biocides' persistence and/or availability because of
settling of large proportion of spores at the bottom of the
mosquito- breeding habitat within 2 h of treatment, in
flux of resistant individuals, changes in the mosquito larval
midgut receptor and physico-chemical factors of the
breeding habitats are important factors that determine
the development of resistance by mosquitoes35’38,39’41,44,71'81.
On the contrary, the factor that could contribute for the
slow process of resistance development is the phenom
enon of back selection (or removal of selection) which
results in lower survivorship ofthe resistant individuals
because of their lower fitness82. However, continued
exposure may improve the fitness of the target popula
tion through co-adaptation of resistant genomes by
modifier genes which lead to a more stable resistance83.'
High and moderate levels of resistance in Cx,
quinquefasciatus to B. sphaericus both in the laboratory
ficity and affinity, thus resulting in moderate degree of
susceptibility of these species to the toxins47,59. In Culex
and under field conditions were attributed to repeated
intense selection pressure with low and slow mixing of
species the toxins bind with high specificity and high
non-selected populations19,35,38.
affinity, thus leading to high susceptibility in these spe
cies47,59,68. In a recent study it was found that there was
no relationship between toxicity and irreversibility of toxin
binding on brush-border membrane69.
Genetics and Mechanism of Resistance
Ingestion
Mosquito larvae are filter feeders and they normally
exclude large particles84. They feed on particulate matters
ranging from microscopic bacteria to clearly visible
particles such as aquactic algae85. Species belonging to
Mosquito pathogenic strains of B. sphaericus have
the genus Anopheles are surface feeders whereas Culex
been assigned to DNA homology group II A70. Resistance
species feed below the water surface as well as at the
to B. sphaericus is monofactorial involving a single sex
bottom84,86. The presence of plenty of bacteria and other
linked recessive gene44 which segregates independently
from the overproduced esterases for organophosphorous
suspended matter in polluted breeding habitats is reported
to influence the rate of ingestion of bacterial spores and
resistance. These esterases are slightly more active in
digestability87 which would account for the wide variation
the B. sphaericus resistant than the susceptible strains
within the genus86,88. Earlier Sun et al.39 showed no effect
2
Table. Selection of resistance in Culex quinquefasciatus to Bacillus sphaericus.
Strain
Mosquito
strain
Selection
(Lab./field)
Selection
Pressure (%)*
Generation
Resistance
level (fold)**
Source/
Country
2362
Lab.
Field
Lab.
Lab.
90
94-98
94-98
F4
F<
F»
LC.,, - 60
LC,0- 1000
LCJ0- 100.000
Georghiou et al,
(1992): USA3’
2362
Lab.
Lab.
Field
Field
Lab.
Lab.
Lab.
Lab.
80
80
80
80
F.<
F„
f3J
f5J
LC«- 43
LCW- 36
LC,„- 12
LCW - 24
Rodcharoen & .
Mu Ila (1991)
USA40
Lab.
Field
Lab.
Lab.
80
80
F„
F.„
LC., - 37
LCJn- 10-30
Lab.
Field •
Lab.
Lab.
80
80
‘■'"‘ino
LCJ0 - 37
LC,„ - 31
Rodcharoen &
Mulla(1994):
USA35
Rodcharoen &
Mulla(1996):
USA39
2362
Field
Field
(8 years)
18 treatments
LC50 - 16.000
Sinegre et al.
(1994) France15
2362
Field
Field
37 treatments
(2 years)
LCJ0- lOt
Silva-Filha et al,
(1995); Brazil”
1593 M
Resistant
field strain
Field
Lab.
75
F„
Rao et al, (1995)
India1’
Field
35 treatements
(2 years)
LCS0 - 6,223
LCW- 31,325
LCJ0- 146
LCW- 180
1593
Lab.
Field
Lab.
Lab.
80
80
»F 100
«F 100
LC„ - 22
LCm-6
Rodcharoen &
Mulla(1996);
USA39’.
2297
Lab.
Field
Lab.
Lab.
80
80
«F.
»F IOO
LC5o - 12
LC„-4
-do-
*
•
-
* Per cent mortality at sublethal dosage.
*• Difference in the susceptibility between the parental and selected population.
t Less susceptible than control Held population
* „
Superscript numbers refer to the si. nos in the reference list.
of water depth between 1 and 5 cm on susceptibility. A
strains could possibly serve as a behaviouristic mechanism
recent study90 is in aggreement with the observation of
of resistance against this microbial larvicide77.
Nugud and White91 that anophelines were less susceptible
as they require higher concentraion of spores or their
Receptor site
surface feeding habit does not allow them to ingest
Two different mechanisms conferring high levels of
sufficient amount of bacterial spores. In a study it was
resistance to B. sphaericus toxin were reported in France
found that larvae of the resistant strains (laboratory and
and California. In the California study it was found that
field-collected) ingested B. sphaericus preparation at
there was an alteration of the toxin binding functionality
significantly lower rates than did larvae of the susceptible
of the receptor31. Whereas, the mechanism of field-se
strains-and yeast was ingested at a higher rate than the
lected resistance in the French study is probably not caused
bacterial preparation when compared to susceptible strains.
by the impairment of the first step of toxin-receptor in
The reduced ingestion rates of B. sphaericus ifl resistant
teractions at the cellular midgut level. It may involve
3
the midgut receptor, but not at the binding step level.
Howeyer, the exact mechanism is not known19. Results
of Brazil and Indian studies showed that B. sphaericus
resistant field strain of Cx. quinquefasciatus was more
susceptible than the normal population to B.4z.18-72, indi
cating that laboratory and field populations have differ
ent receptors. Such disparity is found in all species to
any microbial insecticide. It remains to be seen whether
the B. sphaericus resistance mechanism identified in
values for a susceptible strain from an unsprayed local
ity18. But susceptibility status of Cx quinquefasciatus in
the control and trial areas prior to treatment was not studied
in this trial. Since it is possible that different popula
tions differ in their susceptibility to the pathogen, it is
difficult to conclude that there was emergence of a popu
lation resistant to B. sphaericus. When larvae from a
sprayed area were subjected to selection pressure in the
laboratory, there was 6,223 and 31,325 fold increase in
California and France corresponds to different alleles of
the LC50 and LC90values respectively in the 18th genera
the same gene or to two different genes, and whether
tion in comparison with the susceptible strain.
either of them is the gene conferring high resistance in
India18-19. Binding experiments using isolated midgut brush
border membrane and radio labelled binary toxin of B.
Cross Resistance
sphaericus indicated that there was a direct correlation
between resistance and stable changes in the mosquito
Cross resistance studies showed that resistance to B.
larval midgut receptor71. But various other studies showed
sphaericus strain 2362 did not influence the susceptibility
of the mosquitoes to several other strains as well as towards
no such relationship.between receptor binding and mecha
nism of resistance development44-72-92
toxins of B.t.i.33. Laboratory and field collected strains
of Cx quinquefasciatus that were exposed to B. sphaericus
strain 2362 for about 100 generations and attained 37
Potentiality of Resistance Development
Laboratory and field collected Cx. quinquefasciatus
exposed to B. sphaericus strain 2362 for 35 generations
in the laboratory showed a level of resistance 43 and 12
fold that of parental generation respectively40. Further,
exposure to concentrations that yielded 80% mortality
resulted in 37 and 31 fold increase respectively in the
resistance in the laboratory and field collected colonies
by ~F10039, An extremely high level of resistance of 100,000
fold in the 12 th generation was obtained in the field col
lected colony using 94-98% selection pressure38. How
ever, a field collected population of Cx quinquefasciatus
larvae, which had been subjected to 37 treatments with
and 31 fold resistance respectively, did not show marked
cross resistance to B.t.i.', only 3-4 fold tolerance was noted.
However, they showed significant levels of cross resistance
to strains 1593 (6-22 fold) and 2297 (4-12 fold) of B.
sphaericus39. This showed that any change in receptor.
sites or binding affinities in one of the B. sphaericus strains
would confer cross resistance to other B. sphaericus strains
because these strains produce similar binary toxins which
bind to a single class of specific receptors31 >94. The degree
of cross resistance to strain 2297, which produces a
different type of toxin, may be less pronounced39-94. Further,
the absence of cross resistance between the two microbial
agents B. sphaericus and B.t.i.'3 was due to differences
in the structure and biochemistry of the toxins and
B. sphaericus 2362 for 2 years was found to be ~10 fold
differences in specific receptors for these toxins in Cx
less susceptible than a control field population72. Labo
quinquefasciatus1'.Therefore, Cx. quinquefasciatus larvae.
ratory selection of wild caught Anopheles stephensi with
which were highly resistant (>100,000 fold)38 and no longer
Spherix (a formulation of B. sphaericus) at a concentra
tion 2.8 times that of LC50 for 3 generations resulted in
had functional receptors for B. sphaericus, were still
susceptible to B.t.i.1'. Similarly, B. sphaericus toxin failed
232 times increase in LC50 value than that of the parental
to bind to the midgut brush border membranes (BBM)
population93. It was concluded that the intensive selec
of Ae. aegypti which is refractory to B. sphaericus but
tion of Cx. quinquefas-ciatus with the binary toxin of B.
highly susceptible to B.f.z.31-59. Further, selection with
sphaericus 2362 (51 and 42 kDa proteins) could pro
biocontrol agents had no apparent effect on the resistance
duce high resistance and high degree of specificity af
to organo phosphorus (OP), carbamate and synthetic
fecting members of the same strain (Table).
pyrethroids when compared with the untreated controls38-95.
Field resistance to B. sphaericits 1593M was observed
in a population of Cx quinquefasciatus in Kochi, India,
exposed to 35 rounds of spraying over a 2 year period.
Management of Resistance Development
Resistance management strategy is an important and
Larvae from the sprayed area gave LC50 and LC^ values
integral part of any 1VM programme which focuses on
that were 146 and 180 times greater than the corresponding
minimizing the economic and environmental impacts
4
resulting from pesticide treatments75. It is encouraging
important consideration because of the efficacy of B.
to note that resistance to pathogens by insects takes a
sphaericus against culicines, safety for the environment
longer time than is with chemical insecticides. This may
and usefulness in mosquito control programmes72.
be due to genetic changes at one or more loci against
insect pathogens”" resulting in poor biological fitness and
reversal of resistance
Studies on the biological fitness of laboratory and
field collected strains of C.v. quinquefasciatus. suscep
tible and resistant (37 and 31 fold upon selection) to B.
sphaericus showed that the resistant strains had signifi
cantly lower fecundity and fertility, but significantly higher
survival rates than the susceptible strains. The lower
production of eggs (fecundity) and lower percentage of
hatch (fertility) in resistant population was found to be
due to a lower nutrient reserve in the adults that devel
oped from larvae treated with sublethal concentrations
Among the various resistance management strategies,
the use of biocides in rotation might be an efficient
management measure because it disrupts selection pres
sure from the control agent and enables the insect popu
lation to recover susceptibility to the agent previously
used75. Since the progression of resistance to B.t.i. was
reported to be slow because of its multi-toxin complex107,
this biocontrol agent could be considered a good candi
date for use in rotation with B. sphaericus, larvicidal oils
and insect growth regulators (IGRs)105. Binding experi
ments with B. sphaericus radio labeled toxin in the pres
ence of an excess of un labeled B.t.i. toxin as a competi
tor, indicated that the receptors for B. sphaericus and
of pesticides”8'101. The preadult stages from females of
B.t.i. toxins are different51. Other reports also indicated
^resistant strains developed at slightly faster rates than
"hose of susceptible strains, which could result in a shorter
(150 fold) were 7 fold more susceptible to B.t.i. than the
that C.v. quinquefasciatus larvae resistant to B. sphaericus
generation time. However, lower fecundity was likely to
lead to overall lower population growth rates than in the
untreated population18. Similar results have been reported
susceptible strains. Studies provided evidence that the
resistant strains exhibited fitness disadvantages in the
This allows for the use of B.t.i. for a relatively longer
absence of B. sphaericus13.
and consequently its reintroduction105. The use of syner
Resistance and survival fitness are reported to be
independent102. Studies on the reversal of resistance to
bacterial toxins in insect populations have shown that
low and moderate resistance levels are unstable after the
interruption of the treatment. This is mainly because of
the decline in resistance gene frequencies due to mixing
of a large proportion of susceptible individuals with the
resistant ones due to immigration under field condi-
tion40J05l<M. However, highly resistant populations seem
to reach a plateau of resistance and require a long time
for a population of Plodia interpunctella resistant to B.t.i.I08.
period, allowing the reversal of B. sphaericus resistance
gists with B.t.i. proteins resulted in high toxicity and
expanded host range109. In this respect it should be noted
that studies on the interactions between toxins of B.t.i.
and B. sphaericus and target insects would provide a useful
basis for the appropriate use of these entomopathogens
in large-scale programmes. This would facilitate exploring
the possibility of combining B.t.i mosquitocidal toxin
genes with B. sphaericus genes to increase toxicity and
for delaying the development of resistance44.
Further, there is no report of B. sphaericus linked to
kto recover susceptibility58-7”-”7-105. It was observed that a
esterase overproduction conferring resistance to
larval population of C.v. quinquefasciatus breeding in
organophosphorous insecticides and no cross resistance
habitats treated with B. sphaericus once a month for 26
to certain other B. sphaericus strains producing crystal
months was 10 times less susceptible to B. sphaericus
toxins. This facilitates using this biocontrol agent in
and slightly more susceptible to B.t.i. compared with the
rotation with OP compounds5”.
untreated population. The low level of resistance to B.
sphaericus was unstable in the absence of selection pres
sure but reversed to be susceptible to B. sphaericus when
the treatment was interrupted ofter 16 months105.
The initial overproduction of anophelines in paddy
fields could not be regulated by natural predators and
parasites because of the time lag in the build up of the
population of pests and natural enemies. The maximum
However, the eventual acquisition of acquired
production of mosquitoes occurs during the initial stages
resistance to microbial control agents among field
of plant growth when farmers usually apply fertilizers.
populations of mosquitoes cannot be ruled out10” as
These mosquito breeding habitats are so extensive that
evidenced by recent findings in France and India15-18. This
the use of larvicides as a control method will not be cost
necessitates the formulation of rapid, elaborate and
effective. As biocontrol agents such as B. sphaericus and
practical resistance management strategies. This is a very
B.t.i are non toxic to non-target organisms110111, it has
5
been recommended that they be applied along with fer
resistance management for Cry proteins directly into
bacterial insecticides126127. The production of a bacterium
tilizer for the control of anopheline vectors in agro-ecosystems112. This strategy would check the rapid increase
combining the best attributes of both B.t.i and B. sphaericus
of pest population and subsequent control could be
and studies on the specificity of toxicity was suggested4’.
achieved by naturally occurring parasites and predators.
However, farmers should be educated to avoid contact
Grigorova et al.'28 demonstrated transfer of plasmids from
Streptococcus agulaetine and S. pyrogenes to B.s. but
with the bacillus preparation as recent finding indicated
did not investigate further transfer among B.s. strains.
that B.t.i. might cause serious wound infection in ani
mals and man113.
Orzech and Burke120reported that B.s. 1593 could accept
The combined use of natural enemies such as Gambusia affinisz\ Toxorhynchites rutilus rutilusll\
Mesocyclops longisetus'1* and Notonecta irrorata"** and
B.t.i. has been proved successful in the effective control
of mosquitoes in a cost effective and environmentally
safe manner. The control of Culex tarsalis larvae was
achieved by integrating both B. sphaericus and G. affinis"1.
This approach may slow down resistance development
among the vector populations by inflicting a dispropor
tionately high mortality on resistant individuals118. How
ever, if the natural enemies kill a higher proportion of
susceptible individuals, with the biocides acting equally
on both the populations, there may be an increase in the
population of resistant individuals119.
Future Prospects
Use of control release formulations such as calcium
alginate/Carrageenan microcapsule and encapsulating the
biocontrol agent in a living organism (Tetrahymena
pyriformis) would retain the toxin on the water surface
and act as a phagostimulant5,120. Genes encoding specific
toxicity could be incorporated with the larval food sources,
such as algae and Caulobacter spp. and the response of
mosquito immatures and other aquatic organisms on
modified food sources should be determined121. New toxins
identified from Clostridium bifermentans strain and several
B.t.i strains appeared to be good candidates for use in
combination with B. sphaericus toxin122. However.
Federici54 cautioned that the release of transgenic
insecticidal organisms used as larval food into the breeding
habitat might be counter productive. New genetic
engineering approaches such as recombinant DNA
techniques should be envisaged to overcome these
limitations that will allow stable expression of broad host
range, buoyancy and resistance to ultraviolet light. The
conjugated plasmid pAMpi from Streptococcusfaecalis
JH2.2 and could transfer pAMpi to another strain of B.s.
The plasmid phSP 11 that contained genes coding the binary
toxins of B.s. was introduced into the Colombian strain
of B.t.i. to study its expression and stability. The presence
of the plasmid was detected after 8 h of growth, coinciding
with the movement that the expression of the plasmid-
coded toxins started150. Rodcharoen and Mu Ila77 suggested
that the possibi I ity of encoding toxic genes of B. sphaericus
being transferred to blue green algae if genetically
engineered pathogens are used in a control programme.
be explored. However, a recent observation opined that
it was unlikely that genetically engineered B.s. carrying
a recombinant plasmid could transfer that plasmid to other
bacteria151
Conclusions
Though high levels of resistance in C.v.
quinquefasciatus were reported from laboratory and field
studies, none of the studies have examined the gut con
tent of the larvae for spores. It is possible that due to bad
formulation, the spores might have clumped and could
not have been ingested, or sunk to the bottom. Reversal
of resistance was observed in the field when the treat
ments were withdrawn44. There are no reports of cross
resistance to B./.i58-50-71. and organophosphorous insecti
cides58-05. Cross resistance between strains of B. sphaericus
harbouring different sequences has not been found to
occur58. There have been considerable studies on the
positive interactions of biocontrol agents with other control
agents such as chemicals152. Under normal dosages B.
sphaericus has a minimum impact on non-target organisms,,-57J10-1,,J55. and is non-toxic to mammals154-155. There
fore. there is scope for using B. sphaericus along w ith
other control methods in the integrated vector manage
ment of Culex mosquitoes.
recombinant strains are relatively inexpensive, easy to
mass-produce and possess excellent shelf-life67
The development of CytA/Cry4/Cry 11 and CytA/B..v.
models from a study on the effect of the CytA protein on
resistance to B.t.i proteins and B.s. preparations provided
a potential molecular genetic strategy for engineering
6
References
1.
Federici. B.A. Mosquito control b\ fungi Cuticinomyces.
l.eeenediuni and Caelomoniyees. In: Atierobicd ( oiurot of Pest
and Plant Diseases. I’d. Il.l). Birges. Academic Press. Lon
don. 1981. p. 555.
2.
Chapman. II.C. Ecology and use ol' Coelomomyces in biologi
cal control: A review. In: The Genus Coelomomyces. Eds. J.N.
Couch and C.E. Bland. Academic Press. Newyork and London.
1985. p. 361.
15.
Sinegre. G.. Babinot. M.. Quermel. J.M. and Gaven. B. Field
first occurrence of Culexpipiens resistance to Bacillus sphaericus
in southern France. Proceedings of VI11 European Meeting of
the Soceity for Vector Ecology. Santa Ana. CA. 1994. p. 17.
3.
Lacey. L.A. and Undeen. A.11. Microbial control of black Hies
and mosquitoes. Ann Rev Entomol 31: 265. 1986.
16.
4.
Guillel. P.D.. Kurtak. C.. Philippon. B. and Meyer. R. Use of
Bacillus thuringiensis israelensis for Onchocersiasis control in
West A frica. In: Bacterial ('ontrol of3 losquitoes and Black Flies:
Biochemistry, Genetics and Application ofBacillus thuringiensis
and Bacillus sphaericus. Eds. II. de Barjac and D. J. Sutherland.
Rutgers University Press. New Brunswick. NJ. 1990. p. 187.
Sundararaj. R. and Ragunatha Rao. D. Field evaluation of a
microgel droplet formulaion of Bacillus sphaericus 1593M
(Biocide-S) against Anopheles culicifacies and Anophelessubpictus
in south India. Southeast Asian J Trop Med Pub Health 24:
363. 1993.
17.
Kumar. A.. Sharma. V.P.. Sumdan. P.K.. Thavaselvam. D. and
Kamat. R.H. Malaria control utilizing Bacillus sphaericus against
Anopheles stephensi in Panaji. Goa. J Am Mosq Cont Assoc
10: 534. 1994.
18.
Rao. D.R.. Mani. T.R.. Rajcndran. R.. Joseph. A.S.. Gajanana.
A. and Reuben. R. Development of a high level of resistance to
Bacillus sphaericus in a field population of Culex quiquefasciatus
from Kochi. India. J Am Mosq Cont Assoc 11: 1. 1995.
19.
Nielscn-LcRoux. C.. Pasquicr. F.. Charles. J.F.. Sinegre. G.. Gaven.
B. and Pasteur. N. Resistance to Bacillus sphaericus involves
different mechanisms in Culexpipiens (Diptera: Culicidae) lar
vae. J Med Entomol 34:321. 1997.
20.
Regis. L.. Silva-Filha. M.H.. Oliveira. C.M.F.. Rios. E.M.F..
Silva. S.B. and Furtado. A.F. Integrated control measures against
Culex quinquefasciatus. the vector of filariasis in Recife. Mem
Inst Oswaldo Cruz 90: 115. 1995.
21.
Davidson. E.W. and Youstcn A.A. The mosquito larval toxin
of Bacillus sphaericus. In: Bacterial Control ofMosquitoes and
Black Flies: Biochemistry, Genetics and Application ofBacillus
thuringiensis and Bacillus sphaericus. Eds. H. de Barjac and D.
J. Sutherland. Rutgers University Press. New Brunswick. N.J..
1990. p. 237.
22.
Henlein. B.C.. Levy. R. and Miller .T.W. Jr. Recycling poten
tial and selective retrieval of Bacillus sphaericus from soil in a
mosquito habitat. J Invertebr Pathol 33 : 217. 1979.
23.
Mulligan. F.S. HI. Schaefer. C.H. and Wilder. W.H. Efficacy
and persistence of Bacillus sphaericus and Bacillus thuringiensis
H-14 against mosquitoes under laboratory and field conditions.
J Eco Entomol ~3 : 684. 1980.
5.
Mulla. M.S. Activity, field efficacy and the use of Bacillus
thuringiensis israelensis against mosquitoes. In: Bacterial Con
trol of Mosquitoes and Black Flies: Biochemistry, Genetics and
Application of Bacillus thuringiensis and Bacillus sphaericus
Eds. H. de. Barjac and D. J. Sutherland. Rutgers University Press.
New Brunn ick. NJ. 1990. p. 134.
6.
Balaraman. K.. Gunasekaran. K. and Hoti. S.L. Prospects of
incorporating biological agents in vector control programmes.
IC3IR Bull 25: 97. ? 995?
7.
Singer. S. Bacillus sphaericus (Bacteria). In: Biological Con
trol of Mosquitoes. Ed. I I.C. Chapman. American Mosquito
Control Association Bulletin 6. 1985. p. 123.
8.
Ansari. M.A.. Sharma. V.P.. Mittal. P.K.. Razdan. R.K.. and
Batra. C.P. Evaluation pf Bacillus sphaericus to control breed
ing of malaria vectors. Indian J Malarial 26: 25. 1989.
9.
Chowanadisai. L.. Krairiksh. S. and Thanasripukdikul. S. Mi
crobial control of Anopheles sandaicus. a malaria vector in
Thailand, by Bacillus sphaericus 2362. Mosq Borne Dis Bull 6:
39. 1989.
10.
Yap. I I.H. Field trial of Bacillus sphaericus for mosquito control.
In: Bacteria! Control ofMosquitoes and Black Flies: Biochem
istry. Genetics and Application of Bacillus thuringiensis and
Bacillus sphaericus. Eds. H. de Barjac and D. J. Sutherland.
Rutgers University Press. New Brunswick. NJ. 1990. p. 307.
11.
12.
13.
14.
Karch. S.. Asidi. N.. Manzambi. M. and Salaun. J.J. Efficacy
of Bacillus sphaericus against the malaria vector Anopheles
gambiae and other mosquitoes in swamps and rice fields in Zaire.
J Am Mosq Cont Assoc 8: 376. 1992.
Ragoonanansingh. R.N.. Nijunwa. KJ.. Curtis. C.F. and Becker.
N. A. Field study at'Bacillus sphaericus for the control of culicine
and anophelinc mosquito larvae in Tanzania. Bull Soc lector
Ecol /“: 45. 1992.
Hougard. J.M.. Mbentengam. R.. Lochouarn. L.. Escaffre. IL.
Darriet. F.. Barbazan. P. and Quillevere. D. Control of Culex
quinquefasciatus by Bacillus sphaericus: Results of a pilot cam
paign in a large urban area in equatorial Africa. Bull 111IO "I:
367. 1993.
Sinegre. G.. Babinot. M.. Vigo. G. and Jullien. J.L. Bacillus
sphaericus et demons! ication urbanie. Bilan de cing anneesd'
utilisation experimentale de la speciale 'te Spherimos dans le
sud de la France'. Entente Interde Partementale pour la
Demonstication du Littoral Mediterranean Document E.I.D.L.M.
62 : I. 1993.
24.
Hornby. J.A.. Henlein. B.C.. Levy. R. and Miller. T.W.Jr.
Persistent activity of mosquito larvicidal Bacillussphaericus
1593 in fresh water and sewage. WHO/I'BC/81.830, 1981.
25.
Des Rochers. B. and Garcia. R. Evidence for persistence and
recycling of Bacillus sphaericus. Mosq N’ews 44: 160. 1984.
26.
WHO. Report ofthe 17/ Meeting ofthe Scientific Working Group
on Biological Control. Programme for Research and Training
in Tropical Diseases. UNDP/World Bank. Geneva. Switzerland.
1984.
27.
Charles. J.F. and Nicolas. L. Recycling of Bacillus sphaericus
2362 in mosquito larvae: A laboratory study. Ann Inst Pasteur
Microbiol 13~B: 101. 1986.
28.
Nicolas.!... Dossou-Yovo. J and Hougard. J.M. Persistence and
recycling of Bacillus sphaericus 2362 spores in Culex
quinquefasciatus breeding sites in West Africa. Appl Microbiol
Bidtechnol 25: 341. 1987.
7
29.
30.
31.
32.
33.
de Barjac. H. Classification of Bacillus sphaericus strains and
comparative toxicity to mosquito larvae. In: Bacteria! Control
of Mosquitoes and Black Flies: Biochemistry. Genetics and
Application of Bacillus thuringiensis and Bacillus sphaericus.
Eds. H. de Barjac and D.J. Sutherland. Rutgers University Press.
New Brunswick. NJ. 1990. p. 228..
Lacey. L. A. Persistence and formulation of Bacillus sphaericus.
In: Bacteria! Control ofMosquitoes and Black Flies: Biochem
istry, Genetics and Application of Bacillus thuringiensis and
Bacillus sphaericus. Eds. H. de Barjac and D. J. Sutherland.
Rutgers University Press. New Brunswick. NJ. 1990. p.284..
Nielsen-Leroux. C. and Charles. J.F. Binding of Bacillus
sphaericus binary toxin to a specific receptor on midgut brush
border membranes from mosquito larvae. Eur J Biochem 210:
585. 1992.
Carvalho-Pinto. C.J.. Robinovitch. L.. Alves. R.S.A.. Silva. C.M.B.
and Consoli. R.A.G.B. Fate of Bacillus sphaericus after inges
tion by the predator Belostonia micantulum (Hcmiptcra:
Belostomatidae). Mem Inst Osxvaldo Cru: 90: 329. 1995.
Correa. M. and Yousten. A.A. Bacillus sphaericus spore germi
nation and recycling in mosquito larval cadavers. J Invertebr
Pathol 66: 76. 1995.
34.
Federici. B.A. The future of microbial insecticides as vector control
agents. J Am Mosq Cont Assoc II: 260. 1995.
35.
Rodcharoen. J. and Mulla. M.S. Resistance development in Culex
quinquefasciatus (Diptcra: Culicidae) to Bacillus sphaericus. J
Econ Entomol 87: 1133. 1994.
36.
Beck. M.. Ludwig. M. and Becker. N. Impact of micro organ
isms and water quality in the efficacy of Bacillus sphaericus
(Ncidc) against Culex pipiens larvae in the laboratory. J Vector
Ecol 21: 26. 1996. '
37.
Mulla. M.S.. Rodcharoen. J.. Ngamsuk. W., Tawatsin. A.. PanUrai. P and Thavara. U. Field trials with Bacillus sphaericus
formulations against polluted water mosquitoes in a suburban
area of Bangkok, Thailand. J Am Mosq Cont Assoc. 13: 297.
1997.
38.
39.
Gcorghiou. G.P.. Malik. J.I.. Wirth. M. and Sainato. K. Char
acterization of resistance of Culex quinquefasciatus to the in
secticidal toxins of Bacillus sphaericus (strain 2362). In: An
nual Report. Division of Agriculture and Natural Resources.
University of California. Riverside. CA. 1992. p. 34.
Rodcharoen. J. and Mulla. M.S. Cross-resistance to Bacillus
sphaericus strains in Culex quinquefasciatus. J Am Mosq Com
Assoc 12: 247. 1996.
thuringiensis and Bacillussphaericus. 1*ds. 11, de Barjac and I).
J. Sutherland. Rutgers University Press. New Brunswick. NJ.
1990. p. 221.
43.
Mulla. M.S. Efficacy of the microbial agent Bacillus sphaericus
Neide against mosquitoes (Diptcra: Culicidae) in southern Cali
fornia. Bull Soc Vector Ecol II: 247. 1986.
44.
Charles J.F.. Nielsen-Leroux C. and Deleclusc A. Bacillus
sphaericus toxins: Molecular biology and mode of action. Ann
Rev Entomol 41: 451. 1996.
45.
Liu. J.W.. Porter. A.. Wee. B.Y. and Thanabalu. T. New gene
from nine Bacillus sphaericus strains encoding highly conserved
35.48 kilodalton mosquitocidal toxins. Appl Environ Microbiol
62: 2174. 1996.
46.
Thanabalu T. and Porter A. A Bacillus sphaericus gene encod
ing a novel type pf mosquitocidal toxin of 31.8 kDa. Gene 1 "0:
85. 1996.
47.
Oei. C.. Bindley. J. and Berry. C. Binding of purified Bacillus
sphaericus binary toxin and its deletion
derivatives to Culex
quinquefasciatus gut: Elucidation of functional binding domains,
J Gen MicrobioH38: 1515. 1992.
"
48.
Charles. J.F.. Hauron. S. and Baumann. P. Inclusion bodies and
crystals of Bacillus sphaericus mosquitocidal proteins ex
pressed in various bacterial hosts. Res Microbiol 144: 411. 1993.
49.
Nicolas. L.. Nielsen-Leroux. C.. Charles. J.F. and Deleclusc.
A. Respective role of the 42 and 51 kDa component of the Bacillus
sphaericus toxin overexpressed in Bacillus thuringiensis. FEMS.
Microbiol Lett 106: 275. 1993.
50.
Charles J.F.. Silva-Filha M.B.. Nielsen-Leroux C.. Humphreys
MJ. and Berry C. Binding of the 51 and 42 kDa individual
components from the Bacillus sphaericus crystal toxin to mos
quito larval midgut membranes from Culex and Anopheles sp.
(Diptcra: Culicidae). FEMS. Microbiol Lett 156: 153. 1997.
51.
Davidson. E. W. Biochemistry and mode of action of the Bacil
lus sphaericus toxins. Meni Inst Osxvaldo Crtt: 90: 81. 1995.
52.
Cokmus. C.. Davidson. E.W. and Cooper. K. Electrophysiological
effects of Bacillus sphaericus binary toxin on cultured mosquito
cells. J Invertebr Pathol 69: 197. 1997.
J-x Bern-. C.. Bindley. J.. Ehrhardt. A.F.. Grounds. 1. and de Souza.
I. Genetic determinants of host ranges of Bacillus sphaericus
mosquito larvicidal toxins. J Bacterio! I “5 : 510. 1993.
3-L Singer.S. Bacillus sphaericus for the control of mosquitoes.
Biotechnol Bioeng 22 : 1335. 1980.
^5. Lacey. L.A. and Singer. S. Larvicidal activity of new isolates
40.
41.
Rodcharoen. J. and Mulla. M.S. Resistance in Culex
quinquefasciatus to Bacillus sphaericus. In.- Annual Report.
Division of Agriculture and Natural Resources. University of
California. Riverside. CA. 1991. p. 18..
of Bacillus sphaericus and Bacillus thuringiensis (11-14) against
anopheline and culicinc mosquitoes. Mosq Xews 42: 537.1982.
Silapanuntakul. S.. Pantuwatana. S.. Bhumiratana. A. and
Charoensiri. K. The comparative persistence of toxicity of Bacillus
sphaericus strain 1593 and Bacillus thuringiensis serotype 1114 against mosquito larvae in different kinds of environments.
Davidson. E.W.. Urbina. M.. Payne. J.. Mulla. M.S.. Dulmage.
H.
T..
Darwazeh. H.A. and Correa. J. A. Fate of Bacillus sphaericus
1593 and 2362 spores used as larvicides in the aquatic environ
ment. Appl Environ Microbiol 4~: 125. 1984.
J Invertebr Pathol 42 : 387. 1983.
57.
42.
8
Singer. S. Introduction to the study of Bacillus sphaericus as a
mosquito control agent. In: Bacterial Control ofMosquitoes and
Black Flies: Biochemistry, Genetics and Application ofBacillus
Mulla. M.S.. Darwazeh. H.A.. Davidson. E.W.. Dulmage. H.T.
and Singer. S. Larvicidal activity and field efficacy of Bacillus
sphaericus strains against mosquito larvae and their safety to
non-target organisms. Mosq Xews 44 : 336. 1984.
58.
Mulla. M.S.. Darwazeh. I LA.. Ede. L.. Kennedy. B. and Dulmage.
11.1. Efhcacy and held evaluation of Bacillus ihuringiensis (II14) and Bacillus sphaericus against flood waler mosquitoes in
California. J Am Mosq Cont Assoc / : 310. 1985.
73.
Darwazeh. H.A .. Zgomba. M. and Mulla. M.S. Environmental
factors affecting the efficacy ofmicrobial control agents. In: Annual
Report. Division of Agriculture and Natural Resources. Univer
sity of California. Riverside. CA. 1990. p. .23.
59.
Davidson. EAV. Variation in the binding of Bacillus sphaericus
toxin and wheat germ agglutinin to larval midgut cells of six
species of mosquitoes. J Invertebr Pathol 53 : 251. 1989.
74.
Mulla. M.S.. Darwazeh. ILA. and Zgomba. M. Effects of some
environmental factors on the efficacy of Bacillus sphaericus 2362
and Bacillus ihuringiensis var. israelensis (H-14) against mos
quitoes. Bull Soc Vector Eco! 15 : 166. 1990.
60.
Baumann. P.. Clark. M.A.. Baumann. L. and Broadwell. A.II.
Bacillus sphaericus as a mosquito pathogen: Properties of the
organism and its toxins. Microbiol Rev 55 : 425. 1991.
75.
McGaughey. W.H. and Whalon. M.E. Managing insect resis
tance to Bacillus ihuringiensis toxins. Science 258: 1451. 1992.
76.
Roush. R.T. Occurrence, genetics and management of insecti
cide resistance. Parasito! Today 9 : 174. 1993.
77.
Rodcharoen. .1. and Mulla. M.S. Comparative ingestion rales of
Culex quinquefasciatus (Diptera: Culicidac) susceptible and
resistant to Bacillus sphaericus. J Invertebr Pathol 66 : 242.
61.
Aly. C.. Mulla. M.S. and Federici B./\. Ingestion, dissolution
and proteolysis of the Bacillus sphaericus toxin by mos
quito larvae. J Invertebr Pathol 53 : 12. 1989.
62.
Baumann. P.. Unterman. B.M.. Baumann. L.. Broodwell. A.11.
and Abbene. SJ. Purification of the larvicidal toxin of Bacillus
sphaericus and evidence for high molecular weight precursors.
J Bacterio! 163 : 738. 1985.
63.
Davidson. EAV. Effects of Bacillus sphaericus 1593 and 2362
spores/cryslal toxin on cultured mosquito cells. J Invertebr Pathol
4“ : 21. 1986.
64.
Davidson. E.W.. Bieger. A.L.. Meyer. M. and Sheilabarge. R.C.
Enzymatic activation of the Bacillus sphaericus mosquito lar
vicidal toxin. J Invertebr Pathol 50 : 40. 1987.
65.
Broodwcll. A.H. and Baumann. P. Sporulation associated acti
vation of Bacillus sphaericus larvicides. Appl Environ Microbiol
52 : 758. 1986.
66.
Nicholas. L.. Lecroisey. A. and Charles. J.F. Role of the gut
proteinases from mosquito larvae in the mechanism of action
and the specificity of the Bacillus sphaericus toxin. Can J
Microbiol 36 : 804. 1990.
1995.
78.
Rodcharoen. J. and Mulla. M.S. Biological fitness of Culex
quinquefasciatus (Diptera: Culicidac) susceptible and resistance
to Bacillus sphaericus. J Med Entomo! 34 : 5. 1997.
79.
McGaughey. W.H. and Beeman. R.W. Resistance to Bacillus
ihuringiensis var. israelensis in colonies of Indian meal moth
and almond moth (Lepidoptera: Pyralidae). J Econ Entomol 81:
28. 1988.
80.
Mulla. M.S.. Axelrod. H.A. Darwazeh. H.A. and Matanmi. B.A.
Efficacy and longevity of Bacillus sphaericus 2362 formula
tions for control of mosquito larvae in dairy waste water lagoons.
J Am Mosq Cont Assoc 4: 448. 1988.
81.
Matanmi. B.A.. Federici. B.A. and Mulla. M.S. Fate and per
sistence of Bacillus sphaericus used as mosquito larvicide in
dairy waste water lagoons. J Am Mosq Cont Assoc 4: 1990.
67.
Davidson. E.W. Microbiology, pathology, and genetics ofBacillus
sphaericus: Biological aspects which arc important to field use.
Mosq News 44 : 147. 1984.
82.
Georghiou. G.P. Management of resistance in arthropods. In:
Pest Resistance to Pesticides. Eds. G.P. Georghiou and T. Saito.
Plenum Publishers. New York. 1983. p. 769.
68.
Davidson. E. W. Binding of the Bacillus sphaericus (Eubactcriales:
Bacillaceae) toxin to midgut cells of mosquitoes (Diptera :
Culicidac) larvae: Relationship to host range. J Med Entomol
25:151. 1988.
83.
Georghiou. G.P. and Taylor. C.E. Resistance to pesticides and
antibiotics: How far is it manageable?. In: Combating Resis
tance to Xenobiotics. Eds. M.G. Ford. DAV. Hallowman. P.B.S.
Khambay and R.M. Sawick. Chichester. England. 1986. p. 11.
69.
Silva-Filha. M.H.. Nielsen-Leroux. C. and Charles. J.F. Bind
ing kinetics of Bacillus sphaericus binary toxin to midgut brush
border membranes of Anopheles and Culex sp. mosquito lalrvae.
Ear J Biocheni 24“ : 754. 1997.
84.
Gillett. J.D. The Mosquitoes: Life. Activities and Important Human
Affairs. Richard Clay. Chaucer Press. Bungay. Great Britain:
1971. p. 149.
85.
70.
Krych. V.. Johnson. J. and Yousten. A. Deoxyribonucleic acid
homologies among strains of Bacillus sphaericus. Ini J Syst
Bacterio! 30 : 476. 1980.
Horsfall. W.R. Mosquitoes: Their Bionomics and Relation to
Disease. Ronald Press: New York:. 1955. p. 723.
86.
Aly. C.. Mulla. M.S.. Xu. B. and Schneller. \V. Rate of inges
tion by mosquito larvae (Diptera: Culicidac) as a factor in the
effectiveness of a bacterial stomach toxin../ Med Entomol 25:
191.1988.
87.
King. C.H.. Sanders. R.W.. Sholts. Jr. E.B. and Porter. K.G.
Differential survival of bacteria ingested by zooplankton from
a stratified eutrophic lake. Limnol Oceanogr 36: 829. 1991.
88.
Manasherob. R.. Ben-Dov. E.. Margalit. J.. Zaritsky. A. and Barak.
Z. Raising activity of Bacillus ihuringiensis var. israelensis against
Anopheles stephensi larvae by encapsulation in Tetrahymena
71.
72.
Nielsen-LeRoux. C.. Charles. J.F.. Thiery. I. and Georghiou.
G.P. Resistance in a laboratory population of Culex
quinquefasciatus (Diptera:Culicidae) to Bacillussphaericus binaiy
toxin is due to a change in the receptor on midgut brush border
membranes. Eur J Biocheni 228 : 206. 1995.
Silva-Filha. M.IL. Regis. L.. Nielsen-Leroux. C. and Charles.
J. Low level resistance to Bacillus sphaericus in a Held treated
F.
population of Culex quinquefasciatus
Culicidac). J Econ
Entomo! 88 : 525. 1995.
9
pyriformis (I lymenostomatida: Tetrahy mcnidae). J Am Mosq Cont
Assoc 12: 627. 1996.
89.
90.
Sun. C.. Georghiou. G.P. and Weiss. K. Toxicity of Bacillus
thuringiensis var. israelensis to mosquito larvae variously re
sistant to conventional insecticides, Mosq Xews 40: 614. 1980.
Mahmood. I*. Laboratory bioassay to compare susceptibilities
of Aedes aegypti and Anopheles a Ibiman us to fine Ulus
thuringiensis var. israelensis as affected by their feeding rates.
J Am Mosq Cont Assoc 14: 69. 1998.
91.
92.
Nugud. A.D. and White. G.B. Evaluation of Bacillus ilhtringiensis
serotype 11-14 formulation of larvicides for Anopheles arabiensis
(species B of Anopheles gambiae complex). Mosq Xews 42: 36.
1982.
Gould. F.. Martinez-Ramirez. A.. Anderson. A., Ferre. J.. Silva.
F.J. and Moar. WJ. Broad spectrum resistance to Bacillus
thuringiensis toxins in Heliothis virescens. Proc Xatl Acad
Sci USA 89: 7986. 1992.
93.
Mittal. P.K. and Adak. T. Laboratory selection of resistance in
Anopheles stephensi to Bacillus sphaericus. (/Abstract). J
Parasit Dis 20: 116. 1996.
94.
Baumann. L.. Broodwell. A.H. and Baumann. P. Sequence analysis
ofthe mosquitocidal toxin genes encoding 51.4 and 41.9-kilodalton
proteins from Bacillus sphaericus 2362 and 2297. J Bacterial
l"0: 2045. 1988.
95.
Sun. C.N.. Georghiou. G.P. and Weiss. K. Toxicity of Bacillus
thuringiensis var. israelensis to mosquito larvae variously re
sistant to conventional insecticides. Mosq Xews 40: 614. 1980.
96.
Goldman. I.F.. Arnold. J. and Carlton. B.C. Selection for re
sistance to Bacillus thuringiensis subsp. israelensis in field and
laboratory populations of the mosquito Aedes aegypti. J Invertebr
Pathol 4"-. 317. 1986.
97.
Tabashnik. B.E.. Finson. N. and Johnson. M.W. Managing
resistance to Bacillus thuringiensis: Lessons from the diamond
black moth (Lepidoptera: Plutallidae). J Eco Entomol 84: 49.
1991.
98.
Ferrari. J.A. and Georghiou. G.P. Effects of insecticidal selec
tion and treatment on reproductive potential of resistant, sus
ceptible and heterozygous strains of the southern house mos
quitoes. J Econ Entomol “4: 323. 1981.
99.
Amin. A.M. and White. G.B. Relative fitness of organophos
phate resistant and susceptible strains of Culex quinquefasciatus
Say (Diptera: Culicida'e). Bull Entomol Res “4: 591. 1984.
100.
Lacey. L.A.. Day. J. and lleitzman. C.M. Long-term effects of
Bacillus sphaericus on Culex quinquefasciatus. J Invertebr Pathol
49: 116. 1987.
101.
Rowland. M. Behaviour and fitness ofy-IICII/dieldrin resistant
and susceptible female Anopheles gambiae and Anopheles
stephensi mosquitoes in the absence of insecticide. Med I’et
Entomol 5: 193. 1991.
102.1 leather. N .W. Comparison of population growth rates of malathion
resistant and suseptible populations of the rice weevil. Sitophilus
oryzae (Linnaeus) (Coleoptera: Curculionidae). Queensland J
Agric Anim Sci 39: 61. 1982.
10
103.
Gibson. J.P. Problems in obtaining a description of the evalu
ation of dimethoate resistance in Danish house Hies (Musca
domestica) Pestic Sci 12: 565. 1981.
104.
French-Constant. R.H. and Roush. R.T- Resistance detection
and documentation: The relative roles of pesticidal and biochemical
assays. In: Pesticide Resistance in A arthropods Eds. R.T. Roush
and B.E. Tabashnik. Chapman & Hall. New York. 1990. p. 4.
105.
Silva-Filha. M.H. and Regis. L. Reversal of low-level resis
tance to Bacillus sphaericus in a field population of southern
house mosquito (Diptera: Cuiicidae) from an urban area of Recife.
Brazil. J Econ Entomol 90: 299. 1997.
106.
Rodcharoen. J. and Muila. M.S. Mosquito larval resistance to
the microbial larvicide Bacillus sphaericus. In: Annual Report.
Division of /Agriculture and Natural Resources. University of
California. Riverside. CA. 1990. p. 24.
107.
Georghiou. G.P.. Wirth. M.C.. Delecluse. A. and Klier. A.
Potentiality for development of resistance to single vs. multiple
toxins of bacilli by mosquitoes. In: Proceedings of Interna
tional Conference on Bacillus. Paris Institute Pasteur. Paris. 1993.
p. 68.
108.
Van Rie. J.. Jansens. S.. Hoftc. H.. Degheele. D. and Van Mellaert.
IL Receptors on the brush border membrane of the insect mid
gut as determinants of the specificity of Bacillus thuringiensis
delta endotoxins. Appl Environ Microbiol 56: 1378. 1990.
109.
Federici. B.A.. Ge .B.. Johnson. J J. and Poehner. WJ. Improve
ment of bacterial mosquito larvicides. In: Annual Report. Divi
sion of Agriculture and Natural Resources. University of Cali
fornia. Riverside. CA. 1996. p. 18.
110.
Ali. A. Bacillus thuringiensis serovar. israelensis (ABG-6I08)
against chironomids and some non-target aquatic invertebrates.
J Invertebr Pathol 38: 264. 1981.
III.
Lacey. L.A and Mulla. M.S. Safety of Bacillus thuringiensis
var israelensisis and Bacillus sphaericus to nontarget organ
isms in the aquatic environment. In: Safety of Microbial Insec
ticides. Eds. MvLaird. L.A. Lacey and E.W. Davidson. CRC
Press. Boca Raton. Florida. 1990. p. 169.
112.
Das. P.K and Amalraj. D.D. Biological control of malaria vec
tors. Indian J Med Res 106: 174. 1997.
113.
Caghlon. /\. Friend turns to foe. Xew Scientist. 158 IXo. 2136):
7.1998.
114.
Lacey. L.A. and Dame. D.A. The effect of Bacillus thuringiensis
var. israelensis on Toxorhynchites rutHits rutHus (Diptera: Cu
iicidae) in the presence and absence of prey. J Med Entomol
19: 593. 1982.
U5.Tielze. N.S.. Bestir. P.G.. Shaffer. K.R.. Prescoll. SJ. and
Schreiber. E.T. Integrated management of waste lyre mosqui
toes utilizing Mesocyclops longisetus (Copepoda: Cyclopidae).
Bacillus thuringiensis var. israelensis. Bacillus sphaericus and
methoprene. J .1 m Mosq Cont Assoc 10: 363. 1994.
116. Barbosa. J.F.N.. Quiroz-Martinez. IL. Rodriguez-Tovar. M.L..
Tejada. L.O. and Badii. M.H. Use of BactimosR briquets (B.t.i.
formulation) combined with the backswimmer Xotonecta irrorata
(Hemiptera: Notonectidae) for control of mosquito larvae. J Am
Mosq Cont Assoc 13: 87. 1997.
a 3 fold risk was observed for those who consumed alcohol.
It was observed that the patients who consumed eggs
had nearly 2 fold risk of urolithiasis than those who did
149.7 (±4.7) cm. 42.8 (±5.5) kg and 28.5 (±7.1) yr.
respectively. The percentage of fat. lean body weight and
body surface area was 21.2 (±3.7)%. 34.0 (±0.9) kg and
not consume eggs. Similarly a 2 fold risk of urolithiasis
was observed in patients who consumed flesh food twice
or even less than twice per week compared to vegetarian.
1.5 (±0.1) nr. respectively. The average energy intake
was 1355 kcal. The average walking speed in the field
was 3.4 km/h and the maximum heart rate of 163 beats/
min was observed for women engaged in block carrying.
Heart rates during other activities like carrying concrete
for column and beam, carrying concrete for block making
and mud transferring were between 110-135 beats/min.
Consumption of rajma more than twice a week also resulted
in 2 fold higher risk of urolithiasis.
It was thus concluded that urinary tract infections.
smoking, alcohol intake and consumption of egg. non
vegetarian food items as well as rajma were associated
with increased risk of urolithiasis.
D. Nayar
U. Kapil
Human Nutrition Unit
Department of Gastroenterology
All India Institute of Medical Sciences
New Delhi.
Ergonomic evaluation of different modes of load
carrying and maximum permissible load for Indian
women engaged in manual material handling.
The study was carried out to document the existing
modes of manual load carrying by Indian women and
determine the physiological cost involved during the
process.
Two basic approaches (field and laboratory studies)
were employed for the study. Several parameters like
personal and medical history, sites of pain in the body.
etc. were ascertained through a questionnaire.
Anthropometry, physiological cost of the work, work
rest cycle, walking speed, environmental heat load and
energy consumption were determined by direct
measurements on the subjects in the field.
Women involved in construction activities carried
the load on the head but in case of water carrying four
basic modes like head, shoulder, waist and hand mode
were observed. Besides these, womjen had adopted sev
eral other modes for carrying water like head and hand.
The O,demand varied from 0.77 l/min (45.31% of VO.
max) to 1.41 l/min(82.7% of VO.max.): being lowest at
ground level and maximum after climbing to the 4th floor.
The maximum aerobic power obtained was 1.71 l/min.
A regression equation between heart rate and O,
consumption was derived.
In case of women carrying water the average body
height, weight and age was 150.0 (±5.3) cm. 50.4 (±10.9)
kg and 30.5 (±8.1) yr. respectively. The fat content, lean
body weight and body surface area was 25.6 (±5.9)% of
the whole body weight. 36.6 (±6.0) kg and 1.6 nr. re
spectively. It was evident that women involved in water
carrying had significantly more body weight, fat content
and lean body weight as compared to those involved in
construction activity. The women involved in water car
rying had work experience ranging from a few' months
to 25 years and more. The water was being carried by
HANDA and load varied from 10 to 50 kg. while in case
of construction activity the weight varied from 9-34 kg.
Women carrying water reported the back, elbow, upper
arm. calf and knee joint as the major sites of pain. Simi
lar sites of pain were also reported by the construction
workers. About 6.5% women water carriers reported
miscarriage while miscarriage was reported by 8.3 women
involved in construction activity. The average walking
speed for water carrying was 3.8 km/h which was slightly
higher than that of construction group.- The average
working heart rate was 129 beats/min.
The average maximum heart rate and Onconsumption
for women carrying water during maximum aerobic power
was 182.3 beats/min and 1.47 l/min respectively, tyeing
lower than that of construction workers. This indicates
head and waist, waist and hand etc. Ninety three women
engaged in construction activities and 76 women engaged
that the physical w ork capacity of the construction workers
in water carrying activities were selected at random for
activity.
detailed study. Anthropometric details were recorded on
47 women workers from construction and 25 women
workers from water carrying activities.
In the case of women engaged in construction
activities the average bod\ height, weight and age were
was superior to that of women engaged in water carrying
Vasudha Atreya
G.G. Ray
Industrial Design Centre
Indian Institute of Technology
Mumbai.
ICMR NEWS
The following meetings of various technical groups/
committees of the Council were held:
Meetings of the Scientific Advisory Committee (SAC)/
Project Review Committee (PRC)/Task Forces (TFs)
held:
Sr.Research Officer, ICMR Headquarters, New Delhi,
participated in the workshop on International Research
Ethics at Baltimore (January 5-6, 1999).
Dr. Neena Valecha, Asstt. Director, and Dr. C. Usha
Devi, Sr. Research Officer, Malaria Research Centre,
Delhi, participated in the workshop on Azithromycin in
PRC on Oncology and
December 17-18, 1998
(at New Delhi)
Pathology
TF on Drug Resistance in
Malaria at Kathmandu (January 14, 1999).
December 31, 1998
Malaria
Appointment:
(at New Delhi)
TF on Diagnostics in Malaria
January 11, 1999
Dr. Ashok Mukherjee took over as Director of the
Institute of Pathology, New Delhi w.e.f. January 7,1999.
(at New Delhi)
SAC of the Desert Medicine
January 12-13, 1999
Research Centre, Jodhpur
Participation of ICMR Scientists in Scientific Events:
Dr. S.M. Mehendale, Dy. Director, National AIDS
Research Institute, Pune and Dr. Nandini K. Kumar,
Workshop:
An Indo-US Workshop on Behavioural Prevention
Research on Hl V/AIDS was organized in New Delhi during
January 12-13, 1999 for identification of areas for col
laboration through joint projects.
ICMR AIDED SYMPOSIA/SEMINARS/WORKSHOPS/COURSES/CONFERENCES
Symposium/Seminar/Workshop/
Date & Place
Contact Address
Course/Conference
LXXXVI Session of the Indian Science Congress *
Association
January 3-7. 1999:
(at Chennai)
Prof. Kunthala Jayaraman. Dean of Technology &
Professor of Biotechnology, Anna University.
Chennai-600025.
V International Symposium on Biochemical Roles of
Eukaryotic Cell Surface Macromolecules
January'4-8. 1999:
(at Bangalore)
Dr. A. SuroJia. Convener of the Symposium.
Molecular Biophysics Unit. Indian Institute of
Science. Bangalore-560012.
Workshop on Scientific Communication
, January 9-10. 1999:
(at Mumbai)
Dr. S.R. Naik. Professor & Head. Department of
Gastroenterology. Sanjay Gandhi Postgraduate
Institute of Medical Sciences. Lucknow-226014.
National Workshop on Hclicobactcr/Campylobactcr
January' 19-23. 1999:
(at Lucknow)
Dr. A. Ayyagari. Organising Secretary' of the
Workshop. Department of Microbiology. Sanjay
Gandhi Postgraduate Institute of Medical Sciences..
Lucknow-226014.
National Symposium on Biological Control of Insects in
Agriculture. Forestry'. Medicine and Veterinary’ Science
January 21-22. 1999:
(at Coimbatore)
Dr. K. Murugan. Organising Secretary’ of the
Symposium. Division of Entomology*. Department
of Zoology. Bharathiar University. Coimbatore641046. ~
XVII National Conference of the Society for Reproductive
Biology and Comparative Endocrinology
January'23-25. 1999:
(at Surat)
Dr. Nandita J. Arbatti. Organising Secretary' of the
Conference. Department of Biochemistry'. Govern
ment Medical Collese. Surat-395001.
13
ICMR NEWS
The following meetings of various technical groups/
committees of the Council were held:
Meetings of the Scientific Advisory Committee (SAC)/
Project Review Committee (PRC)/Task Forces (TFs)
held:
Sr.Research Officer, ICMR Headquarters, New Delhi,
participated in the workshop on International Research
Ethics at Baltimore (January 5-6, 1999).
Dr. Neena Valecha, Asstt. Director, and Dr. C. Usha
Devi, Sr. Research Officer, Malaria Research Centre,
Delhi, participated in the workshop on Azithromycin in
PRC on Oncology and
December 17-18, 1998
(at New Delhi)
Pathology
TF on Drug Resistance in
Malaria at Kathmandu (January 14, 1999).
December 31, 1998
Malaria
Appointment:
(at New Delhi)
TF on Diagnostics in Malaria
January 11, 1999
Dr. Ashok Mukherjee took over as Director of the
Institute of Pathology, New Delhi w.e.f. January 7,1999.
(at New Delhi)
SAC of the Desert Medicine
January 12-13, 1999
Research Centre, Jodhpur
Participation of ICMR Scientists in Scientific Events:
Dr. S.M. Mehendale, Dy. Director, National AIDS
Research Institute, Pune and Dr. Nandini K. Kumar,
Workshop:
An Indo-US Workshop on Behavioural Prevention
Research on Hl V/AIDS was organized in New Delhi during
January 12-13, 1999 for identification of areas for col
laboration through joint projects.
ICMR AIDED SYMPOSIA/SEMINARS/WORKSHOPS/COURSES/CONFERENCES
Symposium/Seminar/Workshop/
Date & Place
Contact Address
Course/Conference
LXXXVI Session of the Indian Science Congress *
Association
January 3-7. 1999:
(at Chennai)
Prof. Kunthala Jayaraman. Dean of Technology &
Professor of Biotechnology, Anna University.
Chennai-600025.
V International Symposium on Biochemical Roles of
Eukaryotic Cell Surface Macromolecules
January'4-8. 1999:
(at Bangalore)
Dr. A. SuroJia. Convener of the Symposium.
Molecular Biophysics Unit. Indian Institute of
Science. Bangalore-560012.
Workshop on Scientific Communication
, January 9-10. 1999:
(at Mumbai)
Dr. S.R. Naik. Professor & Head. Department of
Gastroenterology. Sanjay Gandhi Postgraduate
Institute of Medical Sciences. Lucknow-226014.
National Workshop on Hclicobactcr/Campylobactcr
January' 19-23. 1999:
(at Lucknow)
Dr. A. Ayyagari. Organising Secretary' of the
Workshop. Department of Microbiology. Sanjay
Gandhi Postgraduate Institute of Medical Sciences..
Lucknow-226014.
National Symposium on Biological Control of Insects in
Agriculture. Forestry'. Medicine and Veterinary’ Science
January 21-22. 1999:
(at Coimbatore)
Dr. K. Murugan. Organising Secretary’ of the
Symposium. Division of Entomology*. Department
of Zoology. Bharathiar University. Coimbatore641046. ~
XVII National Conference of the Society for Reproductive
Biology and Comparative Endocrinology
January'23-25. 1999:
(at Surat)
Dr. Nandita J. Arbatti. Organising Secretary' of the
Conference. Department of Biochemistry'. Govern
ment Medical Collese. Surat-395001.
13
Symposium/Seminar/Workshop/
Contact Address
Date & Place
Course/Conference
International Workshop on Hypospadias and the Child with
Undetermined Sex
January 30-31. 1999:
(at New Delhi)
Dr. D.K. Gupta. Convenor of the Workshop.
Department of Paediatric Surgery. All India
Institute of Medical Sciences. New Delhi-110029.
Semiqar on Low Level Electromagnetic Field Phenomena
in Biological System
February 3-4. 1999:
(at New Delhi)
Dr. J. Behari. Professor of Environmental Physics,
School of Environmental Sciences. Jawaharlal
Nehru University, New Delhi-110067.
I National Conferen.ee on Hepatitis and Renal Medicine
February 13. 1999:
(at New Delhi)
Dr. Sanjay K. Agarwal. Organising Secretary' of
the Conference, Department of Nephrology. All
India Institute, of Medical Sciences. New
Delhi-110029.
VII Annual Meeting of Molecular Immunology Forum 1999
February 13-15. 1999:
(at Hyderabad)
Dr. Ashok Khar, Convener of the Meeting. Centre
for Cellular and Molecular Biologv. Hvderabad500007.
IV International Symposium on Vectors and Vector-borne
Diseases
February 18-20. 1999;
(at Gwalior)
Dr. P.R. Malhotra. Convener of the Symposium.
Defence Research and Development Establishment
Gwalior-474002.
XVIII Annual Convention of Indian Association for Cancer
Research and National Symposium on Molecular Biology
of Cancer
February' 19-21. 1999:
(at New Delhi)
Dr. Neeta Singh. Addl. Professor. Department of
Biochemistry, All India Institute of Medical
Sciences, New Delhi-110029.
AOCS-OTA1 International Conference and Exhibition on
Oil Seeds*and Vegetable Oil Processing
February' 19-21. 1999:
(at New Delhi)
Sh. K.P. Sharma. Organising Secretary' of the
Conference, Oil Technologists Association of
India. 1205. Nirmal Tower. Barakhambha Road,
New Delhi-110001.
Technical Training Workshop on Industrial Hygiene
Measurements and National Seminar on Occupational
Safety and Health
March 1-6. 1999:
(at Chennai)
Dr. Kalpana Balakrishnan. Organising Secretary'
of the Workshop. Sri Ramachandra Medical
College and Research Institute. Chennai-600016.
COUNCIL’S TRAINING PROGRAMMES FOR 1999-2000
Reproductive Biology
Nutrition '
At the Institutefor Research in Reproduction, Mumbai:
At the National Institute of Nutrition, Hyderabad:
•
•
Training Course in Molecular Techniques in Ovarian
•
Training Course on Cytological Detection of Repro
M.Sc. in Applied Nutrition (June 1, 1999-February 28,
2000).
Function (September 27-October 2, 1999).
•
Postgraduate Certificate Course in Nutrition (Decem
ber 1,1999-February 28,2000).
ductive Tract Infections (December 13-17, 1999).
Endocrinology
Laboratory Animal Technology'
At the National Institute ofNutrition,, Hyderabad:
At the National Centre for Laboratory Animal Science,
National Institute of Nutrition, Hyderabad:
•
Annual Certificate Course on Endocrinological
Techniques and their Application (August 1 -
September 15,1999).
14
•
Training Course for Laboratory Animal Technicians
(June !4-July30,1999).
Symposium/Seminar/Workshop/
Contact Address
Date & Place
Course/Conference
International Workshop on Hypospadias and the Child with
Undetermined Sex
January 30’-31. 1999:
(at New Delhi)
Dr. D.K. Gupta. Convenor of the Workshop.
Department of Paediatric Surgery. All India
Institute of Medical Sciences. New Delhi-110029.
Seminar on Low Level Electromagnetic Field Phenomena
in Biological System
February 3-4. 1999;
(at New Delhi)
Dr. J. Behari. Professor of Environmental Physics,
School of Environmental Sciences. Jawaharlal
Nehru University. New Delhi-110067.
I National Conferen.ee on Hepatitis and Renal Medicine
February 13. 1999:
(at New Delhi)
Dr. Sanjay K. Agarwal. Organising Secretary of
the Conference, Department of Nephrology. All
India Institute, of Medical Sciences. New
Delhi-110029.
VII Annual Meeting of-Molecular Immunology Forum 1999
February 13-15. 1999:
(at Hyderabad)
Dr. Ashok Khar, Convener of the Meeting, Centre
for Cellular and Molecular Biology. Hyderabad500007.
IV International Symposium on Vectors and Vector-borne
Diseases
February 18-20. 1999:
(at Gwalior)
Dr. P.R. Malhotra. Convener of the Symposium.
Defence Research and Development Establishment.
Gwalior-474002.
XVIII Annual Convention of Indian Association for Cancer
Research and National Symposium on Molecular Biology
of Cancer
February' 19-21. 1999;
(at New Delhi)
Dr. Neeta Singh. Addl. Professor. Department of
Biochemistry', All India Institute of Medical
Sciences, New Delhi-110029.
AOCS-OTAI International Conference and Exhibition on
Oil Seeds-and Vegetable Oil Processing
February' 19-21. 1999:
(at New Delhi)
Sh. K.P. Sharma. Organising Secretary' of the
Conference, Oil Technologists Association of
India. 1205, Nirmal Tower. Barakhambha Road.
New Delhi-110001.
Technical Training Workshop on Industrial Hygiene
Measurements and National Seminar on Occupational
Safety and Health
March 1-6. 1999:
(at Chennai)
Dr. Kalpana Balakrishnan. Organising Secretary'
of the Workshop, Sri Ramachandra Medical
College and Research Institute, Chennai-600016.
COUNCIL’S TRAINING PROGRAMMES FOR 1999-2000
Reproductive Biology
Nutrition
At the Institutefor Research in Reproduction, Mumbai:
At the National Institute of Nutrition, Hyderabad:
•
•
Training Course in Molecular Techniques in Ovarian
•
Training Course on Cytological Detection of Repro
M.Sc. in Applied Nutrition (June 1, 1999-February 28,
2000).
Function (September 27-October 2, 1999).
•
Postgraduate Certificate Course in Nutrition (Decem
ber I, 1999-February 28,2000).
ductive Tract Infections (December 13-17, 1999).
Endocrinology
Laboratory Animal Technology
At the National Institute ofNutrition,, Hyderabad:
At the National Centre for Laboratory Animal Science,
National Institute ofNutrition, Hyderabad:
•
Annual Certificate Course on Endocrinological
Techniques and their Application (August 1 -
September 15, 1999).
14
•
Training Course for Laboratory Animal Technicians
(June 14-July30, 1999).
•
Training Course for Laboratory Animal Supervisors
(S eptem ber 1 Nove in ber 30, 1999).
Medical Entomology:
Biomedical Statistics
j
Institutefor Research in Medical Statistics, Chennai:
e
Basic Course in Statistics for Medical Officers (July
At the Vector Control Research Centre, Pondicherry:
•
Postgraduate Diploma Course in Medical Entomol-
1999).
•
Training Course in Field Epidemiology (November,
ICMR PUBLICATIONS
PricefRs.)
Nutritive Value of Indian Foods (1985) by C. Gopalan, B.V. Rama Sastri and
23.00
S.C. Balasubramanian, Revised and Updated (1989) by B.S. Narasinga Rao, Y.G. Deosthale
and K.C. Pant (Reprinted 1996)
Growth and Physical Development of Indian Infants and Children (1972,. Reprinted 1989)
10.00
Studies on .Weaning and Supplementary Foods (1974, Reprinted 1996)
15.00
A Manual of Nutrition (Second Edition 1974, Reprinted 1995)
6.00
Low Cost Nutritious Supplements (Second Edition 1975, Reprinted 1996)
5.00
Menus for Low-Cost Balanced Diets and School-Lunch Programmes Suitable for North India
(Second Edition 1977, Reprinted 1994) *
4.50
Menus for Low-Cost Balanced Diets and School-Lunch Programmes Suitable for South India
(Fourth Edition 1996)
6.00
Some Common Indian Recipes and their Nutritive Value (Fourth Edition 1977, Reprinted 1995)
10.00
by Swaran Pasricha and L.M. Rebello
Nutrition for Mother and Child (Fourth Edition 1994, Reprinted 1996) by P.S. Venkatachalam
11.00
and L.M. Rebello
Japanese Enceph'alitis in India (Revised Edition 1980)
5.00
Some Therapeutic Diets (Fifth Edition 1996) by Swaran Pasricha
7.00
Nutrient Requirements and Recommended Dietary Allowances for Indians (1990,Reprinted 1995)
16.00
Fruits (Second Edition 1996) by Indira Gopalan and M. Mohan Ram
20.00
Count What You Eat (1989, Reprinted 1997) by Swaran Pasricha
15.00
*The Anophelines of India (Revised Edition 1984) by T.Ramachandra Rao
150.00
15
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