|Year : 2020 | Volume
| Issue : 12 | Page : 525-534
Current status and future prospects of bacilli-based vector control
Joleen Savianne Almeida1, Ajeet Kumar Mohanty1, Savita Kerkar2, Sugeerappa Laxmanappa Hoti3, Ashwani Kumar4
1 ICMR-National Institute of Malaria Research, DHS Building, Campal, Panaji, Goa-403001, India
2 Department of Biotechnology, Goa University, Goa-403206, India
3 National Institute of Traditional Medicine, Nehru Nagar, Belagavi-590010, Karnataka, India
4 ICMR-Vector Control Research Centre, Indira Nagar, Puducherry-605 006, UT, India
|Date of Submission||11-Sep-2019|
|Date of Decision||16-Dec-2019|
|Date of Acceptance||11-Jul-2020|
|Date of Web Publication||19-Oct-2020|
ICMR-Vector Control Research Centre, Indira Nagar, Puducherry-605 006, UT
Source of Support: None, Conflict of Interest: None
Mosquito-borne diseases such as malaria, filariasis, dengue, chikungunya, Japanese encephalitis, yellow fever and Zika contribute significantly to health problems of developing as well as developed nations. Vector control is central to control of vector borne diseases. In the last four-five decades, biological control methods have been inducted in the integrated vector management strategy, advocated nationally as well as globally by the World Health Organization. Currently, biological control of vectors is globally acknowledged as the best available strategy in the wake of growing concerns about vector resistance as well as adverse effects of insecticides on the environment and non-target fauna co-inhabiting the same ecological niches as vectors. In India and elsewhere, efforts are ongoing to screen newer isolates to bring forth new biolarvicidal products of public health importance. In this review, by carrying out extensive literature survey, we discuss advances thus far and the prospects of bacilli-based control of vectors and vector borne diseases.
Keywords: Mosquito-borne diseases; Vector control; Biological control; Biolarvicide
|How to cite this article:|
Almeida JS, Mohanty AK, Kerkar S, Hoti SL, Kumar A. Current status and future prospects of bacilli-based vector control. Asian Pac J Trop Med 2020;13:525-34
|How to cite this URL:|
Almeida JS, Mohanty AK, Kerkar S, Hoti SL, Kumar A. Current status and future prospects of bacilli-based vector control. Asian Pac J Trop Med [serial online] 2020 [cited 2022 May 19];13:525-34. Available from: https://www.apjtm.org/text.asp?2020/13/12/525/296720
| 1. Introduction|| |
Mosquitoes are associated with transmission of pathogens to humans and other vertebrates resulting in significant morbidity and mortality due to the difficulty of controlling mosquitoes. The most important disease vectors belong to the subfamily Anophelinae (Anopheles mosquitoes) which transmits malaria; Culicinae i.e. Culex species transmit filariasis; West Nile virus, Japanese encephalitis and Aedes mosquitoes which primarily transmit dengue, chikungunya, yellow fever and Zika. These diseases account for more than 17% of all infectious diseases, causing 7 00 000 deaths annually with 80% of the world’s population at risk of one or more vector-borne diseases. In recent years, changes in public health policy and social factors as well as reports of resistance in both vector mosquitoes and the pathogens transmitted by them have caused a resurgence in the incidence of mosquito borne diseases.
Vector control is a key strategy to control these diseases. In India, vector control is primarily based on the use of long-lasting insecticide treated nets (LLINs) in addition to indoor residual spraying of insecticides in rural areas and anti-larval operations in urban areas. Larval control may be particularly valuable in regions where the eradication or elimination of vector borne diseases is being targeted, as a means of reducing the mosquito larval populations before they emerge to the adult stage.
However, with regards to mosquito control strategies, chemical control agents still play a major role. Insecticides applied with the aim of eliminating mosquitoes have given rise to other serious problems. Not only have mosquitoes developed resistance, but these insecticides also pose threat to human, animal health and the ecosystem as a whole. Chemical insecticide exposure among humans has been linked to immune dysfunction, neurological disorders, various forms of cancer, birth defects, liver damage and infertility. These adverse effects have led to the discovery of alternatives to these insecticides. Microbial control agents are effective and proven to be a method effective against mosquito immatures of both Anophelines and Culicines. Commercial biolarvicide formulations of gram positive and spore forming bacteria, Bacillus (B.) thuringiensis israelensis and B. sphaericus are now being widely used across the globe in the vector control programmes. These strains have been well characterized both at the microbiological and molecular level. Based on these two bacilli, there are several effective and well tested formulations commercially available including the wettable powder, slow release granules, briquettes, tablets and emulsifiable concentrates. These formulations are often deployed as an integral components of the integrated vector management strategy advocated by the World Health Organization and adopted by vector borne disease endemic countries.
In this review article, extensive literature search was done to collect and collate published information on bacilli-based biolarvicides and their control in different parts of the world, especially the articles published on the recent advancements in the field of bacilli-based vector control.
| 2. Bacilli as bio-control agents|| |
In nature, a wide variety of organisms including viruses, protozoans, fungi and bacteria, effectively control mosquitoes. Among many bacteria that have been tested, strains of B. thuringiensis (Bt) and B. sphaericus (Bs) are the most promising for vector control so far. B. thuringiensis var. israelensis (Bti) has an advantage of a broader host range. While B. sphaericus has a narrow spectrum, it has an advantage of increased duration of larvicidal activity against specific mosquito species like Culex (Cx.) quinquefasciatus and possess recycling ability within mosquito cadavers. There are options available for ‘stand-alone’ and combined formulations of these two Bacilli species and their strains for vector control programmes.
2.1. B. thuringiensis
B. thuringiensis is a ubiquitous, gram positive, sporulating aerobic bacterium which can be easily grown and cultured on routinely used media like nutrient agar. It can be isolated from a variety of sources. On sporulation, it produces two types of insecticidal crystal proteins or δ-endotoxins, Cry (for crystal) and Cyt (for cytolytic) proteins and further variations of each of these types. Cry proteins target lepidopteran insects, while few are toxic to dipteran or coleopteran insects. Cyt proteins show moderate toxicity to mosquitoes and black fly larvae occurring mostly in mosquitocidal subspecies e.g. B. thuringiensis subsp. israelensis.
Cyt proteins have been studied less in comparison to Cry proteins. Based mainly on studies of Cyt1Aa, their importance is in the biology of mosquitocidal strains as they synergize with other mosquitocidal Cry proteins (Cry4Aa, Cry4Ba, and Cry11A) resulting in delay in the phenotypic expression of resistance which would require multiple mutations at different loci.
The high degree of host specificity and the complexity of B. thuringiensis mode of action results from the interaction of the mosquitocidal toxins within the complex environment of the insect’s midgut lumen. Although researchers discovered relatively early that the midgut was the primary site of δ-endotoxin activity as seen in [Figure 1]A and [Figure 1]B, the molecular mechanisms of Bt intoxication have continued to be the subject of intensive research.
|Figure 1: (A) Diagrammatic representation of mode of action of Bacillus thuringiensis israelensis toxin; (B) Cartoon showing death of mosquito larvae due to toxin action of biolarvicide.|
Click here to view
Although Bti is proven to be effective against many mosquito species, operational application showed that it is more suitable against Aedes species. Aedes (Ae.) aegypti and Ae. albopictus were most susceptible to B. thuringiensis H-14 in comparison to other vector mosquitoes. More advantages for Aedes control may be due to their feeding behaviour, as most of the Bti toxins sediment to the base of the container during treatment where Aedes larvae frequently feed. On the other hand, Anopheles (An.) balabacensis and Mansonia (Mansonioides) indiana were found comparatively less susceptible to the Bti (H-14).
Cx. quinquefasciatus was also found to be highly susceptible to B. thuringiensis H-14. This mosquito species, however, is more susceptible to B. sphaericus, the latter being more effective in polluted water with high organic contents where Cx. quiquefasciatus prefers to breed as B. sphaericus is known to recycle in polluted water and persists longer than B. thuringiensis H-14.
2.2. B. sphaericus
B. sphaericus is a common aerobic, rod-shaped, endospore forming gram positive soil bacterium with a few entomopathogenic strains. The first discovery of a strain toxic to mosquito larvae was reported by Kellen et al. in 1965. The biolarvicide based on B. sphaericus is unique in that it consists of two binary proteins BinA (42 kDa) and BinB (51 kDa), both of which are required for toxicity to mosquito larval midgut. These binary proteins are cleaved by mosquito gut proteases, forming the active toxin by yielding peptides of 39 kDa and 43 kDa respectively. These associate and bind to the a-glucosidase receptor located on the midgut microvilli, resulting in lysis of midgut cells upon internalization,. It is suspected that reported loss of toxicity i.e. resistance in target mosquito species to B. sphaericus may be due to the reduction or loss of interactions between BinA and BinB or BinB and its receptor. In addition, another 100 kDa mosquitocidal protein appears to be synthesized in lesser toxic and some highly toxic strains. This polypeptide is expressed during the vegetative phase and is not homologous with the 51 and 42 kDa proteins.
2.3. B. subtilis
A B. subtilis strain producing mosquitocidal (larvicidal and pupicidal) toxin was isolated from mangrove forests of Andaman and Nicobar Islands of India and found to kill larval and pupal stages of three species of mosquitoes viz., An. stephensi, Cx quinquefasciatus and Ae. aegypti. It is the first gram-positive bacterium highly toxic to mosquito pupae. Its mosquitocidal activity is associated with an exotoxin identified as surfactin, a cyclic lipopeptide highly active at both acidic and basic pH, temperature range of 25 °C-42 °C, and UV stability, suitable features for the development of a biolarvicide. Preliminary toxicity studies with crude surfactin showed that it is non-toxic to mammals. The arsenal of biocontrol agents is further augmented with this potential mosquitocidal bacterium. The overview of different bio-control agents, their strains, activity profile against target species, toxin genes and strain modifications with recombinant technology is shown in [Table 1].
|Table 1: Overview of different bio-control agents, their strain, activity profile against target species, toxin genes and strain modifications with recombinant technology.|
Click here to view
| 3. Bioassays, isolation, characterization and identification|| |
Microbial isolates are constantly screened and isolated from terrestrial and aquatic environments for mosquito control programmes. The earlier method of isolating mosquito pathogenic Bacillus strains was cumbersome and time consuming, hence Dhindsa et al. in 2002 devised a new soil screening method that could reveal the presence of mosquito pathogenic bacilli in the soil samples. This method involves the use of LB broth (buffered with Sodium acetate) and a heat shock step at 65 °C. Using this method, eight different Bacillus strains, B. pumilus (KSD-1), B. sphaericus (KSD-2), B. brevis (KSD-3), B. sphaericus (KSD-4), B. subtilis (KSD-5), B. stereothermophilus (KSD-6), Bacillus sp. (KSD-7) and B. sphaericus (KSD-8) were successfully isolated, identified and evaluated for their larvicidal activity in Goa, India.
In a screening assay carried out by Radhika et al. in 2011, ten bacilli were isolated from Tamil Nadu, India and tested for larvicidal activity against Ae. aegypti mosquito. Two microbial isolates (B. megaterium and Acinetobacter sp.) effectively caused 97% larval mortality at 48-hour incubation at bacterial concentrations of (4.1±0.39) and (3.6±0.71) mg/L. Another study by Allwin et al. in 2007 showed native strains of B. thuringiensis were isolated from soil samples collected from different locations and characterizations in India. Samples collected from mangroves of Vellar estuary in India yielded mosquitocidal bacteria B. subtilis with increased activity against An. stephensi and Ae. aegypti. Many other reports on the frequent occurrence of mosquito pathogenic bacterial isolates in the natural environment showed high possibility of isolating novel strains.
| 4. Resistance phenomenon and overcoming resistance|| |
Since insecticide resistance can undermine efforts to control vector borne diseases, effective mosquito control can be successfully achieved only by overcoming insecticide resistance. Resistance is a complex genetic, evolutionary and ecological phenomenon. Resistance to microbial insecticides formulations is a serious threat to their success in public health settings.
4.1. Strategies for management of resistance to biolarvicides
Some measures to counter resistance include: (1) rotation or alternation of bacterial biolarvicidal toxins with other toxins, insecticides or biological control strategies; (2) less frequent biocide treatments; (3) use of slow-release, ultra-low volume (ULV) and thermal formulations which are active for longer durations; (4) use of source reduction methods and (5) constant resistance surveillance and monitoring. These principles when combined are essentially a blueprint for integrated pest management which will successfully delay or prevent the development of resistance in vector populations.
Studies have shown that by combining different classes of insecticides or their application by mosaic design can effectively overcome resistance in the target insects. However, unless insecticides of different classes are combined and judiciously used, there is possibility of cross resistance if insecticides induce similar mechanism of resistance and have mode of action in target insect. If mixtures are used, there is inherent risk of resistance build up to multiple classes of insecticides rendering them eventually useless. But there is a drawback in this approach for their practical application mainly due to higher cost and practical difficulty as both compounds need to be present in equally high and persistent concentrations. In such a scenario, the use of two or more interventions has been advocated so that mosquitoes that survive contact with one (e.g. LLINs) are killed due to exposure to the second (e.g. indoor residual spraying). In such a scenario, the use of biolarvicides where feasible, can also delay the onset of resistance and ease selection pressure of insecticide on target vectors.
4.3. ULV and thermal application
The dengue vector Ae. aegypti is a container breeder, hence use of Bti for its control is limited due to difficulty in its effective application. In this respect, ULV cold fogging can be used effectively for larviciding purposes when the agent is applied correctly and under required conditions. Seleena et al. 1996 found that ULV fogging of B. thuringiensis H-14 was highly effective in Aedes larval control and when used together with malathion it induced complete adult mortality.
In addition, the effectiveness of the thermal application of an aqueous suspension of Bti with and without pyrethroids using a thermal fogger has been reported without loss of its larvicidal activity,,.
4.4. Application of ice granules containing endotoxins of microbial agents
A novel method for the aerial delivery of microbial mosquito control agents into vast aquatic sites in the form of ice granules was developed by Becker et al. in 2003. The solutions containing powder formulations of Bti or Bs were transformed into ice pellets (named IcyPearls) using a special ice-making machine and applied aerially. Successful field tests using IcyPearls applied at the rate of 5 and 10 kg/ha containing various dosages of 100, 200, and 400 g of VectoBac® WDG (3 000 ITU/mg) were conducted against larvae of Ae. vexans with mortality rates of 91%-98%.
| 5. Commercial bio-larvicide formulations and their field efficacy|| |
Two biolarvicide formulations-Bacticide® and VectoBac® containing viable endospore and delta endotoxin of Bti H-14 were evaluated in 2001 in Surat city, India against An. subpictus and Cx. quinquefasciatus. Both formulations were equally effective on larvae after second application.
Field testing and evaluation of the efficacy of bio-larvicide, Bactivec® SC (Bti H-14) was carried out in Bengaluru, India. It was found to be operationally feasible and easy to handle.
Kumar et al. 1995 and 1996 tested a formulation of Bactoculicide (Bti strain 164) in construction sites, abandoned overhead tanks and curing waters and a formulation of Spherix (B. sphaericus H5a5b) in Goa, India respectively and found them highly effective,.
The weekly application of biolarvicide B. sphaericus (Strain 101, Serotype H5a5b) in Panaji, Goa, India helped in malaria control and was identified as a useful biocontrol agent of An. stephensi. Similarly, application of biolarvicide Bti strain 164 at 1 g/m2 and introduction of larvivorous fish Aplocheilus blocki in major breeding habitats of An. stephensi was carried out in order to control malaria in Goa, India. This was found to successfully replace DDT and pyrethrum fogging.
In addition, the efficacy of various formulations of Bti (Bactimos®, Teknar®, VectoBac®, Bactisand®, VectoPrime®, VectoMax®) and Bs (HIL-9® & HIL-10®, VectoLex®) in the form of tablets, granules, wettable powder, pellet, aqueous suspension, etc. were tested against mosquito vectors and found to be highly effective.
[Table 2] provides a list of the available commercial bio-larvicide formulations, their type, potency and field evaluation of these formulations.
|Table 2: A list of the available commercial bio-larvicide formulations, their type, potency and field evaluation of these formulations.|
Click here to view
| 6. Future prospects|| |
Future prospects for the use of biolarvicide formulations against mosquito vectors will depend on low cost production and development of cost-effective formulations. Cheaper formulations designed from the seeds of legumes, dried cow blood and mineral salts as well as the use of potato-based culture medium, bird feather waste and de-oiled rice bran waste as culture medium when assessed for growth and production of insecticidal toxins of Bti were shown to be more economical and effective against Ae. aegypti, Cx. quinquefasciatus and An. gambiae,,. This is very important from the point of media optimization for the economical production of Bacillus based insecticides in mosquito control programs.
In addition, enhanced activity of protein toxins by use of recombinant bacteria containing a mixture of endotoxins having different modes of action shows great promise. A few examples include newly discovered mosquitocidal proteins and peptides such as Mtx proteins and trypsin modulating oostatic factor which can be genetically engineered for development and use in vector control programs.
Research is also underway with respect to transgenic algae and cyanobacteria by expressing larvicidal endotoxins of Bti and B. sphaericus to allow the toxins to persist in the feeding zone for a longer duration as well as providing increased protection from sunlight (UV light). The most promising results were obtained when Cry4Aa and Cry11Aa alone or with Cyt1Aa were expressed in the filamentous, nitrogen-fixing cyanobacterium Anabaena PCC 7120. A transgenic strain of Anabaena PCC 7120 was reported to protect the expressed δ-endotoxins of Bti from damage inflicted by UV-B. This organism has an added advantage as it has the ability to multiply in the breeding sites as well as serving as a food source to mosquito larvae.
Recently there has been focus on the development of novel biolarvicides and their applications. The use of entomopathogenic bacteria and fungi mainly ascomycetes fungi such as Metarhizium anisopliae and Beauveria bassiana, for control of both larval and adult stages of mosquito vectors such as Aedes. The use of spatial repellents has been advocated to release volatile chemicals into the air, to induce modifications in insect behaviour and to reduce human-vector contact thereby reducing pathogen transmission.
Although mosquito traps have been used effectively as surveillance tools in order to capture vector mosquitoes for population and disease transmission studies, they have recently been considered a control strategy by the introduction of the lethal ovitrap. These traps such as attractive baited lethal ovitrap are being developed to attract and kill the egg-bearing females. They have shown promise in both lab and field settings for significant reduction in Aedes populations. The use of attractive toxic sugar baits which work by attracting mosquitoes and having them feed on toxic sugar meals could also be a potential vector control tool.
The future vector control includes the use of sterile insect technique (SIT) which has been successfully demonstrated against Ae. albopictus mosquitoes. SIT appears very promising to control mosquito populations and has been recently combined with auto-dissemination i.e., adult females contaminated with dissemination stations of juvenile hormone to treat breeding habitats, especially for the control of Aedes species, but this technique has not been used in large scale at present. Recently, a new control concept has been devised, named “boosted SIT” that might enable the area-wide eradication of mosquitoes. In addition, the exploitation of cytoplasmic incompatibility can be an advantageous mosquito control method. Cytoplasmic incompatibility is induced by the bacterial endosymbiont Wolbachia which is widespread and its use is a promising tool for mosquito control either alone or associated with SIT. Lately, mosquitoes modified with gene drive systems are being proposed as new tools that will complement the existing ones. The synergistic utilization and application of these control measures to protect against mosquito borne diseases could have a major impact on the socio-economic health of populations particularly in developing countries. These methods are currently in the pipeline and could complement the integrated vector management programmes when available.
| 7. Conclusions|| |
Vector borne diseases transmitted by mosquitoes are a major public health concern. Effective vector control requires the deployment of a range of integrated interventions. This review focuses on the current status and future prospects of bacilli-based vector control to explore additional options and potentially augment existing strategies. It is of immense importance to focus on the development, evaluation and deployment of alternative vector control products and strategies. However, for effective control and elimination of the mosquito vector and vector borne diseases, these strategies will have to be locally adapted to account for vector biology and the intensity of disease transmission keeping in mind both human and financial resources. In addition, we are waiting for the discovery of a novel bacterium from nature which could be developed into an ideal biolarvicide having a broad spectrum of activity at very low concentrations without developing resistance in the target mosquito species.
Conflict of interest statement
The authors declare that there is no conflict of interest.
A.K. designed the study, J.S.A and A.K.M. carried out the data collection, data analysis and interpretation. J.S.A and A.K.M. drafted the article. A.K, S.K and S.L.H. edited the article. All authors read and approved the final article.
| References|| |
Shehu-Riskuwa ML, Nataala MK, Baba EE. Biocontrol potential of Bacillus thuringiensis
isolated from soil against mosquito larvae. SAJP
Geetha I, Prabakaran G, Paily KP. Characterization of 3 mosquitocidal Bacillus
strains isolated from mangrove forest. Biol Control
Bukhari T, Takken W, Koenraadt C. Biological tools for control of larval stages of malaria vectors-a review. Biocontrol Sci Technol
Dambach P, Baernighausen T, Traore I, Ouedraogo S, Sie A, Sauerborn R, et al. Reduction of malaria vector mosquitoes in a large-scale intervention trial in rural Burkina Faso using Bti based larval source management. Malar J
Yousef N, Aly N. Effectiveness of Bacillus thuringiensis
and Bacillus sphaericus
isolates against Culex pipens, Aedes aegypti
and Anopheles sergenti
mosquito larvae. PUJ
2014; 6: 59-64.
Nicolopoulou-Stamati P, Maipas S, Kotampasi C, Stamatis P, Hens L. Chemical pesticides and human health: The urgent need for a new concept in agriculture. Front Public Health
Kachhawa D. Microorganisms as a biopesticides. J Entomol Zool Stud
Surendran A, Vennison SJ. Occurrence and distribution of mosquitocidal Bacillus sphaericus
in soil. Acad J Entomol
Palma L, Munoz D, Berry C, Murillo J, Caballero P. Bacillus thuringiensis
toxins: An overview of their biocidal activity. Toxins
Bravo A, Gill SS, Soeron M. Mode of action of Bacillus thuringiensis
Cry and Cyt toxins and their potential for insect control. Toxicon
Tilquin M, Paris M, Reynaud S, Despres L, Ravanel P, Geremia RA. Long lasting persistence of Bacillus thuringiensis
subsp. israelensis (Bti)
in mosquito natural habitats. PLoS One
Bauer L. Resistance: A threat to the insecticidal crystal proteins of Bacillus thuringiensis. Fla Entomol
Lacey LA. Bacillus thuringiensis
serovariety israelensis and Bacillus sphaericus
for mosquito control. J Am Mosq Control Assoc
2007; 23(2): 133-163.
Lee YW, Zairi J. Field evaluation of Bacillus thuringiensis
H-14 against Aedes
mosquitoes. Trop Biomed
Lee HL, Cheong WH. Laboratory evaluation of the potential efficacy of Bacillus thuringiensis israelensis
for the control of mosquitoes in Malaysia. Trop Biomed
Lee HL, Pe TH, Cheong WH. Laboratory evaluation of the persistence of Bacillus thuringiensis
against Aedes aegypti
larvae. Mosq Borne Dis Bull
Lee HL, Chan ST, Cheong WH. Laboratory bioassays of Bacillus sphaericus
1593, 2297 and 2362 against mosquitoes of public health importance in Malaysia. Trop Biomed
Kellen WR, Clark TB, Lindegren JE, Ho BC, Rogoff MH, Singer S. Bacillus sphaericus
neide as a pathogen of mosquitoes. J Invertebr Pathol
Charles JF, Nielsen-LeRoux C, Delécluse A. Bacillus sphaericus
toxins: Molecular biology and mode of action. Annu Rev Entomol
Rahman MA, Khan SA, Sultan MT, Islam MR. Characterization of Bacillus sphaericus
binary proteins for biological control of Culex quinquefasciatus
mosquitoes: A review. Int J Biosci
Boonyos P, Soonsanga S, Boonserm P, Promdonkoy B. Role of cysteine at positions 67, 161 and 241 of a Bacillus sphaericus
binary toxin BinB. BMB
Poopathi S, Ramesh N, Sundaravadivelu K, Samuel P, Tyagi BK. Larvicidal efficacy of various formulations of Bacillus sphaericus
against the resistant strain of Culex quinquefasciatus
(Diptera: Culicidae) from southern India. Trop Biomed
Geetha I, Manonmani AM. Surfactin: A novel mosqitocidal biosurfactant produced by Bacillus subtiliis
(VCRC B471) and influence of abiotic factors on its pupicidal efficacy. Lett Appl Microbiol
Geetha I, Manonmani AM. Mosquito pupicidal toxin production by Bacillus subtilis
subsp. subtilis. Biol Control
Mulla M, Federici B, Darwazeh H, Ede L. Field evaluation of the microbial insecticide Bacillus thuringiensis
serotype H-14 against floodwater mosquitoes. Appl Environ Microbiol
Thanabalu T, Hindley J, Brenner S, Berry C. Expression of the mosquitocidal toxins of Bacillus sphaericus
and Bacillus thuringiensis
by recombinant Caulobacter crescentus
, a vehicle for biological control of aquatic insect larvae. Appl Environ Microbiol
Porter A, Davidson E, Liu J. Mosquito toxins of bacilli and their genetic manipulation for effective biological control mosquitoes. Microbiol Rev
Padua LE, Ohba M, Aizawa K. Isolation of a Bacillus thuringiensis
strain (serotype 8a,8b) highly and selectively toxic against mosquito larvae. J Invertebr Pathol
Delécluse A, Rosso ML, Ragni A. Cloning and expression of a novel toxin gene from Bacillus thuringiensis
subs P. jegathesan
encoding a highly mosquitocidal protein. Appl Environ Microbiol
Ragni A, Thiery I, Delécluse A. Characterization of six highly mosquitocidal Bacillus thuringiensis
strains that do not belong to H-14 serotype. Curr Microbiol
Ramírez-Lepe M, Ramírez-Suero M. Biological control of mosquito larvae by Bacillus thuringiensis
. In: Perveen F. (ed.) Insecticides-pest engineering
. Europe: InTech; 2012, p. 239-264.
Favret M, Yousten A. Insecticidal activity of Bacillus laterosporus
. J Invertebr Pathol
Shida O, Takagi H, Kadowaki K, Komagata K. Proposal for two new genera, Brevibacillus gen nov
. and Aneurinibacillus gen nov. Int J Syst Bacteriol
Orlova MV, Smirnova TA, Ganushkina LA, Yacubovich VY, Arizbekyan RR. Insecticidal activity of Bacillus laterosporus. Appl Environ Microbiol
Huang XW. An extracellular protease from Brevibacillus laterosporus
G4 without parasporal crystals can serve as a pathogenic factor in infection of nematodes. Res Microbiology
Prabakaran G, Balaram K, Hoti SL, Manonmani AM. A cost-effective medium for the large-scale production of Bacillus sphaericus
H5a5b (VCRC B42) for mosquito control. Biol Control
Bar E, Lieman-Hurwitz J, Rahamin E, Keynan A, Sandler N. Cloning and expression of Bacillus thuringiensis
israelensis-endotoxin DNA in B. sphaericus. J Invertebr Pathol
Poncet S, Bernard C, Dervyn E, Cayley J, Klier A, Rapoport G. Improvement of Bacillus sphaericus
toxicity against dipteran larvae by integration, via
homologous recombination, of the Cry11A
toxin gene from Bacillus thuringiensis
subsp. israelensis. Appl Environ Microbiol
Kumar R, Hwang J. Larvicidal efficiency of aquatic predators: A perspective for mosquito biocontrol. Zool Stud
Park HW, Bideshi DK, Wirth MC, Johnson JJ, Walton WE, Federici BA. Recombinant larvicidal bacteria with markedly improved efficacy against Culex
vectors of West Nile virus. Am J Trop Med Hyg
Thiery I, Hamon S, Gaven B, de Barjac H. Host range of Clostridium bifermentans
, a mosquitocidal anaerobic bacterium. J Am Mosq Control Assoc
Khyami Horani H, Katbeh Bader A, Mohsen ZH. Isolation of endospore forming bacilli toxic to Culiseta longiareolata
(Diptera: Culicidae) in Jordan. Lett Appl Microbiol
Darriet F, Hougard JM. An isolate of Bacillus circulans
toxic to mosquito larvae. J Am Mosq Control Assoc
Prabakaran G, Paily KP, Padmanabhan V, Hoti SL, Balaraman K. Isolation of a Pseudomonas fluorescens
metabolite/exotoxin active against both larvae and pupae of vector mosquitoes. Pest Manag Sci
Prabakaran G, Paily KP, Hoti SL. Development of cost-effective medium for the large-scale production of a mosquito pupicidal metabolite from Pseudomonas fluorescens migula. Biol Control
Vankudre M, Balpande A, Athale M, Deshpande SG. Laboratory efficacy of Pseudomonas fluorescens
metabolites on mosquito larvae. Biosc Biotech Res Comm
Geetha I, Manonmani AM, Prabakaran G. Bacillus amyloliquefaciens:
A mosquitocidal bacterium from mangrove forests of Andaman & Nicobar Islands, India. Acta Trop
Ramathilaga A, Murugesan A, Prabu S. Biolarvicidal activity of Peanibacillus
macerans and Bacillus subtilis
isolated from the dead larvae against Aedes aegypti
-vector for chikungunya. Proc Int Acad Ecol Environ Sci
Ramirez JL, Short SM, Bahia AC, Saraiva RG, Dong Y, Kang S, et al. Chromobacterium
CspP reduces malaria and dengue infection in vector mosquitoes and has entomopathogenic and in vitro
anti-pathogen activities. PloS Pathog
Chandra G, Bhattacharya K, Banerjee S, Chatterjee S. Efficacy of a locally isolated strain of Bacillus cereus
as mosquito larvicide. Mol Entomol
Mani C, Selvakumari J, Manikandan S, Thirugnanasambantham K, Sundarapandian SM, Poopathi S. Field evaluation of Bacillus cereus
VCRC B540 for mosquitocidal activity-a new report. Trop Biomed
Lalithambika B, Vani C. Pseudomonas aeruginosa
KUN2, extracellular toxins-a potential source for the control of dengue vector. J Vector Borne Dis
Lee HL. Germ warfare against mosquitoes. What now? In: Lee CY, Robinson WH. (eds.) Proceedings of the Fifth International Conference on Urban Pests
. Malaysia: Perniagaan Ph’ng at P&Y Design Network; 2005, p. 10-18.
Dhindsa K, Sangodkar UMX, Kumar A. Novel cost effective method of screening soils for the presence of mosquito pathogenic bacilli. Lett Appl Microbiol
Radhika D, Ramathilaga A, Prabu CS, Murugesan AG. Evaluation of larvicidal activity of soil microbial isolates (Bacillus
sp.) against Aedes aegypti
(Diptera: Culicidae)-the vector of Chikungunya and Dengue. Proc Int Acad Ecol Environ Sci
Allwin L, Kennedy JS, Radhakrishnan V. Characterization of different geographical strains of Bacillus thuringiensis
from Tamil Nadu. Res J Agric Biol Sci
Balakrishnan S, Indira K, Srinivasan M. Mosquitocidal properties of Bacillus
species isolated from mangroves of Vellar estuary, Southeast coast of India. J Parasit Dis
Ammouneh H, Harba M, Idris E, Makee H. Isolation and characterization of native Bacillus thuringiensis
isolates from Syrian soil and testing of their insecticidal activities against some insect pests. Turk J Agric For
Reid MC, McKenzie FE. The contribution of agricultural insecticide use to increasing insecticide resistance in African malaria vectors. Malar J
Poopathi S, Abidha S. Mosquitocidal bacterial toxins (Bacillus sphaericus
and Bacillus thuringiensis
Mode of action, cytopathological effects and mechanism of resistance. J Physiol Pathophysiol
Raymond B, Wright D, Crickmore N, Bonsall M. The impact of strain diversity and mixed infections on the evolution of resistance to Bacillus thuringiensis. Proc R Soc B
Abbas A, Abbas ZR, Khan JA, Iqbal Z, Bhatti MM, Sindhu Z, et al. Integrated strategies for the control and prevention of dengue vectors with particular reference to Aedes aegypti. Pak Vet J
Seleena P, Lee H, Rohani A, Nazni W, Khadri M. Microdroplet application of mosquitocidal Bacillus thuringiensis
using ultra-low-volume generator for the control of mosquitoes. Southeast Asian J Trop Med Public Health
Seleena P, Lee HL, Chiang YF. Thermal application of Bacillus thuringiensis
for dengue vector control. J Vector Ecol
Chung YK, Lam-Phua SG, Chua YT, Yatiman R. Evaluation of biological and chemical insecticide mixture against Aedes aegypti
larvae and adults by thermal fogging in Singapore. Med Vet Entomol
Yap HH, Lee YW, Zairi J. Indoor thermal fogging against vector mosquitoes with two Bacillus thuringiensis israelensis
formulations, Vectobac ABG 6511 water-dispersible granules and Vectobac 12AS liquid. J Am Mosq Control Assoc
Becker N. Ice granules containing endotoxins of microbial agents for the control of mosquito larvae-a new application technique. J Am Mosq Control Assoc
Haq S, Bhatt RM, Vaishnav KB, Yadav RS. Field evaluation of biolarvicides in Surat city, India. J Vector Borne Dis
Uragayala S, Kamaraju R, Tiwari S, Ghosh SK, Valecha N. Field testing & evaluation of the efficacy & duration of effectiveness of a biolarvicide, Bactivec® SC (Bacillus thuringiensis
SH-14) in Bengaluru, India. Indian J Med Res
Kumar A, Sharma VP, Thavaselvam D, Sumodan PK. Control of Anopheles stephensi
breeding in construction sites and abandoned overhead tanks with Bacillus thuringiensis
var. israelensis. J Am Mosq Control Assoc
Kumar A, Sharma VP, Thavaselvam D, Sumodan PK, Kamat RH, Audi SS, et al. Control of Culex quinquefasciatus
with Bacillus sphaericus
in Vasco City, Goa. J Am Mosq Control Assoc
Kumar A, Sharma VP, Sumodan PK, Thavaselvam D, Kamat RH. Malaria control utilizing Bacillus sphaericus
against Anopheles stephensi
in Panaji, Goa. J Am Mosq Control Assoc
Kumar A, Sharma VP, Sumodan PK, Thavaselvam D. Field trials of bio-larvicide Bacillus thuringiensis
strain 164 and larvivorous fishes Aplocheilus blocki
against Anopheles stephensi
for malaria control in Goa, India. J Am Mosq Control Assoc
Mittal PK, Pant CS, Basil A, Jayaraman K, Sharma VP. Evaluation of the formulations of the mosquito larvicidal agent Biocide-S from Bacillus sphaericus
, 1593 M. Indian J Malariol
Sulaiman S, Jeffery J, Sohadi AR, Yunus H, Busparani V, Majid R. Evaluation of Bactimos wettable
powder, granules and briquettes against mosquito larvae in Malaysia. Acta Trop
Lacey L, Undeen A. Microbial control of black flies and mosquitoes. Annu Rev Entomol
Dua VK, Sharma SK, Sharma VP. Application of Bactoculicide (Bacillus thuringiensis
H-14) for controlling mosquito breeding in Industrial scrap at BHEL, Hardwar (U. P.). Indian J Malariol
Yadav K, Baruah I, Goswami D. Efficacy of Bacillus sphaericus
strain isolated from North East region of India as potential mosquito larvicide. J Cell Tissue Research
Sulaiman S, Pawanchee ZA, Wahab A, Jamal J, Sohadi AR. Field evaluation of Vectobac G, Vectobac 12AS and Bactimos WP against the dengue vector Aedes albopictus
in tires. J Vector Ecol
Tiwari S, Ghosh S, Mittal P, Dash AP. Effectiveness of a new granular formulation of biolarvicide Bacillus thuringiensis
against larvae of malaria vectors in India. Vector Borne Zoonotic Dis
Gunasekaran K, Boopathi PS, Vaidyanathan K. Laboratory and field evaluation of Teknar HP-D, a biolarvicidal formulation of Bacillus thuringiensis
, against mosquito vectors. Acta Trop
Brown ID, Watson TM, Carter J, Purdie DM, Kay BH. Toxicity of VectoLex (Bacillus sphaericus)
products to selected Australian mosquito and nontarget species. J Econ Entomol
Eritja R. Laboratory tests on the efficacy of VBC60035, a combined larvicidal formulation of Bacillus thuringiensis israelensis
(strain AM65-52) and Bacillus sphaericus
(strain 2362) against Aedes albopictus
in simulated catch basins. J Am Mosq Control Assoc
Cetin H, Oz E, Yanikoglu A, Cilek JE. Operational evaluation of Vectomax® WSP (Bacillus thuringiensis
subsp. israelensis Bacillus sphaericus)
against larval Culex pipiens
in Septic Tank. J Am Mosq Control Assoc
Poopathi S, Tyagi B. Mosquitocidal toxins of spore forming bacteria: Recent advancement. Afr J Biotechnol
Poopathi S, Anup Kumar K. Novel fermentation media for production of Bacillus thuringiensis
subsp. israelensis. J Econ Entomol
Poopathi S, Mani C, Rajeswari G. Potential of sugarcane bagasse (agro-industrial waste) for the production of Bacillus thuringiensis israelensis. Trop Biomed
Devidas P, Pandit B, Vitthalrao P. Evaluation of different culture media for improvement in bioinsecticides production by indigenous Bacillus thuringiensis
and their application against larvae of Aedes aegypti. Sci World J
Federici BA, Park HW, Bideshi DK, Wirth MC, Johnson JJ. Recombinant bacteria for mosquito control. J Exp Biol
Ketseoglou I, Bouwer G. Optimization of photobioreactor growth conditions for a cyanobacterium expressing mosquitocidal Bacillus thuringiensis
Cry proteins. J Biotechnol
Manasherob R, Ben-Dov E, Xiaoqiang W, Boussiba S, Zaritsky A. Protection from UV-B damage of mosquito larvicidal toxins from Bacillus thuringiensis
expressed in Anabaena PCC 7120. Curr Microbiol
Scholte E, Knols BGJ, Samson RA, Takken W. Entomopathogenic fungi for mosquito control: A review. J Insect Sci
Achee NL, Bangs MJ, Farlow R, Killeen GF, Lindsay S, Logan JG, et al. Spatial repellents: From discovery and development to evidence-based validation. Malar J
Achee NL, Grieco JP, Vatandoost H, Seixas G, Pinto J, Ching-NG L, et al. Alternative strategies for mosquito-borne arbovirus control. PLoS Negl Trop Dis
Bellini R, Balestrino F, Medici A, Gentile G, Veronesi R, Carrieri M. Mating competitiveness of Aedes albopictus
radio-sterilized males in large enclosures exposed to natural conditions. J Med Entomol
Bouyer J, Lefrançois T. Boosting the sterile insect technique to control mosquitoes. Trends Parasitol
Bourtzis K, Lees RS, Hendrichs J, Vreysen MJ. More than one rabbit out of the hat: Radiation, transgenic and symbiont-based approaches for sustainable management of mosquito and tsetse fly populations. Acta Trop
Yakob L and Walker T. Zika virus outbreak in the Americas: The need for novel mosquito control methods. Lancet Glob Health
James S, Collins FH, Welkhoff PA, Emerson C, Godfray HC, Gottlieb M, et al. Pathway to deployment of gene drive mosquitoes as a potential biocontrol tool for elimination of malaria in sub-Saharan Africa: Recommendations of a scientific working group. Am J Trop Med Hyg
[Table 1], [Table 2]