Larvicidal Activities of Indigenous Bacillus thuringiensis Isolates and Nematode Symbiotic Bacterial Toxins against the Mosquito Vector, Culex pipiens (Diptera: Culicidae)
AbstractBackground: The incidence of mosquito-borne diseases and the resistance of mosquitoes to conventional pesticides have recently caused a panic to the authorities in the endemic countries. This study was conducted to identify native larvicidal biopesticides against Culex pipiens for utilization in the battle against mosquito-borne diseases.Methods: Larvicidal activities of new indigenous Bacillus thuringiensis isolates and crude toxin complexes (TCs) of two nematode bacterial-symbionts, Photorhabdus luminescens akhurstii (HRM1) and Ph. luminescens akhurstii (HS1) that tested against Cx. pipiens. B. thuringiensis isolates were recovered from different environmental samples in Saudi Arabia, and the entomopathogenic nematodes, Heterorhabditis indica (HRM1) and He. sp (HS1) were isolated from Egypt. Larvicidal activities (LC50 and LC95) of the potentially active B. thuringiensis strains or TCs were then evaluated at 24 and 48h post-treatment.Results: Three B. thuringiensis isolates were almost as active as the reference B. thuringiensis israelensis (Bti-H14), and seven isolates were 1.6–5.4 times more toxic than Bti-H14. On the other hand, the TCs of the bacterial symbionts, HRM1 and HS1, showed promising larvicidal activities. HS1 showed LC50 of 2.54 folds that of HRM1 at 24h post-treatment. Moreover, histopathological examinations of the HS1-treated larvae showed deformations in midgut epithelial cells at 24h post-treatment.Conclusion: Synergistic activity and molecular characterization of these potentially active biocontrol agents are currently being investigated. These results may lead to the identification of eco-friend mosquito larvicidal product(s) that could contribute to the battle against mosquito-borne diseases.
Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol. 18: 265–267.
Abulreesh HH, Osman GE H, Assaeedi ASA (2012) Characterization of Insecticidal Genes of Bacillus thuringiensis Strains Isolated from Arid Environments. In- dian J Microbiol. 52(3): 500–503.
Ahmed AM, Taylor P, Maingon R, Hurd H (1999) The effect of Plasmodium yoelii nigeriensis on the reproductive fitness of Anopheles gambiae. Invertebr Re- prod Develop. 36: 217–222.
Ahmed AM, Shaalan EA, Aboul-Soud MAM, Tripet F, AL-Khedhairy AA (2011) Mosquito vectors survey in ALAhsaa district, eastern region, Kingdom ofSaudi Arabia. J Insect Sci. 11: 1–11. Ahmed AM, Abdel Megeed AM, AlQahtaney HM (2014) A Novel Mos- quitocidal Bacterium as a Biocontrol Agent in Saudi Arabia: I - A Promising Larvicide Against Aedes caspius Mos- quito. Pakistan J Zool. 46(1): 191–201.
Al-Ahmed AM (2012) Mosquito fauna (Dip- tera: Culicidae) of the Eastern region of Saudi Arabia and their seasonal abundance. J King Saud Univ Sci. 24 (1): 55–62.
Al-Ghamdi K, Alikhan M, Mahayoub J, Afi- fi ZI (2008) Studies on identification and population dynamics of Anopheline mosquito from Jeddah, Saudi Arabia. Biosci. Biotech Res Comm. 1: 19–24.
Al-Hazmi M, Ayoola EA, Abdurahman M, Banzal S, Ashraf J, El-Bushra A, Hazmi A, Abdullah M, Elamin A, Al- Sammani E, Gadour M, Menon C, Hamza M, Rahim I, Hafez M, Jam- bavalikar M, Arishi H, Aqee A (2003) Epidemic Rift Valley Fever in Saudi Arabia: A Clinical study of severe ill- ness in humans. Clin Infect Dis. 36(3):245–252.
Al-Khuriji AM, Alahmed MA, Kheir SM (2007) Distribution and seasonal activ- ity of mosquitoes (Diptera: Culicidae) in Riyadh Region, Saudi Arabia. J King Saud Univ Agric Sci. 19 (2): 39–55.
Al-Roba AA, Aboul-Soud MAM, Ahmed AM, Al-Khedhairy AA (2011) The gene expression of caspasses is up-regulated during the signaling response of Aedes caspius against larvicidal bacteria. Afr J Biotechnol. 10(2): 225–233.
Al-Sarar AS (2010) Insecticide resistance of Culex pipiens populations (Diptera: Culicidae) from Riyadh City, Saudi Arabia: Status and overcome. Saudi J Biol Sci. 17(2): 95–100.
Al-Thabiani A, Al-Shami SA, Mahyoub JA, Hatabbi M, Ahmad AH, Salmah C (2014) An update on the incidence of dengue gaining strength in Saudi Ara- bia and current control approaches for its vector mosquito. Parasit Vectors. 7:258.
Al-Zahrani HAA, Abuldahab FF (2011) Iso- lation and activity of a Bacillus thurin- giensis Toxin which is Toxic to the Ae- des eagypti. J Ame Sci. 7(11): 269–276.
Ali S, Zafar Y, Ali G, Nazir F (2010) Bacil- lus thuringiensis and its application in agriculture. Afr J Biotechnol. 9: 2022–2031.
Alwafi OM, Scott JM, Ziad AM, Abdullah A (2013) Dengue fever in Makkah, Kingdom of Saudi Arabia, 2008–2012. Am J Res Commun. 1(11): 123–139.
Armengol G, Escobar MC, Maldonado ME, Orduz S (2007) Diversity of Colom- bian strains of Bacillus thuringiensis with insecticidal activity against dip- teran and lepidopteran insects. J Appl Microbiol. 102(1): 77–88.
Aramideh S, Saferalizadeh MH, Pourmirza AA, Bari MR, Keshavarzi M, Mohseniazar M (2010) Characteriza- tion and pathogenic evaluation of Ba- cillus thuringiensis isolates from West Azerbaijan province-Iran. Afr J Mi- crobiol Res. 4(12): 1224–1229.
Assaeedi ASA, Osman GEH, Abulreesh HH (2011) The occurrence and insecticidal activity of Bacillus thuringiensis in the arid environments. Aust J Crop Sci.5(10): 1185–1190.
Ayaad TH, Al Akeel R Kh, Olayan E (2015) Isolation and characterization of mid- gut lectin from Aedes aegypti (L.) (Diptera: Culicidae). Braz Arch Biol Technol. 58: 905–912.
Aziz AT, Al-Shami SA, Mahyoub JA, Hat- abbi M, Ahmad AH, Rawi CS (2014) An update on the incidence of dengue gaining strength in Saudi Arabia and current control approaches for its vec- tor mosquito. Parasites Vectors. 7: 258.
Azmi MA, Naqvi S, Akhtar K, Parveen S, Parveen R, Aslam M (2009) Effect of pesticide residues on health and blood parameters of farm workers from rural Gadap, Karachi. Pak J Environ Biol.30(5): 747–756.
Balenghien T, Vazeille M, Grandadam M (2008) Vector competence of some French Culex and Aedes mosquitoes for West Nile virus. Vector Borne Zo- onotic Dis. 8(5): 589–595.
Balkhy HH, Memish ZA (2003) Rift Valley fever: an uninvited zoonosis in the Arabian Peninsula. Int J Antimicrob Agents. 21: 153–157.
BenDv E (2014) Bacillus thuringiensis subsp. israelensis and its dipteran-spe- cific toxins. Toxins. 6: 1222–1243.
Bernhard K, Jarret P, Meadows M, Butt J, Ellis DJ, Roberts GM, Pauli S, Rodg- ers P, Burges HD (1997) Natural iso- lates of Bacillus thuringiensis: world- wide distribution, characterization, and activity against insect pests. J Invertebr Pathol. 70(1): 59–68.
Bernier UR, Furman KD, Kline DL, Allan SA, Barnard D (2005) Comparison of contact and spatial repellency of catnip oil and N,N-diethyl-3-methylbenzamide (DEET) against mosquitoes. J Med Entomol. 42: 306–311.
Bishop AH (2014) Expression of prtA from Photorhabdus luminescens in Bacillus thuringiensis enhances mortality in lepidopteran larvae by sub-cutaneous but not oral infection. J Invertebr Pathol. 121: 85–88.
Bowen DJ, and Ensign JC (1998) Purifica- tion and characterization of a high mo- lecular weight insecticidal protein complex produced by the entomopath- ogenic bacterium Photorhabdus lumi- nescens. Appl Environ Microbiol. 64:3029–3035.
Boyer S, Paris M, Jego S, Lemperiere G, Ravanel P (2012) Influence of insecti cide Bacillus thuringiensis subsp. is- raelensis treatments on resistance and enzyme activities in Aedes rusticus larvae (Diptera: Culicidae). Biol Con- trol. 62: 75–81.
Bozlagan L, Ayvaz A, Ozturk F, Acik L, Akbulut M, Yilmaz S (2010) Detection of the cry1 gene in Bacillus thurin- giensis isolates from agricultural fields and their bioactivity against two stored product moth larvae. Turk J Agric. 34:145–154.
Bradford M (1976) A rapid and sensitive method for the quantitation of mi- crogram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem. 72: 248–254.
Bravo A, Gill SS, Soberón M (2007) Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon. 49(4): 423–435.
Briassoulis G (2001) Toxic encephalopathy associated with use of DEET insect re- pellents: a case analysis of its toxicity in children. Hum Exp Toxicol. 20: 8–14.
Brouqui P, Parola P, Raoult D (2012) Insec- ticide resistance in mosquitoes and failure of malaria control. Expert Rev Anti Infect Ther 10(12): 1379–1381.
Brown MD, Thomas D, Watson K, Kay BH (1998) Laboratory and field evaluation of efficacy of vectobac 12AS against Culex sitiens (Diptera: Culicidae) lar- vae. J Am Mosq Control Assoc. 14:183–185.
Chatterjee SN, Bhattacharya T, Dangar TK, Chandra G (2007) Ecology and diver- sity of Bacillus thuringiensis in soil en- vironment. Afr J Biotechnol. 6: 1587–1591.
Dulmage HT, Boening OP, Rehnborg CS, Habsen GD (1971) A proposed stand- ardized biossay for formulations of Bacillus thuringiensis based on the in- ternational unit. J Invertebr Pathol. 18:240–245.
El-Kersh TA, Al-sheikh YAA, Al-Akeel R, Alsayed AA (2012) Isolation and char- acterization of indigenous Bacillus thu- ringiensis isolates from Saudi Arabia. Afr J Biotechnol. 11(8): 1924–1938.
El-Kersh TA, Al-akeel RA, Al-sheikh YA, Alharbi SA (2014) Isolation and dis- tribution of mosquito-larvicidal cry genes in Bacillus thuringiensis strains native to Saudi Arabia. Trop Biomed.31(4): 616–632.
El-Kersh TA, Ahmed AM, Al-sheikh YA, Tripet F, Ibrahim MS, Metwalli AAM (2016) Isolation and molecular char- acterization of Bacillus thuringiensis strains native to Saudi Arabia with naturally improved larivicidal toxicity against the mosquito vector Anopheles gambiae s.l. Parasites and Vectors (In Press).
El-Sadawy HA, Forst, S, Abouelhag HA, Ahmed AM, Alajmi RA, Ayaad TH (2016) Molecular and phenotypic char- acterization of two bacteria Photorhab- dus luminescens subsp. akhurstii HRM1 and HS1 isolated from two ento- mopathogenic nematodes Heterorhab- ditis indica RM1 and Heterorhabditis sp. S1. Pak J Zool. 48(1): 51–58.
Eleftherianos I, Joyce S, Ffrench-Constant RH, Clarke DJ, Reynolds SE (2010) Probing the tri-trophic interaction be- tween insects, nematodes and Photo- rhabdus. Parasitol. 137: 1695–1706.
Ffrench-Constant RH, Bowen DJ (2000) Novel insecticidal toxins from nema- tode symbiotic bacteria. Cell Mol. Life Sci. 57: 828–833.
ffrench-Constant, RH, Dowling A, Water- field NR (2007) Insecticidal toxins from Photorhabdus bacteria and their potential use in agriculture. Toxicon.49: 436–451.
Finney DN (1971) Probit Analysis, 3rd Edi- tion. Cambridge University Press, London, p. 318.
Gobatto V, Giani SG, Camassola M, Dillon AJ, Specht A, Barros NM (2010) Ba- cillus thuringiensis isolates entomopath- ogenic for Culex quinquefasciatus (Dip- tera: Culicidae) and Anticarsia gemma- talis (Lepidoptera: Noctuidae). Braz J Biol. 70: 1039–1046.
Guo L, Fatig RO, Orr GL, Schafer BW, Strickland JA, Sukhapinda K, Woodsworth AT, Petell JK (1999) Photorhabdus luminescens W-14 insec- ticidal activity consists of at least two similar but distinct proteins. Purifica- tion and characterization of toxin A and toxin B. J Biol Chem. 274: 9836–9842.
Hawking F (1973) The distribution of hu- man Filariases throughout the world. Mimeograph WHO/FIL/73,114.
Heimpel, AM, Angus TA (1959) The site of action of crystalliferous bacteria in Lepidoptera larvae. J Invertebr Pathol.1: 152–170.
Hernandez CS, Andrew R, Bel Y, Ferre J (2005) Isolation and toxicity of Bacil- lus thuringiensis from potato growing areas in Bolivia. J Invertebr Pathol. 88:8–16.
Hong HA, To E, Fakhry S, Baccigalupi L, Ricca E, Cutting SM (2009) Defining the natural habitat of Bacillus spore- formers. Res Microbiol. 160: 375–379.
Jup PG, Kemp A, Grobbelaar A, Lema P, Burt FJ, Alahmed AM, Mujalli DA, Khamees MA, Swanepoel R (2002) The
epidemic of Rift Valley fever in Saudi Arabia: Mosquito vector studies. Med Vet Entomol. 16(3): 245–252.
Kavitha R, Xavier R, Monica D, Sreeramanan S (2011) Quick isolation and charac- terization of novel Bacillus thurin- giensis strains from mosquito breeding sites in Malaysia. Emirates Journal of Food and Agriculture. 23: 17–26.
Khalil H, El-Badry AA, Eassa AHA, Al- Juhani AM, Al-Zubiany SF, and Ibrahim KD (2008) A Study on Culex spe- cies and Culex transmitted Diseases in Al-Madinah Al-Munawarah, Saudi Ara- bia. Parasitol United J. 1(2): 101–108.
Khan NA, Azhar EI, EL-Fiky S, Madani HH, Abuljadial MA, Ashshi AM, Turki- stani AM, Hamouh EA (2008) Clinical profile and outcome of hospitalized patients during first outbreak of den- gue in Makkah, Saudi Arabia. Acta Trop. 105: 39–44.
Lang AE, Schmidt G, Sheets JJ, Aktories K (2011) Targeting of the actin cyto- skeleton by insecticidal toxins from Photorhabdus luminescens. N-S Arch. Pharmacol. 383: 227–235.
Litchfield JT, Wilcoxin FA (1949) Simpli- fied method of evaluating dose–effect experiments. J Pharmacol Exp Ther.96: 99–103.
Madani TA (2005) Alkhumra virus infec- tion, a new viral hemorrhagic fever in Saudi Arabia. J Infect. 51: 91–97.
Madani TA, Yagob Y, Al-Mazrou MH, Al- Jeffri1AA, Mishkhas AM, Al-Rabeah AM, Turkistani MO, Abodahish SAA, Khan AS, Ksiazek TG, Shobokshi O (2003) Rift Valley Fever Epidemic in Saudi Arabia: Epidemiological, Clinical, and Laboratory Characteristics. Oxford Journals Medicine and Health Clinical Infectious Diseases. 37(8): 1084–1092.
Maguranyi SK, Webb CE, Mansfield S, Rus- sell RC (2009) Are commercially avail- able essential oils from Australian native plants repellent to mosquitoes? J Am Mosq Control Assoc. 25(3): 292–300
Martin PA, Travers RS (1989) Worldwide Abundance and Distribution of Bacil- lus thuringiensis isolates. Appl Environ Microbiol. 55: 2437–2442.
Martin PA, Gundersen-Rindal DE, Black- burn MB (2010) Distribution of phe- notypes among Bacillus thuringiensis strains. Syst Appl Microbiol. 33: 204–208.
Meyer RP, Hardy JL, Presser SB (1983) Comparative vector competence of Culex tarsalis and Culex quinquefasci- atus from the coachella, imperial, and San Joaquin Valleys of California for St. Louis encephalitis virus. Am J Trop Med Hyg. 32(2): 305–311.
Mohammedi S, Subramanian SB, Yan S, Tyagi RD, Valéro JR (2006) Molecu- lar screening of B. thuringiensis strains from wastewater sludge for biopesti- cide production. Process Biochem. 41:829–835.
Morgan JA, Sergeant M, Ellis D, Ousley M, Jarrett P (2001) Sequence analysis of insecticidal genes from Xenorhabdus nematophilus PMFI296. Appl Environ Microbiol. 67: 2062–2069.
Nielsen-LeRoux C, Gaudriault S, Ramarao N, Lereclus D, Givaudan A (2012) How the insect pathogen bacteria Bacillus thuringiensis and Xenorhabdus/ Pho- torhabdus occupy their hosts. Curr Opin Microbiol. 15: 220–231.
Ohba M, Aizawa K (1986) Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J Invertebr Pathol. 47(1): 12–20.
Omar MS (1996) A survey of bancroftian filariasis among South-East Asian ex- patriate workers in Saudi Arabia. Trop Med Int Health. 1(2): 155–160.
Orduz S, Rojas W, Correa MM, Montoya AE, de Barjac H (1992) A new sero- type of Bacillus thuringiensis from Co- lombia toxic to mosquito larvae. J In- vertebr Pathol. 59: 99–103.
Osta MA, Zeinab JR, Pierrick L, Mylène W, Khouzama K (2012) Insecticide re- sistance to organophosphates in Culex pipiens complex from Lebanon. Para- sites Vectors. 5: 132–137.
Padua LE, Ohba M, Aizawa K (1984) Isola- tion of a Bacillus thuringiensis strain (serotype 8a:8b) highly and selectively
toxic against mosquito larvae. J Inver- tebr Pathol. 44: 12–17.
Park HW, Sabrina R, Hayes C, Mangum M (2008) Distribution of mosquitocidal Ba- cillus thuringiensis and Bacillus sphaericus from sediment samples in Florida.J Asia Pac Entomol. 11: 217–220
Ramalakshmi A, Udayasuriyan V (2010) Diversity of Bacillus thuringiensis isolat- ed from Western Ghats of Tamil Nadu State, India. Curr Microbiol. 61: 13–18.
Rampersad J, Ammons D (2005) A Bacillus thuringiensis isolation method utiliz- ing a novel stain, low selection and high throughput produced atypical re- sults. BMC Microbiol. 5: 52–60.
Rey D, Cuany A, Pautou MP, Meyran JC (1999) Differential sensitivity of mos- quito texa to vegetable tannins. J Chem Ecol. 25: 537–548.
Rosen L, Lien JC, Shroyer DA, Baker RH, Lu LC (1989) Experimental vertical transmission of Japanese encephalitis virus by Culex tritaeniorhynchus and other mosquitoes. Am J Trop Med Hyg.40(5): 548–556.
Saferalizadeh AS, Pourmirza AA, Bari MR, Mohseniazar KM (2010) Characteri- zation and pathogenic evaluation of Bacillus thuringiensis isolates from West Azerbaijan province-Iran. Afr J Microbiol Res. 4(12): 1224–1229.
Schünemann R, Knaak N, Fiuza LM (2014) Mode of Action and Specificity of Ba- cillus thuringiensis Toxins in the Con- trol of Caterpillars and Stink Bugs in Soybean Culture. ISRN Microbiology.2014: 1–12.
Seufi AM, Galal FH (2010) Role of Culex and Anopheles mosquito species as po- tential vectors of rift valley fever virus in Sudan outbreak, 2007. BMC Infect Dis. 10: 65–75.
Soberon M, Fernndez LE, Perez C, Gill SS, Bravo A (2007) Mode of action of mosquitocidal Bacillus thuringiensistoxins. Toxicon. 49: 597–600.
Vani C, Lalithambika B (2014) Effect of out- er membrane vesicle proteins of Xenorhabdus bacteria against malarial vectors. Int J Pharm Bio Sci. 5(4):1072–1080.
WHO (2005) Guidelines for laboratory and field testing of mosquito larvicides. (WHO/CDS/WHOPES/GCDPP/2005.
. Available at: http://www.who.int/ whopes/gcdpp/publications/en/index1. html.WHO (2010) World Malaria report. Geneva, Switzerland.
Woodring JL, Kaya HK (1988) Steinernematid and Heterorhabditid nematodes: A hand- book of biology and techniques. Fayette- ville, Ark.: Arkansas Agricultural Experiment Station, Arkansas State Library.
Xavier R, Reena CM, Sreeramanan S (2007) Enviromental distribution and diver- sity of insecticidal proteins of Bacillus thuringiensis Berliner. Malays J Mi- crobiol. 3: 1–6.
Zhu H, Grewal PS, Reding ME (2011) De- velopment of a dessicated cadaver de- livery system to apply entomopatho- genic nem Abul Hab J (1980) A list of arthropoda of medical and veterinary importance rec- orded from Iraq. Bull Biol Res Cent.12(1): 9–40.