Sains Malaysiana 45(4)(2016): 589–594

Bacillus thuringiensis Entomotoxicity Activity in Wastewater Sludge-Culture Medium towards Bactrocera dorsalis and their Histopathological Assessment

(Aktiviti Entomotoksisiti Bacillus thuringiensis dalam Medium Air Sisa Kultur Enap Cemar

ke arah Bactrocera dorsalis dan Penilaian Histopatologinya)

MONA FATIN SYAZWANEE M.G., NOORMASSHELA U.A., NOR AZWADY A.A., RUSEA G. & MUSKHAZLI M.*

Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

Received: 1 April 2014/Accepted: 30 October 2015

ABSTRACT

This study investigates the production of biopesticide based on Bacillus thuringiensis activity in culture media supplemented with semi-solid wastewater sludge as one of the raw ingredient. A series of testing using mixture of sludge and source of protein as B. thuringiensis growth media were carried out and selection of media was based on viable spore count. The entomotoxicity test of B. thuringiensis was carried out against larvae of fruit fly using diet incorporation method. Further impact of entomotoxicity was observed based on histology deformities on columnar epithelial cell and goblet cell of the midgut. A mixture of sludge with 60% wheat bran produced up to 1.64 × 1010 CFU/mL of viable spore count within 10 days of incubation. Based on entomotoxicity test, incorporation of 12 mL of semi-solid wastewater sludge-culture media into fruit fly artificial diet caused the highest fruit fly mortality at 64.8%. The value of semi-solid wastewater sludge-culture media concentration for LC₅₀ was determined at 8.43%. Effect of entomotoxicity can be seen started from 3rd instar larvae where histopathological studies showed that up to 10% of columnar epithelial cells in the intestine were swollen and severe reduction of goblet cell’s size. Thus, it decreases the survivality of the fruit fly larvae. The present study indicated that semi-solid wastewater sludge has the potential to enhance B. thuringiensis entomotoxicity activity.

Keywords: Diet; goblet cell; histopathology; lumen; semi solid; wheat bran

ABSTRAK

Penyelidikan ini mengkaji pengeluaran biopestisid berdasarkan aktiviti Bacillus thuringiensis dalam kultur media yang ditambah dengan enap cemar air sisa separa pepejal sebagai salah satu bahan mentah. Siri ujian menggunakan campuran enap cemar dan sumber protein seperti media pertumbuhan B. thuringiensis telah dijalankan dan pemilihan media adalah berdasarkan kiraan spora berdaya maju. Ujian entomotoksisiti B. thuringiensis telah dijalankan terhadap larva lalat buah menggunakan kaedah penggabungan diet. Satu kesan daripada entomotoksisiti diperhatikan berdasarkan histologi kecacatan sel epitelium turus dan sel goblet daripada perut tengah ini. Campuran enap cemar dengan bran gandum 60% dihasilkan sehingga 1.64 × 1010 CFU/mL kiraan spora berdaya maju dalam tempoh 10 hari dari inkubator. Berdasarkan ujian entomotoksisiti, penubuhan 12 mL media kultur air sisa enap cemar separa pepejal ke dalam diet tiruan lalat buah disebabkan kematian lalat buah yang tertinggi sebanyak 64.8%. Nilai kepekatan kultur media separa pepejal buangan enap cemar PL₅₀ telah ditentukan pada 8.43%. Kesan entomotoksisiti boleh dilihat bermula daripada tiga instar larva dengan kajian histopatologi menunjukkan bahawa sehingga 10% daripada sel-sel epitelium turus dalam usus telah bengkak dan pengurangan saiz sel goblet yang ketara. Oleh itu, ia mengurangkan kemandirian untuk larva lalat buah. Kajian ini menunjukkan bahawa air sisa enap cemar separa pepejal mempunyai potensi untuk meningkatkan aktiviti entomotoksisiti B. thuringiensis.

Kata kunci: Dedak gandum; diet; histopatologi; lumen; sel goblet; separa pepejal

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*Corresponding author; email: muskhazli@upm.edu.my