Toxicity of Lysinibacillus sphaericus Q001 S-Layer Protein Against Culex quinquefasciatus and Anopheles stephensi Larvae | [Pakistan Journal of Zoology • 2024]

Author(s):

1. Hamayun Arshad: School of Biological Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan

2. Muhammad Waris: Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan; Primary and Secondary Healthcare Department, Kasur, Pakistan

3. Qurratulann Afza Gardner: School of Biological Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan

Abstract:

Use of larvicidal proteins based on bacteria is regarded as an effective and environment friendly method of insect control. In this regard, surface layer (S-layer) protein in various strains of Lysinibacillus sphaericus is reported to exhibit bioinsecticidal property. Here, we aimed to search for local strain of L. sphaericus and test its S-layer protein for toxicity against Culex quinquefasciatus and Anopheles stephensi larvae. For this purpose, we isolated various bacterial strains from our local environment using standard microbiological techniques. Among those isolates, we found novel strain of L. sphaericus Q001, identified by ribotyping and MALDI-biotyping. The 16S ribosomal RNA gene of this strain is 96% similar to that of highly entomotoxic WHO reference strain 2362. The gene sequence was submitted to Genbank (OQ701385). We isolated the S-layer protein of this strain by treating it with lithium chloride (LiCl). After SDS-PAGE analysis, the protein was subjected to in-gel digestion and peptide mass fingerprinting for identification. Following the successful identification of this protein, we used it to test its toxicity against mosquito larvae by following WHO guidelines with slight modification. The bioassays were performed in triplicate and LC50 was calculated by applying log probit analysis. In conclusion, we found that S-layer protein of L. sphaericus Q001 was effective against the tested larvae with LC50 values of 8.5 µg/ml and 16.1 µg/ml against Cx. quinquefasciatus and An. stephensi larvae respectively. Furthermore, field trials can reveal the true potential of this protein as bioinsecticide.

Page(s): 2781-2786

DOI: 10.17582/journal.pjz/20230406110459

Published: Journal: Pakistan Journal of Zoology, Volume: 56, Issue: 6, Year: 2024

Keywords:

Culex quinquefasciatus , ribotyping , bioinsecticide , MALDI biotyping , Anopheles stephensi , Lysinibacillus sphaericus

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