Molecular Characterization of Entomopathogenic Fungi Aspergillus amstelodami as a Bio control agent of Tick Infestation in Cattle
DOI:
https://doi.org/10.48165/ijvsbt.20.1.12Keywords:
Acaricide, Bio control , Entomopathogenic fungi (EPF), Molecular marker, ResistanceAbstract
Tick and tick borne diseases affects the production performance of animals ultimately results in economic losses to livestock owners. Tick control using chemicals producing acaricidal resistance and environmental pollution. To overcome this the study was conducted to introduce the biological control agents in the form of entomopathogenic fungi (EPF) isolated from naturally infected ticks. The molecular characterization of EPF species Aspergillus amstelodami isolated from naturally infected ticks of Durg district of Chhattisgarh was carried out by Polymerase Chain reaction (PCR) using internal transcribed spacer (ITS) gene as a molecular marker. The quality of DNA isolated from the culture was evaluated on 1.0% Agarose Gel with a single band of high-molecular weight DNA. Fragment of gene was amplified by PCR. The amplified PCR product was analyzed by agarose gel electrophoresis using 100bp ladder as a marker showed a single intense band corresponding to a 543 bp indicating successful amplification of gene from template DNA. DNA sequencing reaction of PCR amplicon was carried out with ITS1 primer using BDT v3.1 Cycle sequencing kit on ABI 3730xl Genetic Analyzer. The gene sequence was used to carry out BLAST with the NCBI Genbank database. The fungal isolate showed high similarity with Aspergillus amstelodami based on nucleotide homology and phylogenetic analysis. The evolutionary history was inferred using the Neighbor-Joining method. The confidence probability (multiplied by 100) that the interior branch length was greater than 0, as estimated using the bootstrap test (500 replicates was shown next to the branches. The evolutionary distances were computed using the Maximum Composite Likelihood method and were in the units of the number of base substitutions per site. The analysis involved 11 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding.
Downloads
References
Cafarchia, C., Davide, .I, Roberta, .I, Rafael, ANR, Riccardo, .PL. and Daniele, P. (2015). Native strains of Beauveria bassiana for the control of Rhipicephalus sanguineus sensulato. Parasites and Vectors . 8 (1):80
Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, Dufayard J F, Guindon S, Lefort V, Lescot M, Claverie J M, Gascuel O. (2008). Phylogeny fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res. 1:36.
Dich, J., Zahm, S.H., Hanberg, A. and Adami, H.O. (1997). Pesticides and cancer..Cancer Causes & Control. 8(3):420–443.
Dopazo J. (1994). Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach. Journal of Molecular Evolution..38:300-304.
Gardes M and Bruns T D. (1993). ITS primers with enhanced specificity for basidiomycetes: application to the identification of mycorrhizae and rusts. Mol. Ecol. .2: 113–118.
Kolpin, D.W., Barbash, J.E., Gilliom, R.J.(1998). Occurrence of pesticides in shallow groundwater of the United States: Initial results from the National Water-Quality Assessment Program. Environmental Science and Technology. .32(5):558–566.
Kunz, S. E .and Kemp, D.H. (1994). Insecticides and acaricides: resistance and environmental impact. Revue scientifique et technique 13 (4): 1249-1286
Martins, J.R., Correa, B.L., Cereser, V.H. and Arteche, C.C.P. (1995). A situation report on resistance to acaricides by the cattle tick Boophilus microplus in t state of Rio Grande do Sul, southern Brazil. In C. Rodriguez, D. Sergio & H. Fragoso, eds. 3rd International seminary on Animal Parasitology. Acapulco, Guerrero, Mexico.
Rzhetsky A. and Nei M. (1992). A simple method for estimating and testing minimum evolution trees. Molecular Biology and Evolution. 9:945-967
Saitou N. and Nei M. The neighbor-joining method: (1987). A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution . 4:406-425.
Samish, M., Ginsberg, H. and Glazer, I. (2004). Biological control of ticks. Parasitology, 129: 389-403.
Talavera, G., and Castresana, J. (2007). Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Systematic Biology. 56:564-577.
Tamura K., Nei M., and Kumar S. (2004). Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences (USA) .101:11030-11035
Thakur Rupesh and Sandhu Sardul. (2010). Species confirmation of fungal isolates by molecular analysis. Indian J Microbiol .50:280–291
White T J, Bruns T, Lee S, Taylor J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics . 315–322.
Yayan, Sanjaya, Virginia, R. Ocampo and Barbara, L. Caoili. (2016). Identification of potential entomopathogenic fungi of Tetranychus kanzawai Kishida (Tetranychidae: Acarina) using ITS-5.8s rDNA region as molecular marker. Agrivita Journal of Agricultural Science. 38(2):186-19
Downloads
Published
Issue
Section
License
Copyright (c) 2023 Indian Journal of Veterinary Sciences & Biotechnology
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
