Complete mitochondrial genome of Gnathostoma binucleatum

This report describes the mitochondrial genome of the parasite Gna thostoma binucleatum (G.binucleatum), which was obtained from naturally infected freshwater fish in Sinaloa, Mexico (22°46′00.1″N105°40′21.8″W).G. binucleatum is responsible for human gnathostomiasis and is endemic to Mexico.It belongs to the Spirurida order of the Secernentea class of Nematoda.binucleatum) is a parasitic nematode that causes the zoonotic disease gnathostomiasis (1).Little is known about the parasite's molecular and genetic biology, which is necessary for the development of effective anti-parasitic drugs.Only the mitochondrial MT-CO1 gene of G. binucleatum has been characterized (2).Here, we present the complete mitochondrial genome of G. binucleatum (GbMG).Twenty-five advanced third-stage G. binucleatum larvae were isolated from infected freshwater fish in Tecualilla, Sinaloa, Mexico (22°46′00.1″N105°40′21.8″W).All proce dures were conducted in accordance with the International Guiding Principles for Biomedical Research Involving Animals.DNA was extracted by treating the parasites with alkaline lysis buffer (5 M NaCl, 1 M Tris pH 8, 0.5 M EDTA pH 8, 10% SDS, 40-µL proteinase K) at 55°C for 30 min, homogenizing them, and then mixing the homogenate with phenol-Tris-HCl.After centrifugation, the supernatant was transferred to a new tube, and the extraction process was repeated.The final supernatant was mixed with ethanol and centrifuged, and the pellet was then recovered and washed with 70% ethanol by pipette mixing.Following a final centrifugation, the DNA-containing ethanol supernatant was mixed with Tris-EDTA (TE) buffer and prepared for sequencing using the Illumina DNA Prep Kit.Psomagen, Inc. (Rockville MD, USA), sequenced the samples using the Illumina MiSeq platform.A total of 44,235,384 raw reads were produced, which comprised of 22,117,692 paired-end reads (2 × 150 bp).R1 and R2 fastq files were combined and trimmed for adapters, low-quality reads, and short reads.BBDuk (Decontamination Using Kmers) version 38.84, which is part of the BBTools suite, was used for trimming (3).BBNorm version 38.84, which is also a part of the BBTool suite, was used for error correction and normalization to produce a total of 727,338 reads, 97.6% of which had a Phred score of at least Q30 (3).After pre-processing the reads, their mean length was 146 bp (SD 75 bp), with a range between 35 bp and 301 bp long.For all software tools used, the default parameters and native algorithms were chosen unless otherwise noted.Geneious Prime version 2023.0.1 was used for de novo assembly, followed by mapping contigs (4).Fifteen of the de novo assembled contigs (with an N50 of 9.875 kb and a depth of 6) were then subsequently mapped to the mitochondrial genome reference sequence of G. nipponicum (NC_034239.1).However, a consensus sequence with less ambiguity and gaps was produced when all the pre-processed 727,338 reads were mapped, resulting in a consensus sequence made from 13,163 reads.