\documentclass[10pt]{article}

\usepackage{fullpage}
\usepackage{setspace}
\usepackage{parskip}
\usepackage{titlesec}
\usepackage[section]{placeins}
\usepackage{xcolor}
\usepackage{breakcites}
\usepackage{lineno}
\usepackage{hyphenat}





\PassOptionsToPackage{hyphens}{url}
\usepackage[colorlinks = true,
            linkcolor = blue,
            urlcolor  = blue,
            citecolor = blue,
            anchorcolor = blue]{hyperref}
\usepackage{etoolbox}
\makeatletter
\patchcmd\@combinedblfloats{\box\@outputbox}{\unvbox\@outputbox}{}{%
  \errmessage{\noexpand\@combinedblfloats could not be patched}%
}%
\makeatother


\usepackage{natbib}




\renewenvironment{abstract}
  {{\bfseries\noindent{\abstractname}\par\nobreak}\footnotesize}
  {\bigskip}

\titlespacing{\section}{0pt}{*3}{*1}
\titlespacing{\subsection}{0pt}{*2}{*0.5}
\titlespacing{\subsubsection}{0pt}{*1.5}{0pt}


\usepackage{authblk}


\usepackage{graphicx}
\usepackage[space]{grffile}
\usepackage{latexsym}
\usepackage{textcomp}
\usepackage{longtable}
\usepackage{tabulary}
\usepackage{booktabs,array,multirow}
\usepackage{amsfonts,amsmath,amssymb}
\providecommand\citet{\cite}
\providecommand\citep{\cite}
\providecommand\citealt{\cite}
% You can conditionalize code for latexml or normal latex using this.
\newif\iflatexml\latexmlfalse
\providecommand{\tightlist}{\setlength{\itemsep}{0pt}\setlength{\parskip}{0pt}}%

\AtBeginDocument{\DeclareGraphicsExtensions{.pdf,.PDF,.eps,.EPS,.png,.PNG,.tif,.TIF,.jpg,.JPG,.jpeg,.JPEG}}

\usepackage[utf8]{inputenc}
\usepackage[english]{babel}



\usepackage{float}








\begin{document}

\title{Stability of environmental DNA methylation and its utility in tracing
reproductive activities of fish}



\author[1]{Itsuki Hirayama}%
\author[1]{Toshifumi Minamoto}%
\author[1]{Luhan Wu}%
\affil[1]{Kobe University}%


\vspace{-1em}



  
  \date{May 23, 2024}


\begingroup
\let\center\flushleft
\let\endcenter\endflushleft
\maketitle
\endgroup





\selectlanguage{english}
\begin{abstract}
1. The use of environmental DNA (eDNA) as a new method of ecological
monitoring is widely applied. Although eDNA can provide important
information on the distribution and biomass of particular taxa, an
organism's DNA sequences remain unaltered throughout its existence,
which complicates identifying crucial events, including reproduction,
with high accuracy. We thus examined DNA methylation as a novel source
of information from eDNA, considering that methylation patterns of eggs
and sperm released during reproduction differ from those of somatic
tissues. 2. Despite its potential applications, little is known about
eDNA methylation, including its stability and methods for detection and
quantification. Therefore, we conducted tank experiments and performed
methylation analysis targeting 18S rDNA through bisulfite amplicon
sequencing. 3. Methylation of eDNA was not affected by degradation and
was equivalent to the rate of genomic DNA from somatic tissues.
Unmethylated DNA, which is abundant in the ovary, was detected in eDNA
during reproductive activity of fish. 4. These results indicate that
eDNA methylation is a stable signal reflecting genomic methylation and
demonstrate that germ cell-specific methylation patterns can be used as
markers for detecting reproductive activity.%
\end{abstract}%



\sloppy


\textbf{Hosted file}

\verb`eDNA methylation_title&main_ver1.1.docx`  available at \url{https://authorea.com/users/753389/articles/723683-stability-of-environmental-dna-methylation-and-its-utility-in-tracing-reproductive-activities-of-fish}

\selectlanguage{english}
\FloatBarrier
\end{document}

