Detection of changes occurring during recovery from the dauer stage in Heterorhabditis bacteriophora
Abstract
Nematodes of the genus Heterorhabditis are insect parasites that are widely used as biological control agents. When conditions are unfavourable for reproduction in H . bacteriophora, a long-lived, non-feeding, survival and dispersal stage, the dauer juvenile (DJ), is formed. This DJ stage is also adapted for host finding and infection. When it infects a suitable host, the DJ recovers and resumes growth and development. We describe a series of methods for improved detection of recovery in H . bacteriophora. We also describe some of the physiological changes that occur immediately after the onset of recovery in these nematodes as revealed using fluorescent nucleic acid binding SYTO dyes. Although recovery could be monitored using morphological changes, we found that observation of the uptake of fluorescent latex microspheres by recovering nematodes was a far more sensitive and efficient means of detecting recovery. SYTO dyes were also found to be useful indicators of recovery, binding to the pharyngeal glands and genital primordia as little as 3 h after the onset of recovery. The use of SYTO dyes also indicated that the pharyngeal glands produce large quantities of RNA following the onset of recovery, implying that these structures may produce proteins important in the infection and/or feeding process of H . bacteriophora.
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}
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background: no-repeat url(data:image/png;base64,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);
}
}
/* Fancy CSS END */
.ff0_c00001{font-family:sans-serif;visibility:hidden;}
.sc__c00001{text-shadow:none;}
@media screen and (-webkit-min-device-pixel-ratio:0){
.sc__c00001{-webkit-text-stroke:0px transparent;}
}
.y0_c00001{bottom:0.000000px;}
.h0_c00001{height:1263.599985px;}
.h1_c00001{height:1263.750000px;}
.w0_c00001{width:918.000000px;}
.x0_c00001{left:0.000000px;}
@media print{
.y0_c00001{bottom:0.000000pt;}
.h0_c00001{height:1123.199987pt;}
.h1_c00001{height:1123.333333pt;}
.w0_c00001{width:816.000000pt;}
.x0_c00001{left:0.000000pt;}
}
/* vim: set shiftwidth=2 tabstop=2 autoindent cindent expandtab filetype=css: */
/*!
* Base CSS for pdf2htmlEX
* Copyright 2012,2013 Lu Wang
* https://github.com/pdf2htmlEX/pdf2htmlEX/blob/master/share/LICENSE
*/
/* Part 1: Web Page Layout: Free to modify, except for a few of them which are required by pdf2htmlEX.js, see the comments */
#sidebar { /* Sidebar */
position:absolute;
top:0;
left:0;
bottom:0;
width:250px;
padding:0;
margin:0px;
overflow:auto;
}
#page-container { /* PDF container */
position:absolute; /* required for calculating relative positions of pages in pdf2htmlEX.js */
top:0;
left:0px;
margin:0;
padding:0;
border:0; /* required for lazy page loading in pdf2htmlEX.js (page visibility test) */
}
@media screen {
/* for sidebar */
#sidebar.opened + #page-container { left:250px; }
#page-container {
/* `bottom' and `right' are required for lazy page loading in pdf2htmlEX.js (page visibility test)
* alternatively you may set width and height
*/
bottom:0;
right:0;
overflow:auto;
}
.loading-indicator_c00002 {
display:none;
}
.loading-indicator_c00002.active {
display:block;
position:absolute;
width:64px;
height:64px;
top:50%;
left:50%;
margin-top:-32px;
margin-left:-32px;
}
.loading-indicator_c00002 img {
position:absolute;
top:0;
left:0;
bottom:0;
right:0;
}
}
@media print {
@page { margin:0; }
html { margin:0; }
body {
margin:0;
-webkit-print-color-adjust:exact; /* enable printing background images for WebKit */
}
#sidebar { display:none; }
#page-container {
width:auto;
height:auto;
overflow:visible;
background-color:transparent;
}
.d_c00002 { display:none; }
}
/* Part 2: Page Elements: Modify with caution
* The followings are base classes, some of which are meant to be override by PDF specific classes
* So do not increase the specificity (e.g. ".classname_c00002" -> "#page-container .classname_c00002")
*/
.pf_c00002 { /* page */
position:relative;
background-color:white;
overflow: hidden;
margin:0;
border:0; /* required by pdf2htmlEX.js for page visibility test */
}
.pc_c00002 { /* content of a page */
position:absolute;
border:0;
padding:0;
margin:0;
top:0;
left:0;
width:100%;
height:100%;
overflow:hidden;
display:block;
/* set transform-origin for scaling */
transform-origin:0% 0%;
-ms-transform-origin:0% 0%;
-webkit-transform-origin:0% 0%;
}
.pc_c00002.opened { /* used by pdf2htmlEX.js, to show/hide pages */
display:block;
}
.bf_c00002 { /* images that occupies the whole page */
position:absolute;
border:0;
margin:0;
top:0;
bottom:0;
width:100%;
height:100%;
-ms-user-select:none;
-moz-user-select:none;
-webkit-user-select:none;
user-select:none;
}
.bi_c00002 { /* images that cover only a part of the page */
position:absolute;
border:0;
margin:0;
-ms-user-select:none;
-moz-user-select:none;
-webkit-user-select:none;
user-select:none;
}
@media print {
.pf_c00002 {
margin:0;
box-shadow:none;
page-break-after:always;
page-break-inside:avoid;
}
@-moz-document url-prefix() {
/* fix page truncation for FireFox */
.pf_c00002 {
overflow:visible;
border:1px solid #FFFFFF;
}
.pc_c00002 {overflow:visible;}
}
}
.c_c00002 { /* clip box */
position:absolute;
border:0;
padding:0;
margin:0;
overflow:hidden;
display:block;
}
.t_c00002 { /* text line */
position:absolute;
white-space:pre;
font-size:1px;
transform-origin:0% 100%;
-ms-transform-origin:0% 100%;
-webkit-transform-origin:0% 100%;
unicode-bidi:bidi-override;/* For rtl languages, e.g. Hebrew, we don't want the default Unicode behaviour */
-moz-font-feature-settings:"liga" 0;/* We don't want Firefox to recognize ligatures */
}
.t_c00002:after { /* webkit #35443 */
content: '';
}
.t_c00002:before { /* Workaround Blink(up to 41)/Webkit bug of word-spacing with leading spaces (chromium #404444 and pdf2htmlEX #412) */
content: '';
display: inline-block;
}
.t_c00002 span { /* text blocks within a line */
/* Blink(up to 41)/Webkit have bug with negative word-spacing and inline-block (pdf2htmlEX #416), so keep normal span inline. */
position:relative;
unicode-bidi:bidi-override; /* For rtl languages, e.g. Hebrew, we don't want the default Unicode behaviour */
}
.__c00002 { /* text shift */
/* Blink(up to 41)/Webkit have bug with inline element, continuous spaces and word-spacing. Workaround by inline-block. */
display: inline-block;
color: transparent;
z-index: -1;
}
/* selection background should not be opaque, for fallback mode */
::selection{
background: rgba(127,255,255,0.4);
}
::-moz-selection{
background: rgba(127,255,255,0.4);
}
.pi_c00002 { /* info for Javascript */
display:none;
}
.l_c00002 { /* annotation links */
}
/* transparent color - WebKit */
.d_c00002 { /* css drawing */
position:absolute;
transform-origin:0% 100%;
-ms-transform-origin:0% 100%;
-webkit-transform-origin:0% 100%;
}
/* for the forms */
.it_c00002 {
border: none;
background-color: rgba(255, 255, 255, 0.0);
}
.ir_c00002:hover {
cursor: pointer;
}
/* Base CSS END */
/* vim: set shiftwidth=2 tabstop=2 autoindent cindent expandtab filetype=css: */
/*!
* Fancy styles for pdf2htmlEX
* Copyright 2012,2013 Lu Wang
* https://github.com/pdf2htmlEX/pdf2htmlEX/blob/master/share/LICENSE
*/
@keyframes fadein { from { opacity:0;} to { opacity:1;} }
@-webkit-keyframes fadein { from { opacity:0;} to { opacity:1;} }
@keyframes swing {
0% { transform: rotate(0deg); }
10% { transform: rotate(0deg); }
90% { transform: rotate(720deg); }
100%{ transform: rotate(720deg); }
}
@-webkit-keyframes swing {
0% { -webkit-transform: rotate(0deg); }
10% { -webkit-transform: rotate(0deg); }
90% { -webkit-transform: rotate(720deg); }
100%{ -webkit-transform: rotate(720deg); }
}
@media screen {
#sidebar {
background-color:#2f3236;
/* modified from http://philbit.com/svgpatterns/#crossstripes */
background-image:url("data:image/svg+xml;base64,PHN2ZyB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciIHdpZHRoPSI0IiBoZWlnaHQ9IjQiPgo8cmVjdCB3aWR0aD0iNCIgaGVpZ2h0PSI0IiBmaWxsPSIjNDAzYzNmIj48L3JlY3Q+CjxwYXRoIGQ9Ik0wIDBMNCA0Wk00IDBMMCA0WiIgc3Ryb2tlLXdpZHRoPSIxIiBzdHJva2U9IiMxZTI5MmQiPjwvcGF0aD4KPC9zdmc+");
}
#outline {
font-family:Georgia,Times,"Times New Roman",serif;
font-size:13px;
margin:2em 1em;
}
#outline ul {
padding:0;
}
#outline li {
list-style-type:none;
margin:1em 0;
}
#outline li > ul {
margin-left: 1em;
}
#outline a,
#outline a:visited,
#outline a:hover,
#outline a:active {
line-height:1.2;
color:#e8e8e8;
text-overflow:ellipsis;
white-space:nowrap;
text-decoration:none;
display:block;
overflow:hidden;
outline:0;
}
#outline a:hover {
color:rgb(0,204,255);
}
#page-container {
background-color:#9e9e9e;
/* http://philbit.com/svgpatterns/#thinstripes */
background-image: url("data:image/svg+xml;base64,PHN2ZyB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciIHdpZHRoPSI1IiBoZWlnaHQ9IjUiPgo8cmVjdCB3aWR0aD0iNSIgaGVpZ2h0PSI1IiBmaWxsPSIjOWU5ZTllIj48L3JlY3Q+CjxwYXRoIGQ9Ik0wIDVMNSAwWk02IDRMNCA2Wk0tMSAxTDEgLTFaIiBzdHJva2U9IiM4ODgiIHN0cm9rZS13aWR0aD0iMSI+PC9wYXRoPgo8L3N2Zz4=");
-webkit-transition:left 500ms;
transition:left 500ms;
}
.pf_c00002 {
margin: 13px auto;
box-shadow: 1px 1px 3px 1px #333;
/* Needed by IE to make box-shadow works * https://developer.mozilla.org/en-US/docs/Web/CSS/box-shadow */
border-collapse: separate;
}
.pc_c00002.opened { /* used by pdf2htmlEX.js, to show/hide pages */
-webkit-animation: fadein 100ms;
animation: fadein 100ms;
}
.loading-indicator_c00002.active {
/*
* use 0.01s instead of 0s,
* since YUI Compressor will change 0s to 0,
* which is not recognized by Firefox
*/
-webkit-animation: swing 1.5s ease-in-out 0.01s infinite alternate none;
animation: swing 1.5s ease-in-out 0.01s infinite alternate none;
}
.checked_c00002 {
background: no-repeat url(data:image/png;base64,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);
}
}
/* Fancy CSS END */
.ff0_c00002{font-family:sans-serif;visibility:hidden;}
.sc__c00002{text-shadow:none;}
@media screen and (-webkit-min-device-pixel-ratio:0){
.sc__c00002{-webkit-text-stroke:0px transparent;}
}
.y0_c00002{bottom:0.000000px;}
.h0_c00002{height:1263.599985px;}
.h1_c00002{height:1263.750000px;}
.w0_c00002{width:918.000000px;}
.x0_c00002{left:0.000000px;}
@media print{
.y0_c00002{bottom:0.000000pt;}
.h0_c00002{height:1123.199987pt;}
.h1_c00002{height:1123.333333pt;}
.w0_c00002{width:816.000000pt;}
.x0_c00002{left:0.000000pt;}
}