نشكر مانيل بوليفار لمساعدته في العمل الميداني. ونحن ممتنون لل&quot;بارك الطبيعية ديل مونتغري، ليه ميديس ط شرم BAIX تير&quot; على دعمهم لدينا أذونات البحوث وأخذ العينات. تم تصميم مناور تحت الماء عن طريق ايليت دادون-Pilosof وملفقة من قبل السيد Pilosof. وأيد هذا العمل من قبل مشروع الحكومة الاسبانية CSI-كورال [عدد المنح CGL2013-43106-R إلى الصليب الأحمر وMR] وزمالة فبو من &quot;MINISTERIO دي EDUCACION، ثقافة ذ ديبورتي (MECD)&quot; لTM. وهذه مساهمة من الكيمياء الحيوية البحرية ومجموعة التغيير الأبحاث العالمية التي تمولها الحكومة الكاتالانية [منحة عدد 2014SGR1029] وقوى الأمن الداخلي منحة 1280-1213 والبنك السعودي الفرنسي منحة 2012089 لG. ياهيل.

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The authors have nothing to disclose.

الخطوات التحضيرية قارورة جامعي لDOM وتحليل المواد الغذائية منذ السفن جامع قد تتفاعل مع حل المكونات الدقيقة والجدران العينات قد تكون ركيزة لنمو البكتيريا 30-34، تم ?...

مغذيات تعليق القاعية تلعب أدوارا أساسية في أداء النظم الإيكولوجية البحرية 1. من خلال تصفية كميات كبيرة من المياه 2،3، وإزالة وتفرز الجسيمات (العوالق والمخلفات) والمركبات الذائبة 1 (والمراجع فيها) والتي تشكل عاملا مهما من بين القاعية البحرية اقتران 4،5 والمغذيات الدراجات 6،7. قياس بدقة الجسيمات وحلت المركبات إزالتها والتي تفرز مغذيات تعليق القاعية (مثل الإسفنج، المزقييات، الأشواك، وذوات الصدفتين) هو أمر أساسي لفهم علم وظائف الأعضاء، والتمثيل الغذائي، والبيئة التغذية الخاصة بهم. جنبا إلى جنب مع ضخ قياسات معدل، فإنه يمكن أيضا تقدير حجم تدفقات المواد الغذائية بوساطة هذه الكائنات والتأثير البيئي على نوعية المياه فضلا عن العمليات على نطاق والنظام البيئي. اختيار الطريقة المناسبة لقياس إزالة والإنتاج معدلات الجسيمات وكوم المنحلجنيه فلتر مغذيات تعليق أمر حاسم للحصول على بيانات موثوق بها بشأن نشاط التغذية من 8. كما أشار Riisgård وغيرها، غير لائقة النتائج منهجيات التحيز وتشويه ظروف تجريبية، إنتاج تقديرات غير صحيحة من الابتلاع وإفراز مواد معينة، ويمكن أن يؤدي إلى تقدير خاطئ لتدفقات المواد الغذائية المصنعة من قبل هذه الكائنات. طريقتين المستخدمة في معظم الأحيان لقياس جسيمات وتدفقات المغذيات الذائبة في مغذيات الترشيح تنطوي إما حضانة (تقنيات غير المباشرة) أو جمع في وقت واحد من المحيط والمياه الزفير (تقنيات المباشرة). وتعتمد تقنيات الحضانة على قياس معدل التغير في تركيز الجسيمات والمواد الغذائية الذائبة في الماء المحتضنة، وتقدير معدلات الإنتاج أو إزالة مقارنة مع الضوابط الكافية 8. ومع ذلك، أرفق كائن حي في غرفة الحضانة يمكن أن يغير feedin لهاز وضخ السلوك نتيجة للتغيرات في النظام التدفق الطبيعي، وذلك بسبب انخفاض الأكسجين و / أو في تركيز المواد الغذائية، أو بسبب تراكم مركبات افرازه في 7،9 المياه الحضانة (والمراجع فيها). بالإضافة إلى آثار الولادة وإمدادات المياه المعدلة، وجود تحيز كبير من التقنيات حضانة ينبع من آثار إعادة الترشيح (انظر على سبيل المثال 10). على الرغم من أن بعض هذه المشاكل المنهجية التي تم التغلب عليها باستخدام حجم المناسب وشكل سفينة الحضانة 11 أو مع إدخال نظام جرس جرة إعادة تدوير في الموقع 12، هذا الأسلوب غالبا ما يقلل إزالة والإنتاج أسعار الفائدة. قياس استقلاب المركبات الذائبة مثل النيتروجين المذاب العضوية (DON) والكربون (DOC) أو المواد الغذائية غير العضوية، وقد ثبت أن تكون عرضة بشكل خاص لالتحيزات الناجمة عن تقنيات الحضانة 13. في أواخر 60s و 70s في وقت مبكر، هنري Reiswig9،14،15 رائدا في تطبيق تقنيات المباشرة لتحديد إزالة الجسيمات من الإسفنج العملاقة في منطقة البحر الكاريبي، عن طريق أخذ عينات بشكل منفصل الماء استنشاق والزفير من قبل الكائنات الحية في الموقع. نظرا لصعوبة تطبيق تقنية Reiswig على مغذيات تعليق أصغر وفي ظروف تحت الماء أكثر تحديا، واقتصر الجزء الأكبر من البحوث في هذا المجال إلى المختبر (في المختبر) استخدام تقنيات حضانة غير المباشرة في الغالب 16. أصلحت ياهيل وزملاؤه مباشرة في Reiswig في تقنية الموقع على العمل في ظروف نطاق أصغر. طريقتهم، وصف INEX 16، يقوم على أخذ العينات تحت الماء في وقت واحد من الماء استنشاقه (في) والزفير (خروج) من قبل الكائنات الحية دون عائق. وتراكيز مختلفة من مادة (على سبيل المثال، والبكتيريا) بين زوج من عينات (INEX) يوفر قدرا من الاحتفاظ (أو الإنتاج) من هذه المادة من قبل الحيوان. تقنية INEX توظف أنابيب مفتوحة وتعتمد على طائرة إفراغي التي تنتجها النشاط ضخ الكائن درس ليحل محل سلبي على المياه المحيطة في أنبوب جمع. في حين ياهيل وزملاؤه طبقت بنجاح هذه التقنية في دراسة شملت أكثر من 15 تعليق مختلف مغذيات الأصناف (على سبيل المثال، 17)، يتم تقييد طريقة من مستوى عال من الممارسة والخبرة اللازمة، حسب حجم ضئيلة من بعض فتحات إفراغي، و حالة البحر. للتغلب على هذه العقبات، قمنا بتطوير تقنية بديلة تقوم على شفط رقابة من الماء عينات من خلال أنابيب دقيقة (القطر الخارجي &lt;1.6 مم). وكان لدينا هدف إلى إنشاء جهاز بسيط وموثوق بها، وغير مكلفة من شأنها أن تسمح نظيفة وتسيطر عليها في عينات المياه خارج الوضع من وجهة نظر محددة جدا، مثل فتحة إفراغي من مغذيات تعليق القاعية. أن تكون فعالة، وطريقة يجب أن يكون غير تدخلية حتى لا تؤثر على نظام تدفق المحيطة أو تعديل بehavior من الكائنات التي شملتها الدراسة. ويطلق على الجهاز المعروضة هنا VacuSIP. هو تبسيط نظام SIP التي وضعتها ياهيل وآخرون. (2007) 18 لأخذ العينات نقطة أساس ROV في أعماق البحار. وVacuSIP هو أرخص بكثير من SIP الأصلي وتم تكييفه للعمل على أساس الغوص. وقد صمم هذا النظام وفقا للمبادئ التي قدمت واختبارها من قبل رايت وستيفنز (1978) 19 وMøhlenberg وRiisgård (1978) (20) لبيئة معملية. وعلى الرغم من تصميم نظام VacuSIP لفي دراسات الموقع من عملية التمثيل الغذائي للمغذيات تعليق القاعية، ويمكن أن تستخدم أيضا لدراسات مخبرية وحيثما هو مطلوب، عينة المياه نقطة مصدر التحكم ونظيفة. هذا النظام هو مفيد خصوصا عندما يطلب من التكامل على فترات طويلة (مين ساعة) أو في الفلترة الموقع. وقد استخدم VacuSIP بنجاح في المختبر ياهيل منذ عام 2011، ولديه أيضااستخدمت في اثنين من الدراسات التي أجريت مؤخرا من تدفقات المواد الغذائية بوساطة أنواع الإسفنج البحر الكاريبي والبحر الأبيض المتوسط ​​21 (مورجانتي آخرون المقدمة). استخدام عينات محددة، ومدة أخذ العينات لفترات طويلة، والظروف الميدانية، التي يتم تطبيقها VacuSIP، يستلزم بعض الانحرافات من بروتوكولات المحيطات القياسية لجمع وتصفية وتخزين العينات لمدة التحاليل الحساسة. للحد من مخاطر التلوث من قبل النظام VacuSIP أو احتمال تعديل المياه عينات من النشاط البكتيري بعد جمع، نحن اختبار مختلفة في إجراءات الترشيح وتخزين الموقع. تم فحص الأجهزة المختلفة الترشيح، والسفن جمع، وإجراءات تخزين من أجل تحقيق هذه التقنية الأكثر مناسبة لتحليل حل غير العضوية (PO 4 3-، أكاسيد النيتروجين - NH 4 +، شافي 4) والعضوية (DOC + DON) مركبات، وفائقة العوالق (&lt;1081؛ م) والجسيمات العضوية (POC + PON) أخذ العينات. لمزيد من خفض خطر التلوث، خاصة في ظل الظروف الميدانية، تم تخفيض عدد من الخطوات التعامل إلى الحد الأدنى. يتم توجيه شكل البصرية التي يتم عرضها في طريقة لتسهيل التكاثر ويقلل من الوقت اللازم لتطبيق كفاءة هذه التقنية. نبذة عن النظام لأخذ عينات من الماء في الموقع التي يتم ضخها من مغذيات تعليق مع فتحات زافر صغيرة مثل 2 مم، هو تصور النشاط ضخ كل عينة لأول مرة عن طريق الإفراج عن تصفيتها فلوريسئين مصبوغ مياه البحر بالقرب من فتحة المستنشق (ق) ومراقبة التدفق من إفراغي الفتحة 16 (انظر أيضا الشكل 2B في 18). الماء استنشاق والزفير من قبل عينة الدراسة (incurrent وإفراغي) ثم يتم أخذ عينات في وقت واحد باستخدام زوج من أنابيب دقيقة مثبتة على مناور مبنية خصيصا أو على اثنين من &quot;عالسيدة &quot;لترايبود المحمولة مرنة رأسا على عقب (الشكل 1 والتكميلية فيديو 1). ويتم جمع المياه استنشاقه من قبل كائن الدراسة عن طريق وضع بعناية نهاية القريبة من أنبوب واحد داخل أو بالقرب من فتحة المستنشق للكائن الحي الدراسة. متطابقة ثم يتم وضع أنبوب داخل فتحة إفراغي. وتتطلب هذه العملية رعاية جيدة لتجنب الاتصال أو اضطراب الحيوان، على سبيل المثال، من خلال إعادة تعليق الرواسب. للبدء في أخذ العينات، غطاس يخترق الحاجز في وعاء جمع بإبرة حقنة تعلق على النهاية البعيدة من كل أنبوب، والسماح للضغط المياه الخارجية لإجبار المياه عينات في الإناء من خلال أنبوب أخذ العينات. وتبدأ شفط من فراغ إنشاؤها مسبقا في قارورة واختلاف الضغط بين الماء الخارجي وعاء العينة إجلاء . لضمان جمع نظيفة من المياه الزفير وتجنب شفط عرضي AMBIوالأنف والحنجرة المياه 16، يحتاج معدل أخذ العينات الماء إلى أن يبقى على معدل أقل بكثير (&lt;10٪) من معدل تدفق إفراغي. يتم التحكم في معدل الامتصاص من طول الأنبوب وقطره الداخلي (ID). كما يضمن القطر الداخلي صغير حجم القتلى لا يكاد يذكر (&lt;200 ميكرولتر لكل متر من الأنابيب). أخذ العينات على فترات طويلة (دقائق إلى ساعات) يجعل من الممكن دمج patchiness المتأصل في معظم المواد المثيرة للاهتمام. للتأكد من أن العينات يتم الاحتفاظ بشكل كاف في جلسات أخذ العينات تحت الماء لفترات طويلة فضلا عن النقل إلى المختبر، وهو في الترشيح الموقع يستحب للالتحاليل الحساسة. وأملت اختيار السفن أخذ العينات، والتجمع، والترشيح، وأنابيب من الكائنات دراسة ومسألة بحثية محددة. البروتوكول هو موضح أدناه يفترض أن ملف تعريف التمثيل الغذائي الكامل هو من مصلحة (لمحة عامة انظر الشكل 2). ومع ذلك، فإن طبيعة وحدات من هذا البروتوكول يسمح وأو تعديل من السهل استيعاب مخططات أخذ العينات أبسط أو حتى مختلفة جدا. لمحة التمثيل الغذائي الكامل، ينبغي أن يتضمن بروتوكول أخذ العينات الخطوات التالية: (1) التصور التدفق. (2) التغذية أخذ العينات فائقة العوالق (العوالق &lt;10 ميكرون). (3) أخذ العينات غير العضوية المغذيات امتصاص وإفراز (باستخدام مرشحات في خط)؛ (4) أخذ العينات حل امتصاص العضوية وإفراز (باستخدام مرشحات في خط)؛ (5) التغذية الجسيمات وإفراز (باستخدام مرشحات في خط)؛ (6) كرر الخطوة 2 (فائقة العوالق التغذية كما فحص الجودة)؛ (7) تدفق التصور. عندما يكون ذلك ممكنا من الناحية اللوجستية، فمن المستحسن أن القياسات الشخصية الأيضية هي مجتمعة مع معدل الضخ (على سبيل المثال، الأسلوب سرعة صبغ الجبهة، في 16) وكذلك مع القياسات التنفس. من الأفضل أخذ هذه القياسات في بداية ونهاية الدورة أخذ العينات. لقياس التنفس، optodes تحت الماء أو الأقطاب الكهربائية الصغيرة هي الافضل.

<table border="0" cellpadding="0" cellspacing="0" width="1096">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">GorillaPod, Original</td>
			<td style="width:191px;">Joby</td>
			<td style="width:167px;">GP000001</td>
			<td style="width:459px;">flexible portable tripod&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Flangeless Ferrule</td>
			<td>IDEX Health &#38; Science&#160;</td>
			<td>P-200X</td>
			<td>1/16&#34; in Blue/pk</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Male Nut</td>
			<td>IDEX Health &#38; Science&#160;</td>
			<td>P-205X&#160;</td>
			<td>1/16&#34; in Green/10pk</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Female to Female Luer</td>
			<td>IDEX Health &#38; Science&#160;</td>
			<td>P-658</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Female-Male Luer</td>
			<td>IDEX Health &#38; Science&#160;</td>
			<td>P-655</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Peek Tubing (250&#181;m ID)</td>
			<td>IDEX Health &#38; Science&#160;</td>
			<td>1531</td>
			<td>1/16&#34; OD x 0.01in ID x 5ft lenght. Alternative ID can be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Two component resin epoxy</td>
			<td>IVEGOR</td>
			<td>9257</td>
			<td>Mix well the two component resin before use</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">(TOC) EPA VIALS</td>
			<td>Cole -Parmer&#160;</td>
			<td>03756-20</td>
			<td>40 ml glass vials. Manifactured also by Thomas Scientific (ref. number 9711F09)&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HDPE VIALS</td>
			<td>Wheaton</td>
			<td>986701 (E78620)</td>
			<td>20 ml high-density polyethylene vials</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vacuette Z no additive</td>
			<td>Greiner bio-one</td>
			<td>455001</td>
			<td>pre-vacuum by the manufacturer&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Septum Sample Bottles</td>
			<td>Thomas Scientific</td>
			<td>1755C01</td>
			<td>250 ml glass bottles&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Septum Cap 1</td>
			<td>Wheaton</td>
			<td>W240844SP (E7865R)</td>
			<td>22-400 for HDPE vials&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Septum Cap 2&#160;</td>
			<td>Wheaton</td>
			<td>W240846 (1078-5553)</td>
			<td>24-400 for glass vials and bottles. Also manufactured by Thermo Scientific National (ref. 03-377-42)</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:19px;">In-line stainless steel Swinney Filter holders</td>
			<td>Pall&#160;</td>
			<td>516-9067</td>
			<td>13mm of diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PTFE Seal Washer</td>
			<td>Pall&#160;</td>
			<td>516-8064</td>
			<td>ring for stainless steel filter holders</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TCLP Glass Filters</td>
			<td>Pall&#160;</td>
			<td>516-9126</td>
			<td>binder-free glass fiber filters, 13mm of diameter,&#160; pore size 0.7&#181;m</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polycarbonate Filter Holders&#160;</td>
			<td>Cole -Parmer&#160;</td>
			<td>17295</td>
			<td>13mm of diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isopore Membrane Filters</td>
			<td>Millipore</td>
			<td>GTTP01300</td>
			<td>13mm of diameter, pore size 0.2 &#181;m&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Contrad 2000 Solution&#160;</td>
			<td>Decon Labs</td>
			<td>E123FH</td>
			<td>highly soluble basic mix of anionic and non-ionic surfactant solution&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile Syringe Filters</td>
			<td>VWR International Eurolab S.L.</td>
			<td>514-0061P</td>
			<td>25mm of diameter , pore size 0.2 &#181;m&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluorescein</td>
			<td>Sigma-Aldrich</td>
			<td>(old ref.28802) 46955-100G&#160;</td>
			<td>100g&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Holdex, disposable,sterile</td>
			<td>Greiner bio-one</td>
			<td>450263</td>
			<td>sterile, single-use tube holder with off-center luer for Vacuette</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile Needles</td>
			<td>IcoGammaPlus</td>
			<td>5160</td>
			<td>0.7mm x 30mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cryovials Nalgene</td>
			<td>Nalgene</td>
			<td>V5007(Cat. No.5000-0020)</td>
			<td>2ml&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cryobox carton&#160;</td>
			<td>Rubilabor</td>
			<td>M-600</td>
			<td>145x145x55mm p/microtube 1.5 ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Orthophosphoric Acid</td>
			<td>Sigma</td>
			<td>79617</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>Sigma</td>
			<td>P6148</td>
			<td>500g</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glutaraldehyde</td>
			<td>Merck</td>
			<td>8,206,031,000</td>
			<td>25%, 1 L</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hand Vacuum Pump&#160;</td>
			<td>B&#252;rkle&#160;</td>
			<td>5620-2181</td>
			<td></td>
		</tr>
	</tbody>
</table>

vacusip, an improved inex method for in situ measurement of particulate and dissolved compounds processed by active suspension feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

تحسين أساليب جمع مياه البحر اختيار قارورة جامع وإجراء التنظيف يجب أن يكون السفن جمع VacuSIP المتوافقة مع الحاجز الذ?...

Watch this Scientific Journal Video about VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders at JoVE.com

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP، وتحسين أسلوب INEX ل<em&gt; في الموقع</em&gt; قياس الجسيمات والمذاب المركبات التي تتم معالجتها بواسطة مغذيات تعليق النشطة

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

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Research

JoVE Journal

Environment

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

10.8K Views.  Centre d’Estudis Avançats de Blanes (CEAB-CSIC). We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

Video: VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female&#39;s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George&#39;s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Fishery-induced changes to exploited crustacean fisheries, such as the American lobster fishery, could potentially influence their reproductive dynamics, leading to a reduction in mating success. This study&#39;s goal was to develop a noninvasive method for ascertaining the mating success of female lobsters that may be physiologically or functionally mature.

والطريقة موسع لل<em&gt; في الوضع الطبيعي</em&gt; تحديد النجاح في التزاوج أنثى الكركند الأمريكية (<em&gt; Homarus americanus</em&gt;)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA الطريقة 1615. قياس الفيروسة المعوية ونوروفيروس حدوثها في المياه عن طريق الثقافة وRT-QPCR. I. جمع عينات الفيروسات

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. II. Total Culturable Virus Assay

A standardized method is required when national studies on virus occurrence in environmental and drinking waters utilize multiple analytical laboratories. The U.S Environmental Protection Agency&#8217;s (USEPA) Method 1615 was developed with the goal of providing such a standard for measuring Enterovirus and Norovirus in these waters. Virus is concentrated from water using an electropositive filter, eluted from the filter surface with beef extract, and then concentrated further using organic flocculation. Herein we present the protocol from Method 1615 for filter elution, secondary concentration, and measurement of total culturable viruses. A portion of the concentrated eluate from each sample is inoculated onto ten replicate flasks of Buffalo Green Monkey kidney cells. The number of flasks demonstrating cytopathic effects is used to quantify the most probable number (MPN) of infectious units per liter. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. Laboratories must meet defined performance standards. Method 1615 was evaluated by examining virus recovery from reagent-grade and ground waters seeded with Sabin poliovirus type 3. Mean poliovirus recoveries with the total culturable assay were 111% in reagent grade water and 58% in groundwaters.

Here we present a procedure to measure total culturable viruses using the Buffalo Green Monkey kidney cell line. The procedure provides a standardized tool for measuring the occurrence of infectious viruses in environmental and drinking waters.

وكالة حماية البيئة الطريقة 1615. قياس الفيروسة المعوية وقوع نوروفيروس في المياه عن طريق الثقافة وRT-QPCR. II. مجموع زروع الفحص الفيروسات

A standardized method is required when national studies on virus occurrence in environmental and drinking waters utilize multiple analytical laboratories. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA الطريقة 1615. قياس الفيروسة المعوية ونوروفيروس التواجد في المياه عن طريق الثقافة وRT-QPCR. الجزء الثالث. كشف الفيروس عن طريق RT-QPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

Spotting Cheetahs: Identifying Individuals by Their Footprints

The cheetah (Acinonyx jubatus) is Africa's most endangered large felid and listed as Vulnerable with a declining population trend by the IUCN1. It ranges widely over sub-Saharan Africa and in parts of the Middle East. Cheetah conservationists face two major challenges, conflict with landowners over the killing of domestic livestock, and concern over range contraction. Understanding of the latter remains particularly poor2. Namibia is believed to support the largest number of cheetahs of any range country, around 30%, but estimates range from 2,9053 to 13,5204. The disparity is likely a result of the different techniques used in monitoring.
Current techniques, including invasive tagging with VHF or satellite/GPS collars, can be costly and unreliable. The footprint identification technique5 is a new tool accessible to both field scientists and also citizens with smartphones, who could potentially augment data collection. The footprint identification technique analyzes digital images of footprints captured according to a standardized protocol. Images are optimized and measured in data visualization software. Measurements of distances, angles, and areas of the footprint images are analyzed using a robust cross-validated pairwise discriminant analysis based on a customized model. The final output is in the form of a Ward's cluster dendrogram. A user-friendly graphic user interface (GUI) allows the user immediate access and clear interpretation of classification results.
The footprint identification technique algorithms are species specific because each species has a unique anatomy. The technique runs in a data visualization software, using its own scripting language (jsl) that can be customized for the footprint anatomy of any species. An initial classification algorithm is built from a training database of footprints from that species, collected from individuals of known identity. An algorithm derived from a cheetah of known identity is then able to classify free-ranging cheetahs of unknown identity. The footprint identification technique predicts individual cheetah identity with an accuracy of &gt;90%.

The cheetah (Acinonyx jubatus) is an iconic, endangered species, but conservation efforts are challenged by habitat shrinkage and conflict with commercial farmers. The footprint identification technique, a robust, accurate and cost-effective image classification system, is a new approach to monitoring cheetahs.

اكتشاف الفهود: الأفراد تحديد من آثارهم

The cheetah (Acinonyx jubatus) is Africa's most endangered large felid and listed as Vulnerable with a declining population trend by the IUCN1. It ranges ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

بورصة القاعية من O<sub&gt; 2</sub&gt;، N<sub&gt; 2</sub&gt; والمغذيات الذائبة عن طريق الأساسية حضانات الصغيرة

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

A Simple Approach to Manipulate Dissolved Oxygen for Animal Behavior Observations

The ability to manipulate dissolved oxygen (DO) in a laboratory setting has significant application to investigate a number of ecological and organismal behavior questions. The protocol described here provides a simple, reproducible, and controlled method to manipulate DO to study behavioral response in aquatic organisms resulting from hypoxic and anoxic conditions. While performing degasification of water with nitrogen is commonly used in laboratory settings, no explicit method for ecological (aquatic) application exists in the literature, and this protocol is the first to describe a protocol to degasify water to observe organismal response. This technique and protocol were developed for direct application for aquatic macroinvertebrates; however, small fish, amphibians, and other aquatic vertebrates could be easily substituted. It allows for easy manipulation of DO levels ranging from 2 mg/L to 11 mg/L with stability for up to a 5 min animal-observation period. Beyond a 5 min observation period water temperatures began to rise, and at 10 min DO levels became too unstable to maintain. The protocol is scalable to the study organism, reproducible, and reliable, allowing for rapid implementation into introductory teaching labs and high-level research applications. The expected results of this technique should relate dissolved oxygen changes to behavioral responses of organisms.

This article describes a simple and reproducible protocol to manipulate dissolved oxygen conditions in a laboratory setting for animal behavior studies. This protocol may be used in both teaching and research laboratory settings to evaluate organismal response of macroinvertebrates, fishes, or amphibians to changes in dissolved oxygen concentration.

مقاربة بسيطة لالتلاعب الأوكسجين المذاب لملاحظات سلوك الحيوان

The ability to manipulate dissolved oxygen (DO) in a laboratory setting has significant application to investigate a number of ecological and organismal ...

Understanding Dissolved Organic Matter Biogeochemistry Through In Situ Nutrient Manipulations in Stream Ecosystems

Dissolved organic matter (DOM) is a highly diverse mixture of molecules providing one of the largest sources of energy and nutrients to stream ecosystems. Yet the in situ study of DOM is difficult as the molecular complexity of the DOM pool cannot be easily reproduced for experimental purposes. Nutrient additions to streams however, have been shown to repeatedly alter the in situ and ambient DOM pool. Here we demonstrate an easily replicable field-based method for manipulating the ambient pool of DOM at the ecosystem scale. During nutrient pulse experiments changes in the concentration of both dissolved organic carbon and dissolved organic nitrogen can be examined across a wide-range of nutrient concentrations. This method allows researchers to examine the controls on the DOM pool and make inferences regarding the role and function that certain fractions of the DOM pool play within ecosystems. We advocate the use of this method as a technique to help develop a deeper understanding of DOM biogeochemistry and how it interacts with nutrients. With further development this method may help elucidate the dynamics of DOM in other ecosystems.

Understanding Dissolved Organic Matter Biogeochemistry Through In Situ Nutrient Manipulations in Stream Ecosystems

Dissolved organic matter provides an important source of energy and nutrients to stream ecosystems. Here we demonstrate a field-based method to manipulate the ambient pool of dissolved organic matter in situ through easily replicable nutrient pulses.

فهم الذائب المواد العضوية الكيمياء الحيوية من خلال<em&gt; في الموقع</em&gt; المغذيات التلاعب في تيار النظم الإيكولوجية

Dissolved organic matter (DOM) is a highly diverse mixture of molecules providing one of the largest sources of energy and nutrients to stream ecosystems. ...

Production and Measurement of Organic Particulate Matter in a Flow Tube Reactor

Organic particulate matter (PM) is increasingly recognized as important to the Earth's climate system as well as public health in urban regions, and the production of synthetic PM for laboratory studies have become a widespread necessity. Herein, experimental protocols demonstrate approaches to produce aerosolized organic PM by &#945;-pinene ozonolysis in a flow tube reactor. Methods are described for measuring the size distributions and morphology of the aerosol particles. The video demonstrates basic operations of the flow tube reactor and related instrumentation. The first part of the video shows the procedure for preparing gas-phase reactants, ozonolysis, and production of organic PM. The second part of the video shows the procedures for determining the properties of the produced particle population. The particle number-diameter distributions show different stages of particle growth, namely condensation, coagulation, or a combination of both, depending on reaction conditions. The particle morphology is characterized by an aerosol particle mass analyzer (APM) and a scanning electron microscope (SEM). The results confirm the existence of non-spherical particles that have grown from coagulation for specific reaction conditions. The experimental results also indicate that the flow tube reactor can be used to study the physical and chemical properties of organic PM for relatively high concentrations and short time frames.

This paper describes the operation procedure for the flow tube reactor and related data collection. It shows the protocols for setting the experiments, recording data and generating the number-diameter distribution as well as the particle mass information, which gives useful information about chemical and physical properties of the organic aerosols.

الإنتاج وقياس الجسيمات العضوية في مفاعل أنبوب التدفق

Organic particulate matter (PM) is increasingly recognized as important to the Earth's climate system as well as public health in urban regions, and the ...

Production and Measurement of Organic Particulate Matter in the Harvard Environmental Chamber

The production and the evolution of atmospheric organic particulate matter (PM) are insufficiently understood for accurate simulations of atmospheric chemistry and climate. The complex production mechanisms and reaction pathways make this a challenging research topic. To address these issues, an environmental chamber, providing enough residence time and close-to-ambient concentrations of precursors for secondary organic materials, is needed. The Harvard Environmental Chamber (HEC) was built to serve this need, simulating the production of gas and particle phase species from volatile organic compounds (VOCs). The HEC has a volume of 4.7 m3 and a mean residence time of 3.4 h under typical operating conditions. It is operated as a completely mixed flow reactor (CMFR), providing the possibility of indefinite steady-state operation across days for sample collection and data analysis. The operation procedures are described in detail in this article. Several types of instrumentation are used to characterize the produced gas and particles. A High-Resolution Time-of-Fight Aerosol Mass Spectrometer (HR-ToF-AMS) is used to characterize particles. A Proton-Transfer-Reaction Mass Spectrometer (PTR-MS) is used for gaseous analysis. Example results are presented to show the use of the environmental chamber in a wide variety of applications related to the physicochemical properties and reaction mechanisms of organic atmospheric particulate matter.

This paper describes operation procedures for the Harvard Environmental Chamber (HEC) and related instrumentation for measuring gaseous and particle species. The environmental chamber is used to produce and study secondary organic species produced from the organic precursors, especially related to atmospheric organic particulate matter.

قياس الجسيمات العضوية في قاعة جامعة هارفارد البيئية والإنتاج

The production and the evolution of atmospheric organic particulate matter (PM) are insufficiently understood for accurate simulations of atmospheric ...