Onların işbirliği çok takdir - Yazarlar olan tekneler biz örnekleme için izin verildi, tüm New England ticari lobstermen teşekkür etmek istiyorum. Buna ek olarak, biz Little Bay Lobster Şirketi (Newington, NH) ve bize kendi tesislerinde örnekleme yapmak izin Champlains Seafood (Point Judith, RI) teşekkür ederim. Aşağıdaki UNH öğrenciler bu çalışma boyunca değerli yardım sağladı: Haley Beyaz, Françoise Morrison, Sarah HAVENER, Audra CHAPUT ve Mayıs grose. Bu proje whw (proje # R/CFR-11) ve JSG bir UNH Deniz Programı Hibe NH SeaGrant hibe tarafından desteklenmiştir.

Kısım A: Alan Örnekleme Tekniği 1.. Kadın Ölçümler <ol><li> Her dişi ıstakoz, karapas uzunluğu (CL) ve kaliperler bir çift kullanarak en yakın 1.0 mm, ikinci karın segmentinin genişliğini ölçmek. NOT: (dişiler yumurta taşıyan) karın genişliği cinsel olgunluk göstergesidir çünkü ikinci karın segmenti ölçülür. Buna ek olarak, (adım 1.2), daha sonra üreme aşaması ile, bu veriler için gerekli dökücü sahne tespit edilmesi avantajlı olabilir. </li><li> Karın pleopods birinde (İstakozlarda standart bir metodolojik protokol) kapalı bir küçük distal bölümünü Klip ve daha sonra bir diseksiyon mikroskobu 9 görüntüleme için temiz deniz suyunda pleopod saklayın. NOT: Bu adım aynı zamanda dişiler yumurtalarını 10 a&#39;ya hazırlanıyor bir göstergesidir pleopods, çimento bezlerinin varlığını ortaya çıkarabilir. </li></ol> 2. İğne Insertion <ol><li> (18, G Tablo 1 e bakınız), şırınga iğnesi kaydedin ~ 45 ° (Şekil 1) ° &#39;de, kuyruk doğru açılı seminal deponun içine 1 cm. Not: önce seminal deponun altındaki sperm en yüksek konsantrasyonu (Şekil 1) ulaşmak için sperm tamponundan itilir gibi dokunulmamış sperm fişli ıstakoz, iğne, sokma esnasında bir direnç karşılayabilir. NOT: fiş varlığı, kendi içinde, ıstakozlar evlendirilen olduğunu güvenilir bir göstergedir. Bazı hayvanlarda hiçbir spermatofor mevcut olabilir, çünkü çok az direnç olabilir, spermatofor henüz bir fiş içine sertleştirilmiş edilmemiştir, ya da daha önce yumurta bir veya daha fazla kavramaları dölleme için kullanılmış olabilir küçüktür. </li></ol><img alt="Şekil 1" fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig1highres.jpg" src="/files/ftp_upload/50498/50498fig1.jpg" width = &quot;600px&quot; /&gt; Şekil 1. İğne ekleme ve ıstakoz seminal kabın görüntüle. Sperm örneği ayıklamak için ıstakoz seminal yuvasına künt uçlu iğne takma. İğnenin açısı ve derinliği de tutarlı bir numune elde etmek için kritiktir. Sağ: disseke ve dikey olarak bölünmüş olan bir seminal deponun göster. Spermatozoa deponun dibinde (derin noktası) seviyesinde yer alan ise sperm fiş, kabın en üst ya da en dış kısmını kaplar. Bu bir derinlikten numune kabını delinerek kan kirlenmiş numunesi neden olabilir dikkat etmek önemlidir. <a href="https://www.jove.com/files/ftp_upload/50498/50498fig1highres.jpg" target="_blank">resmi büyütmek için buraya tıklayın</a> . 3. Sperm Tak Penetran <ol><li> Başlangıçtaki yerleştirme sonrası, (vücut eksenine dik olarak) yaklaşık dikey iğne açısını yükselmek ve çok yavaş çalışmaya başlaraşağı kabın içine. </li><li> Uç kuyruk doğru hafifçe işaret edilmektedir, böylece belirli bir açıda eğimli iğne tutun. Sonunda matris içine nüfuz kadar bu durumda, iğne ucu, tıpanın üst sıkıştırma edilir. </li><li> Direnç kez daha hissedilir kadar yavaş yavaş aşağıya doğru itmek için iğne devam - bu kabın alt ve numune alınır burada. </li></ol> 4. Sperm Kaldırma <ol><li> Haznenin dibinden bir örnek (çoğunlukla katı fiş madde, sıvı, sperm ve diğer sıvı) çıkarın ve 2.0 ml işaretlenmiş bir plastik tüpe örnek yatırmak. </li><li> Soğuk deniz suyu ~ 0.1-0.5 ml yuvasını yıkayın. </li><li> Aynı tüp içerisinde sıvı ve yeri, yaklaşık 0.3 ml çıkarın. </li><li> Buz üzerinde mağaza sperm numuneleri laboratuvarda incelenir kadar. Bölüm B: Lab Sperm Örneklerinin İncelenmesi Not: Bireysel l seminal kapların kaldırılır Örnekler<html> obsters sperm hücreleri mevcut veya yok olmadığını belirlemek için muayene edilmelidir. </li><li> Her plastik tüp, bir kapak kayma ile bir cam slayt üzerinde bir yerde, ve bir bileşik mikroskop ile 100X at görünümünde ~ sıvının 50 ul çıkarın. </li><li> Gibi örnekler Puan: kıt sayıda, ya da hiç sperm (bu çalışmada gerçek sperm sayıları, Şekil 2 ölçmek vermedi). </li></ol><img alt="Şekil 2," fo:content-width="5in" fo:src="/files/ftp_upload/50498/50498fig2highres.jpg" src="/files/ftp_upload/50498/50498fig2.jpg" width="600px" /> Şekil 2. Istakoz sperm örneği görüntüler. 40X büyütmede bir bileşik mikroskop ile bakıldığında, bir dişinin seminal priz alınan ıstakoz sperm örneklerinin Görüntüler. Soldaki resim, diğer (sağ), seyrek bir örneği (ölçek bar = 10 mikron) göstermektedir ise çok sayıda sperm bulunan bir örnek gösterir.. Jpg &quot;target =&quot; _blank &quot;&gt; büyük resmi görebilmek için buraya tıklayın.

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> FAO and Agriculture Organization of the United Nations). Fisheries and Aquaculture Department. , (2012).</li><li>MacDiarmid, A., Butler, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sperm+economy+and+limitation+in+spiny+lobsters.">Sperm economy and limitation in spiny lobsters.</a> Behav. Ecol. Sociobiol. 46, 14-24 (1999).</li><li>Kendall, M. S., Wolcott, D. L., Wolcott, T. G., Hines, A. 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Bull. 201, 204-217 (2001).</li><li>Waddy, S. L., Aiken, D. E., Bauer, R. T., Martin, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mating and insemination in the Ameri can lobster,Homarus americanus. Crustacean sexual behavior. , 126-144 (1990).</li><li>Johnson, K. J., Goldstein, J. S., Watson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+methods+for+determining+the+fertility+status+in+early-stage+American+lobster,+Homarus+americanus,+eggs.">Two methods for determining the fertility status in early-stage American lobster, Homarus americanus, eggs.</a> J. Crust. Biol. 31, 693-700 (2011).</li><li>Aiken, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proecdysis,+setal+development,+and+molt+prediction+in+the+American+lobster+(Homarus).">Proecdysis, setal development, and molt prediction in the American lobster (Homarus).</a> 30, 1337-1344 (1973).</li><li>Aiken, D. E., Waddy, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cement+gland+development,+ovary+maturation,+and+reproductive+cycles+in+the+American+lobster+Homarus+americanus.">Cement gland development, ovary maturation, and reproductive cycles in the American lobster Homarus americanus.</a> J. Crust. Biol. 2, 315-327 (1982).</li><li>Pugh, T. L., Goldstein, J. S., Lavalli, K. L., Clancy, M., Watson, W. 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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproduction+and+embryonic+development.">Reproduction and embryonic development.</a> Biology of the Lobster: Homarus americanus. , 177-216 (1995).</li><li>Cooper, R. A., Uzmann, J. R., Cobb, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ecology of juvenile and adult Homarus. The biology and management of lobsters. 2, 97-142 (1980).</li><li>Estrella, B. T., McKiernan, D. 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Yazarlar çıkar çatışması beyan ederim.

Bu video ve kağıt anahat ve dişi ıstakoz başarıyla evlendirilen varsa belirlemek için bir yöntem ortaya koymaktadır. Bu yaklaşım kullanıcıların çeşitli denizde ıstakoz büyük ölçekli örnekleme için kullanılabilir olması yeterince basittir. Bu yöntem çiftleşme sırasında erkek ikinci dişi ıstakoz geçirilen seminal hazne içinde bir spermatofor varlığını tespit dayanmaktadır. Bu spermatophores kısmen da hazne içinde yatırılır kısa bir süre sonra, bir sperm fiş halinde sertleşme...

<html> Amerikan ıstakoz (Homarus americanus) Kuzey Atlantik (~ $ 390,000,000 üzerinde değerli 2011 yılında 56.000 mt) 1 en verimli balıkçılık biridir. Ancak, vahşi popülasyonlar bu türün üreme dinamikleri konusunda anlayış genel bir eksikliği vardır. Aktif üreme katılan bireylerin sayısı ve boyutları da dahil olmak üzere üreme çıkış daha doğru tahminler, üreten bir stok değerlendirme sürecini iyileştirmek olabilir. Dikenli ıstakoz 2, mavi yengeç 3, kral yengeç 4, taş yengeçler 5 ve kar yengeç: Örneğin, nedeniyle sperm sınırlaması tam üreme potansiyelini elde engellenir kadın dahil olmak üzere birçok ticari istismar deniz kabuklular için bir endişe olarak tespit edilmiştir 6. Genel amaç sperm sınırlama yanı sıra Amerikan ıstakoz balıkçılığın belirli bölgelerinde bir faktör olabilir eğer belirlemektir. nt &quot;&gt; (gözle görülür sağlam, son zamanlarda sıkılır yumurta) ovigerous kadın ıstakoz ile alakasız bir etiketleme çalışması yapılırken İstakozlarda potansiyel bir sperm sınırlama sorununun ilk belirtileri gözlendi. ~ bu hayvanların% 15 düşmüş olduğu belirtilen balıkçıların raporları Recapture onların yumurta kavramaları sadece 1-2 ay sonra. çalışma hipotezi bazı ıstakoz döllenmiş olmasaydı yumurta taşıma ve, sonuç olarak, &#39;düştü&#39; 7., bir sonraki çalışmasının ön veriler gerçeği doğruladı olduğunu olduğunu ıstakoz olacak başarıyla evlendirilen değil ve bu döllenmemiş yumurta, sadece ~ 1 ay 8 uygulansa bile yumurta a&#39;ya. Dolayısıyla, doğal popülasyonlarının döllenmemiş yumurta taşıyan dişilerin gözlem verilen, biz sperm sınırlama katkıda olabilir için kapsamını belirlemek için çalıştı cinsel olgun ıstakozlarına üreme bir submaksimal düzeyde. Bu hedefe ulaşmak için, teknik spermatositlerde algılamak için geliştirilmiştirPhore çiftleşme sırasında erkek tarafından yatırılır. Bu kağıt ve video kadın ıstakoz çiftleşme başarısını tespit etmek için geliştirilen bir invaziv olmayan, basit bir yöntem. Teknik denizde, ya araştırma ya da ticari balıkçı teknelerinde hızlı ve güvenilir şekilde kullanılabilir. Bu numune alma yöntemin ayrıntıları yanı sıra bazı temsili bulgular tekniğinin uygulanmasını göstermek için sunulmuştur.

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a noninvasive method for in situ determination of mating success in female american lobsters (homarus americanus)

Kuzeybatı Atlantik&#39;te en verimli balıkçılık biri olmasına rağmen, çok Amerikan ıstakoz doğal üreme dinamikleri hakkında bilinmemektedir. Sömürülen kabuklu nüfus (yengeç ve ıstakoz) son çalışmalar olgun kadın nedeniyle sperm sınırlaması tam üreme potansiyeline ulaşmak mümkün durumlar vardır olduğunu göstermektedir. Amerikan ıstakoz balıkçılık farklı bölgelerinde bu olasılığı incelemek için, güvenilir ve ağrısız bir yöntem denizde kadın ıstakoz çok sayıda örnekleme için geliştirilmiştir. Bu yöntem, bir sperm fiş varlığını veya yokluğunu belirlemek için ve sperm varlığı açısından incelenebilir bir örnek geri alma dişinin seminal deponun içine bir küt uçlu bir iğne ekleme içerir. Kontrol çalışmalarında bir dizi bu tekniğin güvenilirliğini test etmek için dock ve laboratuarda yapılmıştır. Bu çabalar 294 kadın ıstakoz onaylamak için örnekleme getirdiğini varlığı ofa sperm fiş güvenilir bir kap içinde sperm göstergesidir ve dolayısıyla, çiftleşme oldu. Bu kağıt metodoloji ve örneklenen toplam kadın bir alt elde edilen sonuçları göstermektedir. George Bankası ve Cape Ann, MA örneklemeyi 230 kadın ıstakoz (boyut aralığı = Kabuk uzunluğu olarak 71-145 mm),% 90.3 sperm için pozitiftir. Bazı kadınlarda sperm yokluğu için potansiyel açıklamalar şunlardır: gelişmemişlik (fizyolojik olgunluk eksikliği), yumurtası döllemek için kullanılan sonra sperm fiş dökümünü ve çiftleşme aktivite eksikliği. Anketler kadın ıstakoz çiftleşme başarısını incelemek için bu teknik doğal popülasyonlarının üreme aktivitesini belgelemek için alanda kullanılabilecek güvenilir bir vekil olduğunu göstermektedir.

<html> Burada bildirilen veriler nearshore ve offshore yerlerde hem kökenli kadın ıstakoz bir alt kümesi için vardı. Nearshore grup Cape Ann, Massachusetts yakın kıyı sularında yakalanan ve inceleme için laboratuvara soğutucularda taşınan 44 ıstakoz oluşuyordu. 186 deniz ıstakoz toplam George Bank yakalanan ve New Hampshire&#39;da bir ticari tutma tesiste örneklenmiştir. Sperm numuneleri yukarıda tarif edilen yöntemlere göre tüm hayvanların elde edilmiş ve hemen sperm...

Watch this Scientific Journal Video about A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus at JoVE.com

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

Böyle Amerikan ıstakoz balıkçılık gibi sömürülen kabuklu balıkçılık için balıkçılık kaynaklı değişimler, potansiyel çiftleşme başarısında düşüşe neden üreme dinamiklerini etkileyebilir. Bu çalışmanın amacı, fizyolojik veya işlevsel olgun olabilir dişi ıstakoz çiftleşme başarısını tespit etmek için non-invaziv bir yöntem geliştirmek için oldu.

A Noninvaziv Yöntemi<em&gt; In situ</em&gt; Kadın American Istakoz&#39;ların çiftleşme Başarı (Belirlenmesi<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 ...

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JoVE Journal

Environment

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

10.9K Görüntüleme.  University of New Hampshire.  Böyle Amerikan ıstakoz balıkçılık gibi sömürülen kabuklu balıkçılık için balıkçılık kaynaklı değişimler, potansiyel çiftleşme başarısında düşüşe neden üreme dinamiklerini etkileyebilir. Bu çalışmanın amacı, fizyolojik veya işlevsel olgun olabilir dişi ıstakoz çiftleşme başarısını tespit etmek için non-invaziv bir yöntem geliştirmek için oldu. 

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

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

Kayın Ölçek Böcek Direnç Amerikan kayını (Screen A Tekniği<em&gt; Cryptococcus fagisuga</em&gt; Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

Çevre

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.

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, bir Geliştirilmiş INEX Yöntemi için<em&gt; In Situ</em&gt; Partikül Ölçümü ve Çözünmüş bileşikler Aktif Süspansiyon besleme üniteleri İşlenmiş

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

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide (GFH), with little effort and high reliability. The phosphonate, e.g., nitrilotrimethylphosphonic acid (NTMP), is brought into contact with the GFH in a rotator in a solution buffered by an organic acid (e.g., acetic acid) or Good buffer (e.g., 2-(N-morpholino)ethanesulfonic acid) [MES] and N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid [CAPSO]) in a concentration of 10 mM for a specific time in 50 mL centrifuge tubes. Subsequently, after membrane filtration (0.45 &#181;m pore size), the total phosphorus (total P) concentration is measured using a specifically developed determination method (ISOmini). This method is a modification and simplification of the ISO 6878 method: a 4 mL sample is mixed with H2SO4 and K2S2O8 in a screw cap vial, heated to 148-150 &#176;C for 1 h and then mixed with NaOH, ascorbic acid and acidified molybdate with antimony(III) (final volume of 10 mL) to produce a blue complex. The color intensity, which is linearly proportional to the phosphorus concentration, is measured spectrophotometrically (880 nm). It is demonstrated that the buffer concentration used has no significant effect on the adsorption of phosphonate between pH 4 and 12. The buffers, therefore, do not compete with the phosphonate for adsorption sites. Furthermore, the relatively high concentration of the buffer requires a higher dosage concentration of oxidizing agent (K2S2O8) for digestion than that specified in ISO 6878, which, together with the NaOH dosage, is matched to each buffer. Despite the simplification, the ISOmini method does not lose any of its accuracy compared to the standardized method.

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric hydroxide, with little effort and high reliability. In a buffered solution, the phosphonate is brought into contact with the adsorbent using a rotator and then analyzed via a miniaturized phosphorus determination method.

Fosfonat adsorpsiyon taneli ferrik minyatürize fosfor belirleme yöntemini kullanarak hidroksit üzerine belirlemek için en iyi duruma getirilmiş yordamı

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ocean temperatures and ocean acidification, corals are increasingly under threat. To tackle these challenges, advances in cell and molecular biology have proven to be crucial for diagnosing the health of corals. Modifying some of the techniques commonly used in human medicine could greatly improve researchers&#39; ability to treat and save corals. To address this, a protocol for in situ hybridization used primarily in human medicine and evolutionary developmental biology has been adapted for use in adult corals under stress.
The purpose of this method is to visualize the spatial expression of an RNA probe in adult coral tissue that has been embedded in paraffin and sectioned onto glass slides. This method focuses on removal of the paraffin and rehydration of the sample, pretreatment of the sample to ensure permeability of the sample, pre-hybridization incubation, hybridization of the RNA probe, and visualization of the RNA probe. This is a powerful method when using non-model organisms to discover where specific genes are expressed, and the protocol can be easily adapted for other non-model organisms. However, the method is limited in that it is primarily qualitative, because expression intensity can vary depending on the amount of time used during the visualization step and the concentration of the probe. Furthermore, patience is required, as this protocol can take up to 5 days (and in many cases, longer) depending on the probe being used. Finally, non-specific background staining is common, but this limitation can be overcome.

In Situ Hybridization Techniques for Paraffin-Embedded Adult Coral Samples

The goal of this protocol is to perform in situ hybridization on adult coral samples that have been embedded in paraffin and sectioned onto glass slides. This is a qualitative method used to visualize the spatial expression of an RNA anti-sense probe in paraffin-embedded tissues.

Situ Parafin gömülü yetişkin mercan örnekleri için hibridizasyon teknikleri

Corals are important ocean invertebrates that are critical for overall ocean health as well as human health. However, due to human impacts such as rising ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

Bir döngü-aracılı izotermal amplifikasyon (lamba) tahlil hızlı teşhis edilmesi için kullanılan Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

Implantation of a New Micro Acoustic Tag in Juvenile Pacific Lamprey and American Eel

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology has been identified as a way to obtain more detailed information on movement and behavior across a broader spatial scale than is possible with other known technology. The purpose of this method is to provide a detailed step by step instruction on tag implantation for both lampreys and eel. For laboratory studies using actively migrating juvenile Pacific Lamprey (120-160 mm), we determined that the presence of the tag did not alter the swimming ability between tagged and untagged Individuals or have any significant tag loss (&lt;3%). Similar results were determined during laboratory testing of Yellow phase American Eels (113-175 mm). No mortality occurred during a 38-day holding period and there was minimal tag loss (3.8%). The presence of the tag did not have any significant effect on the swimming ability or survival of tagged eels compared to untagged controls and there was minimal tag loss.

This article illustrates the procedure for implanting a micro acoustic tag in juvenile Pacific Lamprey and American Eel. Our application is based on laboratory trials, which indicate that acoustic tags can be successfully implanted with no apparent effects on swimming ability or survival and minimal tag loss.

Yeni bir mikro akustik etiketi Juvenil Pasifik taşemen ve Amerikan yılan balığı implantasyonu

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology ...

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&ndash;Environment Interactions

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating a knowledge gap. To meet the challenge of improving crops to feed the growing global population, this gap must be bridged.
Physiological traits are considered key functional traits in the context of responsiveness or sensitivity to environmental conditions. Many recently introduced high-throughput (HTP) phenotyping techniques are based on remote sensing or imaging and are capable of directly measuring morphological traits, but measure physiological parameters mainly indirectly.
This paper describes a method for direct physiological phenotyping that has several advantages for the functional phenotyping of plant&#8211;environment interactions. It helps users overcome the many challenges encountered in the use of load-cell gravimetric systems and pot experiments. The suggested techniques will enable users to distinguish between soil weight, plant weight and soil water content, providing a method for the continuous and simultaneous measurement of dynamic soil, plant and atmosphere conditions, alongside the measurement of key physiological traits. This method allows researchers to closely mimic field stress scenarios while taking into consideration the environment&#8217;s effects on the plants&#8217; physiology. This method also minimizes pot effects, which are one of the major problems in pre-field phenotyping. It includes a feed-back fertigation system that enables a truly randomized experimental design at a field-like plant density. This system detects the soil-water-content limiting threshold (&#952;) and allows for the translation of data into knowledge through the use of a real-time analytic tool and an online statistical resource. This method for the rapid and direct measurement of the physiological responses of multiple plants to a dynamic environment has great potential for use in screening for beneficial traits associated with responses to abiotic stress, in the context of pre-field breeding and crop improvement.

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&#8211;Environment Interactions

This high-throughput, telemetric, whole-plant water relations gravimetric phenotyping method enables direct and simultaneous real-time measurements, as well as the analysis of multiple yield-related physiological traits involved in dynamic plant&#8211;environment interactions.

Bitki-Çevre Etkileşimlerinin Gerçek Zamanlı Fizyolojik Fenotimi için Telemetrik, Gravimetrik Platformu

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating ...

Mistletoe Eradicator - A Novel Tool for Simultaneous Mechanical and Chemical Control of Mistletoe

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic and topic associations with various organisms and at the same time acting as biological pirates, exhausting the host trees by stealing water and nutrients via the haustorium. Many crop and timber trees are attacked by mistletoes, for example, dwarf mistletoes parasitize conifers, representatives of genus Tapinanthus target guava, and Phoradendron mistletoe target pecans. Recently, an increasing infestation of the European mistletoe (Viscum album subsp. album L.) in monoculture stands of common walnuts (Juglans regia L.)&#160;was recorded for the first time during 2018 in different regions of Kashmir Valley, India. The common chemical approach based on the spraying of mistletoe exophytes with the growth regulators, ethephon and glyphosate, was not efficient since V. album resprouted in the next vegetation season. Hand pruning of mistletoe was time-consuming, and a polyethylene covering resulted in the localized rotting of host tissues. The most efficient approach to eradicate mistletoe was the use of a novel tool called Mistletoe Eradicator, which allows to use mechanical and chemical means of mistletoe control simultaneously. It consists of a manipulating pole with the terminal pruning saw having a container filled with the growth regulator, from which the liquid slowly drops onto the saw blade through the nozzle. It allows working both from the ground and in the tree canopy. After the targeted application of 5%-10% ethephon or glyphosate to the base of the just pruned V. album stems (stubs), exophyte resprouting was significantly reduced 9 months after the pruning as compared to the only pruned or sprayed exophytes. An alternative variant of Mistletoe Eradicator was constructed in Ukraine filled just with a food color additive to track the liquid dropping and was successfully tested.

The protocol describes an easy-to-handle tool called Mistletoe Eradicator which combines mechanical and chemical approaches and is efficient in the control of mistletoe infestation on the host trees. The manuscript uses the European mistletoe (Viscum album subsp. album L.) as an example.

Ökse Otu Eradikatörü - Ökse Otunun Eşzamanlı Mekanik ve Kimyasal Kontrolü için Yeni Bir Araç

Some species of mistletoes, an aerial plant hemiparasites, play a Janus-like role in ecosystems by being rich biodiversity hotspots due to their trophic ...

In Situ Soil Moisture Sensors in Undisturbed Soils

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture data has been slow due to inconsistent data collection, poor standardization, and typically short record duration. Soil moisture, or quantitatively volumetric soil water content (SWC), is measured using buried, in situ sensors that infer SWC from an electromagnetic response. This signal can vary considerably with local site conditions such as clay content and mineralogy, soil salinity or bulk electrical conductivity, and soil temperature; each of these can have varying impacts depending on the sensor technology.
Furthermore, poor soil contact and sensor degradation can affect the quality of these readings over time. Unlike more traditional environmental sensors, there are no accepted standards, maintenance practices, or quality controls for SWC data. As such, SWC is a challenging measurement for many environmental monitoring networks to implement. Here, we attempt to establish a community-based standard of practice for in situ SWC sensors so that future research and applications have consistent guidance on site selection, sensor installation, data interpretation, and long-term maintenance of monitoring stations.
The videography focuses on a multi-agency consensus of best-practices and recommendations for the installation of in situ SWC sensors. This paper presents an overview of this protocol along with the various steps essential for high-quality and long-term SWC data collection. This protocol will be of use to scientists and engineers hoping to deploy a single station or an entire network.

In Situ Soil Moisture Sensors in Undisturbed Soils

The determination of soil water content is a critical mission requirement for many state and federal agencies. This protocol synthesizes multi-agency efforts to measure soil water content using buried in situ sensors.

Yerinde Bozulmamış Topraklarda Toprak Nemi Sensörleri

Soil moisture directly affects operational hydrology, food security, ecosystem services, and the climate system. However, the adoption of soil moisture ...

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers (mainly phosphorus concentration and light intensity) and species composition on primary productivity. Primary productivity is mainly driven by light intensity, temperature, availability of nutrients, and distribution of the ionic species of the carbonate system in the respective depths of the euphotic zone. It is a complex system that is very difficult to simulate in the laboratory. This cheap, transportable, and easy-to-build floating barge allows measuring the primary productivity accurately-directly under the actual natural conditions. The methodology is based on measuring the primary productivity in real time using noninvasive oxygen sensors integrated into tightly sealed glass jars, enabling online oxygen flux monitoring and providing new insights into metabolic activities. Detailed seasonal in situ measurements of gross primary productivity of microbial mats (or other benthic organisms) can improve current knowledge of the processes controlling primary productivity dynamics in lentic waters.

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Presented here is a cost-effective and transportable method/facility for measuring the primary productivity of microbial mats under actual in situ environmental temperature and light conditions. The experimental setup is based on widely available materials and can be used under various conditions while offering the advantages of laboratory-based models.

Lentik Suların Perifiton Topluluklarının In Situ Birincil Verimliliğini Ölçmenin Düşük Maliyetli Bir Yöntemi

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers ...