野生生物復元法 (W-48-61) 連邦政府の援助からの資源を活用した金融支援、ミシシッピ州野生生物部、水産と公園 (MDWFP) に感謝します。我々 はデータ コレクションの MDWFP 生物学者・ w ・ マッキンリー、A. ブレイロック、A. ゲイリー ・ l ・ ウィルフ広範な関与を感謝します。我々 はまた、施設コーディネーターとして s. タッカーと複数の大学院生、技術者データを収集、彼らの助けのために感謝します。この原稿は、ミシシッピ州立大学森林野生動物研究センターの貢献 WFA427 です。

Ethics Statement: The Mississippi State University Institutional Animal Care and Use Committee approved all capture, handling, and marking techniques under protocols 04-068, 07-036, 10-033 and 13-034.
1. Establish Capture Sites, Immobilize and Transport wild White-tailed Deer
<ol>
	<li>Identify public and private properties that are enrolled in the Deer Management Assistance Program22 and establish &#8805;29 capture sites throughout three source regions in Mississippi, USA.
	<ol>
		<li>Identify several capture locations within each source region to ensure that the range of genetic variation present in the regional population is captured.</li>
		<li>Note: Here, source regions included the Delta, which comprises almost 17% of total land area in Mississippi, USA, and is considered a high-quality soil region with agriculture as the primary land use23,24. All study animals were captured from this region within the distribution of O. v. virginianus25. Other regions included the Thin Loess region (upper and lower Thin Loess combined), which comprises almost 14% of total land area in Mississippi, USA and is considered a medium quality soil region. Agriculture is also a primary land use in the Thin Loess region, though not as prevalent as in the Delta23,24. All study animals were captured from this region within the distribution of O. v. virginianus21. Lastly, the Lower Coastal Plain (LCP) soil region comprises nearly 22% of Mississippi and is classified as a low quality soil region. Primary land uses in the LCP are pine timber production and livestock grazing23,24. The LCP also overlaps the geographical distribution of O. v. osceola; four of the six source populations were within 21 km of this distribution25. This subspecies was described as smaller than O. v. virginianus26.</li>
	</ol>
	</li>
</ol>
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56059/56059fig1.jpg" /> 
Figure 1: Source Populations. Physiographic regions where pregnant dams and fawns were caught in Mississippi, USA. This figure has been modified from reference31. <a href="//cloudfront-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<ol start="2">
	<li>Select potential capture sites that meet the following criteria; habitat characteristics conducive to deer movement, proximity to roads for access, and distribution across the study area. 
	NOTE: Capture sites must allow for concealment of the capture technician.
	<ol>
		<li>Bait sites with about 10 kg of shelled corn to entice deer to visit and evaluate use based on bait consumption and deer photographed by motion-sensitive cameras. Relocate to alternate sites if deer do not attend baited sites within 5-7 days.</li>
	</ol>
	</li>
	<li>During capture events, sit in a concealed &#34;stand.&#34;
	<ol>
		<li>Place stand about 20 m downwind from the bait pile, taking the prevailing wind direction into consideration so that deer approaching the bait are less able to smell the capture technician. 
		NOTE: Elevated stands are strongly preferred, and safety harnesses are required. There are several variations of stands with a variety of commercial sources and use varies by personal preference. For example, a lock-on stand would include seating with a ladder for access attached to a tree with straps. Portable climbing stands can be carried in by the capture technician and allow for increased mobility as the technician can choose a specific tree once they arrive at the capture site. Portable climbing stands are limited to use in straight trees without branches up the chosen height.</li>
	</ol>
	</li>
	<li>Use a dart gun coupled with a 3 cc radio-telemetry dart to deliver a mixture of teletamine HCl (4.4 mg/kg) and xylazine HCl (2.2 mg/kg).
	<ol>
		<li>Schedule capture efforts to coincide with the typical crepuscular activity cycle of deer27. Begin each capture attempt 2-3 h prior to sunset.</li>
		<li>Continue capture events for 2-3 h after sunset using night-vision goggles and a red dot laser for shot placement if deer are not captured during daylight hours.</li>
		<li>Take shots at deer when they are broadside and stationary. 
		NOTE: The hind quarter of the deer is the target because it has significant muscle tissue and is located away from the heart and lungs.</li>
		<li>Wait about 15 min for darted target animals (six-month-old fawns of either sex or pregnant adult females) to become fully immobilized before locating it with directional radio-telemetry equipment.</li>
		<li>Confirm individuals are sedated by checking for eye reflexes (blinking). Then apply ophthalmic ointment to the eyes and blindfold deer to reduce stress. 
		NOTE: Loss of thermoregulation is a consequence of immobilization.</li>
		<li>Use a rectal thermometer to assess body temperature after recovery. Warm deer with heated blankets if the animal&#39;s temperature is below 37.7 &#176;C. Cool deer with ice packs if the animal&#39;s temperature is above 40.0 &#176;C.</li>
		<li>Place deer in a sternal position on a military style gurney and transport deer from the capture location to an enclosed trailer.</li>
		<li>After placing the deer into the trailer, reverse the effects of xylazine HCl with 0.125 mg/kg yohimbine HCl28.</li>
		<li>Transport all captured deer to the desired captive facility (e.g., Mississippi State University Rusty Dawkins Memorial Deer Unit; MSU Deer Unit) and keep them separated by source region.</li>
	</ol>
	</li>
</ol>
2. Captive Facilities and General Husbandry Practices of Research Animals
NOTE: The MSU Deer Unit is subdivided into five 0.4 to 0.8 ha pens.
<ol>
	<li>Cover every side of each pen with shade cloth to act as a visual and physical barrier between pens. Shade cloth helps reduce injuries and provides shade during summer months.</li>
	<li>Place 1-2 elevated box blinds at one end of each pen to facilitate darting events during data collection.</li>
	<li>Place two trough style feeders at separate ends of each pen to reduce competition for food among deer. Also provide a water trough in each pen.</li>
	<li>Provide deer with a high-quality diet (20% crude protein deer pellets) ad libitum. 
	NOTE: Here, additional available forages within pens included (Trifolium spp) and fescue (Festuca spp) along with volunteer grasses and forbs.</li>
	<li>If present, maintain available forages within pens using a mixture of herbicides to control broadleaf weeds and grasses using mixture rates found on respective labels. 
	NOTE: Using off-campus facilities to house &#8805;5.5 month old males will likely be needed. These facilities consisted of two 0.7 ha pens on each of three properties with husbandry practices similar to the MSU Deer Unit.</li>
</ol>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56059/56059fig2.jpg" /> 
Figure 2: Captive Facility Locations. Study area where satellite facilities and the Mississippi State University (MSU) Deer Unit were located. Shaded areas indicate Oktibbeha (A), Noxubee (B), Attala (C), Scott (D), and Copiah (E), counties, Mississippi, USA.This figure has been modified from reference34. <a href="//cloudfront-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
3. Parasite and Disease Control
<ol>
	<li>Monitor research animals for roundworm parasites (Strongyloides spp) using fecal floatation with parasites measured as eggs per gram (EPG).
	<ol>
		<li>If present at high levels, provide parasite control by administering a pelleted wormer (active ingredient fenbendazole) at a rate of about 0.77 kg of pelleted wormer per 22.7 kg of feed during the month of May.</li>
		<li>If parasite levels remain high after initial treatment, use an ivermectin pour-on treatment (5 mg/mL)29, mixed at a rate of 2 mL/0.45 kg and administer to animals at a rate of 0.45 kg of treated feed per 45.4 kg of animal mass. 
		NOTE: Epizootic hemorrhagic disease is sometimes lethal viral disease spread by a biting midge (Culicoides spp) during summer and fall months. If present, treat the research facility with insecticide (5% ultra-low volume insecticide) 2-3 times per week from July 1 to October 1 to decrease prevalence of the vectors among research animals. Spray this insecticide within each pen and around the perimeter of the facility about 90 min before official sunset via fogger. Preferred methods to control for parasites and diseases are unique to each captive facility. Veterinarians must be consulted during any captive wildlife research to ensure animal health and safety.</li>
	</ol>
	</li>
</ol>
4. Data Collection
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56059/56059fig3.jpg" /> 
Figure 3: Data Collection of Newborn Fawns. Measuring hind foot length from a new born fawn at the Mississippi State University Rusty Dawkins Memorial Deer Unit in Oktibbeha County, Mississippi, USA. <a href="//cloudfront-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<ol>
	<li>Search the captive facility daily for fawns during the parturition season.
	<ol>
		<li>Uniquely mark newborn fawns with medium plastic ear tags using an ear tagger with antibiotic applied to the male end of the tag to prevent potential infection. Place ear tag about the center of the fawn&#39;s ear.</li>
		<li>Measure body mass (nearest 0.01 kg) with a digital hanging scale and measure hind foot length and total body length (nearest mm).</li>
		<li>Collect hair samples and send them to a remote site for parentage assignment (see the Table of Materials). 
		NOTE: Parentage assignment was made using DNA based on a proprietary, non-statistical custom structured query language database. In the pairwise allele comparison, the remote parentage assignment site assigned parentage when they excluded all but one sire and one dam based upon a shared allele from each parent at all loci tested (B. G. Cassidy, personal communication). This method of parentage assignment was also used in previous research conducted on captive white-tailed deer30,31.</li>
		<li>Administer 2 cc of Clostridium perfringens types C and D toxoid and Clostridium perfringens types C and D antitoxin subcutaneously and administer 0.3 cc/kg of ivermectin in propylene glycol (Mississippi State University Veterinarian School, Mississippi, USA) orally to each fawn.</li>
	</ol>
	</li>
	<li>Chemically immobilize adult males (&#8805;1.5 years-old) during October and November for data collection.
	<ol>
		<li>Immobilize penned adults using the same combination of teletamine HCl and xylazine HCl used for capture of wild animals (step 1.4).</li>
		<li>During sedation events, walk the technician who will be darting to the end of the pen where the elevated blinds are located. Have a single technician in each of two blinds.</li>
		<li>Have the individual who walked the technician to the blind walk back to the opposite end of the pen. 
		NOTE: Deer move away from these technicians and locate themselves in front of the blinds where technicians are in position to take ethical shots on each deer.</li>
		<li>Take shots at deer when they are broadside and stationary (section 1.4).</li>
		<li>Wait about 15 min for darted animals to become fully immobilized before approaching it.</li>
		<li>Confirm individuals are sedated by checking for eye reflexes (blinking). Apply ophthalmic ointment to the eyes and blindfold deer to reduce stress.</li>
		<li>After the darters successfully sedate an individual deer, monitor the deer&#39;s vital rates.
		<ol>
			<li>Use a rectal thermometer to assess body temperature after recovery. Warm deer with heated blankets if the animal&#39;s temperature is below 37.7 &#176;C. Cool deer with ice packs if the animal&#39;s temperature is above 40.0 &#176;C.</li>
		</ol>
		</li>
		<li>Load the deer on a military-style gurney, and transport it via utility task vehicle to a predetermined data collection area.</li>
		<li>Once transported, record the same morphometric measurements recorded at birth (step 4.1).
		<ol>
			<li>Measure body mass (nearest 0.01 kg) with a digital hanging scale and measure hind foot length and total body length (nearest mm). 
			NOTE: Individual deer react differently to the combination of drugs used during sedation events so administer about 0.1-0.3 cc of the teletamine HCl and xylazine HCl mixture (depending on body mass of an individual deer) if an individual comes out of sedation before data collection is completed.</li>
		</ol>
		</li>
		<li>Administer size-appropriate amounts of antibiotic, ivermectin, a clostidrial vaccine, and a leptospirosis vaccine to all deer after they are transported to the data collection area (see sections 1 and 3).</li>
	</ol>
	</li>
	<li>Take three antler measurements from adult males using an antler measuring tape while the animal is sedated.
	<ol>
		<li>Measure the inside spread (widest distance between main beams), basal circumference (smallest diameter located between the burr and G1 tine), and main beam length (distance from antler base to the tip of the main beam) of antlers prior to antler removal.</li>
		<li>Remove antlers about 3 cm above the burr using a reciprocating saw. Do not remove antlers less than 3 cm.</li>
	</ol>
	</li>
</ol>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56059/56059fig4.jpg" /> 
Figure 4: Data Collection of Adult Males. Antler removal via reciprocating saw from a captive adult male white-tailed deer. <a href="//cloudfront-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<ol start="4">
	<li>After all data is collected from the sedated individual, place the deer into the appropriate pen and administer either 0.125 mg/kg yohimbine HCl28 or 4.0 mg/kg tolazoline HCl32 to reverse the effects of xylazine HCl. Monitor individuals to ensure they remain in a sternal position until they come out of sedation and are fully alert. 
	NOTE: If complications occur and animals must be euthanized, then euthanasia by cerebral dislocation via bolt stunner and severing of the jugular vein are ethical means to dispatch the animal.</li>
	<li>Bring the antlers to a designated area to finish measuring antler size.
	<ol>
		<li>Measure each individual tine protruding from the main beam (G1, G2, G3, etc.) and additional abnormal points by aid of wire. 
		NOTE: Points that do not have a matching counterpart on the opposite main beam or are not consistent with the definition of a typical antler set are defined by the Boone and Crockett Club33.</li>
		<li>Wrap the wire around where the tine intersects the main beam and mark that point for reference.</li>
		<li>Measure from this reference point to the tip of the tine and repeat for each tine.</li>
		<li>Collect remaining circumference measurements by identifying the smallest point between the G1 and G2 tines (H2 circumference), the G2 and G3 tines (H3 circumference), and the G3 and G4 tine if present (H4 circumference).</li>
		<li>If the G4 tine is not present, measure the distance between the midpoint of the G3 tine and the end of the main beam and measure the H4 circumference at the midway point.</li>
		<li>Measure less than four circumferences when antlers contain less than three tines. 
		NOTE: For example, a main beam with two typical points include only three circumference measurements. Individuals may use other guidelines (Safari Club International) for calculating antler size; however, consistent methods must be used for each animal for valid comparison.</li>
		<li>After making all measurements, calculate an antler score similar to the gross nontypical Boone and Crockett score33.</li>
		<li>Weigh antlers to the nearest 0.1 g using a scientific digital scale and assign a minimal critical antler mass of 1 g for first-year animals with antlers shorter than 3 cm.</li>
	</ol>
	</li>
	<li>Chemically immobilize penned juveniles at approximately 5.5 months of age using the same methods for adults (section 4.2) and mark juveniles with a large plastic ear tag (step 4.1.1).
	<ol>
		<li>Use the same drug mixture rates to immobilize captive adults as used for immobilizing wild deer (section 1).</li>
		<li>Collect the same measurements collected at birth (step 4.1.2) and administer the same prophylactics as adults (section 4.2). 
		NOTE: After all data is collected, transport each juvenile male to its randomly assigned satellite facility via trailer.</li>
	</ol>
	</li>
</ol>
5. Producing First- and Second-generation Offspring
<ol>
	<li>Classify six-month-old wild-caught fawns and offspring born at the captive facility from wild-caught mothers as first-generation individuals.</li>
	<li>During the breeding season, place two males with 7-16 females for an average breeding sex ratio of one male to eight females. 
	NOTE: Select breeding males from satellite facilities based on physical appearance, because the healthiest males (largest antlers and body size) are most likely to service females for the entirety of the breeding season without suffering from injury due to the aggressive nature of males during the breeding season.
	<ol>
		<li>Only allow deer to breed with other individuals from the same source region (e.g., Delta males breed Delta females, Thin Loess males breed Thin Loess females, and LCP males breed LCP males).</li>
	</ol>
	</li>
	<li>Classify deer conceived by first-generation parents as second-generation offspring. Raise these individuals in captivity from birth and feed the same high-quality diet as their parents. 
	NOTE: Females may produce offspring multiple years but typically with different sires each year. Collect the same data on second-generation offspring as collected on first-generation and wild-caught individuals.</li>
</ol>

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B., Demarais, S., Strickland, B. K., Wang, G., Dacus, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+nutrition+cues+switch+from+efficiency+to+luxury+phenotypes+for+a+long-lived+ungulate.">Improved nutrition cues switch from efficiency to luxury phenotypes for a long-lived ungulate.</a> Ecol Evol. 6 (20), 7276-7285 (2016).</li><li>Miller, B. F., Muller, L. I., Doherty, T., Osborn, D. A., Miller, K. V., Warren, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+antagonists+for+tiletamine-zolazepam/xylazine+immobilization+in+female+white-tailed+deer.">Effectiveness of antagonists for tiletamine-zolazepam/xylazine immobilization in female white-tailed deer.</a> J Wildl Dis. 40 (3), 533-537 (2004).</li><li>Nesbitt, W. H., Wright, P. L., Buckner, E. L., Byers, C. R., Reneau, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Measuring and scoring North American big game trophies. 3rd edn. , (2009).</li><li>Michel, E. S., Demarais, S., Strickland, B. K., Smith, T., Dacus, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antler+characteristics+are+highly+heritable+but+influenced+by+maternal+factors.">Antler characteristics are highly heritable but influenced by maternal factors.</a> J Wildl Manage. 80 (8), 1420-1426 (2016).</li><li>Severinghaus, C. 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V., Manikkam, M., Savenkova, M. I., Skinner, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+transgenerational+inheritance+of+altered+stress+responses.">Epigenetic transgenerational inheritance of altered stress responses.</a> Proc Natl Acad Sci. 109 (23), 9143-9148 (2012).</li><li>Maher, J. M., Werner, E. E., Denver, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+hormones+mediate+predator-induced+phenotypic+plasticity+in+amphibian+tadpoles.">Stress hormones mediate predator-induced phenotypic plasticity in amphibian tadpoles.</a> Proc R Soc Lond B Biol Sci. 280 (1758), 20123075 (2013).</li></ol>

著者が明らかに何もありません。

我々 のプロトコルに関連付けられているいくつかのステップがあります。ただし、このプロトコルの成功を保障するために取られなければならない 4 つの重要なステップがあります。まず、野生の鹿のキャプチャ中に単一のソース領域 (ステップ 1.1.1) 各地キャプチャする必要があります。複数のキャプチャ場所によってソース領域に関連付けられているすべての遺伝の可変性は鹿の間で表されることになります。第二に、鹿保たれなければならない区切られた震源域で繁殖期 (手順 1.4.9 と 5.2.1)。動物はソースで区切られますを確保する繁殖の際に地域制限、同じ震源域から他の鹿でのみ繁殖する鹿です。各鹿を一意に識別することができるされても重要なステップ正確な年代測定 (ステップ 4.1.1) のことができ 。最後に、栄養はすべて集団 (ステップ 2.4) の定数握られなければなりません。
拘束オジロジカの勉強を評価および/または野生で変わることは困難ではないいくつかの変数を制御することができました。たとえば、老化鹿歯の交換と着用方法35に従ってまたは身体機能36はしばしば困難と精度が低下するとともに年齢37。正確な年齢を知ることは、表現型に影響を与える要因を評価する重要なコンポーネントです。人骨は年齢12で一般に増加するため、異なる高齢動物の間で比較を行うが正しくありません。栄養価はまた野生13,14で変わることができますや目標が栄養源に域内の変動を観察する複数の集団の代償性発育するかどうかを評価するときに特に、一定に保たれる必要があります。年齢と栄養、有効な比較および年齢の相対的な効果についての結論を制御せず、栄養とオジロジカの表現型の遺伝学を確立できません。潜在的な世代別効果に関してより決定的な結果飼育施設でこれらの要因の定数を保持できます。野生の観測の調査、効果だけに栄養制限3,17のプロキシに関連付けられています。したがって、飼育下の研究デザインは、複数の要因が交絡 (例えば栄養、遺伝学、および年齢) 状況で使用するより適切かもしれないこれらの変数は一定に保たれることはできません/または知られていない無料の至るまで人口の実施された試験と比較して。
飼育下の研究に関連するいくつかの利点がある、また制限があります。私たちの設計の 1 つの制限が適切なサンプル サイズを取得します。在庫あり動物野生から直接必要数生産、子孫の総数だけでなくそれらを生産するのにかかる時間の量に影響を与えます。野生の動物をターゲットとしてセックスと妊娠中の女性の 1 歳 6 か月の子鹿を判断しました。のみ 6 ヶ月歳子鹿をターゲットがさらに変動が減少に関する母性効果すべての個人が育てられた捕われの身で同じ持続期間のためクラスを減らしたと思います 1 つだけ年齢をターゲットとして野生の私たちのサンプル サイズの動物をキャッチ、最終的に本研究で使用する母親の数を減らしていると思います。したがって、6 ヶ月齢の子鹿とサンプル サイズを増加する妊娠中の女性をターゲットにしました。さらに、真のコントロールを犠牲にすること言うことが多い飼育下の大型哺乳類を操作する場合の生成サンプル サイズ統計的推論に対して十分な大きさ。理想的には、我々 だろうが飼料をコントロール (栄養の質と量のそれぞれの震源域での国会代表) オジロジカのサブセットに比較のため各震源域から各世代で。しかし、論理的な制約制限のコントロールの使用、我々 としてこうして使用角サイズと本体質量収穫データから収集された、比較私たちの最初の世代の子孫に。完全な比較については、我々 だけの収穫データがあったが角サイズのデータを収穫し、本体質量;測定した他の人骨はよく収穫された個人から収集されません。
捕獲作業に関連付けられた論理的な制約は、またそれが多くの調査官の不可能にみなすことができます。この研究に含まれている個々 の動物の多数は MSU 鹿ユニットの容量を超えたしたがって、衛星設備の家成人男性の使用とし、研究を行った。設備にミシシッピ州、米国など、各サテライト植生の地域変異の可能性もあった、ミシシッピ州、米国13全体自然植生の栄養的な品質の地域変動に類似を発見しました。しかし、我々 はこの潜在的な変化では、自然発生する植生は、シカの高密度のため各施設各ペン内に限られて、我々 の結果が影響を受けることを感じていません。小球形にされた蛋白質は主要な源として鹿の食事を提供しています。すべての施設表示と同様の飼育方法が、MSU 鹿単位ですべての研究動物の住宅であろう理想的です;したがって、衛星設備の使用は、研究設計制限です。また、飼育下の研究は、研究動物の不自然に高い密度を通常表示されます。高密度は、表現型40,41に影響も可能性がある慢性的なストレス38,39を引き起こす可能性があります。しかし、慢性的なストレスの効果場合は、表示されませんでした最初から各人口のための第二世代に増加した人骨として我々 の結果に影響を与える。これは、存在する場合、すべての人口は、慢性的なストレスに反応同様に、示唆しています。
飼育下の研究の過程で、全体で必要がありますいくつかの変更があります。鹿はよく立つペン内の配置に慣れるし、イベントを投げ掛ける中にスタンドの前に移動するには消極的になることがあります。技術者は、(例えば、ユーティリティビークル タスクを介してスタンドにそれらを運転対スタンドにそれらを歩いて) のスタンドにシューティング ゲームと付き合う方法を変える必要があります。各ショットの試行が最大化されるので、正しいブラインド配置とトリミングの木の枝や明確なショットの植え込みすることまた重要作業です。また、出血性疾病などの潜在的な病気の発生する鹿を公開研究施設で人工的に高密度で鹿を維持します。出血性疾病について受けることができますが生じる他の病気は無いかもしれません。獣医師は、病気の問題やそれらに対処するために必要な適切な方法を識別できます。野生動物捕獲作業中に獣医師に相談する必要があります。時々 捕虜の野生動物研究を行う研究者が発生した問題を解決するために固有のメソッドを使用する必要があります。たとえば、施設場所によって出血性疾患などの病気は可能性がありますいくつかの年で普及している、研究動物への影響を減らすために取られるアクションを追加が必要な場合があります。
我々 のプロトコルを操作および様々 な放し飼いの人口を使用して解決困難な哺乳類の生活史に関する栄養の影響に関する質問を評価するために使用できます。生殖栄養品質や数量の違いに妊娠の長さや牛乳生産の変化がどのように関係を評価するなどの複数の側面を含む質問に対処する可能性があります。評価方法栄養制限の優位性などの動作に影響を与えるも対処できます。飼育研究を任意の分類群に適用できる私たちの質問がミシシッピ州、米国からのオジロジカに特有しかし、研究の種に応じて施設改修が必要。

母親の妊娠および授乳期に経験する環境要因は彼女の子孫の表現型は、遺伝子型1,2,3の独立に影響を与えます。質の低い環境に生息する母親が子供を産む効率表現型 (小さな角と体サイズ4) を示すに対し可能性が高い高品質な環境に生息する母親 (大きな枝角と体サイズ4)、高級表現型を示す子孫が生成されます。したがって、高品質の環境で永続化が子孫の生殖機会5,6,7、8に直接影響を与えるし、母の包括適応度に直接な影響を及ぼす可能性のある大規模な表現型特性を持つ男性の子孫を産む母できます。
栄養イチイ (熊の胆、 Ursus arctos9; 全体表現型特性に直接影響がLiasis fuscusI10;都 michahellis11) ミシシッピ州、米国におけるオジロジカの表現型に影響を与える可能性がありますいくつかの要因。角と本体のサイズが12他の人と比較して約 3 分の 1 いくつかの人口のために大きいです。このバリエーションの飼料品質13,14; と相関が強い最も大きい男性は、飼料の最高の品質と地域で発見されます。ただし、ミシシッピ州のオジロジカの歴史的な復興は遺伝的ボトルネックや創始者効果15,16オジロジカの表現型で観測された局所的な値変動のいくつかを説明するかもしれないまた、部分的につながっている可能性があります。
我々 は野生で捕獲されたオジロジカ、男性表現型が遺伝学人口レベルによって制限されているかどうかを評価することができたの栄養の品質管理に使用するプロトコルを提供します。このプロトコルも母性効果を遅れているかどうかを評価することができました私たちの集団に存在していた。私たちの制御設計は栄養制限3,17プロキシとして環境変数の使用に限定されている無料の至るまで人口に調査に優先されます。私たちの制御設計は、潜在的な慢性的なストレス社会に関連する相互作用を似たような住宅と畜産の実践にさらされているすべての個人と一定に保たれるよう、その他の変数もできます。さらに、ため栄養生活史の他の側面に直接影響に至る再生サバイバル18,19栄養の制御により哺乳類の生活史の側面に影響を与える他の変数を評価するために調査官。類似したプロトコルは、(例えば、 20,21) 北アメリカを渡る他の有蹄類の生活史の側面に関連する質問を評価するために記載されています。

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	<tbody>
		<tr height="20">
			<td height="20" style="width:416px;height:20px;">Shelled Corn</td>
			<td style="width:155px;"></td>
			<td style="width:163px;"></td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Elevated Stand</td>
			<td></td>
			<td></td>
			<td></td>
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			<td height="20" style="height:20px;">Safety Harness</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
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			<td height="20" style="height:20px;">Ground Blind</td>
			<td></td>
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		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Model 196 Projector</td>
			<td style="width:316px;">Pneu-Dart, Pennsylvania, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">3cc Radio-Telemetry Darts</td>
			<td style="width:316px;">(Pneu-Dart, Pennsylvania, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Various Sized Darts&#160;</td>
			<td style="width:316px;">(Pneu-Dart, Pennsylvania, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Teletamine HCl&#160;</td>
			<td style="width:316px;">(Telazol, Fort Dodge Animal Health, Iowa, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine HCl&#160;</td>
			<td style="width:316px;">(West Texas Rx Pharmacy, Amarillo, Texas, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Yhoimbine HCl</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tolazoline HCl</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Military Style Gurney</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Rectal Thermometer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Shade Cloth</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20% Crude Protein Deer Pellets&#160;</td>
			<td style="width:380px;">(Purina AntlerMax Professional High Energy Breeder 59UB, Purina, Missouri, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trough Style Feeders</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Commercial Clover&#160;</td>
			<td style="width:380px;">(Durana Clover, Pennington Seed Co., Georgia, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Commercial Fescue&#160;</td>
			<td style="width:380px;">(Max-Q Fescue, Pennington Seed Co., Georgia, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Blankets</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ice Packs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Broadleaf Weed Control (2, 4-DB Herbacide, Butyrac 200)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Grass Control&#160;</td>
			<td style="width:316px;">(Poast Herbacide, BASF Co.)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Pelleted Wormer</td>
			<td>Safeguard Co.,&#160;</td>
			<td></td>
			<td style="width:227px;">active ingredient fenbendazole</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Parasite Pour-on Treatment&#160;</td>
			<td>(Ivomec, Merial Co.)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Insecticide</td>
			<td style="width:316px;">Riptide, McLaughlin Gormley King Co.)&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Medium and Large Plastic Ear Tags&#160;</td>
			<td>(Allflex, Texas, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Remote site that assigned parentage</td>
			<td style="width:380px;">DNA Solutions Animal Solutions Manager (DNA Solutions, Oklahoma, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Digital Hanging Scale&#160;</td>
			<td style="width:316px;">(Moultrie, EBSCO Industries, Inc.)&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tape Measure</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Clostridium Perfringens Types C and D Toxoid Essential 3&#160;</td>
			<td>(Colorado Serum Co.)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Clostridium Perfringens Types C and D Antitoxin Equine Origin</td>
			<td>(Colorado Serum Co.)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ivermectin in propylene glycol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Antibiotic</td>
			<td style="width:380px;">(Nuflor, Schuering-Plough Animal Health Corp., New Jersey, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ivermectin&#160;</td>
			<td style="width:380px;">(Norbrook Labratories, LTD., Down, Northern Ireland, UK)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Clostidrial vaccine</td>
			<td style="width:316px;">(Vision 7 with SPUR, Ivesco LLC, Iowa, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Leptospirosis vaccine&#160;</td>
			<td style="width:316px;">(Leptoferm-5, Pfizer, Inc., New York, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trailer for transport</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Reciprocating saw&#160;</td>
			<td style="width:316px;">(DEWALT, Maryland, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Scientific Digital Scale&#160;</td>
			<td style="width:316px;">(Global Industrail, Global Equipment Company Inc)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Antler Measuring Tape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fogger</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Plastic Ear Tags&#160;</td>
			<td>(Allflex, Texas, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Plastic Ear Tagger</td>
			<td>(Allflex, Texas, USA)</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

protocol for assessing the relative effects of environment and genetics on antler and body growth for a long-lived cervid

シカの表現型は、2 つのカテゴリのいずれかに配置できる: 効率、贅沢な形態成長、高級、大型武器と体の成長を促進する生存を促進します。同じ種の個体数は、環境条件によって各々 の表現型を表示します。男性オジロジカ (ジューテリウムで子鹿) の枝と体サイズ ミシシッピ州、米国の地形の地域によって異なりますが、栄養的な品質の地域変動と相関が強くがネイティブ株式、返品前の努力からの人口レベルの遺伝学の影響は無視できません。このプロトコルでは、年齢や栄養などの表現型に影響を与えるその他の要因が制御されます制御研究の設計方法について説明します。我々 は、ミシシッピ州、米国ミシシッピ州大学さびたドーキンス記念鹿単位に 3 つの地形領域から 1 歳 6 か月フォーンズ野生で捕獲された妊娠した女性をもたらした。子孫の世代別応答と母性効果を評価することの第二世代を生成する同じ地域から鹿を飼育されました。すべての鹿は、同じ高品質 (20% 粗蛋白質鹿ペレット) ダイエット広告自由を食べた。我々 は一意に各新生児をマークし、体の質量、後ろ足の足と全身の長さを記録しました。各後続の秋はリモート注射を介して個人を鎮静し、同じ津雲プラス大人の鹿の角をサンプリングします。最初から角のサイズ (地域的変動はもはや存在しない) の完全な補償と体重 (地域的変化がいくつかの証拠) 第二世代の明らかの部分的な補償の第二世代のサイズにすべて津雲が増加したことがわかった。表示角サイズの 40% の増加、および野生の収穫された相手に比較すると体の質量の 25% の増加について私たちの最も貧しい品質土壌地域に由来する 2 番目の世代の男性。ワイルド男性オジロジカ ミシシッピ州での表現型変異はより人口レベルの遺伝学より栄養価の違いに関連が示唆されました。

個々 の年齢、栄養的な品質、および遺伝学男性オジロジカの表現型に影響を与えます。私たち研究デザインができました栄養鹿の品質を制御するために使用していたし、年クラス内で有効な比較の各鹿の年齢を識別することができました。栄養と私たちの研究デザインと年齢を制御してができますより人口レベルの遺伝学は 2 つの学習集団から男性の表現型が制限されたかどうかを理解することができます。改善された栄養 3.5 歳男性オジロジカ世代変数 (テーブル 1の重要な効果に代表される各ソース領域からのすべての地形計測に肯定的な効果を持っていた図 5および6)。アントラー質量とサイズに改良された栄養は、(図 6)、角サイズの人口レベルの遺伝によって制限されていないことを示唆しているの二世代後 3 つの震源域の間で変わらないでした。本体質量大幅に増加最初から第二世代に野生のすべての集団の栄養制限があることを示唆しかし、第 2 世代の男性 (図 5) の間でまだ地域差があった。したがって、私たちはミシシッピ州のオジロジカの究極体の質量を制限する人口レベルの遺伝学の可能性を排除できません。
栄養と男性オジロジカの年齢を制御することにより、いくつかの形態が他のものより優先は明らかに成長の階層も識別されます。たとえば、アントラー質量がすべての地域 (表 2) から最初に 2 番目の世代の最大の増加を表示します。角のサイズには、ボディの質量と最後に骨格のメトリックに続いて、次の最大の増加が表示されます。この成長階層では、骨格のメトリックが高い運河をつけたが、少なくとも栄養の変化に対応する可能性が示唆しています。さらに、このパターンは男性オジロジカ、津雲と栄養量と質の向上を目指して管理体制に最初応答どれがマネージャーに通知の 3 つの集団で一貫しています。完全な結果は報告とや参照31で説明。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56059/56059fig5.jpg"> 
図 5: 大人の男性の体のメトリックの世代が増加。キャプティブ 3.5 歳男性オジロジカ Noxubee、Attala、Copiah、スコット郡、ミシシッピー、米国で建物の本体質量、総ボディ長さ (TBL) と後部のフィート長さ (HFL) の中央の世代の改善。Y 軸上の破線はミシシッピ州、米国から収集された収穫データの平均体重を表し、最初と 2 番目の世代と比較するため使用されます。ブラック ダイヤモンドは、予測の平均を表します。ひげは最小値を示し、白丸の最大値は外れ値を示します。この図は、参照31から変更されています。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig5large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56059/56059fig6.jpg"> 
図 6: 大人の男性の角特性の世代が増加。キャプティブ 3.5 歳男性オジロジカ Noxubee、Attala、Copiah、スコット郡、ミシシッピー、米国で建物の中央枝特性世代別改善。Y 軸上の破線は、ミシシッピ州、米国から収集された収穫データの角の平均スコアを表します、最初と 2 番目の世代と比較するために使用されます。ブラック ダイヤモンドは、予測の平均を表します。ひげは最小値を示し、白丸の最大値は外れ値を示します。この図は、参照31から変更されています。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig6large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Table 1" src="/files/ftp_upload/56059/56059table1.jpg"> 
表 1: 各モデル パラメーターの効果: MCMCglmm モデル表現型特性の生成 (F2)、年齢と地域 (regionLoess、regionLCP) の影響を記述します。我々 は、カテゴリカルな変数と連続変数として年齢世代と地域をコーディングしました。切片は、第一世代 (F1)、1 歳デルタ男性を表し、世代・年齢・地域土壌ソース人口の比較のための参照用語と見なされます。この表は、参照31から変更されています。
<img alt="Table 2" src="/files/ftp_upload/56059/56059table2.jpg"> 
表 2: 成長の優先順位の階層: パーセント増加キャプティブ 3.5 歳男性オジロジカ Noxubee、Attala、Copiah、スコット郡、ミシシッピー、米国で収容の第 2 世代に最初から人骨。この表は、refefence31から変更されています。

Watch this Scientific Journal Video about Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid at JoVE.com

長命のシカ科動物の角と体の成長に対する環境と遺伝学の相対的な影響を評価するためのプロトコル |環境 |ビデオ

Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

アントラーとシカの長命の体の成長の遺伝と環境の相対的な効果の評価のためのプロトコル

人口レベルの遺伝学や栄養物; に関連するシカの集団間の表現型の違い肥えた野生では困難であります。このプロトコルでは、栄養の変化が除去された制御研究の設計方法について説明します。男性オジロジカの表現型変異が遺伝学よりより栄養によって限られたがわかった。

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which ...

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7.3K ビュー.  Mississippi State University. 人口レベルの遺伝学や栄養物; に関連するシカの集団間の表現型の違い肥えた野生では困難であります。このプロトコルでは、栄養の変化が除去された制御研究の設計方法について説明します。男性オジロジカの表現型変異が遺伝学よりより栄養によって限られたがわかった。

ビデオ: Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

トウモロコシとテオシンテ線における ウスチラゴ の病原性を評価するための迅速かつ効率的な方法

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

環境学

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of natural proteins have a molecular mass between 40 - 200 kDa1,2, a robust and high-throughput method with a nanometer resolution capability is needed. Negative staining (NS) electron microscopy (EM) is an easy, rapid, and qualitative approach which has frequently been used in research laboratories to examine protein structure and protein-protein interactions. Unfortunately, conventional NS protocols often generate structural artifacts on proteins, especially with lipoproteins that usually form presenting rouleaux artifacts. By using images of lipoproteins from cryo-electron microscopy (cryo-EM) as a standard, the key parameters in NS specimen preparation conditions were recently screened and reported as the optimized NS protocol (OpNS), a modified conventional NS protocol 3 . Artifacts like rouleaux can be greatly limited by OpNS, additionally providing high contrast along with reasonably high&#8208;resolution (near 1 nm) images of small and asymmetric proteins. These high-resolution and high contrast images are even favorable for an individual protein (a single object, no average) 3D reconstruction, such as a 160 kDa antibody, through the method of electron tomography4,5. Moreover, OpNS can be a high&#8208;throughput tool to examine hundreds of samples of small proteins. For example, the previously published mechanism of 53 kDa cholesteryl ester transfer protein (CETP) involved the screening and imaging of hundreds of samples 6. Considering cryo-EM rarely successfully images proteins less than 200 kDa has yet to publish any study involving screening over one hundred sample conditions, it is fair to call OpNS a high-throughput method for studying small proteins. Hopefully the OpNS protocol presented here can be a useful tool to push the boundaries of EM and accelerate EM studies into small protein structure, dynamics and mechanisms.

More than half of proteins are small proteins (molecular mass &#60;200 kDa) that are challenging for both electron microscope imaging and three-dimensional reconstructions. Optimized negative staining is a robust and high-throughput protocol to obtain high contrast and relatively high resolution (~1 nm) images of small asymmetric proteins or complexes under different physiological conditions.

最適化された負の染色：電子顕微鏡による小型·非対称蛋白質構造を調べるためのハイスループットプロトコル

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

CO<sub&gt; 2</subテスト用のCO&gt;濃度勾配施設<sub&gt; 2</sub&gt;濃縮および草地生態系の機能に及ぼす土壌の影響

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

A Protocol for Collecting and Constructing Soil Core Lysimeters

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the landscape has complex and varied soil morphology, an accompanying disparity in flow paths for leachate through the soil macropore and matrix structure is present. The rate of flow through these paths is further affected by antecedent soil moisture. Lysimeters are used to quantify flow rate, volume of water and concentration of nutrients leaching downward through soils. While many lysimeter designs exist, accurately determining the volume of water and mass balance of nutrients is best accomplished with bounded lysimeters that leave the natural soil structure intact. 
Here we present a detailed method for the extraction and construction of soil core lysimeters equipped with soil moisture sensors at 5 cm and 25 cm depths. Lysimeters from four different Coastal Plain soils (Bojac, Evesboro, Quindocqua and Sassafras) were collected on the Delmarva Peninsula and moved to an indoor climate controlled facility. Soils were irrigated once weekly with the equivalent of 2 cm of rainfall to draw down soil nitrate-N concentrations. At the end of the draw down period, poultry litter was applied (162 kg TN ha-1) and leaching was resumed for an additional five weeks. Total recovery of applied irrigation water varied from 71% to 85%. Nitrate-N concentration varied over the course of the study from an average of 27.1 mg L-1 before litter application to 40.3 mg L-1 following litter application. While greatest flux of nutrients was measured in soils dominated by coarse sand (Sassafras) the greatest immediate flux occurred from the finest textured soil with pronounced macropore development (Quindocqua).

A detailed method for extraction and assembly of intact soil core lysimeters and their use for study of leachate and associated loss of nutrients from surface applied poultry litter is demonstrated.

土壌コアLysimetersを収集し、構築するためのプロトコル

Leaching of nutrients from land applied fertilizers and manure used in agriculture can lead to accelerated eutrophication of surface water. Because the ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

水生生物に浮遊堆積物の影響を評価するためのフロースルー露光装置

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

海面とサンプル分析にMicroplasticsサンプリングのためのプロトコル

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several limitations that have resulted in the exclusion of many species. Advances in the field over the last two decades have led occupational health researchers to turn to molecular-based approaches for identifying fungal hazards. These methods have resulted in the detection of many species within indoor and occupational environments that have not been detected using traditional methods. This protocol details an approach for determining fungal diversity within air samples through genomic DNA extraction, amplification, sequencing, and taxonomic identification of fungal internal transcribed spacer (ITS) regions. ITS sequencing results in the detection of many fungal species that are either not detected or difficult to identify to species level using culture or microscopy. While these methods do not provide quantitative measures of fungal burden, they offer a new approach to hazard identification and can be used to determine overall species richness and diversity within an occupational environment.

Determining the fungal diversity within an environment is a method utilized in occupational health studies to identify health hazards. This protocol describes DNA extraction from occupational air samples for amplification and sequencing of fungal ITS regions. This approach detects many fungal species that can be overlooked by traditional assessment methods.

コレクションと真菌 DNA の分析のための産業大気サンプルの抽出

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at capturing and quantifying the dynamics and impact of this invasive insect pest as well as the development of its host plant across a heterogeneous landscape. The cornerstone of this comprehensive monitoring system is timely georeferenced data collection on CBB movement, coffee berry infestation, mortality by the fungus Beauveria bassiana, and coffee plant phenology via a mobile electronic data recording application. This electronic data collection system allows field records to be georeferenced through built-in global positioning systems, and is backed by a network of weather stations and records of farm management practices. Comprehensive monitoring of CBB and host plant dynamics is an essential part of an area-wide project in Hawaii to aggregate landscape-level data for research to improve management practices. Coffee agroecosystems in other parts of the world that experience highly variable environmental and socioeconomic factors will also benefit from implementing this protocol, in that it will drive the development of customized integrated pest management (IPM) to manage CBB populations.

Comprehensive monitoring of coffee berry borer and host plant dynamics is essential for aggregating landscape-level data to improve management of this invasive pest. Here, we present a protocol for scientific monitoring of coffee berry borer movement, infestation, mortality, coffee plant phenology, weather, and farm management via a mobile electronic data recording application.

コーヒーのベリーの穴あけ器の科学的なモニタリングのためのハワイ プロトコル: コーヒー生態系世界のモデル

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...

Assessing the Particulate Matter Removal Abilities of Tree Leaves

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on collecting various sized particulate matter (PM) retained on leaf surfaces. We further characterized the retention efficiency of leaves to various sized PM, which will help to assess the abilities of urban trees to remove PM from ambient air quantitatively. 
Taking three broadleaf tree species (Ginkgo biloba, Sophora japonica, and Salix babylonica) and two needleleaf tree species (Pinus tabuliformis and Sabina chinensis) as the research objects, leaf samples were collected 4 days (short PM retention period) and 14 days (long PM retention period) after the latest rainfall. PM retained on the leaf surfaces was collected by means of WC, BC, and UC in sequence. Then, retention efficiencies of leaves (AEleaf) to three types of the various sized PM, including easily removable PM (ERP), difficult-to-remove PM (DRP), and totally removable PM (TRP), were calculated. Only around 23%-45% of the total PM retained on leaves could be cleaned off and collected by WC. When the leaves were cleaned through WC+BC, the underestimation of the PM retention capacity of different tree species was in the range of 29%-46% for various sized PM. Almost all PM retained on leaves could be removed if UC was supplemented to WC+BC. 
In conclusion, if the UC was complemented after the conventional cleaning methods, more PM on leaf surfaces could be eluted and collected. The procedure developed in this study can be used for assessing the PM removal abilities of different tree species.

The ultrasonic cleaning method was applied to elute the particulate matter (PM) retained on leaf surfaces after PM was eluted by the conventional cleaning methods (water cleaning only or water cleaning plus brush cleaning). The methodology can help to improve the estimation accuracy for PM retention capacity of leaves.

葉の木の粒子状物質の除去能力を評価します。

Based on the conventional cleaning methods (water cleaning (WC) + brush cleaning (BC)), this study evaluated the influence of ultrasonic cleaning (UC) on ...

An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations

Organic-mineral interactions are widely occurring in hydrothermal environments, such as hot springs, geysers on land, and the hydrothermal vents in the deep ocean. Roles of minerals are critical in many hydrothermal organic geochemical processes. Traditional hydrothermal methodology, which includes using reactors made of gold, titanium, platinum, or stainless-steel, is usually associated with the high cost or undesired metal catalytic effects. Recently, there is a growing tendency for using the cost-effective and inert quartz or fused silica glass tubes in hydrothermal experiments. Here, we provide a protocol for carrying out organic-mineral hydrothermal experiments in silica tubes, and we describe the essential steps in the sample preparation, experimental setup, products separation, and quantitative analysis. We also demonstrate an experiment using a model organic compound, nitrobenzene, to show the effect of an iron-containing mineral, magnetite, on its degradation under a specific hydrothermal condition. This technique can be applied to study complex organic-mineral hydrothermal interactions in a relatively simple laboratory system.

Earth-abundant minerals play important roles in the natural hydrothermal systems. Here, we describe a reliable and cost-effective method for the experimental investigation of organic-mineral interactions under hydrothermal conditions.

有機水熱変換のミネラル効果を研究するための実験プロトコル

Organic-mineral interactions are widely occurring in hydrothermal environments, such as hot springs, geysers on land, and the hydrothermal vents in the ...