我们感谢密西西比州野生动物部门、 渔业和公园 (MDWFP) 在野生动物恢复法 (W-48-61) 中使用从联邦援助资源的金融支持。我们感谢 MDWFP 生物学家 · 麦金利、 A.Blaylock、 A.Gary L.维尔夫为它们广泛参与了数据集合中。我们还感谢美国塔克作为设施协调员和多个研究生和技术人员对他们的帮助下收集数据。这份手稿是密西西比州大学森林和野生动物研究中心贡献 WFA427。

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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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. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tooth+development+and+wear+as+criteria+of+age+in+white-tailed+deer.">Tooth development and wear as criteria of age in white-tailed deer.</a> J Wildl Manage. 13 (2), 195-216 (1949).</li><li>Gee, K. L., Webb, S. L., Holman, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+and+implications+of+visually+estimating+age+of+male+white-tailed+deer+using+physical+characteristics+from+photographs.">Accuracy and implications of visually estimating age of male white-tailed deer using physical characteristics from photographs.</a> Wild Soc Bull. 38, 96-102 (2014).</li><li>Storm, D. J., Samuel, M. D., Rolley, R. E., Beissel, T., Richards, B. J., Van Deelen, T. 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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>

作者没有透露。

有几个步骤与我们的协议;然而，有必须采取确保与本议定书的成功的四个关键步骤。第一，期间捕获的野鹿，必须有几个捕获地点 （步骤 1.1.1） 单一来源地区。有多个捕获位置可确保鹿表示任何与源区域关联的遗传变异性。第二，鹿必须保持分离的源区在繁殖季节 （1.4.9 和 5.2.1 的步骤）。确保动物被隔开源区域在繁殖期间限制鹿育种只有与其他鹿从他们相同的源区。正在能够唯一地标识每个鹿也是关键的一步，因为这样做允许准确年龄测定 （步骤 4.1.1）。最后，营养必须保持恒定的所有人口 （步骤 2.4）。
研究圈养白尾鹿使我们能够控制的否则难以评估和/或在野外会发生变化的几个变量。例如，老化鹿牙更换和磨损方法35或身体功能36往往是困难的和准确性随年龄37。知道确切的年龄是一个关键的组件时评估表型影响的因素。因为合肥工业大学一般随年龄12，制作不同老年动物之间的比较是无效的。营养质量还可以改变在野生13，14，和必须保持恒定不变，特别是当目标是评估是否补偿生长观察变化在营养源区域内的多个种群间发生。无需控制年龄和营养、 有效的比较和年龄的相对影响的结论，不能作营养和白尾鹿表型遗传学。持有这些因素保持不变在圈养设施还允许更具决定性的结果，关于世代的潜在影响。在野外进行的观测研究，效果也只能与营养限制3，17代理关联。因此，圈养研究设计可能是更适合应用于多因素干扰 （例如，营养、 遗传学和年龄） 的情况相比，开展免费测距人口这些变量在哪里不能保持恒定不变和/或不详的研究。
尽管有几个好处与圈养研究相关联，也有局限性。我们设计的一个限制获得足够的样本。股票直接需要从野外的动物的数量不仅影响的后代产生，总数也交出他们所需的时间量。我们视为目标动物在野外的六个月岁小鹿的性别和怀孕的女性。虽然只针对六个月岁小鹿可能有进一步减少的变异与母体效应作为所有个人会有被都笼养的同一时间，针对只有一个年龄班将会减少我们样本大小的野外捕获的动物，亦会最终减少母亲在我们的研究中使用的人数。因此，我们选择了以六个月岁小鹿和怀孕的女性增加样本量为目标。此外，生成样本足够大的统计推断与圈养的大型哺乳动物工作往往就意味着牺牲真正的控件时。理想情况下，我们会有美联储控制饮食 （饮食代表在各自的源地区发现的营养质量） 子集的白尾鹿从比较每个源区域在每一代人。然而，后勤方面的限制限制控制，我们使用，因此我们使用鹿茸大小和身体大量收集从收获数据作为比较给我们的第一代后代。虽然收获数据更全面的比较，我们只允许有收获鹿茸大小的数据及体质量;不会从收获个人通常收集我们测量其他补益。
后勤方面与俘虏工作相关联的限制还可能认为不可行，许多调查员。大量的包含在本研究中的动物个体超过容量的 MSU 鹿单位;因此，我们进行这项研究用卫星设施到房子成年男性。设施被横跨密西西比州，美国，因此，每个卫星设施内的植被区域变化的潜力，类似于中营养品质的天然植被区域的变化发现横跨密西西比河，美国13。但是，我们不觉得这个潜在的变化影响我们的结果，因为自然发生的植被是有限内每一笔每一处设施由于高鹿密度。丸粒化的蛋白鹿的饮食担任的主要来源。虽然所有设施都显示类似的饲养方法，房屋在 MSU 鹿单位的所有研究动物本来理想;因此，使用卫星设施是我们的研究设计的局限性。此外，圈养的研究一般显示高得不正常密度的研究的动物。高的密度可能诱发慢性应激38，39 ，还可能影响表型40，41。然而，任何慢性应激的影响，如果存在，并不影响我们的结果作为补益从第一次增加到每个人口的二代。这表明，是否存在，所有人口的都反应同样对慢性应激。
在整个过程中俘虏的研究，有可能需要的几个修改。鹿经常成为习惯站在笔中的位置，并可能成为愿意在站期间窜事件面前移动。技术人员必须改变他们陪射手到看台上 （例如，他们走到看台上与通过实用任务车辆驾驶他们向看台） 的方式。正确的盲安置和修剪树枝和灌木，用于拍摄清晰的镜头也是关键所以每次尝试投篮最大化。此外，保持鹿人工高密度在研究设施暴露潜在的疾病暴发流行性出血性疾病如鹿。虽然可以接种流行性出血性疾病，可能不会出现其他疾病。兽医可以帮助确定疾病问题和解决他们所需的适当方法。必须在野生动物圈养工作咨询兽医。有时进行圈养野生动物研究要求研究人员用独特的方法来解决问题，因为他们出现。例如，根据设施的位置，疾病，如流行性出血性疾病可能普遍在某些年份和可能需要额外为减少其对研究动物的影响而采取的行动。
我们的协议可以操纵，用来评估各种问题关于影响营养对哺乳动物的生活史，很难解决使用自由放养的人群。可能处理的问题涉及多个方面的再生产等评估妊娠期长度或牛奶生产中的变化如何相关的营养质量和 （或） 数量的差异。评估如何营养限制影响，优势度的行为也可以解决。虽然我们的问题是特定于白尾鹿从密西西比州，美国，圈养的研究可以应用于任何类群;然而，将根据研究物种需要设施修改。

一位母亲在妊娠期和哺乳经验的环境因素可能会影响她的后代的表型，基因型1，，23的独立。母亲居住高质量环境可能会产生的后代表现出豪华表型 (大鹿角和身体大小4)，而母亲居住在低质量环境可能产生的后代表现出效率 (小鹿角和身体大小4) 表型。因此，坚持高质量的环境可能让母亲产下的后代以大的表型特征，可能直接影响到后代的生殖机会5，，67，8和间接影响母亲的包容性健身。
虽然营养直接影响表型特征跨分类群 （美洲黑熊，熊9;Liasis fuscusI10;黑 michahellis11)，几个因素可能会影响白尾鹿表型在密西西比州，美国。鹿茸和体型都大为一些人口约三分之一的人比较12。这种变化强烈关联与牧草质量13，14;最大的男性发现在区域与最优质的饲料。然而，白尾鹿在密西西比州的历史恢复努力可能导致遗传瓶颈和 （或） 创始人影响15，16，这也部分解释一些中白尾鹿表型观察区域的变化。
我们提供我们用来控制营养品质的野生白尾鹿，使我们能够评估是否男性表型受人口水平遗传学的协议。此协议还允许我们评估是否落后母体效应在我们人口在场。我们控制的设计是优惠到土著居民免费测距仅限于为营养限制3，17作为代理使用环境变量进行研究。我们控制的设计还允许其他变量如潜力慢性压力有关社会的相互作用，如所有个人都遭受相似的住房和畜牧业做法保持恒定不变。此外，因为营养直接影响其他生活史方面从繁殖到生存18，19，控制营养允许调查人员评估影响哺乳动物生活史方面的其他变量。介绍了类似的协议，以评估 （例如， 20，21） 在北美与其他有蹄类动物的生活史方面有关的问题。

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			<td height="20" style="width:416px;height:20px;">Shelled Corn</td>
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			<td height="20" style="height:20px;">Elevated Stand</td>
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			<td height="20" style="height:20px;">Safety Harness</td>
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			<td height="20" style="height:20px;">Ground Blind</td>
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			<td height="20" style="height:20px;">Model 196 Projector</td>
			<td style="width:316px;">Pneu-Dart, Pennsylvania, USA</td>
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			<td height="20" style="height:20px;">3cc Radio-Telemetry Darts</td>
			<td style="width:316px;">(Pneu-Dart, Pennsylvania, USA)</td>
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			<td height="20" style="height:20px;">Various Sized Darts&#160;</td>
			<td style="width:316px;">(Pneu-Dart, Pennsylvania, USA)</td>
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			<td height="20" style="height:20px;">Teletamine HCl&#160;</td>
			<td style="width:316px;">(Telazol, Fort Dodge Animal Health, Iowa, USA)</td>
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			<td height="20" style="height:20px;">Xylazine HCl&#160;</td>
			<td style="width:316px;">(West Texas Rx Pharmacy, Amarillo, Texas, USA)</td>
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			<td height="20" style="height:20px;">Yhoimbine HCl</td>
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			<td height="20" style="height:20px;">Tolazoline HCl</td>
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			<td height="20" style="height:20px;">Military Style Gurney</td>
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		</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

鹿表型可以放入两个类别之一: 效率，促进生存在奢侈的形态生长和促进增长的大型武器装备和身体大小的奢侈品。同一物种的种群显示每个表型取决于环境条件。虽然鹿茸和身体大小的雄性白尾鹿 （Odocoileus 鹑） 因在密西西比州，美国的地理区域而异，与营养品质的区域变化密切相关，不能无视人口水平遗传学从本机股票和以前重新进货努力的影响。此协议说明我们如何设计在哪里控制影响表型，如年龄和营养，其他因素的对照的研究。我们从三个明显的地理区域，在密西西比州，美国密西西比州大学生锈道金斯纪念鹿单位到带野外捕捉的怀孕雌兽和六个月岁幼鹿。鹿从同一区域是被培养来生产一二代的后代，使我们能够评估代反应及孕产妇的影响。所有的鹿吃了相同的高质量 （20%粗蛋白鹿丸） 饮食随意。我们唯一标记每个新生儿，并记录身体质量，后腿脚和身体总长度。每个后续的秋天，我们个人通过远程注射镇静剂和采样的同一补益，加上成年人的鹿角。我们发现所有合肥工业大学都增加的大小从第一到二代，与全额赔偿的鹿茸大小 （区域变化不再存在） 和体重 （区域变化一些证据） 明显在二代的部分补偿。二代源于我们贫穷的质量的男性土壤区域显示有关鹿茸大小增加了 40%和体重时与他们野生的收获相比增加了 25%。我们的结果表明，表型变异的野生雄性白尾鹿在密西西比州更涉及营养质量的差异，比人口水平遗传学。

个人年龄、 营养品质和遗传学影响雄性白尾鹿表型。我们允许我们的研究设计的营养鹿质量控制消耗和使我们能够识别每个鹿进行有效比较内年班的年龄。通过控制营养和年龄与我们的研究设计，我们得以更好地理解人口水平遗传学是否限制了从两个研究群体表型的男性。改善营养状况对 3.5 岁雄性白尾鹿从每个源区域作为代表由我们代变量 (表 1; 显著疗效有积极影响所有形态测量图5和图6）。经过两代的改善营养状况，(图 6)，这表明鹿茸大小不受人口水平遗传学，鹿茸质量和大小没有不异三个源区域。身体体积大大增加从第一到二代，建议为所有人群; 野生的营养限制然而，有人仍然区域变化二代男性 (图 5)。因此，我们不能排除限制最终体质量的白尾鹿在密西西比州的人口水平遗传学的可能性。
通过控制营养和年龄为男性白尾鹿，我们还确定增长层次结构，揭示了一些补益优先于其他人。例如，鹿茸质量显示的最大增幅从第一到第二代在所有地区 (表 2)。鹿茸大小显示下一增长最快，其次是身体质量和最后骨骼度量。这种增长层次结构表明骨骼肌度量高度开凿运河和最不可能在营养中的变化作出反应。此外，这种模式是一致的三个群体的雄性白尾鹿，通知经理何种补益会回应第一次旨在改善营养的数量和质量的管理制度。全部结果进行报告和讨论在参考31。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56059/56059fig5.jpg"> 
图 5: 代增加的成年男性的身体指标。代改进的中位数为圈养 3.5 岁雄性白尾鹿位于诺克苏比，阿塔拉 · 斯科特月县日，密西西比州，美国，Copiah，身体质量、 总的身体长度 (TBL) 和后肢长度 （高频）。在 y 轴上的虚线表示收获收集的数据来自密西西比州，美国的平均体重和用于向第一、 二代的比较。黑钻石代表的预测的均值。晶须表明最小和最大值同时开放圆圈指示异常值。此图的参考31已经被修改。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig5large.jpg" target="_blank">请点击这里查看此图的大版本。</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56059/56059fig6.jpg"> 
图 6: 代增加鹿茸特性的成年男性。中位数鹿茸特性为圈养 3.5 岁雄性白尾鹿位于诺克苏比，阿塔拉 · 斯科特月县日，密西西比州，美国，Copiah，代改进。Y 轴上的虚线表示平均鹿茸分收获收集的数据来自密西西比州，美国，和用于比较对第一、 二代。黑钻石代表的预测的均值。晶须表明最小和最大值同时开放圆圈指示异常值。此图的参考31已经被修改。<a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56059/56059fig6large.jpg" target="_blank">请点击这里查看此图的大版本。</a>
<img alt="Table 1" src="/files/ftp_upload/56059/56059table1.jpg"> 
表 1: 每个模型参数的影响: MCMCglmm 模型对表型特征描述代 (F2)、 年龄和地区 （regionLoess，regionLCP） 的影响。我们编码生成和地区为分类变量和连续变量的年龄。截距代表第一代 (F1)，一位年岁三角洲男性和被认为是一代、 年龄和区域土壤源人口比较参考术语。此表的参考31已经被修改。
<img alt="Table 2" src="/files/ftp_upload/56059/56059table2.jpg"> 
表 2: 增长确定优先级层次结构: %增加到第二代的俘虏 3.5 岁雄性白尾鹿位于诺克苏比，阿塔拉 · 斯科特月县日，密西西比州，美国，Copiah，从第一补益。Refefence31，这张桌子已经被修改。

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

评估环境和遗传学对长寿鹿茸和身体生长的相对影响的协议|环境 |视频

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

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书 》

鹿族群间的表型差异可能涉及人口水平遗传学或营养;雪亮的很难在野外。此协议说明我们如何设计消除营养成分变化了对照的研究。我们发现，雄性白尾鹿的表型变异更受到限制营养比遗传学。

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 次观看.  Mississippi State University. 鹿族群间的表型差异可能涉及人口水平遗传学或营养;雪亮的很难在野外。此协议说明我们如何设计消除营养成分变化了对照的研究。我们发现，雄性白尾鹿的表型变异更受到限制营养比遗传学。

视频: 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.

评估 乌西拉戈 病原性的快速高效方法，可在玉米和特奥辛特线上使用

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

科研

环境科学

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.

优化负染:通过电子显微镜检查小和不对称蛋白结构的高通量协议

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.

一个外线<sub&gt; 2</sub&gt;浓度梯度用于测试CO<sub&gt; 2</sub&gt;浓度与土壤对草地生态系统功能的影响

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.

一种用于收集和构建土芯协议测渗计

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.

对水生生命值评价悬浮泥沙影响的流通系统曝光

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.

协议塑料微粒采样的海面和样品分析

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.

真菌 DNA 分析用职业空气标本的收集与提取

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.

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

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.

树叶颗粒物去除能力的评价

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.

研究矿物对有机热液转化影响的实验协议

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

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书 》

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书》