우리 야생 동물 복원 법 (W-48-61)에 연방 원조에서 리소스를 사용 하 여 재정 지원에 대 한 야생 동물의 미시시피 부, 수 산업 및 공원 (MDWFP) 감사 합니다. 우리는 데이터 수집에서 MDWFP 생물학자를 W. 매킨리, A. Blaylock, A. 게리, 그리고 L. Wilf 그들의 광범위 한 참여에 대 한 감사합니다. 우리는 또한 시설 코디 네이 터로 터 커 미와 여러 대학원 학생 및 기술자 데이터를 수집 하는 그들의 도움에 대 한 감사 합니다. 이 원고는 미시시피 주립 대학 숲과 야생 동물 연구 센터의 기여 WFA427 있습니다.

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/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/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/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/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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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cooperative+management+of+white-tailed+deer+on+private+lands+in+Mississippi.">Cooperative management of white-tailed deer on private lands in Mississippi.</a> Wildl Soc Bull. 11 (3), 211-214 (1983).</li><li>Pettry, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soil+resource+areas+of+Mississippi.">Soil resource areas of Mississippi.</a> Mississippi Agricultural and Forestry Experiment Station. , (1977).</li><li>Snipes, C. E., Nichols, S. P., Poston, D. H., Walker, T. W., Evans, L. P., Robinson, H. 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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. 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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저자는 공개 없다.

우리의 프로토콜;와 관련 된 몇 가지 단계가 그러나,이 프로토콜 성공을 보장 하기 위해 수행 해야 하는 4 개의 중요 한 단계가 있다. 첫째, 야생 사슴의 캡처, 동안 단일 소스 (1.1.1 단계)의 지역에 걸쳐 여러 캡처 위치 되어야 합니다. 여러 개의 캡처 위치를 갖는 소스 영역과 관련 된 모든 유전 가변성 사슴 중에서 대표 될 것 이다 보장 합니다. 둘째, 사슴 유지 되어야 한다 분리 된 소스 지역별 사?...

어머니는 임신과 수 유 하는 동안 경험 하는 환경 요인 그녀의 자손의 표현 형, 유전자 형1,2,3의 독립에 영향을 미칠 수 있습니다. 고품질 환경에 서식 하는 어머니는 효율 형 (작은 뿔과 신체 크기4) 하는 자손을 생산 수 있습니다 반면 가능성이 높은-품질 환경에 서식 하는 어머니는 럭셔리 형 (큰 몸과 뿔 크기4), 자손을 생산할 예정 이다. 따라서, 높은-품질 환경에서 유지는 자손의 생식 기회5,6,,78 에 직접 영향을 미칠 수 있습니다 그리고 어머니의 포함 피트 니스를 직접 영향 큰 phenotypic 특성을 가진 남성 자손을 생산 하는 어머니를 허용할 수 있습니다.
비록 영양 taxa (우수 스 있습니다, 우수 스 arctos9;에 걸쳐 직접 영향을 phenotypic 특성 Liasis fuscusI 10; Larus michahellis 11), 여러 가지 요인 미시시피, 미국에서에서 흰 꼬리 사슴 고기에 영향을 미칠 수 있습니다. 뿔과 몸 크기는 약 1 / 3 일부 집단에 대 한 더 큰12다른 사람에 비해. 이 변화는 사료 품질13,14;와 강한 상관 관계가 가장 큰 남성은 마 초의 최고의 품질을 가진 지역에서 발견 된다. 그러나, 미시시피에 흰 꼬리 사슴의 역사 복원 노력 수 있습니다 끌고있다 유전자 병목 또는 창시자 효과15,16을 또한 부분적으로 흰 꼬리 사슴 표현 형에서 관찰된 지역 변이의 일부를 설명할 수 있습니다.
우리는 우리가 야생 흰 꼬리 사슴, 남성 표현 형 유전자 인구 수준에 의해 제한 됩니다 여부를 평가 하는의 영양 품질을 제어 하는 데 사용 하는 프로토콜을 제공 합니다. 이 프로토콜 모성 효과 후행을 봐도 우리의 인구에 참석 했다. 우리의 제어 디자인 연구는 영양 제한3,17에 대 한 프록시 환경 변수를 사용 하 여 제한 됩니다 무료 이르기까지 인구에 게 우선 이다. 우리의 제어 디자인 또한 수 있습니다 다른 변수 같은 잠재력 만성 스트레스와 관련 된 사회적 상호 작용 모든 개인 비슷한 주택 및 농업 관행을 복종 된다 지속적인 개최 되. 또한, 영양 직접 영향을 미치는 다른 생활 역사 측면 때문에 복제에서 생존18,에 이르는19, 영양 제어 수 있습니다 포유류 생활 역사 측면에 영향을 주는 다른 변수를 평가 하는 조사. 비슷한 프로토콜 (예를 들어, 20,21) 북미 지역에 걸쳐 다른 ungulates 위한 생활사 측면에 관련 된 질문을 평가 하기 위해 설명 했습니다.

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

Cervid 형 두 가지 범주 중 하나에 배치 될 수 있습니다: 효율, 사치 스러운 형태학 성장, 및 큰 무기와 몸 크기의 성장을 촉진 럭셔리 생존을 승진 시키는. 같은 종족의 인구는 환경 조건에 따라 각 표현 형을 표시합니다. 남성 흰 꼬리 사슴 (Odocoileus virginianus)의 몸과 뿔 크기 미시시피, 미국에서에서 지형 지역 다릅니다 영양 품질에 지역 편차와 강한 상관 관계가 있지만, 기본 주식 및 이전 re-stocking 노력에서 인구 수준 유전학의 효과 무시 수 없습니다. 이 프로토콜은 우리가 나이 및 영양, 같은 표현 형에 영향을 주는 다른 요인 제어 됩니다 제어 연구를 설계 하는 방법을 설명 합니다. 우리는 미시시피, 미시시피 주립 대학 녹슨 도킨스 기념 사슴 단위를 미국에서에서 3 가지 지형 지역에서 야생 임신 암컷과 6 개월 마리의 가져왔다. 같은 지역에서 사슴 세대 반응과 모성 효과 평가 하기 위해 수 있도록 자손의 2 세대 생산 자란 했다. 모든 사슴 같은 높은-품질 (20% 조 단백질 사슴 펠 릿) 다이어트 광고 libitum먹었다. 우리는 유일 하 게 각 신생아를 표시 하 고 몸 질량, 뒷 발, 그리고 몸 전체 길이 기록. 각 연속적인가, 우리 개인 원격 사출을 통해 진정 하 고 동일한 morphometrics 플러스 성인의 뿔 샘플링. 우리는 모든 morphometrics 뿔 크기 (지역 변이 이상 존재)의 완전 한 보상 및 신체 질량 (일부 지역 변이의 증거) 2 세대에서 분명 부분 보상 2 세대에 첫 번째에서 크기가 증가 발견. 2 세대에서 우리의 가난한 품질 남성 뿔 크기에서 40% 증가 신체 질량 그들의 야생 수확에 비해 25% 증가 대해 표시 하는 지역 토양. 우리의 결과 야생 남성 흰 꼬리 사슴 미시시피에서의 phenotypic 변이 더 인구 수준 유전학 보다 영양 품질 차이에 관련 된 것이 좋습니다.

개인 나이, 영양 질, 그리고 유전학 남성 흰 꼬리 사슴 표현 형에 영향. 우리의 연구 우리가 영양 사슴의 품질 제어를 소모 했다 디자인과 비교 년 클래스 내에서 각 사슴의 나이 식별할 수 있었습니다. 제어 함으로써 영양 및 우리의 연구 설계와 나이, 우리 더 이해 여부 인구 수준 유전학 두 연구 인구에서 남성의 표현 형을 제한 했다 수 있었다. 향상 된 영양 3.5 세 남성 흰...

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

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

뿔과 긴 Cervid에 대 한 신체 성장에 환경 및 유전학의 상대적 효과 평가 하기 위한 프로토콜

Cervid 인구 가운데 phenotypic 차이 관련이 있을 수 있습니다 인구 수준 유전학 또는 영양; 분별 있는 야생에서 어렵습니다. 이 프로토콜은 우리가 영양 변형 삭제 되었다 제어 연구를 설계 하는 방법을 설명 합니다. 우리는 남성 흰 꼬리 사슴의 phenotypic 변이 유전학 보다 영양에 의해 더 제한 되었다 발견.

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 조회수.  Mississippi State University. Cervid 인구 가운데 phenotypic 차이 관련이 있을 수 있습니다 인구 수준 유전학 또는 영양; 분별 있는 야생에서 어렵습니다. 이 프로토콜은 우리가 영양 변형 삭제 되었다 제어 연구를 설계 하는 방법을 설명 합니다. 우리는 남성 흰 꼬리 사슴의 phenotypic 변이 유전학 보다 영양에 의해 더 제한 되었다 발견.

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

Instrumentation of Near-term Fetal Sheep for Multivariate Chronic Non-anesthetized Recordings

The chronically instrumented pregnant sheep has been used as a model of human fetal development and responses to pathophysiologic stimuli such as endotoxins, bacteria, umbilical cord occlusions, hypoxia and various pharmacological treatments. The life-saving clinical practices of glucocorticoid treatment in fetuses at risk of premature birth and the therapeutic hypothermia have been developed in this model. This is due to the unique amenability of the non-anesthetized fetal sheep to the surgical placement and maintenance of catheters and electrodes, allowing repetitive blood sampling, substance injection, recording of bioelectrical activity, application of electric stimulation and in vivo organ imaging. Here we describe the surgical instrumentation procedure required to achieve a stable chronically instrumented non-anesthetized fetal sheep model including characterization of the post-operative recovery from blood gas, metabolic and inflammation standpoints.

The chronically instrumented non-anesthetized fetal sheep model is used to study human fetal development in health and disease, because it permits surgical placement and maintenance of catheters and electrodes, repetitive blood sampling, substance injection, recording of bioelectrical activity, and in vivo imaging. We describe the procedures required to establish this model.

다변량 만성 비 마취 녹음을위한 단기적 태아 양의 계측

The chronically instrumented pregnant sheep has been used as a model of human fetal development and responses to pathophysiologic stimuli such as ...

연구

발생학

A Method to Define the Effects of Environmental Enrichment on Colon Microbiome Biodiversity in a Mouse Colon Tumor Model

Several recent studies have illustrated the beneficial effects of living in an enriched environment on improving human disease. In mice, environmental enrichment (EE) reduces tumorigenesis by activating the mouse immune system, or affects tumor bearing animal survival by stimulating the wound repair response, including improved microbiome diversity, in the tumor microenvironment. Provided here is a detailed procedure to assess the effects of environmental enrichment on the biodiversity of the microbiome in a mouse colon tumor model. Precautions regarding animal breeding and considerations for animal genotype and mouse colony integration are described, all of which ultimately affect microbial biodiversity. Heeding these precautions may allow more uniform microbiome transmission, and consequently will alleviate non-treatment dependent effects that can confound study findings. Further, in this procedure, microbiota changes are characterized using 16S rDNA sequencing of DNA isolated from stool collected from the distal colon following long-term environmental enrichment. Gut microbiota imbalance is associated with the pathogenesis of inflammatory bowel disease and colon cancer, but also of obesity and diabetes among others. Importantly, this protocol for EE and microbiome analysis can be utilized to study the role of microbiome pathogenesis across a variety of diseases where robust mouse models exist that can recapitulate human disease.

Environmental Enrichment (EE) is an animal housing environment that is used to reveal mechanisms that underlie the connections between lifestyle, stress, and disease. This protocol describes a procedure that uses a mouse model of colon tumorigenesis and EE to specifically define alterations in microbiota biodiversity that may impact animal mortality.

마우스 결 장 종양 모델에서 콜론 미생물 생물 다양성에 환경 농축의 효과 정의 하는 방법

Several recent studies have illustrated the beneficial effects of living in an enriched environment on improving human disease. In mice, environmental ...

암 연구학

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical data that may be used in predictive modelling to forecast evolutionary responses of natural ecosystems to future environmental changes. However, excluding a few exceptional cases, long-term studies are scarce because of logistic difficulties associated with accessing temporal samples. Temporal dynamics are frequently studied in the laboratory or in controlled mesocosm experiments with exceptional studies that reconstruct the evolution of natural populations in the wild.
Here, a standard operating procedure (SOP) is provided to revive or resurrect dormant Daphnia magna, a widespread zooplankton keystone species in aquatic ecosystems, to dramatically advance the state-of-the-art longitudinal data collection in natural systems. The field of Resurrection Ecology was defined in 1999 by Kerfoot and co-workers, even though the first attempts at hatching diapausing zooplankton eggs date back to the late 1980s. Since Kerfoot&#39;s seminal paper, the methodology of resurrecting zooplankton species has been increasingly frequently applied, though propagated among laboratories only via direct knowledge transfer. Here, an SOP is described that provides a step-by-step protocol on the practice of resurrecting dormant Daphnia magna eggs.
Two key studies are provided in which the fitness response of resurrected Daphnia magna populations to warming is measured, capitalizing on the ability to study historical and modern populations in the same settings. Finally, the application of next generation sequencing technologies to revived or still dormant stages is discussed. These technologies provide unprecedented power in dissecting the processes and mechanisms of evolution if applied to populations that have experienced changes in selection pressure over time.

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

부활의 휴면 벼룩 마그나: 프로토콜 및 응용 프로그램

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical ...

환경과학

Using a Comparative Species Approach to Investigate the Neurobiology of Paternal Responses

A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors. Using animal models to accomplish this goal, many methodological strategies require invasive techniques to manipulate the intensity of the behavior of interest (e.g., lesion methods, pharmacological manipulations, microdialysis techniques, genetically-engineered animal models). The utilization of a comparative species approach allows researchers to take advantage of naturally occurring differences in response strategies existing in closely related species. In our lab, we use two species of the Peromyscus genus that differ in paternal responses. The male California deer mouse (Peromyscus californicus) exhibits the same parental responses as the female whereas its cousin, the common deer mouse (Peromyscus maniculatus) exhibits virtually no nurturing/parental responses in the presence of pups. Of specific interest in this article is an exploration of the neurobiological factors associated with the affiliative social responses exhibited by the paternal California deer mouse. Because the behavioral neuroscience approach is multifaceted, the following key components of the study will be briefly addressed: the identification of appropriate species for this type of research; data collection for behavioral analysis; preparation and sectioning of the brains; basic steps involved in immunocytochemistry for the quantification of vasopressin-immunoreactivity; the use of neuroimaging software to quantify the brain tissue; the use of a microsequencing video analysis to score behavior and, finally, the appropriate statistical analyses to provide the most informed interpretations of the research findings.

The comparative species approach allows behavioral neuroscientists to explore various neurobiological factors associated with specific behaviors viewed as characteristic of a specific animal model. Taking advantage of naturally occurring differences in behavior between closely related species, this technique doesn&#x2019;t require invasive techniques to manipulate the expression of the behavior.

아버지의 응답의 신경 생물학을 맡은 이후로 비교 종의 접근법을 사용

A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors.  Using animal models to accomplish ...

신경과학

Transabdominal Ultrasound for Pregnancy Diagnosis in Reeves' Muntjac Deer

Reeves' muntjac deer (Muntiacus reevesi) are a small cervid species native to southeast Asia, and are currently being investigated as a potential model of prion disease transmission and pathogenesis. Vertical transmission is an area of interest among researchers studying infectious diseases, including prion disease, and these investigations require efficient methods for evaluating the effects of maternal infection on reproductive performance. Ultrasonographic examination is a well-established tool for diagnosing pregnancy and assessing fetal health in many animal species1-7, including several species of farmed cervids8-19, however this technique has not been described in Reeves' muntjac deer. Here we describe the application of transabdominal ultrasound to detect pregnancy in muntjac does and to evaluate fetal growth and development throughout the gestational period. Using this procedure, pregnant animals were identified as early as 35 days following doe-buck pairing and this was an effective means to safely monitor the pregnancy at regular intervals. Future goals of this work will include establishing normal fetal measurement references for estimation of gestational age, determining sensitivity and specificity of the technique for diagnosing pregnancy at various stages of gestation, and identifying variations in fetal growth and development under different experimental conditions.

Transabdominal ultrasound is described as an effective, noninvasive means for assessing reproductive status in Reeves' muntjac deer. These methods can be used to achieve early pregnancy diagnosis and to evaluate fetal viability. Future applications of this technique include estimation of gestational age and effects of maternal disease on fetal development.

리브스의 문작 사슴에서 임신 진단을 위한 복부 초음파

Reeves' muntjac deer (Muntiacus reevesi) are a small cervid species native to southeast Asia, and are currently being investigated as a potential model of ...

의학

Application of Long-term cultured Interferon-&#947; Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle

Effector and memory T cells are generated through developmental programing of na&#239;ve cells following antigen recognition. If the infection is controlled up to 95 % of the T cells generated during the expansion phase are eliminated (i.e., contraction phase) and memory T cells remain, sometimes for a lifetime. In humans, two functionally distinct subsets of memory T cells have been described based on the expression of lymph node homing receptors. Central memory T cells express C-C chemokine receptor 7 and CD45RO and are mainly located in T-cell areas of secondary lymphoid organs. Effector memory T cells express CD45RO, lack CCR7 and display receptors associated with lymphocyte homing to peripheral or inflamed tissues. Effector T cells do not express either CCR7 or CD45RO but upon encounter with antigen produce effector cytokines, such as interferon-&#947;. Interferon-&#947; release assays are used for the diagnosis of bovine and human tuberculosis and detect primarily effector and effector memory T cell responses. Central memory T cell responses by CD4+ T cells to vaccination, on the other hand, may be used to predict vaccine efficacy, as demonstrated with simian immunodeficiency virus infection of non-human primates, tuberculosis in mice, and malaria in humans. Several studies with mice and humans as well as unpublished data on cattle, have demonstrated that interferon-&#947; ELISPOT assays measure central memory T cell responses. With this assay, peripheral blood mononuclear cells are cultured in decreasing concentration of antigen for 10 to 14 days (long-term culture), allowing effector responses to peak and wane; facilitating central memory T cells to differentiate and expand within the culture.

Long-term cultured interferon-&#947; enzyme-linked immunospot assay is used as a measure of central memory responses and correlates with protective anti-mycobacterial vaccine responses. With this assay, peripheral blood mononuclear cells are stimulated with mycobacterial antigens and interleukin-2 for 14 days, enabling differentiation and expansion of central memory T cells.

소에 이펙터 및 메모리 T 세포 반응을 평가하기위한 장기 배양 인터페론-γ 효소 링크 된 Immunospot 분석의 응용 프로그램

Effector and memory T cells are generated through developmental programing of na&#239;ve cells following antigen recognition. If the infection is ...

면역학 및 감염병학

Modeling Early-Life Adversity in Mice Using Limited Bedding and Nesting

Limited Bedding and Nesting as a Model for Early-Life Adversity in Mice

Early-life adversity (ELA), such as abuse, neglect, lack of resources, and an unpredictable home environment, is a known risk factor for developing neuropsychiatric disorders such as depression. Animal models for ELA have been used to study the impact of chronic stress on brain development, and typically rely on manipulating the quality and/or quantity of maternal care, as this is the major source of early-life experiences in mammals, including humans. Here, a detailed protocol for employing the Limited Bedding and Nesting (LBN) model in mice is provided. This model mimics a low-resource environment, which provokes fragmented and unpredictable patterns of maternal care during a critical developmental window (postnatal days 2-9) by limiting the amount of nesting materials given to the dam to build a nest for her pups and separating the mice from the bedding via a mesh platform in the cage. Representative data are provided to illustrate the changes in maternal behavior, as well as the diminished pup weights and long-term changes in basal corticosterone levels, that result from the LBN model. As adults, offspring reared&#160;in the LBN environment have been shown to exhibit an aberrant stress response, cognitive deficits, and anhedonia-like behavior. Therefore, this model is an important tool to define how the maturation of stress-sensitive brain circuits is altered by ELA and results in long-term behavioral changes that confer vulnerability to mental disorders.

저자 스포트라이트: Exploring Microglial Interaction and Stress-Response Circuitry Using the Limited Bedding and Nesting Model (제한된 침구 및 중첩 모델을 사용하여 미세아교세포 상호 작용 및 스트레스-응답 회로 탐색)

This protocol describes an animal model for studying how early-life adversity, provoked by an impoverished environment and unpredictable maternal care during the early postnatal period, affects brain development and the future risk of mental disorders.

제한된 침구와 둥지는 생쥐의 초기 삶의 역경에 대한 모델입니다.

Early-life adversity (ELA), such as abuse, neglect, lack of resources, and an unpredictable home environment, is a known risk factor for developing ...

An Experimental Approach to Investigating Effects of Artificial Light at Night on Free-Ranging Animals: Implementation, Results, and Directions for Future Research

Animals have evolved with natural patterns of light and darkness. However, artificial light is being increasingly introduced into the environment from human infrastructure and recreational activity. Artificial light at night (ALAN) has the potential to have widespread effects on animal behavior, physiology, and fitness, which can translate into broader-scale effects on populations and communities. Understanding the effects of ALAN on free-ranging animals is non-trivial due to challenges such as measuring levels of light encountered by mobile organisms and separating the effects of ALAN from those of other anthropogenic disturbance factors. Here we describe an approach that allows us to isolate the effects of artificial light exposure on individual animals by experimentally manipulating light levels inside nest boxes. To this end, a system can be used consisting of light-emitting diode (LED) light(s) adhered to a plate and connected to a battery and timer system. The setup allows exposure of individuals inside nest boxes to varying intensities and durations of ALAN while simultaneously obtaining video recordings, which also include audio. The system has been used in studies on free-ranging great tits (Parus major) and blue tits (Cyanistes caeruleus) to gain insight into how ALAN affects sleep and activity patterns in adults and physiology and telomere dynamics in developing nestlings. The system, or an adaptation thereof, could be used to answer many other intriguing research questions, such as how ALAN interacts with other disturbance factors and affects bioenergetic balance. Furthermore, similar systems could be installed in or near the nest boxes, nests or burrows of a variety of species to manipulate levels of ALAN, evaluate biological responses, and work towards building an interspecific perspective. Especially when combined with other advanced approaches for monitoring the behavior and movement of free-living animals, this approach promises to yield ongoing contributions to our understanding of the biological implications of ALAN.

Artificial light at night (ALAN) has wide-reaching biological effects. This article describes a system for manipulating ALAN inside nest boxes while monitoring behavior, consisting of LED lights coupled to a battery, timer, and audio-capable infrared video camera. Researchers could employ this system to explore many outstanding questions regarding the effects of ALAN on organisms.

야간에 인공 조명이 자유 범위 동물에 미치는 영향을 조사하는 실험적 접근 방식 : 향후 연구를위한 구현, 결과 및 방향

Animals have evolved with natural patterns of light and darkness. However, artificial light is being increasingly introduced into the environment from ...

뿔과 긴 Cervid에 대 한 신체 성장에 환경 및 유전학의 상대적 효과 평가 하기 위한 프로토콜

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다변량 만성 비 마취 녹음을위한 단기적 태아 양의 계측

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