우리는 희석 및 농약 제형 지원 몰리 굳게 감사합니다. 우리는 또한 실험 설계에 도움을 프로 빗 분석 지침 트로이 앤더슨과 재커리 아델 감사합니다. 이 연구는 부분적으로 곤충학 재단 상으로 버지니아 해충 관리 장학금 기금에 의해 지원되었다.

참고 :이 프로토콜은 별도로 침대 버그 계란과 첫 령충을 치료하기위한 두 개의 살충제 분석을 포함한다. 두 프로토콜은 동일한 살충제를 사용하여 수행 하였다 그러나, 프로토콜은 살충제 노출을 보장하기 용이 시료를 조작하도록 구성 할 수 있었다. 1. 계란 디핑 살충제 분석 <ol><li> 식민지에서 성관계를 잘 자란 성인 여성 침대 버그를 잡아 당겨 필터 종이 페트리 접시로 여성 (5-10)를 배치합니다. 살충제 분석을위한 충분한 계란이 될 수 있도록하기 위해 적어도 10 복제가 있습니다. 암컷 매일 계란을 마련했다 있는지 확인합니다. <ol><li> 그들이 알을 낳는되면, 페트리 접시 (6cm × 5cm)에서 암컷을 매일 새 필터 종이 새로운 배양 접시에 넣어 및 이전 및 새로운 배양 접시에 날짜를 기록. </li><li> 분석을 시작하기 전에 오일에 4 세에 계란을 허용합니다. 배아 개발하는 동안 살충제의 변화에 ​​대한 민감도는이야오 계란 나이를 통제하는 것이 중요하다. D의 광주, 60 % RH, 27 ° C : 12시 12분 L 모든 삶의 단계를 잡으십시오. </li></ol></li><li> 계란이 적절하게 숙성 한 후, 소프트 - 팁 집게를 사용하여 필터 논문에서 조심스럽게 침대 버그 알을 제거합니다. 부드럽게 집게를 사용하여 여과지에 부착 된 계란의 바닥을 긁어 부드럽게 던지고 침대 벌레 알을 제한하기 위해 계란 많은 압력을 가하지 않고 잡아. </li><li> 내부 플라스틱 배양 접시의 각각의 그룹으로 제거 침대 버그 알을 놓습니다. 직물 연화제 시트와 외부에 각각 페트리 접시의 바닥을 문질러 침대 버그 계란 마리의 정전기를 제한합니다. 한 번에 침대 버그 계란의 치료 그룹 (그룹당 5 ~ 10 알). 한 그룹에게 복제를 고려하고이 분석을위한 복제 적어도 다섯 개 (또는 그 이상)를 사용합니다. </li><li> 5 가지 농도 (의 범위를 얻기 위해 살충제의 연속 희석을 준비 0.21-21 μL / mL의 이미다클로 프리드 (0.1 %) / β-cyfluthrin (0.5 %) 및 제어를위한용제 (수돗물)와 솔루션 만. 침지 분석을위한 충분한 볼륨을 각 농도의 20 ㎖를 확인합니다. </li><li> 35 mL의 회선 절단 원심 분리 튜브 (50 mL)의 내측 차단 휠 액세서리와 회전 공구를 사용하고 원심 분리 튜브 (직경 = 2.54 cm)는 알루미늄과 같은 회전 공구를 사용하여 뚜껑에 구멍을 잘라 산화 돌 ​​액세서리를 연마. 캡 안전하게 튜브 (도 1)에 나사되도록 절단 할 때, 캡의 가장자리에 5cm 남기기. </li><li> 캡에 맞게 충분히 큰 사각형으로 미세 메쉬 컷 (~ 5cm 2). 원심 분리 튜브의 캡을 사용하여 원심 분리기 튜브에 어떠한 충돌을 제거 단단히 메쉬 스크류. </li><li> 원심 분리기 튜브의 내부에 메쉬에 페인트 브러시를 사용하여 침대 버그 계란의 그룹 (하나의 복제)를 놓습니다. 5 초 동안 살충제 농도로 절단 원심 분리기 튜브의 내부에 침대 버그의 그룹을 찍어. </li><li> 일에서 원심 분리기 튜브를 제거전자 살충제 솔루션입니다. 내부 기관 조직에 직접 원심 관을 배치하여 계란 메쉬를 건조. </li><li> 작은 미세 강모 페인트 브러시를 사용하여 원심 분리 관에서 계란의 그룹을 제거하고 깨끗한 종이 필터와 깨끗한 페트리 접시에 배치합니다. </li><li> 기록 달걀 사망률 모든 계란을 보장하기 위해 14 일 동안 (부화 실패는) 부화 할 수있는 충분한 시간이 있었다. </li><li> 애보트의 식 (- % 제어 사망 / 100 - 제어 사망률 × 100 % 테스트 사망률)를 사용하여 제어 치료 사망률에 대한 올바른. </li><li> LC (50) 값을 계산하는 분석 소프트웨어 (15)에 대응하는 농도 기록 사망률을 삽입합니다. </li></ol> 2. 먼저 탈피 살충제 분석 <ol><li> 바람직하게는 1 일전 공급, 잘 공급 침대 버그 (5 성인 남성 5 성인 여성)의 그룹을 분리하고, MATIN 제공 여과지와 페트리 접시 (6cm × 5cm)로 배치g, 계란 누워. 성인 침대 버그가 짝을 약 7-9일에 대한 알을 낳기 할 수 있습니다. 수유, 혈액 (16)을 둘러싸는 유리관에 온수를 순환 인공 공급 시스템을 사용한다. 이 연구를 위해, defibronated 토끼의 혈액을 사용합니다. </li><li> 페트리 접시에서 성인 (5cm × 6cm)를 제거하고 계란이 배양 접시 내에서 첫 번째 령충에 부화 할 수 있습니다. 침대 버그 계란은 실온에서 부화하는 것은 약 7-9일 소요됩니다. </li><li> 페인트 브러시를 사용하여 같은 나이의 unfed 첫 번째 령충를 수집합니다. 연령은 빗금 하루 매일 제 령기를 제거하고 기록함으로써 결정될 수있다. 살충제 분석까지 새로운, 깨끗한 페트리 접시에 배치합니다. 살충제 분석이 가능한 한 빨리 완료해야하므로 먼저 령 약충은, 저항하는 매우 건조하지 않습니다. </li><li> 분석을 위해, 5 가지 농도 및 제어 SOLU의 범위를 얻을 이미다클로 / β-cyfluthrin의 희석액을 제조용매 만에 기. 미국에서 베드 버그 제어 표지 배합 제품에 따라서 용매 (라벨에 따라) 물이고, 이러한 연구에 사용되었다. </li><li> 피펫 개별 여과지 상 대조군에 대한 각 살충제 농도 및 물 150 μL 씩 (# 1; 4.2 cm 직경). 여과지의 전체 가장자리와 중심을 적을 적용하여 피펫을 사용하여 전체 여과지 위에 살충제 배포. </li><li> 개별 널빤지 패널 (7cm 2)의 상부에 개별적으로 처리 필터 종이를 놓습니다. </li><li> 반면 여전히 살충제와 습식 및 페트리 접시의 반전 바닥 첫 번째 령충을 커버 개별 여과지 위에 첫 번째 령충의 릴리스 그룹 (그룹당 10-5). </li><li> 처리 된 표면을 탈출 첫 번째 령 침대 버그를 방지하기 위해 페트리 접시 위에 무게를 놓습니다. </li><li> 모두 24 시간 노출 후 처음 령 침대 버그의 기록 사망살충제 농도. </li><li> 애보트의 식 (- % 제어 사망 / 100 - 제어 사망률 × 100 % 테스트 사망률)를 사용하여 제어 치료 사망률에 대한 올바른. </li><li> LC (50) 값을 계산하는 분석 소프트웨어 (15)에 기록 사망률과 농도를 넣습니다. </li></ol>

<ol>
	<li>Sabou, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bed+bugs+reproductive+life+cycle+in+the+clothes+of+a+patient+suffering+from+Alzheimer's+disease+results+in+iron+deficiency+anemia.">Bed bugs reproductive life cycle in the clothes of a patient suffering from Alzheimer's disease results in iron deficiency anemia.</a> Parasite. 20, (2013).</li><li>Reinhardt, K., Kempke, D., Naylor, R. A., Siva-Jothy, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitivity+to+bites+by+the+bedbug,+Cimex+lectularius.">Sensitivity to bites by the bedbug, Cimex lectularius.</a> Med. Vet. Entomol. 23, 163-166 (2009).</li><li>Leverkus, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bullous+allergic+hypersensitivity+to+bed+bug+bites+mediated+by+IgE+against+salivary+nitrophorin.">Bullous allergic hypersensitivity to bed bug bites mediated by IgE against salivary nitrophorin.</a> J. invest. entomol. 126, 91-96 (2006).</li><li>Fletcher, C. L., Ardern-Jones, M. R., Hay, 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=Widespread+bullous+eruption+due+to+multiple+bed+bug+bites.">Widespread bullous eruption due to multiple bed bug bites.</a> Clin. Exp. Dermatol. 27, 74-75 (2002).</li><li>Pritchard, M. J., Hwang, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Severe+anemia+from+bedbugs.">Severe anemia from bedbugs.</a> Can. Med. Assoc. J. 181, 287-288 (2009).</li><li>Goddard, J., de Shazo, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychological+effects+of+bed+bug+attacks+(Cimex+lectularius+L).">Psychological effects of bed bug attacks (Cimex lectularius L).</a> Am. J. Med. 125, 101-103 (2012).</li><li>Harlan, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bed+bugs+101:+the+basics+of+Cimex+lectularius.">Bed bugs 101: the basics of Cimex lectularius.</a> Am. Entomol. 52, 99-101 (2006).</li><li>Pinto, L. J., Cooper, R., Kraft, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Bed Bug Handbook: The Complete Guide to Bed Bugs and Their Control. , (2007).</li><li>Toloza, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+patterns+of+insecticide+resistance+in+eggs+and+first+instars+of+Triatoma+infestans+(Hemiptera:+Reduviidae)+from+Argentina+and+Bolivia.">Differential patterns of insecticide resistance in eggs and first instars of Triatoma infestans (Hemiptera: Reduviidae) from Argentina and Bolivia.</a> J. Med. Entomol. 45, 421-426 (2008).</li><li>Cueto, G. M., Zerba, E. N., Picollo, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+pyrethroid+resistance+in+eggs+of+Pediculus+humanus+capitis+(Phthiraptera:+Pediculidae)+from+Argentina.">Evidence of pyrethroid resistance in eggs of Pediculus humanus capitis (Phthiraptera: Pediculidae) from Argentina.</a> J. Med. Entomol. 45, 693-697 (2008).</li><li>Ho, S. H., Goh, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deltamethrin+as+a+potential+ovicidal+pyrethroid+against+Plutella+xylostella+L.">Deltamethrin as a potential ovicidal pyrethroid against Plutella xylostella L.</a> Toxicol. let. 22, 161-164 (1984).</li><li>Bell, C. H., Hole, B. D., Evans, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+occurrence+of+resistance+to+phosphine+in+adult+and+egg+stages+of+strains+of+Rhyzopertha+dominica+(F.)(Coleoptera:+Bostrichidae).">The occurrence of resistance to phosphine in adult and egg stages of strains of Rhyzopertha dominica (F.)(Coleoptera: Bostrichidae).</a> J. Stored Prod. Res. 13, 91-94 (1977).</li><li>Campbell, B. E., Miller, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insecticide+Resistance+in+Eggs+and+First+Instars+of+the+Bed+Bug,+Cimex+lectularius+(Hemiptera:+Cimicidae).">Insecticide Resistance in Eggs and First Instars of the Bed Bug, Cimex lectularius (Hemiptera: Cimicidae).</a> Insects. 6, 122-132 (2015).</li><li>Price, L. A., Mills, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+toxicity+of+phosphine+to+the+immature+stages+of+resistant+and+susceptible+strains+of+some+common+stored+product+beetles,+and+implications+for+their+control.">The toxicity of phosphine to the immature stages of resistant and susceptible strains of some common stored product beetles, and implications for their control.</a> J. Stored Prod. Res. 24, 51-59 (1988).</li><li>Robertson, J. L., Preisler, H. K., Russel, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> PoloPlus: Probit and Logit Analysis User's Guide. , (2003).</li><li>Montes, C., Cuadrillero, C., Vilella, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+a+laboratory+colony+of+Cimex+lectularius+(Hemiptera:+Cimicidae)+using+an+artificial+feeding+technique.">Maintenance of a laboratory colony of Cimex lectularius (Hemiptera: Cimicidae) using an artificial feeding technique.</a> J. Med. Entomol. 39, 675-679 (2002).</li><li>Brogdon, W. G., McAllister, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplification+of+adult+mosquito+bioassays+through+use+of+time-mortality+determinations+in+glass+bottles.">Simplification of adult mosquito bioassays through use of time-mortality determinations in glass bottles.</a> J. Am. Mosq. Control. Assoc. 14, 159-164 (1998).</li><li>Katarzyna, K., Saddler, A., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+age+and+larval+nutrition+on+phenotypic+expression+of+insecticide-resistance+in+Anopheles+mosquitoes.">Effects of age and larval nutrition on phenotypic expression of insecticide-resistance in Anopheles mosquitoes.</a> PLoS One. 8 (3), (2013).</li><li>Outram, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+the+resistance+of+insect+eggs+to+sulphuryl+fluoride-II:+the+distribution+of+sulphuryl-35+S+fluoride+in+insect+eggs+after+fumigation.">Factors affecting the resistance of insect eggs to sulphuryl fluoride-II: the distribution of sulphuryl-35 S fluoride in insect eggs after fumigation.</a> J. Stored. Prod. Res. 3, 353-358 (1967).</li></ol>

저자는 공개 아무것도 없어.

A critical step in this assay is to ensure that no eggs that are removed from a surface are damaged prior to the assay. Many insects cement their eggs to a substrate, therefore, a preliminary test may be needed to ensure that removal does not cause mortality. This test can be conducted with several replications of a treated group (bed bugs removed from filter paper) compared to a control group (bed bugs not removed). Similar low rates of mortality (or no mortality) between the treated and control group will ensure that removal did not cause egg mortality. Thus, any mortality recorded for the insecticide assay should be due to the insecticide treatment. The same preliminary experiment can be conducted using first instars to ensure that methods for collecting do not kill the experimental insects. All experiments conducted later should include control replications that do not receive insecticide but are treated with the solvent (for these experiments, tap water was the solvent). 
Preliminary experiments should be conducted with the recommended label rate for formulated products and then serially diluted to determine the correct concentrations to produce a concentration response curve of mortality ranging from 20-80%. Previous insecticide studies on the same insects or compounds may also be insightful when trying to determine the appropriate concentration. Concentrations will need to be modified dependent upon the insect, life stage, and compound used. New solutions should be made each time an experiment is conducted. 
The exposure of first instars to wet residues was necessary to get high mortality. Although this life stage is considered more susceptible to insecticides than older nymphs and adults, high mortality was still difficult to obtain with formulated products. Submerging the first instars into insecticides similar to the eggs was not plausible because of the resultant high mortality, thus this was a limitation to the study. The alternative first instar bioassay was manipulated to maintain first instars on surfaces to force constant contact with the treatment.
Dry residue assays and bottle assays commonly used for insecticide efficacy and resistance studies do not provide methodology for testing against multiple insect life stages17. These assays are often used to determine resistance in insects, and not to test insecticide efficacy. There are limited studies on egg dipping toxicological studies and none that have reported a standard methodology previously that can be repeated for multiple insect species.
This study provides a protocol for testing insecticide efficacy against bed bug eggs and first instars. Efficacy should be evaluated in different life stages, if not all, for a more holistic view of the efficacy of a treatment program. This consideration is important because an insect's susceptibility to insecticides changes with age and development 18. While age is an important factor in insecticide susceptibility, a factor in egg insecticide susceptibility is the eggshell. The eggshell provides a barrier to insecticides and makes insect eggs particularly difficult to control19. 
These toxicological assays provide researchers a way to evaluate insecticide toxicity against various life stages of insects. Insecticidal solutions are prepared to evaluate mortality and to determine LC50 concentrations. While this protocol was used for formulated products, the steps can be manipulated for technical grade compounds and for a variety of insects and life stages to evaluate toxicity.

빈대는 인간 호스트 1, 2, 3, 4, 5, 6에서 혈액 손실, 피부 자극, 불면증, 우울증, 불안을 일으킬 수있는 중요한 도시 해충이다. 침대 버그를 제거하고 제어하는 비용은 높은 종종 시간은 해충 방제 회사 (7)에 의해 집에 여러 개의 방문이 필요합니다. 여러 방문은 보통 때문에 침대 버그의 비밀 행동을 죽이는과 관련된 어려움 필요합니다. 특히, 침대 벌레 알은 그들의 작은 크기와 껍질로 태아 보호의 제어가 곤란하다. 현재 미국 전역의 많은 해충 방제 기업에 대한 일반적인 관행은 화학 살충제를 사용하여 침대 버그에 대한 가정을 치료하는 것입니다. 이것은 잘 알려져계란을 제어하기 어려운 것으로, 많은 회사들은 재 치료 8 2 주 기간을 구현 하였다. 첫 번째 령이 등장하고 소문 살충제로 죽일 쉽게 될 수 있도록이 침대 버그 달걀을 부화 할 수있는 충분한 시간이 있습니다. 그러나, 침대 버그 계란, 첫 번째 령충에 대한 액체 살충제의 효능을 평가하는 연구의 부족이있다. 곤충 계란 많은 농업 해충 이외에 빈대 이외 도시 해충으로 제어 할 수있는 가장 어려운 수명 단계로 설명되었다. 몇 가지 연구가 살충제 저항성을보고하면서 이러한 어려움의 대부분은 껍질에 기인하고있다. 계란 단계에서 저항, 배추 11 xylostella, 10 capitis Triatoma의 infestans 9 Pediculus humanus에 Rhyzopertha의 도미니카 (12)를 기록하고있다 <em&gt; CIMEX lectularius (13), 복수 저장 제품 딱정벌레 (즉 Oryzaephilus surinamensis, Tribolium castaneum, Cryptolestes ferrugineus입니다 및 Rhyzopertha의 도미니카). 14 껍질 통해 살충제 침투와 관련된 계란 스테이지 미성숙 단계뿐만 아니라 합병증 저항성의 발달은 이러한 성장 단계에 대한 살충 효과 생물 검정에 대한 요구를 필요로한다. 이 프로토콜은 일반적인 침대 버그의 계란과 첫 번째 령 단계에서 살충제의 효과를 결정하기위한 단계별 절차를 제공하고, CIMEX는 lectularius. 이들 프로토콜 모두는 LC (50) 값의 정량화를 허용하는 농도 - 반응 분석이다. 농도 - 반응 분석은 일반적으로 그러나이 프로토콜은 쉽게 베드 버그 계란, 첫 번째 령충 그룹을 치료에 적용되어, 독성 연구에 사용됩니다. 이러한 분석은 적용 할 수있다다양한 곤충 종의 계란과 미성숙 한 삶의 단계에 대한.

<table border="0" cellpadding="0" cellspacing="0" width="1067">
	<tbody>
		<tr>
			<td>Petri dishes</td>
			<td>Fisher Scientific Inc.</td>
			<td>08-757-105</td>
			<td>Plastic</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>Whatman</td>
			<td>1001-042</td>
			<td></td>
		</tr>
		<tr>
			<td>Featherweight forceps</td>
			<td>Fisher Scientific Inc.</td>
			<td align="right">4750</td>
			<td></td>
		</tr>
		<tr>
			<td>Temprid SC (imidacloprid/bifenthrin)</td>
			<td>Bayer CropScience</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Transport GHP (acetamiprid/bifenthrin)</td>
			<td>FMC Corp.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge tube</td>
			<td></td>
			<td>06-443-18</td>
			<td>50 mL; flat-top threaded</td>
		</tr>
		<tr>
			<td>Fine mesh</td>
			<td></td>
			<td>7250C</td>
			<td></td>
		</tr>
		<tr>
			<td>KimWipe</td>
			<td></td>
			<td>06-666A</td>
			<td>11 cm X 21 cm</td>
		</tr>
		<tr>
			<td>Small paint brush</td>
			<td></td>
			<td></td>
			<td>Any small paint brush</td>
		</tr>
		<tr>
			<td>Hardboard panels</td>
			<td></td>
			<td></td>
			<td>Composite wood or tile would be sufficient from a home improvement store</td>
		</tr>
		<tr>
			<td>Metal weights</td>
			<td></td>
			<td></td>
			<td>Any type of metal screw that will hold the weight of the Petri dish down onto a surface</td>
		</tr>
	</tbody>
</table>

a method for evaluating insecticide efficacy against bed bug, cimex lectularius, eggs and first instars

해충에 대한 살충제의 표준 독성 평가는 주로 성인 곤충에 실시하고 있습니다. 평가는 용량 또는 살충제의 농도 증가 사망률 증가 용량 - 반응 또는 농도 - 반응 곡선에 기초한다. 표준 치사 농도 (LC 50)과 치사량 (LD 50) 테스트 인구의 50 %의 사망률을 초래할 테스트는 계란과 조기 령 또는 nymphal 단계와 같은 비 성인 곤충 생활 단계에 대한 살충제의 독성을 평가하기 위해 도전 할 수있다 . 그러나이 정보는 제어 및 치료 노력에 영향을 미치는 모든 침대 버그 삶의 단계에서 살충제의 효과를 이해하는 데 필수적이다. 이 프로토콜은 침대 버그 계란에 대한 수정 된 표준 찍기 생물 검정 및 nymphal 첫 번째 령충 치료에 대한 연락처 살충 분석을 사용합니다. 이러한 분석은 또한 살충제 평가를 위해 50 LC 값을 정량화하는 농도 - 반응 곡선을 생성한다.

계란은 이미다클로 프리드 / β-cyfluthrin의 5 가지 농도 (μL / ㎖)로 감소 하였다. 첫 번째 령충은 이미다클로 프리드 / β-cyfluthrin의 다섯 가지 농도 (그림 2)의 처리 표면에 배치했다. 할란, 리치몬드, 에픽 센터 (표 1) : 우리는 C. lectularius의 세 가지 다른 집단을 사용했다. 할란 변형은 피레스 로이드 계 살충제에 민감 박사 해롤드 할란에서 2005 년에 인수되었다. 리치몬드와 에픽 센터 균주는 델타 메트 린에 강한 상표입니다. 리치몬드 균주는 2008 년 에픽 센터 균주에서 리치몬드, 버지니아에서 노인 그룹 홈에서 수집되었다 OH, 신시내티에서 2008 년에 수집 하였다. LC 50 높은 값은 더 높은 농도의 50 %를 죽이는 데 필요한 것을 지시리치몬드와 에픽 센터 균주에서 테스트 인구 계란입니다. 계란과 유사하게, 리치몬드와 에픽 센터 인구의 첫 번째 령충은 테스트 인구의 50 %를 죽이는 살충제의 높은 농도 필요합니다. 다른 문자 다음에 LC (50) 값은 크게 다르다. <img alt="그림 1" class="xfigimg" src="/files/ftp_upload/55092/55092fig1.jpg" /> 변형 된 원심 분리 튜브 1. 사진도. 메쉬의 사각형 조각 뚜껑에 나사 결합 될 수 있도록 홀이 캡으로 절단되었다. 메쉬 편 파기 다른 살충제를 사용할 때 교체 될 수있다. 침대 버그 계란 침지 분석에 대한 살충제 솔루션에 페인트 브러시를 사용하여 메시에 직접 배치하고 침수된다. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55092/55092fig1large.jpg" target="_blank">이 figur의 더 큰 버전을 보려면 여기를 클릭하십시오</a>이자형. <img alt="그림 2" class="xfigimg" src="/files/ftp_upload/55092/55092fig2.jpg" /> 용량 - 반응 그래프 2. 표현한 도표. 이미다클로 프리드 / β-cyfluthrin에 할란 균주 침대 벌레 계란 백분율 응답 (사망률)은 Y 축과 빈대는 x 축 상에 표현되는 노출되었다고 로그 농도에 도시된다. 용량이 증가하고 사망률이 시험에서 가장 높은 농도에서 100 %에 도달로 계란 사망률은 증가했다. <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 라이프 스테이지 / 스트레인 </td><td colspan="2"> 엔 </td><td> LC (50) (95 % CI) </td><td> 경사 ± SE </td><td> X 2 (DF) </td></tr><tr><td> 달걀 </td><td colspan="2"></td><td></td><td></td><td></td></tr><tr><td> 할란 </td><td colspan="2"> (250) </td><td> 0.41a (<html> 0.276-0.548) </td><td> 1.86 ± 0.24 </td><td> 33.42 (23) </td></tr><tr><td> 리치몬드 </td><td colspan="2"> (320) </td><td> 1.23b (0.59-2.10) </td><td> 1.13 ± 0.14 </td><td> 82.57 (30) </td></tr><tr><td> 에픽 센터 </td><td colspan="2"> (400) </td><td> 2.10b (1.049-4.587) </td><td> 0.95 ± 0.10 </td><td> 149.91 (38) </td></tr><tr><td> 먼저 령충 </td><td colspan="2"></td><td></td><td></td><td></td></tr><tr><td> 할란 </td><td colspan="2"> (150) </td><td> 0.04a (0.030-0.063) </td><td> 2.16 ± 0.34 </td><td> 38.17 (28) </td></tr><tr><td> 리치몬드 </td><td colspan="2"> 195 </td><td> 4.81b (1.94-10.26) </td><td> 0.66 ± 0.12 </td><td> 45.87 (37) </td></tr><tr><td> 에픽 센터 </td><td colspan="2"> (190) </td><td> 19.72b (8.18-184.48) </td><td> 0.75 ± 0.17 </td><td> 45.39 (36) </td></tr></tbody></table>&quot;&gt; 침대 버그 계란과 첫 번째 령 LC 캠벨과 밀러 2015 (13) 수정 (50) 값의 표 1. 비교.

Watch this Scientific Journal Video about A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars at JoVE.com

A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars

살충제의 평가는 종종 성인 곤충보다는 미성숙 단계에 대해 목표로하고 있습니다. 여기, 우리는 첫 번째 nymphal 침대 버그 단계에 대한 비교와 함께 침대 버그 계란에 대한 살충제를 평가하기위한 프로토콜을 제시한다. 이러한 프로토콜은 비 성인의 삶의 단계에서 살충제의 효능을 평가하기 위해 다른 곤충에 따라 조정 될 수있다.

베드 버그에 대해 살충제 효능을 평가하는 방법에있어서,<em&gt; CIMEX lectularius</em&gt;, 계란과 첫 령충

Standard toxicity evaluations of insecticides against insect pests are primarily conducted on adult insects. Evaluations are based on a dose-response or ...

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A Method for Evaluating Insecticide Efficacy against Bed Bug, Cimex lectularius, Eggs and First Instars

11.2K 조회수.  University of Florida.  살충제의 평가는 종종 성인 곤충보다는 미성숙 단계에 대해 목표로하고 있습니다. 여기, 우리는 첫 번째 nymphal 침대 버그 단계에 대한 비교와 함께 침대 버그 계란에 대한 살충제를 평가하기위한 프로토콜을 제시한다. 이러한 프로토콜은 비 성인의 삶의 단계에서 살충제의 효능을 평가하기 위해 다른 곤충에 따라 조정 될 수있다. 

비디오: A Method for Evaluating Insecticide Efficacy against Bed Bug,  Cimex lectularius, Eggs and First Instars

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.

옥수수와 테오신테 라인에 우스틸라고 성 실증의 병원성을 평가하기위한 신속하고 효율적인 방법

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

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환경과학

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and enables to perform rapid, automated and simple SPE. The pre-concentrated solution is compatible with analysis by immunoassay, with a low organic solvent content. A method is described for the extraction and pre-concentration of natural hormone 17&#946;-estradiol in 100 ml water samples. Reverse phase SPE is performed with octadecyl-silica sorbent and elution is done with 200 &#181;l of methanol 50% v/v. Eluent is diluted by adding di-water to lower the amount of methanol. After preparing manually the SPE column, the overall procedure is performed automatically within 1 hr. At the end of the process, estradiol concentration is measured by using a commercial enzyme-linked immune-sorbent assay (ELISA). 100-fold pre-concentration is achieved and the methanol content in only 10% v/v. Full recoveries of the molecule are achieved with 1 ng/L spiked de-ionized and synthetic sea water samples.

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

작은 오염 물질의 면역 학적 분석을위한 물 샘플의 자동화 된 고체상 추출을위한 간단한 방법

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in a watch list of environmental pollutants under the European Water Framework Directive. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. Much of the estrogenic potency of wastewater treatment plant (WWTP) effluents can be attributed to the discharge of steroid estrogens including estradiol (E2), EE2 and E1 due to incomplete removal of these substances at the treatment plant. An evaluation of the efficacy of wastewater treatment processes requires the quantitative determination of individual substances most often undertaken using chemical analysis methods. Most frequently used methods include Gas Chromatography-Mass Spectrometry (GCMS/MS) or Liquid Chromatography-Mass Spectrometry (LCMS/MS) using multiple reaction monitoring (MRM). Although very useful for regulatory purposes, targeted chemical analysis can only provide data on the compounds (and specific metabolites) monitored. Ecotoxicology methods additionally ensure that any by-products produced or unknown estrogenic compounds present are also assessed via measurement of their biological activity. A number of in vitro bioassays including the Yeast Estrogen Screen (YES) are available to measure the estrogenic activity of wastewater samples. Chemical analysis in conjunction with in vivo and in vitro bioassays provides a useful toolbox for assessment of the efficacy and suitability of wastewater treatment processes with respect to estrogenic endocrine disrupting compounds. This paper utilizes a battery of chemical and ecotoxicology tests to assess conventional, advanced and emerging wastewater treatment processes in laboratory and field studies.

Endocrine Disrupting Compounds (EDC) pose a substantial risk to the aquatic environment. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. The methodology provided in this paper allows for an assessment of the efficacy and suitability of wastewater treatment processes with respect to EDC removal.

폐수 처리 프로세스의 효능의 평가를위한 화학 및 생체 독성 방법의 배터리를 사용하면 에스트로겐 효능을 제거하는 방법

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in ...

A Gusseted Thermogradient Table to Control Soil Temperatures for Evaluating Plant Growth and Monitoring Soil Processes

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a series of incubators. A temperature gradient is passively established across the surface of the table between the heated and cooled ends and is lost quickly at distances above the surface. Since temperature is only controlled on the table surface, experiments are restricted to shallow containers, such as Petri dishes, placed on the table. Welding continuous aluminum vertical strips or gussets perpendicular to the surface of a table enables temperature control in depth via convective heat flow. Soil in the channels between gussets was maintained across a gradient of temperatures allowing a greater diversity of experimentation. The gusseted design was evaluated by germinating oat, lettuce, tomato, and melon seeds. Soil temperatures were monitored using individual, battery-powered dataloggers positioned across the table. LED lights installed in the lids or along the sides of the gradient table create a controlled temperature chamber where seedlings can be grown over a range of temperatures. The gusseted design enabled accurate determination of optimum temperatures for fastest germination rate and the highest percentage germination for each species. Germination information from gradient table experiments can help predict seed germination and seedling growth under the adverse soil conditions often encountered during field crop production. Temperature effects on seed germination, seedling growth, and soil ecology can be tested under controlled conditions in a laboratory using a gusseted thermogradient table.

Traditional thermogradient tables create a range of temperatures across the surface. Welding gussets perpendicular to the surface of a thermogradient table will control temperature in depth increasing possible research applications.

거싯 Thermogradient 표는 토양 프로세스 식물 성장 평가 및 모니터링을위한 토양 온도를 제어하는

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

이 필드 조건에서 종자 발아, 휴면 및 사망률을 평가하기위한 신뢰할 수있는 방법

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

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.

수산 생명에 일시 중지를 평가하면 퇴적물 효과에 대한 플로우를 통해 노출 시스템

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

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition.
An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. 
This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step.

The article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples.

많은 샘플을 이용한 실험에서 토양 미생물을 특성화하기위한 지질 추출 및 분석 방법

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial ...

Image-based Lagrangian Particle Tracking in Bed-load Experiments

Image analysis has been increasingly used for the measurement of river flows due to its capabilities to furnish detailed quantitative depictions at a relatively low cost. This manuscript describes an application of particle tracking velocimetry (PTV) to a bed-load experiment with lightweight sediment. The key characteristics of the investigated sediment transport conditions were the presence of a covered flow and of a fixed rough bed above which particles were released in limited number at the flume inlet. Under the applied flow conditions, the motion of the individual bed-load particles was intermittent, with alternating movement and stillness terms. The flow pattern was preliminarily characterized by acoustic measurements of vertical profiles of the stream-wise velocity. During process visualization, a large field of view was obtained using two action-cameras placed at different locations along the flume. The experimental protocol is described in terms of channel calibration, experiment realization, image pre-processing, automatic particle tracking, and post-processing of particle track data from the two cameras. The presented proof-of-concept results include probability distributions of the particle hop length and duration. The achievements of this work are compared to those of existing literature to demonstrate the validity of the protocol.

The manuscript presents a protocol for the conduction of bed-load sediment transport experiments where the moving particles are tracked by image analysis. The experimental facility, the procedures for run realization and data processing, and finally some proof-of-concept results are presented here.

베드로드 실험에서 이미지 기반 라그랑주 입자 추적

Image analysis has been increasingly used for the measurement of river flows due to its capabilities to furnish detailed quantitative depictions at a ...

A Novel Method for the Pentosan Analysis Present in Jute Biomass and Its Conversion into Sugar Monomers Using Acidic Ionic Liquid

Recently, ionic liquids (ILs) are used for biomass valorization into valuable chemicals because of their remarkable properties such as thermal stability, lower vapor pressure, non-flammability, higher heat capacity, and tunable solubility and acidity. Here, we demonstrate a method for the synthesis of C5 sugars (xylose and arabinose) from the pentosan present in jute biomass in a one-pot process by utilizing a catalytic amount of Br&#248;nsted acidic 1-methyl-3-(3-sulfopropyl)-imidazolium hydrogen sulfate IL. The acidic IL is synthesized in the lab and characterized using NMR spectroscopic techniques for understanding its purity. The various properties of BAIL are measured such as acid strength, thermal and hydrothermal stability, which showed that the catalyst is stable at a higher temperature (250 &#176;C) and possesses very high acid strength (Ho 1.57). The acidic IL converts over 90% of pentosan into sugars and furfural. Hence, the presenting method in this study can also be employed for the evaluation of pentosan concentration in other kinds of lignocellulosic biomass.

We present a protocol for the synthesis of C5 sugars (xylose and arabinose) from a renewable non-edible lignocellulosic biomass (i.e., jute) with the presence of Br&#248;nsted acidic ionic liquids (BAILs) as the catalyst in water. The BAILs catalyst exhibited better catalytic performance than conventional mineral acid catalysts (H2SO4 and HCl).

당 분석에 대 한 새로운 방법을 황 바이오 매스 및 산 성 이온 성 액체를 사용 하 여 설탕 단위체로 그것 변환에

Recently, ionic liquids (ILs) are used for biomass valorization into valuable chemicals because of their remarkable properties such as thermal stability, ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

The efficacy of public health pesticides targeting nuisance and disease-vector insects is not uniform across different ecological zones. Here we present a system of techniques using captive vector insects as sentinels for pesticide efficacy to derive electronic maps supporting the standard evaluation of pesticides across multiple environments.

시각화 분야에서 질병 벡터 모기에 대 한 살충제의 효능

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...