identifier	taxonID	type	CVterm	format	language	title	description	additionalInformationURL	UsageTerms	rights	Owner	contributor	creator	bibliographicCitation
921187C58B59FFBC198DFE8AFC4AFE9D.text	921187C58B59FFBC198DFE8AFC4AFE9D.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Phlebotomus perniciosus	<html xmlns:mods="http://www.loc.gov/mods/v3">
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            <p> Phlebotomus perniciosus colony maintenance </p>
            <p> Phlebotomus perniciosus larvae and adult females were obtained from a colony originating from Murcia, Spain, maintained for two decades at the Department of Parasitology, Charles University, Prague, Czech Republic, under controlled conditions as described previously [47]. </p>
            <p> Aza and ecdysone treatment in  P. perniciosus larvae  Phlebotomus perniciosus larvae were fed a diet containing composted and powdered rabbit feces, as described previously [47]. Aza (Sigma-Aldrich, Darmstadt, Germany) and α-ecdysone (Sigma-Aldrich) (the precursor of 20E) were first dissolved in 250 μl of 99% ethanol P.A. and then diluted in 750 μl of 0.9% NaCl (1:4 ethanol/saline), resulting in final concentrations of 1 and 2.5 μg/ml, respectively. Different volumes of diluted Aza were initially tested to determine the appropriate method for incorporating the liquid Aza into the powdered sieved larval food suitable for first-instar larvae (L1). </p>
            <p> To standardize the volume of Aza diluent that could be mixed into the L1 food, we aimed for a pasty texture that would allow the larvae to feed and develop normally. Te chosen mixture consisted of 35 mg of larval food plus 125 μl of 0.9% NaCl. Consequently, Aza and ecdysone final concentrations were 1 μg and 2 μg per milligram of L1 food, respectively. Tese concentrations were based on previous studies with  Lutzomyia longipalpis [42]. Tis mixture was sufficient to feed approximately 200 L 1 larvae in a small plaster pot for 2 days. </p>
            <p>Te experimental groups were as follows: (a) control group: L1 food plus 0.9% NaCl; (b) Aza group: L1 food plus 1 μg/mg of Aza; (c) Aza + Ecd group: L1 food containing 1 μg/mg of Aza plus 2 μg/mg of ecdysone; (d) Ecd group: L1 food containing 2 μg/mg of ecdysone. Each group consisted of three pots, each containing approximately 200 larvae. Each pot received L1 food equivalent to 35 mg of the dry powder mixture containing the specified chemicals, provided twice until the first molting to L 2 larvae was observed in the control group (around 15 days after larval hatching week from eggs).</p>
            <p>Larval development was monitored daily, considering the size of larvae and the number of caudal setae (two in L1 and four in L2). Adult emergence was observed following standard colony maintenance. Te larvae were collected in pools of 15 individuals from each group at 1 and 7 days after first feeding (DAF) for subsequent gene expression analysis. Te larval collection was random, without distinction of larval stage, ensuring that the results reflected the overall impact of the treatments on the larval populations. Te experiments were conducted in triplicate.</p>
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	https://treatment.plazi.org/id/921187C58B59FFBC198DFE8AFC4AFE9D	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Vieira, Cecilia Stahl;Bisogno, Sara;Salvemini, Marco;Telleria, Erich Loza;Volf, Petr	Vieira, Cecilia Stahl, Bisogno, Sara, Salvemini, Marco, Telleria, Erich Loza, Volf, Petr (2024): Azadirachtin disrupts ecdysone signaling and alters sand fly immunity. Parasites & Vectors (526) 17 (1): 1-11, DOI: 10.1186/s13071-024-06589-8, URL: https://doi.org/10.1186/s13071-024-06589-8
921187C58B58FFBD198DFBF3FBADFD1B.text	921187C58B58FFBD198DFBF3FBADFD1B.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Phlebotomus perniciosus	<html xmlns:mods="http://www.loc.gov/mods/v3">
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            <p> Phlebotomus perniciosus gene expression </p>
            <p> Te expression of AMPs att, def 1, def 2, and the ecdysone-related genes EcR, Eip74EF, and Eip75B and serpent was evaluated through quantitative PCR. Quantitative PCR (qPCR) was prepared with 1 ng of cDNA samples, gene-specific primers, and SYBR Green PCR Master mix (Roche) in a 384-well plate to detect the expression of target genes using a LightCycler 480 thermocycler (Roche) in a final volume of 10 μl. Relative gene expression was calculated relative to  P. perniciosus endogenous control genes GAPDH and GPDH [48] using the following cycling conditions: 95 °C for 10 min enzyme activation, initial denaturation at 95 ºC for 10 s, annealing at 60 °C for 20 s; extension at 72 °C for 45 s, repeated 45 times. Expression levels were expressed as the fold change compared to the control groups, using the Pfaffl method [52]. Te three independent experiments were performed with two pools of each sample in technical duplicates (n = 6). </p>
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	https://treatment.plazi.org/id/921187C58B58FFBD198DFBF3FBADFD1B	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Vieira, Cecilia Stahl;Bisogno, Sara;Salvemini, Marco;Telleria, Erich Loza;Volf, Petr	Vieira, Cecilia Stahl, Bisogno, Sara, Salvemini, Marco, Telleria, Erich Loza, Volf, Petr (2024): Azadirachtin disrupts ecdysone signaling and alters sand fly immunity. Parasites & Vectors (526) 17 (1): 1-11, DOI: 10.1186/s13071-024-06589-8, URL: https://doi.org/10.1186/s13071-024-06589-8
921187C58B5FFFBA198DFEF0FE10FD7C.text	921187C58B5FFFBA198DFEF0FE10FD7C.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Phlebotomus perniciosus	<html xmlns:mods="http://www.loc.gov/mods/v3">
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            <p> Development of  P. perniciosus larvae after hormonal modulation </p>
            <p> Phlebotomus perniciosus L 1 larvae were fed L1 food containing Aza, Aza plus ecdysone, or ecdysone, to observe the effect of these components on the molting process from L1 to L2. </p>
            <p>Te group fed with Aza showed significant inhibition of ecdysis when compared to the control group (Fig. 1, P &lt;0.001). Tis effect of Aza on molting was partially rescued by the simultaneous addition of ecdysone in the L1 food (Fig. 1, P &lt;0.01). Te proportion of L 2 in the control group was similar to the ecdysone group but significantly higher than in the Aza and Aza plus ecdysone groups (Fig. 1, P &lt;0.01, Fig. 1, P &lt;0.05).</p>
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	https://treatment.plazi.org/id/921187C58B5FFFBA198DFEF0FE10FD7C	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Vieira, Cecilia Stahl;Bisogno, Sara;Salvemini, Marco;Telleria, Erich Loza;Volf, Petr	Vieira, Cecilia Stahl, Bisogno, Sara, Salvemini, Marco, Telleria, Erich Loza, Volf, Petr (2024): Azadirachtin disrupts ecdysone signaling and alters sand fly immunity. Parasites & Vectors (526) 17 (1): 1-11, DOI: 10.1186/s13071-024-06589-8, URL: https://doi.org/10.1186/s13071-024-06589-8
