<documentid="5C9EFABED8EFCA6446DF2CD9AC9490B2"ID-CLB-Dataset="41381"ID-DOI="10.11646/zootaxa.3949.3.1"ID-GBIF-Dataset="0d89e1d1-ae71-438f-8fdc-8d48de0aa8d6"ID-ISSN="1175-5326"ID-Zenodo-Dep="254206"ID-ZooBank="43B43A7C-644C-4B46-BAAF-522C3C1FDC7D"IM.illustrations_approvedBy="admin"IM.materialsCitations_approvedBy="felipe"IM.metadata_approvedBy="felipe"IM.tables_requiresApprovalFor="operationResults"IM.taxonomicNames_approvedBy="admin"IM.treatments_approvedBy="admin"checkinTime="1461045905660"checkinUser="plazi"docAuthor="Borkent, Art & Brown, Brian V."docDate="2015"docId="BE4D120DED07B525FF26FCDBBB2E7919"docLanguage="en"docName="zt03949p322.pdf"docOrigin="Zootaxa 3949 (3)"docStyle="DocumentStyle:647186512141C8FC8976D5BCC54AEB7D.9:Zootaxa.2013-.journal_article"docStyleId="647186512141C8FC8976D5BCC54AEB7D"docStyleName="Zootaxa.2013-.journal_article"docStyleVersion="9"docTitle="Diptera Linnaeus 1758"docType="treatment"docVersion="12"lastPageNumber="318"masterDocId="42746A75ED03B534FFB1FF9FBA49787C"masterDocTitle="How to inventory tropical flies (Diptera) — One of the megadiverse orders of insects"masterLastPageNumber="322"masterPageNumber="301"pageNumber="305"updateTime="1720401617231"updateUser="ExternalLinkService">
survey of a mid-elevation tropical cloud forest to the National Science Foundation (
<collectingCountryid="4EF3E38BED07B530FE4DFC31B87E7BBA"box="[508,567,942,966]"name="United States of America"pageId="4"pageNumber="305">USA</collectingCountry>
) (see http://phorid.net/zadbi/ and YouTube link https://www.youtube.com/ watch?v=HkROS6-K02U). We chose a study site at Zurquí de Moravia (hereafter "Zurquí), San José Province,
, for a number of reasons. From previous prospecting, this site was known among some entomologists as remarkably rich in insect species (
<bibRefCitationid="5275DEEAED07B530FD9EFB85B8867C4E"author="Hanson"box="[559,719,1050,1074]"pageId="4"pageNumber="305"refString="Hanson, P. (2000) 4. Insects and Spiders. In: Nadkarni, N. M. & Wheelwright, N. T. (Eds.), Monteverde, ecology and conservation of a tropical cloud forest. Oxford University Press, New York, Oxford, 95 - 147."type="book chapter"year="2000">Hanson 2000</bibRefCitation>
). The site is also easily accessible, about a 30 minute drive northeast of San José and, although privately owned, we had assurances from the owner, Jorge Arturo Lizano, that it will remain protected. Our study site, at 1585 meters elevation, was limited to a 150 x
<figureCitationid="AEDFBF9EED07B530FA90FBFEBF297C06"box="[1313,1376,1121,1146]"captionStart="FIGURE 1"captionStartId="7.[151,250,2010,2032]"captionTargetBox="[151,1426,807,1973]"captionTargetId="figure@7.[151,1436,807,1985]"captionTargetPageId="7"captionText="FIGURE 1. Map of study site at Zurquí de Moravia, Costa Rica, with primary features indicated."httpUri="https://zenodo.org/record/254207/files/figure.png"pageId="4"pageNumber="305">Fig. 1</figureCitation>
) that included a small ridge, a permanent stream, a temporary stream and a variety of vegetation, including some disturbed habitat (pasture) (
<figureCitationid="AEDFBF9EED07B530FE62FB36B8087CBE"box="[467,577,1193,1218]"captionStart-0="FIGURE 2"captionStart-1="FIGURE 3"captionStartId-0="8.[151,250,1934,1956]"captionStartId-1="9.[151,250,1953,1975]"captionTargetBox-0="[151,1436,511,1904]"captionTargetBox-1="[151,1436,194,1930]"captionTargetId-0="figure@8.[151,1436,193,1913]"captionTargetId-1="figure@9.[151,1436,193,1932]"captionTargetPageId-0="8"captionTargetPageId-1="9"captionText-0="FIGURE 2. Details of study site at Zurquí de Moravia, Costa Rica. A. Primary cloud forest with bordering pasture. B. Primary Malaise trap set on ridge indicated with black arrow; red arrow points to temporary black light set over pan with soapy water; large white mass in middle of photo was a piece of plastic garbage."captionText-1="FIGURE 3. Details of study site at Zurquí de Moravia, Costa Rica. A. Supplementary Malaise trap beside narrow, permanent stream. B. Permanent stream."httpUri-0="https://zenodo.org/record/254208/files/figure.png"httpUri-1="https://zenodo.org/record/254209/files/figure.png"pageId="4"pageNumber="305">Figs. 2–3</figureCitation>
). It is abutting the extensive and virtually pristine Braulio Carillo National Park. Our baseline sampling was a single Malaise trap set on the ridge at the edge of the forest (
<figureCitationid="AEDFBF9EED07B530FB43FB52BF717C9A"box="[1266,1336,1229,1254]"captionStart="FIGURE 2"captionStartId="8.[151,250,1934,1956]"captionTargetBox="[151,1436,511,1904]"captionTargetId="figure@8.[151,1436,193,1913]"captionTargetPageId="8"captionText="FIGURE 2. Details of study site at Zurquí de Moravia, Costa Rica. A. Primary cloud forest with bordering pasture. B. Primary Malaise trap set on ridge indicated with black arrow; red arrow points to temporary black light set over pan with soapy water; large white mass in middle of photo was a piece of plastic garbage."httpUri="https://zenodo.org/record/254208/files/figure.png"pageId="4"pageNumber="305">Fig. 2</figureCitation>
B), to be employed for one year, the results of which were to be studied by all cooperating systematists (
<tableCitationid="7B6696A0ED07B530FA81FB6EBFC27D76"box="[1328,1419,1265,1290]"captionStart="TABLE 1"captionStartId="4.[151,239,1372,1395]"captionTargetBox="[159,1108,1454,1972]"captionText="TABLE 1. Families present at Zurquí de Moravia, Costa Rica with associated collaborating systematists. Families in partial phyletic sequence."httpUri="http://table.plazi.org/id/629BF393ED07B530FF26FAC3BBEE7DEF"pageId="4"pageNumber="305"tableUuid="629BF393ED07B530FF26FAC3BBEE7DEF">Table 1</tableCitation>
<tdid="CB05C825ED074ACBFD82F8D3BE1D7F1E"box="[563,1108,1868,1890]"gridcol="1"gridrow="10"pageId="4"pageNumber="305">Art Borkent, Gustavo R. Spinelli, Maria M. Ronderos</td>
<tdid="CB05C825ED064ACBFD82FEF1BF2479F8"box="[563,1389,366,388]"gridcol="1"gridrow="4"pageId="5"pageNumber="306">Gregory R. Curler, Sergio Ibáñez-Bernal, Gunnar M. Kvifte</td>
<tdid="CB05C825ED064ACBFD82FCAABF247B16"box="[563,1389,821,874]"gridcol="1"gridrow="15"pageId="5"pageNumber="306">Marc Pollet (coord.), Daniel J. Bickel, Scott E. Brooks, Renato Capellari, Neal L. Evenhuis, Stefan Naglis, Justin Runyon</td>
<tdid="CB05C825ED064ACBFD82FCE2BF247BEF"box="[563,1389,893,915]"gridcol="1"gridrow="16"pageId="5"pageNumber="306">Jeffrey M. Cumming, Bradley J. Sinclair</td>
<tdid="CB05C825ED064ACBFD82FC4FBF247B9A"box="[563,1389,976,998]"gridcol="1"gridrow="18"pageId="5"pageNumber="306">Christian Thompson, Manuel Zumbado</td>
<tableNoteid="CB02A295ED06B531FBE8F82FBFD97FBA"box="[1113,1424,1968,1990]"pageId="5"pageNumber="306"targetBox="[159,1389,201,1948]"targetPageId="5">......continued on the next page</tableNote>
To ensure that our samples would be fully identified we approached over 50 fellow systematists. Nearly all had been involved in the previously published MCAD (
<bibRefCitationid="5275DEEAED05B532FCBCFBCBB9B27C10"author="Brown"box="[781,1019,1108,1132]"pageId="6"pageNumber="307"refString="Brown, B. V., Borkent, A., Cumming, J. M., Wood, D. M., Woodley, N. E. & Zumbado, M. A. (Eds.) (2009) Manual of Central American Diptera. Vol. 1. National Research Council Press, Ottawa, xi + 714 pp. [Canada]"type="book"year="2009">
<bibRefCitationid="5275DEEAED05B532FBBFFBCBBE0E7C10"author="Brown"box="[1038,1095,1108,1132]"pageId="6"pageNumber="307"refString="Brown, B. V., Borkent, A., Cumming, J. M., Wood, D. M., Woodley, N. E. & Zumbado, M. A. (Eds.) (2011) Manual of Central American Diptera. Vol. 2. National Research Council Press, Ottawa, xvi + 727 pp. [Canada, pp. 715 - 1442.]"type="book chapter"year="2011">2011</bibRefCitation>
) and they gave enthusiastic endorsement of this new project when we approached them.
<tableCitationid="7B6696A0ED05B532FB25FB03BEAE7CC8"box="[1172,1255,1180,1205]"captionStart="TABLE 1"captionStartId="4.[151,239,1372,1395]"captionTargetBox="[159,1108,1454,1972]"captionText="TABLE 1. Families present at Zurquí de Moravia, Costa Rica with associated collaborating systematists. Families in partial phyletic sequence."httpUri="http://table.plazi.org/id/629BF393ED07B530FF26FAC3BBEE7DEF"pageId="6"pageNumber="307"tableUuid="629BF393ED07B530FF26FAC3BBEE7DEF">Table 1</tableCitation>
expected, many of the discovered species proved to be undescribed and, as such, are for the present given a morphospecies code number, entered into the database, and, if not described soon, will be housed in the established collections at the Instituto Nacional de Biodiversidad (INBio),
and Natural History Museum of Los Angeles County, California,
<collectingCountryid="4EF3E38BED05B532FE69FA23B8597DA8"box="[472,528,1468,1492]"name="United States of America"pageId="6"pageNumber="307">USA</collectingCountry>
. The institution provides logistical support and serves as a center for incorporating results into their database and collection. To assist with sampling and curation of material we hired five technicians, all of whom were trained and had extensive previous field and lab experience at INBio (
<figureCitationid="AEDFBF9EED05B532FC92F9D3B9D97E19"box="[803,912,1612,1637]"captionStart-0="FIGURE 4. A"captionStart-1="FIGURE 5. A"captionStartId-0="10.[151,250,1913,1935]"captionStartId-1="11.[151,250,1934,1956]"captionTargetBox-0="[166,1408,193,1874]"captionTargetBox-1="[151,1435,193,1905]"captionTargetId-0="figure@10.[165,1409,193,1876]"captionTargetId-1="figure@11.[151,1436,193,1913]"captionTargetPageId-0="10"captionTargetPageId-1="11"captionText-0="FIGURE 4. A. The ZADBI team: Back row, left to right: Manual Zumbado (Coordinator of Biosciences at INBio), Brian Brown (co-PI), Art Borkent (co-PI). Front row, left to right: Elena Ulate (technician), Wendy Porras (technician), Anna Holden (project manager), Carolina Avila (technician), Annia Picado (technician), Marco Moraga (technician). B. Annia Picado (technician) preparing slide mounts."captionText-1="FIGURE 5. A. Wendy Porras (technician) pinning freshly collected specimens. B. Brian Brown teaching Carolina Avila (technician) how to dry specimens using ethyl acetate."httpUri-0="https://zenodo.org/record/254210/files/figure.png"httpUri-1="https://zenodo.org/record/254211/files/figure.png"pageId="6"pageNumber="307">Figs. 4–5</figureCitation>
). The individual talents of our team—Carolina Avila, Marco Moraga (who moved away during this study), Annia Picado, Wendy Porras, Elena Ulate, and Elvia Zumbado (more recently hired)—made our project possible. At the start of our project, most could already identify
to at least family level and there was a high level of skill in sorting and preparing both slide-mounted and pinned material. Hiring locals had huge advantages in lower costs, less damage to specimens during transport from the field and also contributed to local structure and economy.
We also hired a project manager at the Natural History Museum of Los Angeles County (Anna Holden for the first portion of our project and Estella Hernandez in the latter part) to oversee the mechanical details of our project including ordering supplies, helping to organize the technicians, assisting with the initial set up of sampling equipment, ensuring that specimens flowed well throughout our curatorial system and loans were arranged correctly. Our project manager also provided educational outreach at the Natural History Museum of Los Angeles County.
a strong educational component of our project, the museum promoted flies in educational programming, offered resources for teachers, collaborated with the Encyclopedia of Life and encouraged the use of social media.
To ensure the highest levels of curation we approached each of our collaborators to ask for their exact requirements. Because of the nature of their groups and the history of their preparation, many had different curatorial requests. Some systematists, in addition, had their own idiosyncratic method for handling material. The result was the preparation of a 33 page protocol manual, detailing the curatorial needs for each family (summarized in
<figureCitationid="AEDFBF9EED04B533FF06FEF0BB4479F4"box="[183,269,367,392]"captionStart="FIGURE 10"captionStartId="17.[151,250,1270,1292]"captionTargetBox="[161,1429,420,1232]"captionTargetId="figure@17.[151,1436,411,1248]"captionTargetPageId="17"captionText="FIGURE 10. Sorting protocol after a sample has been collected and databased (location, date, method of collection). Many families required further specific manipulation or arrangement, depending on the needs of the systematist (e. g. arrangement of parts on a microscope slide; size of pin or specific position on a pin). HMDS (hexamethyldisilazane) is a liquid used to dry insects through immersion and subsequent evaporation (Heraty & Hawks, 1998)."httpUri="https://zenodo.org/record/254216/files/figure.png"pageId="7"pageNumber="308">Fig. 10</figureCitation>
). Collaborators needing slide preparations, for example, differed in how they wanted dissected parts arranged and whether they wanted specimens in Euparal or
Balsam as the final mounting medium. Individuals requesting pinned specimens wanted their specimens either on points (white or gray paper), glued directly to the pin, pinned through the thorax or on secondary minuten pins. We needed to teach the technicians these various additional protocols. To ensure highest quality, we sent out an initial small batch of prepared specimens to each collaborator for them to give us their appraisal and further instructions. Our goal was to produce "perfect" specimens as far as possible. Generally, the resulting specimens were in very good to excellent condition and for collaborators, pleasurable to study. Each specimen was fully labeled, including a unique barcode number, and entered into a database, to be housed at LACM and duplicated at INBio (ATTA system). The care given to all specimens produced material allowing our collaborating systematists to immediately begin employing their expertise in distinguishing species.
Details of study site at Zurquí de Moravia, Costa Rica. A. Primary cloud forest with bordering pasture. B. Primary Malaise trap set on ridge indicated with black arrow; red arrow points to temporary black light set over pan with soapy water; large white mass in middle of photo was a piece of plastic garbage.
A. The ZADBI team: Back row, left to right: Manual Zumbado (Coordinator of Biosciences at INBio), Brian Brown (co-PI), Art Borkent (co-PI). Front row, left to right: Elena Ulate (technician), Wendy Porras (technician), Anna Holden (project manager),
Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Emergence trap over wet vegetation. B. Black light over tray with soapy water.
Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. bucket light trap (with Wendy Porras and Art Borkent). B. CDC light trap. C. Sweeping at the site (left to right, Marco Moraga, Annia Picado, Art Borkent).
Our sampling with a single Malaise trap at Zurquí (
<figureCitationid="AEDFBF9EED13B524FCB2FF08B90F78CC"box="[771,838,151,176]"captionStart="FIGURE 2"captionStartId="8.[151,250,1934,1956]"captionTargetBox="[151,1436,511,1904]"captionTargetId="figure@8.[151,1436,193,1913]"captionTargetPageId="8"captionText="FIGURE 2. Details of study site at Zurquí de Moravia, Costa Rica. A. Primary cloud forest with bordering pasture. B. Primary Malaise trap set on ridge indicated with black arrow; red arrow points to temporary black light set over pan with soapy water; large white mass in middle of photo was a piece of plastic garbage."httpUri="https://zenodo.org/record/254208/files/figure.png"pageId="16"pageNumber="317">Fig. 2</figureCitation>
B) was proposed to our collaborators as the minimum requirement for identifying species. We also placed an additional continuous Malaise trap near the permanent stream at the bottom of the ravine, a habitat appearing substantially different to us (
<figureCitationid="AEDFBF9EED13B524FBECFF40BEEB7884"box="[1117,1186,223,248]"captionStart="FIGURE 3"captionStartId="9.[151,250,1953,1975]"captionTargetBox="[151,1436,194,1930]"captionTargetId="figure@9.[151,1436,193,1932]"captionTargetPageId="9"captionText="FIGURE 3. Details of study site at Zurquí de Moravia, Costa Rica. A. Supplementary Malaise trap beside narrow, permanent stream. B. Permanent stream."httpUri="https://zenodo.org/record/254209/files/figure.png"pageId="16"pageNumber="317">Fig. 3</figureCitation>
A). Although Malaise traps were our primary collecting method, we were keenly aware that some flies would not be collected by this method alone. We therefore had a team of two technicians go to the site once a month for three days (including three nights) to utilize various other methods of collecting (
<figureCitationid="AEDFBF9EED13B524FCC4FED3B9A07919"box="[885,1001,332,357]"captionStart-0="FIGURE 6"captionStart-1="FIGURE 7"captionStart-2="FIGURE 8"captionStart-3="FIGURE 9"captionStartId-0="12.[151,250,1926,1948]"captionStartId-1="13.[151,250,1926,1948]"captionStartId-2="14.[151,250,1910,1932]"captionStartId-3="15.[151,250,1953,1975]"captionTargetBox-0="[152,1436,194,1899]"captionTargetBox-1="[152,1436,194,1899]"captionTargetBox-2="[151,1436,194,1879]"captionTargetBox-3="[151,1436,193,1930]"captionTargetId-0="figure@12.[151,1436,193,1904]"captionTargetId-1="figure@13.[151,1436,193,1904]"captionTargetId-2="figure@14.[151,1436,193,1889]"captionTargetId-3="figure@15.[151,1436,193,1932]"captionTargetPageId-0="12"captionTargetPageId-1="13"captionTargetPageId-2="14"captionTargetPageId-3="15"captionText-0="FIGURE 6. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Emergence trap over wet vegetation. B. Black light over tray with soapy water."captionText-1="FIGURE 7. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. bucket light trap (with Wendy Porras and Art Borkent). B. CDC light trap. C. Sweeping at the site (left to right, Marco Moraga, Annia Picado, Art Borkent)."captionText-2="FIGURE 8. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Flight intercept trap. B. Bat (Sturnira ludovici Anthony) trapped with mist net and examined for bat flies (Streblidae) by Carl Dick."captionText-3="FIGURE 9. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A-B. Bait traps. C. Brian Brown observing fly behaviour."httpUri-0="https://zenodo.org/record/254212/files/figure.png"httpUri-1="https://zenodo.org/record/254213/files/figure.png"httpUri-2="https://zenodo.org/record/254214/files/figure.png"httpUri-3="https://zenodo.org/record/254215/files/figure.png"pageId="16"pageNumber="317">Figs. 6–9</figureCitation>
). We rented a cabin at the site for accommodation and storage of some traps when these were not in use (e.g. light traps). This effort resulted in numerous additional samples from techniques including hand collecting (
<figureCitationid="AEDFBF9EED13B524FC68FE0BBE5479D0"box="[985,1053,404,429]"captionStart="FIGURE 9"captionStartId="15.[151,250,1953,1975]"captionTargetBox="[151,1436,193,1930]"captionTargetId="figure@15.[151,1436,193,1932]"captionTargetPageId="15"captionText="FIGURE 9. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A-B. Bait traps. C. Brian Brown observing fly behaviour."httpUri="https://zenodo.org/record/254215/files/figure.png"pageId="16"pageNumber="317">Fig. 9</figureCitation>
C), sweep netting (
<figureCitationid="AEDFBF9EED13B524FB44FE0BBF7379D0"box="[1269,1338,404,429]"captionStart="FIGURE 7"captionStartId="13.[151,250,1926,1948]"captionTargetBox="[152,1436,194,1899]"captionTargetId="figure@13.[151,1436,193,1904]"captionTargetPageId="13"captionText="FIGURE 7. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. bucket light trap (with Wendy Porras and Art Borkent). B. CDC light trap. C. Sweeping at the site (left to right, Marco Moraga, Annia Picado, Art Borkent)."httpUri="https://zenodo.org/record/254213/files/figure.png"pageId="16"pageNumber="317">Fig. 7</figureCitation>
of light trapping (CDC, bucket traps, black light over pans of soapy water) (
<figureCitationid="AEDFBF9EED13B524FB39FE28BE9F79AC"box="[1160,1238,439,464]"captionStart="FIGURE 6"captionStartId="12.[151,250,1926,1948]"captionTargetBox="[152,1436,194,1899]"captionTargetId="figure@12.[151,1436,193,1904]"captionTargetPageId="12"captionText="FIGURE 6. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Emergence trap over wet vegetation. B. Black light over tray with soapy water."httpUri="https://zenodo.org/record/254212/files/figure.png"pageId="16"pageNumber="317">Figs. 6</figureCitation>
B, 7A–B), baiting with various attractants (fruit, carrion) (
<figureCitationid="AEDFBF9EED13B524FDEAFE43B8E37988"box="[603,682,476,501]"captionStart="FIGURE 9"captionStartId="15.[151,250,1953,1975]"captionTargetBox="[151,1436,193,1930]"captionTargetId="figure@15.[151,1436,193,1932]"captionTargetPageId="15"captionText="FIGURE 9. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A-B. Bait traps. C. Brian Brown observing fly behaviour."httpUri="https://zenodo.org/record/254215/files/figure.png"pageId="16"pageNumber="317">Figs. 9</figureCitation>
A–B), emergence traps (
<figureCitationid="AEDFBF9EED13B524FC70FE43BE4D7989"box="[961,1028,476,501]"captionStart="FIGURE 6"captionStartId="12.[151,250,1926,1948]"captionTargetBox="[152,1436,194,1899]"captionTargetId="figure@12.[151,1436,193,1904]"captionTargetPageId="12"captionText="FIGURE 6. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Emergence trap over wet vegetation. B. Black light over tray with soapy water."httpUri="https://zenodo.org/record/254212/files/figure.png"pageId="16"pageNumber="317">Fig. 6</figureCitation>
A), a flight intercept trap (
<figureCitationid="AEDFBF9EED13B524FA82FE43BF3F7988"box="[1331,1398,476,501]"captionStart="FIGURE 8"captionStartId="14.[151,250,1910,1932]"captionTargetBox="[151,1436,194,1879]"captionTargetId="figure@14.[151,1436,193,1889]"captionTargetPageId="14"captionText="FIGURE 8. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Flight intercept trap. B. Bat (Sturnira ludovici Anthony) trapped with mist net and examined for bat flies (Streblidae) by Carl Dick."httpUri="https://zenodo.org/record/254214/files/figure.png"pageId="16"pageNumber="317">Fig. 8</figureCitation>
southeast of Zurquí, respectively, for the same one year period as that at Zurquí. This additional sampling substantially increased the amount of material of most groups. Even so, every collaborator wanted to study all the specimens from the supplementary trapping at Zurquí and nearly all also wanted the additional material from Tapantí National Park and Las Alturas.
Collecting by specialists can often strikingly increase the number of species obtained. A few examples illustrate this point. In the summer of 2013, we held the "
Blitz" at Zurquí (see YouTube link https:// www.youtube.com/watch?v=HkROS6-K02U). Nineteen systematists were invited to use their specialized knowledge to obtain species that our trapping program might otherwise miss. Even with the plethora of collecting methods employed, it is often the case that a specialist in a particular group can zero in on those special microhabitats and obtain species not (or rarely) collected by mechanical means. One of these experts, retired Smithsonian Dipterist Wayne Mathis, raised the number of shore flies (family
) sampled at the site from three to 26 species. Additionally, he added three entire families to the inventory that our various traps had not collected (
<figureCitationid="AEDFBF9EED13B524FC48FBFBBE777C00"box="[1017,1086,1124,1149]"captionStart="FIGURE 8"captionStartId="14.[151,250,1910,1932]"captionTargetBox="[151,1436,194,1879]"captionTargetId="figure@14.[151,1436,193,1889]"captionTargetPageId="14"captionText="FIGURE 8. Some supplementary collecting methods utilized at Zurquí de Moravia, Costa Rica. A. Flight intercept trap. B. Bat (Sturnira ludovici Anthony) trapped with mist net and examined for bat flies (Streblidae) by Carl Dick."httpUri="https://zenodo.org/record/254214/files/figure.png"pageId="16"pageNumber="317">Fig. 8</figureCitation>
) larvae were collected from the permanent stream (
<figureCitationid="AEDFBF9EED13B524FA8DFB18BF367CDC"box="[1340,1407,1159,1184]"captionStart="FIGURE 3"captionStartId="9.[151,250,1953,1975]"captionTargetBox="[151,1436,194,1930]"captionTargetId="figure@9.[151,1436,193,1932]"captionTargetPageId="9"captionText="FIGURE 3. Details of study site at Zurquí de Moravia, Costa Rica. A. Supplementary Malaise trap beside narrow, permanent stream. B. Permanent stream."httpUri="https://zenodo.org/record/254209/files/figure.png"pageId="16"pageNumber="317">Fig. 3</figureCitation>
B) and preserved in Carnoy's solution, allowing their chromosomes to be examined and thereby the species identified by Peter Adler.
The handling of material is a crucial component of a successful inventory (
<figureCitationid="AEDFBF9EED13B524FBA5FB6BBE217D70"box="[1044,1128,1268,1293]"captionStart="FIGURE 10"captionStartId="17.[151,250,1270,1292]"captionTargetBox="[161,1429,420,1232]"captionTargetId="figure@17.[151,1436,411,1248]"captionTargetPageId="17"captionText="FIGURE 10. Sorting protocol after a sample has been collected and databased (location, date, method of collection). Many families required further specific manipulation or arrangement, depending on the needs of the systematist (e. g. arrangement of parts on a microscope slide; size of pin or specific position on a pin). HMDS (hexamethyldisilazane) is a liquid used to dry insects through immersion and subsequent evaporation (Heraty & Hawks, 1998)."httpUri="https://zenodo.org/record/254216/files/figure.png"pageId="16"pageNumber="317">Fig. 10</figureCitation>
). Samples in the field were nearly entirely preserved in 95% ethanol; only hand-swept specimens of some larger and/or robust specimens were pinned directly after being collected. Alcohol material in variously sized containers was completely topped up with ethanol in the field to ensure that no sloshing occurred during transport to the laboratory. Such groups as
, amongst others, are often badly damaged in the initial stages of sample collection in other projects, meaning that these taxa are generally in too poor condition to study. Even slight jostling in alcohol can break the legs and antennae of the more sensitive taxa and great care in transport and subsequent handling is critically important. In the laboratory, samples were first databased and separated into fractions: non-Diptera and each of the different families of
(some uncommon families were kept as a group and separated after further curation) (
<figureCitationid="AEDFBF9EED13B524FD8AF9A8B8C67E2C"box="[571,655,1591,1616]"captionStart="FIGURE 10"captionStartId="17.[151,250,1270,1292]"captionTargetBox="[161,1429,420,1232]"captionTargetId="figure@17.[151,1436,411,1248]"captionTargetPageId="17"captionText="FIGURE 10. Sorting protocol after a sample has been collected and databased (location, date, method of collection). Many families required further specific manipulation or arrangement, depending on the needs of the systematist (e. g. arrangement of parts on a microscope slide; size of pin or specific position on a pin). HMDS (hexamethyldisilazane) is a liquid used to dry insects through immersion and subsequent evaporation (Heraty & Hawks, 1998)."httpUri="https://zenodo.org/record/254216/files/figure.png"pageId="16"pageNumber="317">Fig. 10</figureCitation>
). Material which was to be further prepared as dried, pinned material using either ethyl acetate for larger specimens or HMDS (
<bibRefCitationid="5275DEEAED13B524FC98F9C3BE7A7E08"author="Heraty"box="[809,1075,1628,1653]"pageId="16"pageNumber="317"refString="Heraty, J. & Hawks, D. (1998) Hexamethyldisilazane-A chemical alternative for drying insects, Entomological News, 109, 369 - 374."type="journal article"year="1998">Heraty & Hawks, 1998</bibRefCitation>
) for smaller, more delicate taxa (
<figureCitationid="AEDFBF9EED13B524FF2EF9E0BABB7EE4"box="[159,242,1663,1688]"captionStart="FIGURE 10"captionStartId="17.[151,250,1270,1292]"captionTargetBox="[161,1429,420,1232]"captionTargetId="figure@17.[151,1436,411,1248]"captionTargetPageId="17"captionText="FIGURE 10. Sorting protocol after a sample has been collected and databased (location, date, method of collection). Many families required further specific manipulation or arrangement, depending on the needs of the systematist (e. g. arrangement of parts on a microscope slide; size of pin or specific position on a pin). HMDS (hexamethyldisilazane) is a liquid used to dry insects through immersion and subsequent evaporation (Heraty & Hawks, 1998)."httpUri="https://zenodo.org/record/254216/files/figure.png"pageId="16"pageNumber="317">Fig. 10</figureCitation>
) included all specimens in the samples. Further specifics of pinning protocol followed our collaborators' wishes, as indicated above. Other taxa, to be dissected and placed on microscope slides, were generally more abundant and were first examined in ethanol and differing morphotypes selected for subsequent mounting in either
<bibRefCitationid="5275DEEAED13B524FE7FF973BECC7F79"author="Pinder"box="[462,1157,1772,1797]"pageId="16"pageNumber="317"refString="Pinder, L. C. V. (1989) The adult males of Chironomidae (Diptera) of the Holarctic Region-Introduction. In: Wiederholm (Ed.), Chironomidae of the Holarctic Region. Keys and diagnoses. Part 3 - Adult males. Entomologica Scandinavica, 34 (Supplement), pp. 1 - 532."type="journal article"year="1989">Pinder 1989; specific processing varying somewhat by family</bibRefCitation>
). For some of these latter families, specimens were selected for slide-mounting by the specialist (all
), in others by the technicians who had been taught to mount every specimen even suspected of being different. Finally, a few researchers wished to receive their specimens in alcohol, ultimately preparing the specimens themselves, or returning these as identified specimens to be pinned or slide mounted at a later date. Each prepared specimen was databased when it was labeled (including a unique barcode number) and subsequently tracked as batches of specimens were sent to our specialists for identification. With each group of fully prepared specimens sent (and particularly for the first, small batch of specimens) we received feedback on the quality of specimen preparation and in some instances, fine-tuned the subsequent preparation by the technicians.
To track identifications, collaborators were directed to a dedicated website in which collecting data were indicated and species identification and gender could be inserted for each of their specimens. Unnamed morphotypes were indicated with a prefaced number (e.g. "
