AQUATIC INSECTS (ENT 5361) Collecting Aquatic Insects

COLLECTING AQUATIC INSECTS There is an amazing diversity of aquatic insects, but these organisms often go unnoticed because many are secretive and spend most of there life well hidden. However finding and collecting many aquatic insect species is not difficult even with limited equipment. This is a brief overview of techniques you can use for collecting aquatic insects with an emphasis on qualitative methods for collecting as many species as possible using a few inexpensive items. For additional information see: Merritt, R.W., K.W. Cummins, and V.H. Resh. 1996. Design of aquatic insect studies: collecting, sampling and rearing procedures. Pages 12-28 in An introduction to the aquatic insects of North America. Merritt, R.W., K.W. Cummins, editors. Kendall Hunt Publishing, Dubuque, Iowa. Voshell, J. R., Jr. 2002. A guide to common freshwater invertebrates of North America. McDonald & Woodward Publishing, Blacksburg, VA. 442 pp. SAMPLING LOCATIONS Aquatic insects can be found in nearly any type of aquatic environment including lakes, streams, springs, wetlands, puddles, and tree holes. By collecting from as many different water body types as possible, you can maximize the number of species you find since different types support different insect communities. Within a site you can also sample different habitats. For example in streams, there may be riffles, pools, over hanging terrestrial vegetation, aquatic vegetation, leaf packs, and fine sediment. Habitats that are more heterogeneous will have the greater diversity. In general, the most diverse habitats are midsized streams flowing through forested regions with alternating riffle and pool habitats. PRESERVING SPECIMENS You will need to preserve or fix specimens as soon as possible to prevent decomposition. The safest fluids to use are ethanol or isopropyl alcohol. However, isopropyl is poisonous and the ethanol sold by most drugstores contains acetone so you should avoid long term exposure. You can purchase these from a local drugstore for only a few dollars. Depending on which fluid you use, you will need different percentages for preservation. Insects should be preserved in ~75% ethanol or ~30% isopropyl (too much and specimens become brittle, too little and they will rot). However, be aware that these percentages are final amounts and when you pick specimens from water into a vial of alcohol, the preservative percentage will decrease. If a large number of specimens are picked into a vial, it is best to change the alcohol after 24 to 48 hours to prevent decomposition. SAMPLING TECHNIQUES First of all, it is usually best to collect with other people for safety reasons and because several collectors tend to be more effective. For example, several collectors can use different collecting techniques or collect in different areas to maximize sampling diversity. In general, it is best to use as many methods as possible since different techniques will collect different types of organisms. In the field you can either pick individual specimens into vials or you can grab material to pick back at the

lab. If you pick them in the field, at the minimum you will need a vial of alcohol, a pair of forceps, and a white pan. You can find good picking pans at a photography store that sells film developing supplies. Light colored pans may also be sold at department stores as drawer organizers. Place the collected material in a pan with a small amount of water and pick insects into a vial of alcohol. If you grab some wetted leaves or grass and place them into a ziplock bag, you can pick them in the lab at your leisure (a plus if air temperatures are low) and often you will be able to find more species. If you are going to bring the material back to the lab, place the material in a ziplock bag with only a little water and keep it cool in the refrigerator until you plan to pick the sample. Try to grab material from different habitats. Collect terrestrial leaves or aquatic vegetation from pools and edge habitats. To collect insects from riffle areas, you can look for leaf packs caught among the rocks or you can try to gently scrub organisms from rocks. There may also be macroalgae (e.g., Cladophora) attached to substrates in riffles that can be removed and placed into a bag. You can grab material by hand, but the best way to collect concentrated samples is using a dip net (see below). Useful collecting items: - dip net (or aquarium net or kitchen sieve) - forceps - turkey baster - light colored pan - vials with alcohol - waders or hip boots thermometer field notebook field bag for equipment camera GPS unit

Hand collecting: You can collect many aquatic insects with only a pair of forceps and a vial of alcohol. The best way to do this is by picking up substrates such as rocks, logs, sticks, leaves, or even human trash (e.g., tires, buckets) and picking insects directly from the substrate into a vial of alcohol. When removed from the water, many insects will crawl from the substrate so it is sometimes a good idea to place it in a white pan. Many organisms will crawl actively making them easy to identify. Also look for slow moving insects or insect cases attached to substrates. If you find a piece of wood, check the surface and then peel away bark or break open weakened areas where insects will often be hiding. Dip Netting: The best way to qualitatively collect as many species of aquatic insect as possible from the water is by using a dip net. The best type of net to use is one specifically made for collecting aquatic insects. These usually have fine mesh (often 500 m), a sturdy frame, and a long handle. They tend to be expensive so your best option may be to borrow one from someone. There are other, cheaper options that will work nearly as well. A large, fine mesh aquarium net works nicely although they are usually not very rugged. You can also use a fine mesh wire kitchen strainer. Nets used for fishing or collecting minnows are not very effective because of the large mesh. A dip net is particularly effective because you can sample a variety of locations. A dip net can be used in riffles by holding the net downstream and disturbing the substrate with your hands or feet so that the insects are dislodged and washed downstream into the net. A dip net is also useful in pools or edge habitats where the net can be scooped through or under the substrate while being careful not to collect too much material. This is effective in areas where there is overhanging terrestrial vegetation or submerged tree roots. Where ever you collect, it is best to take several small samples or else is can be difficult to sort insects from large amounts of collected material.

Collecting Adults: Most aquatic insects have terrestrial or aerial adults and you will often need to use different techniques to collect this life stage. At the family level, many aquatic insects are more difficult to identify as adults. However, many adults can be more reliably identified to the genus or species level so they may be important for obtaining better taxonomic resolution in studies. Many adults aquatic insects are attracted to lights so you can collect them at porch lights. However, you need to be careful since both non-aquatic and aquatic insects are attracted to lights. To improve your chances to collect aquatic insects, collect at lights near water bodies or set up your own light near a water body. To collect these insects, the easiest way is to use an aspirator. You can also pick small insects directly into a vial by wetting your finger with alcohol and touching the insects wings so that it sticks to you finger and dipping your finger into the vial. For larger insects, it is sometimes possible to knock them down into a vial of alcohol. Most aquatic insect adults are not active during the day and are often resting on vegetation near the water body (although in the cooler months, a greater percentage of adults are active during the day when it is warmer). You can collect adults using an aerial insect net and sweeping the vegetation along the edge of a water body and carefully picking specimens into a vial. Rearing: Many immatures can not be identified to the species or even genus level without associated material (i.e., pupae and/or adults). Rearing can be done by collecting live mature late instar larvae or pupae and maintaining them in vials (one organism per vial). Different insects have different requirements so specific techniques may be required to successfully rear insects. Some species can be reared in small vials with a small amount of water and a cotton stopper. Others can be more difficult and will require food, current, or specific temperatures. Vials should be checked frequently for emergence or mortality. Once emergence or death has occurred, preservative and a label should be added to the vial. Other Techniques: Check Merritt and Cummins (1996) for other techniques including quantitative methods. There are a variety of quantitative methods used for biological monitoring however most are overkill for making an insect collection. TAKING NOTES In many cases it is useful to have a field notebook to take notes on when you collected, how you collected, and anything else that might be interesting or pertinent. For example, you may take notes on anything you saw emerging, air/water temperature, or detailed information of what habitat types were present and sampled at the locality. It is not uncommon to finally get around to processing a sample years after it was collected or you may want to know additional information about a specimen that is not included on the locality label. LABELING SAMPLES When placing labels into specimen vials, put them in so that they can be read when the vial is held by the cap (see Figure 1). Locality labels should be placed so that they are facing out and the determination label should be facing in. It also helps if the labels are wide enough so that they curl around the edge of the vial rather than float around with the specimen(s). These tips may seem picky and ridiculous, but after looking at a few hundred vials with the labels arranged differently, it starts to make more sense.

Figure 1: Sample label placement. Locality Labels: An aquatic insect sample is useless without proper locality information. There are three important elements to a locality label: location, date, and collector. In some cases you may want to include more information such as time of day, collecting method, or what habitat. Usually this information will be included in your field notes (see above), but in some cases it may be particularly important to include additional information on the locality label since in a hundred years, a researcher may not have access to your notes. Locality information should be specific enough for someone to find that site again from only the information on the label. It is useful to include country, state (or province or district depending on the country), and county. The rest of the data should provide the remaining detail needed to allow someone to find the site again. Latitude and longitude is not essential in developed countries, but it is still very useful especially since it is not difficult to obtain with a handheld GPS unit. If you can not get a GPS unit, you can get lat/long info from maps. If I dont have a GPS with me when I sample, I get my coordinates from www.topozone.com. This website has searchable USGS quadrangle maps so you can find your site, click on it, and the coordinates are displayed at the top of the page. If you include this information it will make finding that location much easier in the future. The date the sample was collected is also extremely important and should be included on all locality labels. The month should be spelled out (e.g., Jan) or represented by roman numerals. Never write the date with numbers only (e.g., 5/6/04). This is ambiguous since there is no standard for which number goes first in Europe this reads as the 5th of June and the 6th of May in the United States. Also write out the entire year (e.g., 2004). It may not seem important now, but in a hundred years it may not be clear if a specimen was collected in 2004 or 2104. So a sample collected on the 6th of May in 2004 would be written as 6-May-2004 or 6.v.2004. It is also important to include who collected the sample. If there was more than one person collecting, include their names as well. Include first and middle initials with the last name spelled out or only the last name spelled out (e.g., R.W. Bouchard, Jr. or Bouchard) Specimen vials should always have a label inside. Dont write locality information on a piece of tape and attach it to the outside of the vial. These labels can and will fall off and then you are left with

useless specimens. In the field, you will probably need to use a temporary label. The best way to do this is to bring several small pieces of paper with you in the field and then using a pencil or alcohol proof pen, write the locality information on these pieces of paper and place them in the vial with the specimens. Once back in the lab, you will want to print out permanent labels for you specimens. This also makes it easier if you have many different species from the same locality. Rather than hand writing the label many times you can just print out many copies. For alcohol labels, I usually use 8 pt. font and Times New Roman although any type of font is acceptable as long as it is easily read. I type the label out once and then copy it as many times as I need into columns. When printing alcohol labels, make sure you use a laser printer (since an ink jet will bleed). If you dont have access to a laser printer, you can print it using an ink jet and then photocopy the sheet (provided it is a good photocopier) and the photo copied sheet should not bleed. If you are not sure if your printing technique will work, run a test by dropping some test labels into alcohol. However, it is best to find out from others what is the best technique since without a long term test you wont know if your labels will survive for decades.
USA: MN; Dakota Co. Trout Brook; Miesville Ravine Park Reserve 44.5453N, 92.8057W 24-Jan-2005 coll: R.W. Bouchard, Jr.

Locality label example.

Determination Labels: Once you have determined what your insect is, you need to label it so that you or anyone else who looks at that specimen in the future knows what it is without having to key it out again. Identified specimens are also important as vouchers for comparison with future collections. Determination labels should have at least two elements: identification and who determined the specimen. It may also be useful to include the date the specimen was identified. For your collections, you should have Order and Family listed.
EPHEMEROPTERA Baetidae ID date: det. R.W. Bouchard, Jr.

Determination label example.