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Collecting and Preserving Insects

P. M. Choate

Insects provide many opportunities for biological studies, ranging from detailed research to casual field observations. Frequently the need arises for collecting specimens to accurately identify the species of interest. Collecting techniques and methods of preservation of insects vary considerably. A few of these techniques are described here. Key elements to successful insect collecting - Collectors frequently wish to find as many different insects as possible. In order to accomplish this, the greater the diversity of collection techniques employed and habitats explored, the greater the diversity of insects that will be collected. Most entomology textbooks present an overview of collecting and preserving insects for study. There are two major categories of information dealing with these subjects, collecting techniques and preservation techniques (how to deal with the insects once collected). Collecting Techniques Collecting techniques are limited only by the imagination of the student or researcher. Practically any technique will yield a few insects. Here are a few: For insects on the soil surface - Hand-collecting, turning over logs and rocks and other objects - Unbaited pitfall traps - Pitfall traps baited with carrion, dung, yeast, or fermented fruits For insects on plant surfaces - Hand-collecting, or use of sweep nets, beating trays, aspirators For insects within living or dead plants - Cutting sections of plants to place into containers within which the insects will later emerge, breaking logs apart with an ax or chisel, stripping bark of dead trees to find subcortical insects For flying insects - Hand-held nets - Passive intercept traps Malaise and window traps - Attractant traps with mercury-vapor or ultraviolet light, or carbon dioxide, or specific insect pheromones as bait For subterranean insects - Sifting soil or debris with a sieve - Placing debris into a Berlese or Tullgren funnel to extract insects by heat, or light, or a chemical repellent For aquatic insects - Aquatic nets

Living insects If your objective is to collect insects alive for study alive, then your best option to prevent damage is to chill them as soon as they are collected. To do this, place them into suitable containers in an insulated chest with ice or other refrigerant, and transport them as soon as you can. If your objective is to kill them to obtain preserved specimens, kill rapidly to prevent damage. Killing methods Large specimens are typically killed in a jar with closely fitting lid using a volatile toxicant such as ethyl acetate absorbed onto plaster or sawdust or cotton. Small specimens may be killed in vials using the same toxicant. Alternatively, if they will not be damaged by immersion in liquid, they may be killed in 70% alcohol or soapy water. Recording Vials or other containers should be labeled in the field as the specimens are collected. Never use a ball-point pen for insect labels if you do not have a pen with India ink available in the field, make a temporary label with pencil. Record at least all the information you may eventually need (for which see section below on Labeling specimens). Preserving Insects Small and soft-bodied insects should be stored in 70% alcohol (either ethanol or isopropanol) in tightly sealed glass vials. Isopropanol (rubbing alcohol) is inexpensive and readily obtained in most countries. Ethanol may be subject to excise laws because it may be used for human consumption, so typically is more expensive and more difficult to obtain and there seems to be no point in going to that trouble if isopropanol can be obtained. A label should be placed in the vial, with information printed or written in insoluble ink. An additional label may be taped to the outside of the vial. Insect larvae may need brief insertion in boiling water before they are preserved in alcohol, without which they will blacken. Vials These are available in many shapes and sizes. Plastic vials are useful in the field because they are less subject to breakage than are glass vials, but glass vials are very much preferable for long-term storage. When specimens are stored in alcohol in vials, be aware that the type of vial and type of stopper is important. Straight-sided glass vials (shell vials) are thinnerwalled and more fragile than other types. Vials stoppered with screw-caps or corks will sooner or later allow the liquid contents to evaporate. Thick-walled glass vials with necks (homeopathic vials), when fitted with natural rubber stoppers, seem to offer the greatest permanence, but the rubber will eventually crack and decompose. Stoppers of neoprene or other synthetic materials have not yet shown an advantage over natural rubber. If using homeopathic vials with rubber or neoprene stoppers, it is important to seat

the stopper firmly; this may be done by inserting a straightened-out steel paper clip alongside the stopper as the latter is inserted, then withdrawing the paper clip to release trapped air. All vials should be checked periodically for leakage and loss of preservative. Mounting techniques - Different insects require different mounting techniques. Large moths and butterflies and some other winged insects are mounted with wings neatly spread on a spreading board. Pinning - Insects of average size are usually mounted on pins. In many countries, steel pins of 3.5 cm length are used; the best quality is stainless steel with nylon heads. Lacquered steel pins may rust, and brass pins may corrode (become covered with green verdigris) in humid climates. Pin widths are numerically-designated, with standard sizes 0-3 increasing in thickness with increase in numerical value. Extremely narrow pins are of size 00, and extremely thick pins are of sizes 4-7 but are longer. Most insects large enough to be pinned may be pinned on sizes 2 or 3. Insects are pinned in specific locations depending on their classification. Large beetles are pinned in the middle of their right wing cover, close to the front margin. Other examples of pin positions are included in the diagrams provided here. [In the United Kingdom, white brass pins were once used; their length varied with width]. Minuten pins (from a German adjective meaning minute) are very small, fine pins used for pinning tiny insects into a small block of pinning material, which is in turn pinned by a standard pin. Storage in transparent envelopes - Some Odonatologists (dragonfly collectors) stuff their specimens into semi-transparent ( Glassine) envelopes. This is an alternative to pinning specimens, saving much museum space. However, this is not necessarily the most desirable method of storage method because the specimens are prone to breaking up. Pointing - Insects too small to be pinned safely may be mounted on cardboard points, glued to the point on their right side. Points should be bent down slightly to better attach to the side of each specimen. Such points may be purchased, or may be made by a hand-held device called a point press, which cuts points from a sheet of cardboard. Care should be taken not to cover the underside of the insect with glue, obscuring key characters. This is the standard mounting technique in the USA. It is claimed to be better because underside characters are visible. There are two major problems: (1) insects thus mounted are very susceptible to physical damage. (2) adhesives used to obtain secure adhesion to the small surface may be soluble only in harsh solvents ( benzene, toluene, xylene, acetone, etc.]. Subsequent attempts to dissect soft body tissues are thwarted because these are destroyed.

Card mounts Some groups of insects are better mounted on tiny rectangular cards. Cards of standard sizes are sold by some supply houses. This technique is popular with European entomologists. It is claimed to be better because the specimens are better-protected from physical damage and because they may be viewed in one plane against the white background of the card (as contrasted with specimens that may be drooped over card points), and because they may later be removed easily for dissection. Only watersoluble adhesives should be used. There are perhaps two problems: (1) The underside characters are not visible without removal from the card. However, preparators with a series of specimens available will mount some dorsal side up, and some ventral side up. (2) It is said that such a method of preparation is more timeconsuming. This may be true, but it varies with the expertise of the preparator. More importantly, if the specimen is rare and from a distant locality, it makes far more sense to spend a few seconds more to prepare it this way (better-protected) than to face the days or weeks and expense necessary to collect more specimen from the distant locality, even assuming that a replacement specimen can be found for a rare species. Adhesives Many kinds of adhesives are used to secure insects to card points. The objective is to secure the insect firmly to the point. We recommend that only water-soluble or alcohol-soluble adhesives be used. When other kinds of adhesives are used, there will arise questions about the necessary solvents to remove the specimens (it is virtually impossible to guess what adhesive some collector may have used and what its solvent is). For specimens adhered to card mounts, only water-soluble adhesives should be used. When card-mounting, the strength of the adhesive is less important because much more surface area is used. Many water-soluble adhesives are available, although the two most commonly sold in stationery stores in the USA are unsuitable -- one because it dries hard, and cracks and turns brown the other because it is not truly transparent and dries from the outside inward, forming first a sticky skin. Card mounts, card points, and labels Use only a heavy weight, high quality, non-yellowing, 100% rag card stock for making these items. Some years ago, Bristol board, a laminated card, was recommended. We see no advantage to such laminated card stock. Genitalia It is often desirable to dissect specimens to make the genitalia evident for identification purposes. With card-mounted specimens, the genitalia (if large enough and well-sclerotized) may be mounted on the same card as the specimen, a great advantage. If, however, they are feebly sclerotized or too small, they may be mounted (after dehydration through

alcohols to xylene) in a drop of Canada balsam on a small celluloid rectangle pinned directly below the specimen (no cover slip is needed). When specimens are card-pointed, the usual method is to place them in genitalia vials pinned through the stopper below the specimen. Such genitalia vials are tiny, of glass or plastic, and contain a cork or stopper of some synthetic material; typically, they contain alcohol, glycerine, or a mixture of the two. When genitalia almost fill the vial, and when such vials are checked frequently for leakage, the method may have merit. Otherwise they can be disastrous: extremely small genitalia in a vial can be lost when the stopper is pulled, or the vial can leak and desiccate the parts. Such genitalia vials are to be avoided wherever possible. Slides Some insect specimens are best mounted on microscope slides. These include very small and soft-bodied insects as well as dissections of insect parts. The most permanent mountant available is Canada balsam. Some insects (scale insects, aphids and lice, tiny parasitic Hymenoptera, and many insect larvae) are typically prepared by clearing and differential staining before being mounted on microscope slides. Soft-bodied specimens Because of their hard exoskeleton, very many insects will maintain their shape when dead and dry. Some, however, will not they tend to shrivel. Techniques have been developed for maintaining the shape of dried specimens (one alternative was to preserve them in alcohol, another to mount them on microscope slides). For decades, the preferred method for preserving large Orthoptera (grasshoppers and crickets), Phasmida (stick-insects) and Mantodea (mantids) was to eviscerate them and stuff their abdomen with cotton before pinning and drying. For Odonata (dragonflies and damselflies) the method was to eviscerate them and stuff a toothpick-shaped object into the abdominal cavity. For larvae of butterflies and moths (caterpillars) the method was to roll the abdominal contents out through the anus, then inflate the abdomen (by blowing into it) while drying the inflated skin in a tiny oven. Mosquitoes and some other flies, and Trichoptera, having feeble exoskeletons, could not be dealt with by such rough physical methods because they are covered in scales, and the scales would be lost by such treatment; they were destined to shrivel as they dried. Many of these methods have now been abandoned because of the availability of critical-point drying. A critical-point drier is an expensive piece of equipment and may be unavailable in some institutions and to amateur collectors. Labeling specimens - Specimens without accurate and complete collection data are of little value. A proper label contains locality data, date of collection, collector's name, method of collection, and identification label with determiner's name. Additional information may be included (host plant, behavior, etc.). Computer-generated laser-printed labels in general seem
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entirely adequate. Such labels with a font size of 3 or 4 are suitable for pinned specimens. Labels with larger font (10 or 12) can be used in vials with alcohol. Most laser-printer inks seem stable in alcohol (you may want to test labels made with the laser printer available to you before relying upon them), and printed labels are far easier to read than are most hand-written labels, Use them wherever you can for pinned and alcohol-preserved specimens (provided that the ink is not alcohol-soluble). Information on labels - Labels for pinned insects should be placed beneath specimens, aligned parallel with the insect's body. The top label should contain (1) Country, (2) Province or State, etc. (3) Township, etc. (4) route information when known if the collection was a result of a route followed, (5) date of collection, and (6) collector(s) names. A second label should contain method of collection (bait, technique, trap) and habitat. Dates should be written in order day-month-year (the international standard) with month either in letters or in Roman numerals and the year with four digits. A date of the 4th of July 2002 should thus be written as for example 4-JUL-2002 or 4VII-2002 not as 7/04/02 (as in commonly done in the USA) or as 4/7/02 (as is commonly done in most of the world) or as 2002/7/4 as is commonly done in the Far East. A third label may contain rearing information. A fourth label may contain determination information, species name, determiner, and date of determination. Pinning blocks Supply houses sell blocks on which specimens and labels may be pinned. These achieve a standard height for each specimen and label and make a resultant collection appear more uniform. Unfortunately, we have not seen a block from a supply house with more than three steps. If the steps are (1) specimen, (2) dissections of genitalia from the specimen, (3) locality information, (4) habitat information, and (5) identification information, then five steps are needed. The only current option seems to be for collectors to manufacture their own pinning blocks (one of us has done so). Storing collections - Insect collections require protection from atmospheric conditions such as humidity and pests capable of destroying specimens. Such pests include cockroaches, silverfish, ants, dermestid and anobiid beetles, booklice, and mice. Tight containers, with a fumigant, will deter most pests. Be sure not to use chemicals toxic to humans or pets. The safest method is to store in or donate a collection to a recognized museum where it will be maintained safely. Museum curators often will provide space for your materials, hoping that at some time in the future you will donate your collection to them. Storage containers and cabinets Insect specimens are typically housed in wooden storage boxes (in the USA called Schmit or Schmitt or Schmidt

boxes) or in cabinets with drawers. The storage boxes have tightly-fitting lids, and the floor is lined with a material into which pins can easily be thrust. Traditionally, this material was cork, usually lined with paper, but now is normally some white synthetic foam. European cabinets were and are of numerous sizes, each drawer lined with a pinning material. The disadvantage of the European system is that dozens or hundreds of specimens may have to be moved one by one to make way for a few new specimens. Additionally, drawers of some cabinets are exceptionally shallow, allowing vertical space only for short pins. In the USA, another system arose in which drawers of standard size were designed to accept a standard number of cardboard unit trays, the unit trays (not the drawers themselves) are lined with a pinning material, and all drawers accept standard 3.5 cm pins. Unit trays are manufactured in a range of complementary sizes so that a combination of small and large trays, as need arises, will fit precisely into a drawer. The trays make it easy to move groups of specimens about a drawer or from drawer to drawer, and protect specimens from physical damage. The US system was a good idea, but unfortunately it is at least three systems not one developed at competing museums - which differ in dimensions of drawers and size of unit trays so that they are not interchangeable. Supply houses Several companies maintain an inventory of entomological supplies and sell by mail order. They sell nets, beating sheets, light traps, vials, pins, forceps, lenses, pinning blocks, storage boxes, cabinets, microscopes, microscope illuminators, microscope slides, preservatives, light traps, numerous other items, and and assorted entomological literature. Permits A very few species of insects have been declared to be endangered species. They may not be collected, even on your own property, without a very hard-to-obtain permit. In many countries, you may collect any other insect (apart from endangered species) or your own property, or (with permission from the landowner) on someone elses property. If the landowner is a government agency and the property is a park or preserve, you may need a written permit from that agency. In a few countries you may not collect insects of any kind without a government permit; even if you use a pesticide to kill pests on your property, and along with the pests you kill harmless or beneficial insects, you may not collect any of these insects without a permit. So, be aware of the laws in your country.

Spreading board used to prepare moths and butterflies.

Pointed insect showing position of point against right side of insect, and arrangement of labels.

Position of pins in various kinds of insects.

Examples of how to position mounting point under right side of insect specimen.

Minuten pin used to position small insect above piece of cork. This technique is frequently used for delicate flies and very small moths and butterflies.

Card-mounted specimen (European fashion).

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Large long-legged flies such as crane flies may need to have their body supported with an extra large point. Alternatively, the specimen could be place on a card mount, which will offer much more protection from damage by protecting the legs, too.

Selected References
Hatch, M. 1926. Concerning the insect collection. Entomological News 37: 329-332. Lehker, G. E. and H. O. Deay. 1969. How to collect, preserve, and identify insects. Extension Circular 509. Cooperative Extension Service, Purdue University, Lafayette, Indiana. 43 pp. Ross, H. H. 1962. How to collect and preserve insects. Illinois State Natural History Survey Circular 39. Urbana. 71 pp. Smart, J. 1954. Instructions for collectors no. 4a. Insects. British Museum (Natural History), London, 178 pp. Steyskal, George C., W. L. Murphy, and E. M. Hoover, eds. 1986. Insects and mites: techniques for collection and preservation. U.S. Department of Agriculture, Misc. Pub. No. 1443, 103 pp. Online at http://www.sel.barc.usda.gov/Selhome/collpres/contents.htm

Worldwide Web references online:


Note: the following Internet addresses are transient and may change or disappear without warning. Labeling insects http://www.biology.ualberta.ca/bsc/briefs/brlabelstandards.htm Collecting techniques http://www.eman-rese.ca/eman/ecotools/protocols/terrestrial/arthropods/intro.html Insects on WWW (6000 references) http://www.isis.vt.edu/~fanjun/text/Links.html

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