BIO 322 ENTOMOLOGY LABORATORY 1 FALL 2009

Arthropod Curation Techniques

For this and every subsequent laboratory exercise, you should prepare ahead of time. Review relevant material in the textbook and handouts and prepare to answer questions for labs.

For each laboratory, you will need the full lab time to complete the requirements.

Insects are the largest and most diverse group of organisms on the planet. Because of this diversity and the huge theoretical number of undiscovered species, we need a stable and efficient ways to collect, identify, and preserve them in perpetuity. Proper and consistent curation techniques allow us to preserve specimens in a manner that lets us readily focus on key distinguishing features. Professional taxonomists, collectors, and curators all demand specimens be presented in specific ways and so it is important that we know and are able to apply these techniqes.

The purpose of this lab is to explore the various methods and techniques used in arthropod curation in order to establish a proper reference collection. Students will learn how to prepare specimens, which is essential for taxonomic identification. This is critical for biodiversity studies and any type of natural resources monitoring involoving arthropods. It is also important for any type of education or research activity that involves wild species. Even if one does not become a taxonomist it is important to know how to prepare and curate specimens so that you can work with taxonomists, who can provide you with appropriate identifications. Well curated specimens are a joy to taxonomists!

We will refer to various invertebrate groups, in this lab we introduce Arthropods, Hexapods and Insects. Insects are hexapods and arthropods. Arthropods include other invertebrates besides insects that are characterized by an exoskeleton and jointed appendages. Most notably these non-insect arthropod groups include Crustaceans (e.g., shrimp, crabs, pillbugs) and Arachnida (e.g., spiders, scorpions, mites).

In this lab students will learn the following:

1) Temporary storage

2) Pinning & Mounting

3) Pointing

4) Spreading

5) Labeling

6) Alcohol preservation

7) Slide mounts

8) Storing your collection

Materials:

Insect pins #0, 1, 2, 3, and 4

Card paper

Precut points

Elmers Glue

Pinning blocks

Scissors

Schmit Boxes or other storage boxes with foam bottoms

70% Ethanol

# .5 Mechanical pencil

Spreading boards

Slides & cover slips

Slide glue

1) Temporary storage

Instructor will discuss how to properly store specimens in the freezer.

2) Pinning & Mounting

Various insects will be provided by the instructor for the students to practice pinning and mounting with insect pins and card paper. Each student must know how to correctly pin large beetles, (Coleoptera), true bugs, (Hemiptera), wasps & bees (Hymenoptera), flies (Diptera), crickets & grasshoppers (Orthoptera) and other large insects. Students will be allowed to use the insects for their required collection.

Examples A and D are the correct ways to pin insects. Examples B, C, E, and F are the incorrect ways to pin insects.

3) Pointing

Small insects such as leaf hoppers (Hemiptera), small beetles (Coleoptera), flies (Diptera) and parasitoids (Hymenoptera) will be provided by the instructor for students to practice pointing. Almost all insects with a thorax smaller than 4 mm width should be placed on a point. Students will be allowed to use the insects for their required collection.

The instructor will demonstrate how to pin small specimens with minuten pins. Minuten pins will be available to students who want to pin specimens using this technique. Minuten pinning is most commonly used for tiny delicate specimens such as microlepidoptera and small Diptera, mosquitos (Culicidae) and midges (Chironomidae).

4) Spreading

The instructor will discuss the delicate procedure involved with spreading wings of Lepidoptera and some other flying insects and how to make simple spreading boards. Moths (Lepidoptera) and antlions (Neuroptera) will be provided by the instructor for students to practice spreading. Students will be allowed to use the insects for their required collection.

5) Labeling

Labels will be provided by Jacob Higgins, students will learn how to record collection label data that they will submit to Jacob as MS Word documents.. .

Once your insect has been pinned, pointed or placed in alcohol, it is ready to be labeled. Some collectors use up to three labels, although in this class most specimens will just have one locality-date-collector label. The first label will contain locality information including date collected, state, city and collector. The first line includes state and county (see below). The second and sometimes third line contains the location. We are trying to get everyone to list a named locality and a latitude-longitude. Latitude-Longitude should be in decimal degrees with precision to the 3rd, or 4th decimal point. A lat-long value to the 4th decimal point puts you within 10 meters (e.g., N 35.2162), and a value to the 3rd decimal point puts you within ~100m of where you collected, assuming you collected in just one location. If you collect within a 10-meter radius then use the 4th decimal point, if you sample in a larger area within 100-meter radius of where you set the point then use the 3rd decimal point. Only use a lat-long value that goes to just the 2nd decimal point if you collect within a 1 km radius or you are not sure where you collected.

The two most common ways to get lat-long in decimal degrees is either by using a GPS unit (basic Garmin unit is fine) or pinpointing your location on Google Earth.

The last line contains the collector’s name. For this class the date begins with the month and is followed by the day and the year. You can make a 2nd and 3rd label to indicate the plant species or microhabitat (e.g., under rock) the specimen was collected or the method you used to collect the specimen (e.g., pitfall trap). But these additional labels are not neccesary.

Coconino Co. AZ

N 35.2162 W-111.6430

Flagstaff, Cobb 7/14/09

Coll. Neil Cobb

Coconino Co. AZ

N 34.919 W-111.434

Mormon Lake 8/29/09

Coll. Jacob Higgins

Example of actual size:

Yavapai Co. AZ

T18N R6W sec 25

Walnut Creek 7/8/04

Coll. Neil Cobb

Example of actual size for host vegetation label and trap label:

EX: Pinus

edulis

EX: Malaise Trap

10am-2pm

The label should be pinned through the center in the second deepest hole on the pinning block. The label should be parallel with the insect and be read from top to bottom from the insect’s left side. For pointed insects, the label orientation should be the same, but the label should be pinned on the far right side. The labels should be printed in 3.5 pt. Font using New York Times Roman or Arial. The labels must be printed on 67lb acidfree cardstock. To make sure labels are correct and printed on the right paper, students will be provided with a MS Word document template and will send the completed labels to Jacob for printing.

6) Alcohol Preservation

Many insects that are in their larval or nymph stage are soft bodied and do not pin very well as a dry specimen thus should be preserved in ethanol. In addition all non insect specimens such as arachnids, chilopods, diplopods, isopods and so on should also be preserved in ethanol. Some larval Lepidoptera, Coleoptera and Hymenoptera need to be boiled first before being placed in ethanol so they do not shrivel up. Boiling is generally reserved for larger larvae and only need to be boiled for about 30 seconds to a minute. The instructor will discuss which specimens need to be in ethanol and proper techniques used for alcohol preservation. Remember 90% of the collection must be spiders and/or hexapods, the other 10% can be other arthropods..

7) Slide mounts

Many small insects and hexapods are too small to even be placed on points and therefore must be placed on a slide. Most slide mount arthropods include the primitive hexapods such as Proturans, springtails (Collembola) and japygids (Diplurans). Thrips, lice and nats are some of the other insects which are often put on slides. Most all of these specimens can also be placed in ethanol as well. The instructor will demonstrate how to properly make a slide specimen. Springtails (Collembola) and Thrips (Thysanoptera) will be provided by the instructor for students to practice making slide mounts and may be used in their collection.

8) Storing your collection

Once your specimens are pinned, mounted and labeled, you will be responsible for the quality and safe storage of your collection. Dead insects are delicate and can break easily and must be handled with care. The instructor will discuss proper storage, handling and upkeep of your collection. Trip to the arthropod museum!

Laboratory Exercise

Students will practice pinning and prepping insects provided by the instructor, which will be used for their final collections. Each student must be able to properly pin, point, spread and make a slide mount. We list below the specimens that will be provided for this lab. Unless noted each student will prepare at least one of each specimen listed below as directed. For some groups we may only have enough individuals for one specimen per student

PINNING

1. Scarabs or Tenebrionid Beetles

2. Pentatomid Bugs

3. Grasshopper

4. Bees

5. Moths (Pin and Spread at least 3)

6. Coccinellid, Melyrid, Chrysomelid mix

POINTING
1. Lygeaid Bugs
2. Scolytid Bark Beetles

3. Mordellid Beetles

4. Ants & Parasitoids

5. Leafhoppers & Planthoppers

ALCOHOL

1. Spider (prepare alcohol label and place in vial)

SLIDE MOUNT
1. Collembola

Web Resources

http://www.sel.barc.usda.gov/selhome/collpres/collpres.htm Download the USDA manual for collecting and preserving insects

http://www.bugs.nau.edu/learning_modules.html

Questions:

1) What is the best way to euthanize and store and insects prior to pinning/pointing?

2)      List the conditions under which you would:
                    pin a specimen
                    point a specimen
                    place a specimen in alcohol
                    place a specimen on a slide

3) Which insects must have their wings spread and why?

4) What is the rationale for where you place a pin for any given insect?

5) Name at least 3 groups (Classes or Orders) of non-insect arthropods and 5 orders of Hexapods/Insects that should be placed in ethanol.

Practice using the BIO 322 website to find information on the syllabus for lab and lecture. Print out a copy of next weeks lab, and information on the hexapod collection, hexapod orders and families to know.