Excerpt: Interest in soil biology has been prompted by recent investigations into the action of insecticides on plants and animals. Observations in the field must be supplemented by laboratory investigations conducted under controlled conditions. Consequently, it becomes necessary to rear and handle soil animals under artificial situations for bio-assay and life cycle studies. When large numbers of individuals are required, special problems in maintenance and manipulation arise. Relatively inexpensive and simple methods for such projects are essential and this paper describes some of those techniques which we have found expedient

Butcher, James W

Shaddy, James H

Snider, Richard J

English

Valparaiso University

The Great Lakes Entomologist Volume 1 Number 10 -- March 1969 Number 10 -- March 1969 Article 2 March 1969 Culture Techniques for Rearing Soil Anthropods Richard J. Snider Michigan State University James H. Shaddy Michigan State University James W. Butcher Michigan State University Follow this and additional works at: https://scholar.valpo.edu/tgle  Part of the Entomology Commons Recommended Citation Snider, Richard J.; Shaddy, James H.; and Butcher, James W. 1969. "Culture Techniques for Rearing Soil Anthropods," The Great Lakes Entomologist, vol 1 (10) Available at: https://scholar.valpo.edu/tgle/vol1/iss10/2 This Peer-Review Article is brought to you for free and open access by the Department of Biology at ValpoScholar. It has been accepted for inclusion in The Great Lakes Entomologist by an authorized administrator of ValpoScholar. For more information, please contact a ValpoScholar staff member at scholar@valpo.edu. THE MICHIGAN ENTOMOLOGIST .- 357 CULTURE TECHNIQUES FOR REARING SOIL ARTHROPODS* Richard J.  Snider, James H. Shaddy and James W. Butcher Department of Entomology, Michigan State University East Lansing, Michigan 48823 Interest in soil biology has been prompted by recent investigations into the ac- tion of insecticides on plants and animals. Observations in the field must be supplemented by laboratory investigations conducted under controlled condi- tions. Consequently, it becomes necessary to rear and handle soil animals under artificial situations for bio-assay and life cycle studies. When large numbers of individuals are required, special problems in maintenance and manipuation arise. Relatively inexpensive and simple methods for such projects are essential and this paper describes some of those techniques which we have found expe- dient. Various rearing chambers have been suggested by authors in the past, and from them we developed our own particular method. Wharton (1946) used weighing bottles fitted with ground glass stoppers and a plaster-charcoal sub- strate, in which ratio of plaster to charcoal was 9 to 1. Rohde (1956) described the use of Lucite cylinders set into a plaster-bottomed container. A series of these small vessels can be observed under the dissecting microscope. Goto (1960) improved upon earlier culture methods by using a heat-resistant glass container with a plaster-charcoal base and sealed with a cork. He was able to heat-sterlize the jars, but fungi reinvaded his cultures. This problem was solved by adding a low concentration of Nipagin-M (methyl-p-hydroxybenzoate) solution to the plaster and/or the food. The food supply was yeast. For his isolation chambers, Hale (1964) used Goto's method with 2x314" tubes filled with 1/2" of the top with plaster-charcoal mixture. He sealed the top with a cover slip imbedded in vaseline. Vail (1965) used essentially the same tech- nique with tubes imbedded in a plaster-charcoal base within a fingerbowl. He reviews some of the earlier culturing techniques by other authors. Abbatiello (1965) used a plastic container similar to one described in this paper with a wire cloth bent in the lid. More recently, Kyle and Long (1967) developed a method for rearing Collem- bola on filter paper suspended within vials containing sand set in a water reservbir. Algae were used as a food source in this method. The studies being conducted on Collembola and other arthropods in our lab- oratory required variable numbers of individuals and, therefore, large num- bers of culture containers, For ease of repetition a standardized container was adopted. At first i t  consisted of a small glass jar (5% cm x 5 cm) fitted with a plastic screw cap (fig. 1). A 5:l plaster-charcoal mixture was poured into each jar to a depth of 2 centimeters. This container has the advantage of being air-tight, and water loss is held to a minimum. Brewers yeast in small amounts was supplied as food. *This paper was supported by U.S. Public Health Service Research grant CC00246, from the National Communicable Disease Center, Atlanta, Ga. Facilities were made possible by the Soil Biology Group, Michigan State University, and the Michigan Agricultural Experiment Station, for which this is jour- nal number 4583. 1Snider et al.: Culture Techniques for Rearing Soil AnthropodsPublished by ValpoScholar, 1969358 THE MICHIGAN ENTOMOLOGIST Vol. 1, No. 10 Fig. 1. Glass jar with plaster-charcoal substrate. Fig. 2. Plastic jar with plaster-charcoal substrate. When several hundred replications were made, the disadvantages of screw- cap jars became apparent. The constriction for the screw-cap forced the ob- server to tilt the jar under the microscope in order to make observations near the sidewall. Furthermore, it was necessary to remove the cap every time a check was to be made. Later, straight sided plastic containers (5 cm x 3 314 c m )  were obtained for general culture use (fig. 2 ) .  They were kept in controlled temperature cabinets a t  60, 70 and 80 degrees Fahrenheit. The same 5:l plaster-charcoal substrate was used, but while the mixture was still soft, a 15mm circular coverslip was pushed into the center of the surface to serve a s  a feeding station. The plastic containers had a clear snap-on top which allowed observations to be made without disturbing the culture. The tops were not always air-tight and some water vapor did escape. This was overcome by addition of water to the substrate once a week. A higher humidity and lower water loss could have been maintained in the cultures if a pan of water was placed in the bottom of the temperature cabinet. The reason for using a 5:l plaster-charcoal mixture was based on egg ob- servations. When it is necessary to count, photograph or remove eggs, the black background afforded by the substrate accentuates most arthropod eggs which a r e  white to hyaline in color (fig. 5). Brewer's yeast was used as  a food to support the stock cultures. Most of the soil species of Collembola did very well on such a diet. I t  was added in small quantities along with a drop bf water. Provided the substrate was not too wet, fungal growth was kept a t  a minimum. Using these containers, various feeding experiments with Collembola and other arthropods were undertaken. Many soil species in culture were found to prefer dried corn leaf. I t  was a simple matter to collect a supply of corn leaves and store them in a freezer until needed. A cork borer was used to cut uniform discs from the leaves (fig. 3). The discs in turn were oven dried and then weighed. Once this was accomplished, the discs were placed on the coverslip feeding station within the culture containers. Various num- bers of arthropods were introduced into the containers to ascertain how much 2The Great Lakes Entomologist, Vol. 1, No. 10 [1969], Art. 2https://scholar.valpo.edu/tgle/vol1/iss10/21969 THE MICHIGAN ENTOMOLOGIST 359 Fig. 3. Corn leaf discs cut with a cork borer, ready to be weighed and placed in the culture jars. plant material a species could consume. At the end of a given period of time the remains of the corn leaf disc were removed, dried and weighed (fig. 4) .  Other experiments were then conducted by treating the discs with insecticides. The results of such experiments will be presented elsewhere. However we men- tion them as  an example of what can be done with these relatively simple con- tainers. Early in our studies, manipulation of small arthropods was accomplished after anesthetizing them with ethyl ether. A hole was cut in the lid of a plastic petri dish, and a cotton plug inserted (figs. 6 & 7 ) .  Ether was dropped on the cotton and a cap was placed over the area. The cotton plug was centered over the culture container and after a short exposure, the individuals or eggs could be transferred with a camel's hair brush. Later carbon dioxide gas was used in place of the ether, and the procedure was modified accordingly. A sim- ple glass funnel was attached to the cylinder by means of a plastic hose. The culture was then placed under the funnel and the gas introduced (fig. 8 ) .  C02 has very little long-range effect on most species and has the advantage of being safer to use in the laboratory. The method described in this paper enabled us to rear the following species of Collembola: Protophorura armatus (Tullberg), Folsomia fimetaria (L. ), Proisotoma minuta (Tullberg), Zsotoma albella Packard, Hypogastrura armatus 3Snider et al.: Culture Techniques for Rearing Soil AnthropodsPublished by ValpoScholar, 1969360 THE MICHIGAN ENTOMOLOGIST Vol. 1, No. 10 Fig. 4. Corn leaf disc after one week exposure to Folsornia fimetaria (L.) in a culture jar. The remains can be removed and weighed. (Nicolet), Tomoeerccs Jbvescens (Tullherg), Lepidocyrtus violaceous (Geof- froy), Orehesella hexhsciata Harvey, Pseudosinella violenta (Folsom), and Neanrara rnuscorurn Templeton. Other arthmpds maintained were Entotrophi, Pseradoscorpmns, Isopoda, and Dichpoda. The techniq~~s  involved with these o~ders  varied somewhat fnnn those used for Cd-la. The Enbkophi lived adequately in the con- tainers with f!he adtlition of yeast, as well as some fragments of leaves for shelter. Cod temperatures (around 70 F.) seem to be adequate to maintain the cuItttres. The impads and diplopods are best kept i~n moist containers with corn ieaves a d  a small quantity of chak scrapings. Two species of ID pods were ~ s s f u b l y  cuitured though their life cycles; Cylisticus convexus (DeGeer) and TmBecnrhcus ralhki (Ekandt). When their juveniles first hatcked, yeast was added as a dietary supplement. The Pseudoscorpions were easily reared by the addition of Collembola to their containers. These provided an adequate diet. Further modificatiotts of the above techniques are presently under way and the numbers of species in culture are being increased. 4The Great Lakes Entomologist, Vol. 1, No. 10 [1969], Art. 2https://scholar.valpo.edu/tgle/vol1/iss10/21969 THE MICHIGAN ENTOMOLOGIST 361 Fig. 5. Folsomia eggs photographed on the plaster-charcoal substrate. Fig. 6. Petri dish placed over culture jar ready to receive ether. Fig. 7. Cap in place after ether has been put on cotton wad. 5Snider et al.: Culture Techniques for Rearing Soil AnthropodsPublished by ValpoScholar, 1969THE MICHIGAN ENTOMOLOGIST Vol. 1, No. 10 Fig. 8. C02 technique with culture jar in place. LITERATURE CITED Abbatiello, M. J. 1965. A culture chamber for rearing soil mites. Turtox News 43: 162-163. Goto, H. E .  1960, Simple techniques for the rearing of Collembola and a note on the use of a fungistatic substrate in the cultures. Ent. Month. Mag. 96:138-140. Hale, W. G. 1964. Experimental studies on the taxonomic status of some mem- bers of the Onychiurus armatus species group. Rev. Ecol. Biol. Sol. 1:501- 510. Kyle, M. L. and W. H. Long. 1967. A simple technique for rearing Collembola in the laboratory. J. Econ. Entomol. 60:882-883. Rohde, C. J. 1956. A modification of the plaster-charcoal technique for the rearing of mites and other small arthropods. Ecology 37:843-844. Vail, P .  V. 1965. Colonization of Hypogastrura manubrialis (Collembola: Poduri- dae),  with notes on its biology. Ann. Ent. Soc. Amer. 58:555-561. Wharton, G. W. 1946. Observations on Ascoschorigastia indica (Hirst, 1915) (Acarinida: Trombiculidae). Ecol. Mongr. 16:151-184. 6The Great Lakes Entomologist, Vol. 1, No. 10 [1969], Art. 2https://scholar.valpo.edu/tgle/vol1/iss10/2

Culture Techniques for Rearing Soil Anthropods

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Culture Techniques for Rearing Soil Anthropods

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