Floyd, Martin A.

Heidt, Gary A.

English

UARK (University of Arkansas )

Journal of the Arkansas Academy of Science Volume 33 Article 28 1979 Modifications and Improvements in the Formax Method of Preparing Small Avian Study Specimens Martin A. Floyd University of Arkansas at Little Rock Gary A. Heidt University of Arkansas at Little Rock Follow this and additional works at: https://scholarworks.uark.edu/jaas  Part of the Poultry or Avian Science Commons Recommended Citation Floyd, Martin A. and Heidt, Gary A. (1979) "Modifications and Improvements in the Formax Method of Preparing Small Avian Study Specimens," Journal of the Arkansas Academy of Science: Vol. 33 , Article 28. Available at: https://scholarworks.uark.edu/jaas/vol33/iss1/28 This article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able to read, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author. This General Note is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the Arkansas Academy of Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu. 76 Arkansas Academy of Science Proceedings, Vol.XXXIII,1979Arkansas Academy of ScienceThis survey resulted inthe finding of three specimens. Two specimens were captured inMay on a 65-acre prairie inBoone County (Marsh,1978), and the other specimen was captured in July on a 40-acre prairie inFranklin County, representing the first record for this county. Thusfrom 1909 to 1978 there are reports of the ornate box turtle inten Arkansas counties: Benton, Boone, Fulton, Washington, Franklin, Perry, Gar-land, Prairie, Lafayette, and Columbia (Figure I).Being a species which is restricted by habitat availability, the ornate box turtle has been greatlyaffected by the changes inland use practices occurring in Arkansas, and has been considered a rare species (Reagen, 1974). Conversion tomoreproductive agricultural use has reduced the amount of native prairie and undisturbed grassland available to the ornate box turtle, and this may bethe most important factor limitingthe ornate box turtle, and this may be the most important factor limiting the ornate box turtle's range in Arkan-sas. Inaddition, the Arkansas highway system has taken a tollbecause the turtle seems to exhibit a certain affinity for roadways and a great num-ber are killed bymotor vehicles (Legler, 1960 and Reagan, 1974). Toproperly assess the status of the ornate box turtle (Terrapene ornata ornataland to eliminate confusion concerning its distribution inArkansas a detailed studyis obviously needed.The author expresses appreciation to the following forvarious types of assistance during the course of this project: Dr.R.Baldauf,Dr.R. V.McDaniel, and Valerie Thwing. Dr.M.V. Plummer is gratefully acknowledged for his assistance with literature and forhis confirmation of species. Special thanks is extended to the Arkansas Natural Heritage Commission and staff, for continued sup-port and cooperation.LITERATURE CITEDCAGLE,F. R. 1957. Reptiles. In:Vertebrates of the United States.McGraw-HillBook Company, New York.pp. 213-268.MARSH, M. A. 1978. Baker prairie. The Ozark Soc. Bull. 12(1):10-11.CONANT,R. 1975. A field guide to reptiles and amphibians of east-ern and central North America. Houghton Mifflin Company,Boston. 429 pp.REAGAN,D. P. 1974. Threatened native reptiles of Arkansas. In°Arkansas natural area plan. Arkansas Department of Planning,LittleRock. pp. 101-105.DELLINGER, S. C. and J. D.BLACK. 1938. Herpetology of Arkan-sas: part 1, the reptiles. Occ. Papers Univ. Ark.Mus. no. 1. 47pp.SCHWARDT, H. H. 1938. Reptiles of Arkansas. Univ. Ark. AgriExper. Stat. Bull.357:1-47.WARD,J. P. 1978. Terrapene ornata. In: Catalog of American am-phibians and reptiles. Society for the Study of Amphibians andReptiles, pp. 217.1-217.4.DOWLING,H.G. 1957. Areview of the amphibians and reptiles ofArkansas. Occ. Papers Univ.Ark.Mus. no.3. 51 pp.LEGLER, J.M. 1960. Natural history of the ornate box turtle, Terra-pene ornata ornata, Agassiz. Univ. Ka. Mus. Nat. Hist. Pub.ll(10):527-669.LAWANA ENGLAND, Arkansas Natural Heritage Commission, Suite 500, Continental Building, Main and Markham, Little Rock, Arkansas72201.MODIFICATIONS AND IMPROVEMENTS INTHE FORMAX METHOD OF PREPARING SMALL AVIANSTUDY SPECIMENSTraditional methods of preparing study skins of small avian specimens are often not feasible due to time and expertise required. This may beespecially true when large numbers of specimens, as in the case of tower kills,need to be prepared. Sheridan (Am.Biol. Teacher, January, 1978)reported a method of preparing small avian specimens which entailed injecting formaldehyde saturated with sodium borate (Formax) into thespecimen. However, he didnot present the proportions of the mixture, pinning procedure, or injection amounts in the different areas of the speci-mens. Thispaper describes improvements and standardization ofSheridan's technique.The Formax used in our laboratory consists of 1 gram of sodium borate to 125 mlof standard (37%) formaldehyde. This mixture is easily in-jected into the muscles and internal organs where it diffuses throughout the tissues, drying and preserving them in place. Materials needed forpreserving the specimen are minimal, requiring only syringe, needles, pins, pinning board, ruler and specimen tags. Thus, the preparation ofspecimens is as easy in the field as in the laboratory.Prior to injection, standard measurements of the specimen should be taken (e.g. see Pettingill, Ornithology in laboratory and field, p.447,1970). Formax is then injected into the flightmuscles on each side of the keel, into the abdominal cavity, and into the cranial cavity next to theeye. Larger birds require additional injections in the nape, feet, wings, and/or other parts of the body depending upon specimen size. Table 1summarizes the amounts of Formax injected into various areas forbirds of representative sizes. For additional support of the head, an elongatedS-shaped wire hook isinserted down the mouth and throat (Fig. 1). The use of this wireand the dryingfactor of Formax increases the usefulness ofthe teaching specimen as itis not as fragile as one prepared by the traditional skinning method. To prevent seepage of Formax from the mouth,withmatted feathers as a result, a small piece ofcotton is inserted inthe mouth. The cotton is usually placed inthe mouth prior to injection of thecranial cavity and then replaced by a fresh piece after injection.Figure 1illustrates proper positioning and pinning of the specimen for drying. Insome cases, additional support for the wings may be neces-sary. During the pinning process, feathers, especially on the dorsal side, may be moved outof place. This can be corrected bypushing a pin underthe specimen from the anterior to posterior, several times. By modifying the pinning procedure, mounted specimens may also be prepared usingthis method.Formax-prepared specimens have been particularly useful for teaching purposes. Birds dissected after a one year period still retain excellentpreservation properties, and the Formax-prepared specimens appear to be much more durable than traditional bird skins. This last factor isparticularly important inthe classroom where the specimen is handled a great deal by a large number ofstudents.76Journal of the Arkansas Academy of Science, Vol. 33 [1979], Art. 28Published by Arkansas Academy of Science, 1979Arkansas Academy ofScience Proceedings, Vol.XXXIII,1979 77General NotesTable 1. Approximate Amounts of Formax Required for Birds ofVarious SizesCranial Cavity Each Pectoral Muscle Abdominal Cavi^Short-billedMarsh Wren 0.05cc 0.15cc 0.25ccPurple Finch 0.10 0.40 0.60Sharp-shinnedHawk 0.30 1.30 1.60Bobwhite Quail 0.30 1.40 1.90Roadrunner 0.50 Z.00 3.50Figure 1. Positioning and pinningof a Formax preserved specimen.Note position of S-shaped support inmouth and throat.MARTIND. FLOYDand GARYA.HEIDT, Dept. ofBiology,University ofArkansas atLittle Rock, LittleRock, AR 72204.TRICHOMES OF SOME MEMBERS OF THE LOASACEAE, A SCANNING ELECTRON MICROSCOPE STUDY.The plant familyLoasaceae contains herbs, shrubs, and some woody vines. The leaves are alternate or opposite, entire or variously divided,and generally covered with rough bristly or barbed hairs. Fifteen genera of about 250 species are native to the Americas, and one species toSouthwest Africa.Metcalfe and Chaulk (1950) have presented a summary of the known trichome types; however, most of this seems tohave been from the workof Solereder (1908). Thurston and Lersten (1969) and Thurston (1969, 1974) provided data on the stinging emergences of Loasa tricolor. Thomp-son (1963) and his co-workers (Davis and Thompson, 1967; Ernst and Thompson, 1963; Thompson and Roberts, 1971;and Thompson and Zavor-tink, 1968) have supplied taxonomic data on species ofLoasa of the United States; they have done only limited work with trichome morphology.They worked with trichomes onall parts of the plant, and provided an illustration of the dendritic or candelabra type as described by Payne(1978). Hill(1975, 1976, 1977) worked with the seeds, and Jensen et al(1978) have recently worked with protein chemistry and chromosome num-bers of the group.This report provides some new morphological data concerning the types of trichomes, employing the scanning electron microscope.tOnespecies of Mentzelia was examined from live material, four species of Mentzelia and one of Loasa were rehydrated from herbariumrial, and all other specimens were frompressed herbarium materials. The livingleaf samples were fixedinCRAF V solution, dehydrated withfied DMP,and critical point dried from CO2. The rehydrated specimens were prepared byplacing them inwater which was then heated untilample dropped to the bottom of the container. After soaking there forone week, they were dehydrated withDMP and critical point driedCOi. Allspecimens were then mounted onto stubs and coated with approximately 50 A of carbon and 50 A of 40/60 gold palladium byurnevaporation. They were then examined and photographed in a Cambridge S-600 using Polaroid 665 P/N film.IThe trichomes observed during this study which appear to be different from those described in the literature are numerically listed belowg withthe genus from which they were observed:1. Ananchor trichome withelongated, downwardlydirected barbs alternately arranged on the stalk. Eucnide. Fig. 1.2. Very elongated, pointed trichome covered withsmall, oppositely or whorled upwardly directed spines. Mentzelia. Fig. 2.3. Short conical-shaped with straight barbs inwhorls, and anchor-like cap, and a raised, apparently multicellular base. Mentzelia. Fig. 3.4. Long tapering trichome with spines pointing straight out and seemingly randomly arranged. Petalonyx. Fig. 4. There may be two differ-ent sizes within this group.5. Long tapering trichome withelongated horizontal protuberances randomly arranged. Loasa. Fig. 5.6. Thin tapering trichome withoutward pointing spines which do not extend to the apical end. This type is attached to a large, flat basal oraccessory cell. Mentzelia. Fig.6.These types give onlyapartial presentation of the data gathered. Figures 7-10illustrate apicalportions of trichomes seen onotherwise similartrichomes. Further investigation must be completed before a clear understanding of these observations can be achieved.1Although Loasaceae trichome morphology has been organized into groups ofsix unicellular forms and one multicellular form, this examina-using the scanning electron microscope has revealed trichome morphology that does not satisfy previous criterion.Several other modifications not presented here also have been observed, but it is not yet known whether these are developmental stages ordistinct forms. A study utilizing livingspecimens is inprogress from which it is anticipated that the developmental stages of some of these tri-chomes can be determined. From current morphological data, it appears that a classification system dividingLoasaceae trichomes into at leastthe followingfour (4) majorcategories wouldbe more appropriate:1. Stinging emergences2. Simple acerate to attenuate, of variable length, withor without tuberous swelling3. Branching or candelabra type4. Attenuate anchor hairs, with categorization based upon:a. Apex formb. Barb type and arrangement on shaftc. Length of shaftd. Base type77Journal of the Arkansas Academy of Science, Vol. 33 [1979], Art. 28https://scholarworks.uark.edu/jaas/vol33/iss1/28

Modifications and Improvements in the Formax Method of Preparing Small Avian Study Specimens

ScholarWorks@UARK

Journal of the Arkansas Academy of ScienceVolume 33 Article 281979Modifications and Improvements in the FormaxMethod of Preparing Small Avian Study SpecimensMartin A. FloydUniversity of Arkansas at Little RockGary A. HeidtUniversity of Arkansas at Little RockFollow this and additional works at: http://scholarworks.uark.edu/jaasPart of the Poultry or Avian Science CommonsThis article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able toread, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking priorpermission from the publisher or the author.This General Note is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the ArkansasAcademy of Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu, ccmiddle@uark.edu.Recommended CitationFloyd, Martin A. and Heidt, Gary A. (1979) "Modifications and Improvements in the Formax Method of Preparing Small Avian StudySpecimens," Journal of the Arkansas Academy of Science: Vol. 33 , Article 28.Available at: http://scholarworks.uark.edu/jaas/vol33/iss1/2876 Arkansas Academy of Science Proceedings, Vol.XXXIII,1979Arkansas Academy of ScienceThis survey resulted inthe finding of three specimens. Two specimens were captured inMay on a 65-acre prairie inBoone County (Marsh,1978), and the other specimen was captured in July on a 40-acre prairie inFranklin County, representing the first record for this county. Thusfrom 1909 to 1978 there are reports of the ornate box turtle inten Arkansas counties: Benton, Boone, Fulton, Washington, Franklin, Perry, Gar-land, Prairie, Lafayette, and Columbia (Figure I).Being a species which is restricted by habitat availability, the ornate box turtle has been greatlyaffected by the changes inland use practices occurring in Arkansas, and has been considered a rare species (Reagen, 1974). Conversion tomoreproductive agricultural use has reduced the amount of native prairie and undisturbed grassland available to the ornate box turtle, and this may bethe most important factor limitingthe ornate box turtle, and this may be the most important factor limiting the ornate box turtle's range in Arkan-sas. Inaddition, the Arkansas highway system has taken a tollbecause the turtle seems to exhibit a certain affinity for roadways and a great num-ber are killed bymotor vehicles (Legler, 1960 and Reagan, 1974). Toproperly assess the status of the ornate box turtle (Terrapene ornata ornataland to eliminate confusion concerning its distribution inArkansas a detailed studyis obviously needed.The author expresses appreciation to the following forvarious types of assistance during the course of this project: Dr.R.Baldauf,Dr.R. V.McDaniel, and Valerie Thwing. Dr.M.V. Plummer is gratefully acknowledged for his assistance with literature and forhis confirmation of species. Special thanks is extended to the Arkansas Natural Heritage Commission and staff, for continued sup-port and cooperation.LITERATURE CITEDCAGLE,F. R. 1957. Reptiles. In:Vertebrates of the United States.McGraw-HillBook Company, New York.pp. 213-268.MARSH, M. A. 1978. Baker prairie. The Ozark Soc. Bull. 12(1):10-11.CONANT,R. 1975. A field guide to reptiles and amphibians of east-ern and central North America. Houghton Mifflin Company,Boston. 429 pp.REAGAN,D. P. 1974. Threatened native reptiles of Arkansas. In°Arkansas natural area plan. Arkansas Department of Planning,LittleRock. pp. 101-105.DELLINGER, S. C. and J. D.BLACK. 1938. Herpetology of Arkan-sas: part 1, the reptiles. Occ. Papers Univ. Ark.Mus. no. 1. 47pp.SCHWARDT, H. H. 1938. Reptiles of Arkansas. Univ. Ark. AgriExper. Stat. Bull.357:1-47.WARD,J. P. 1978. Terrapene ornata. In: Catalog of American am-phibians and reptiles. Society for the Study of Amphibians andReptiles, pp. 217.1-217.4.DOWLING,H.G. 1957. Areview of the amphibians and reptiles ofArkansas. Occ. Papers Univ.Ark.Mus. no.3. 51 pp.LEGLER, J.M. 1960. Natural history of the ornate box turtle, Terra-pene ornata ornata, Agassiz. Univ. Ka. Mus. Nat. Hist. Pub.ll(10):527-669.LAWANA ENGLAND, Arkansas Natural Heritage Commission, Suite 500, Continental Building, Main and Markham, Little Rock, Arkansas72201.MODIFICATIONS AND IMPROVEMENTS INTHE FORMAX METHOD OF PREPARING SMALL AVIANSTUDY SPECIMENSTraditional methods of preparing study skins of small avian specimens are often not feasible due to time and expertise required. This may beespecially true when large numbers of specimens, as in the case of tower kills,need to be prepared. Sheridan (Am.Biol. Teacher, January, 1978)reported a method of preparing small avian specimens which entailed injecting formaldehyde saturated with sodium borate (Formax) into thespecimen. However, he didnot present the proportions of the mixture, pinning procedure, or injection amounts in the different areas of the speci-mens. Thispaper describes improvements and standardization ofSheridan's technique.The Formax used in our laboratory consists of 1 gram of sodium borate to 125 mlof standard (37%) formaldehyde. This mixture is easily in-jected into the muscles and internal organs where it diffuses throughout the tissues, drying and preserving them in place. Materials needed forpreserving the specimen are minimal, requiring only syringe, needles, pins, pinning board, ruler and specimen tags. Thus, the preparation ofspecimens is as easy in the field as in the laboratory.Prior to injection, standard measurements of the specimen should be taken (e.g. see Pettingill, Ornithology in laboratory and field, p.447,1970). Formax is then injected into the flightmuscles on each side of the keel, into the abdominal cavity, and into the cranial cavity next to theeye. Larger birds require additional injections in the nape, feet, wings, and/or other parts of the body depending upon specimen size. Table 1summarizes the amounts of Formax injected into various areas forbirds of representative sizes. For additional support of the head, an elongatedS-shaped wire hook isinserted down the mouth and throat (Fig. 1). The use of this wireand the dryingfactor of Formax increases the usefulness ofthe teaching specimen as itis not as fragile as one prepared by the traditional skinning method. To prevent seepage of Formax from the mouth,withmatted feathers as a result, a small piece ofcotton is inserted inthe mouth. The cotton is usually placed inthe mouth prior to injection of thecranial cavity and then replaced by a fresh piece after injection.Figure 1illustrates proper positioning and pinning of the specimen for drying. Insome cases, additional support for the wings may be neces-sary. During the pinning process, feathers, especially on the dorsal side, may be moved outof place. This can be corrected bypushing a pin underthe specimen from the anterior to posterior, several times. By modifying the pinning procedure, mounted specimens may also be prepared usingthis method.Formax-prepared specimens have been particularly useful for teaching purposes. Birds dissected after a one year period still retain excellentpreservation properties, and the Formax-prepared specimens appear to be much more durable than traditional bird skins. This last factor isparticularly important inthe classroom where the specimen is handled a great deal by a large number ofstudents.76Journal of the Arkansas Academy of Science, Vol. 33 [1979], Art. 28Published by Arkansas Academy of Science, 1979Arkansas Academy ofScience Proceedings, Vol.XXXIII,1979 77General NotesTable 1. Approximate Amounts of Formax Required for Birds ofVarious SizesCranial Cavity Each Pectoral Muscle Abdominal Cavi^Short-billedMarsh Wren 0.05cc 0.15cc 0.25ccPurple Finch 0.10 0.40 0.60Sharp-shinnedHawk 0.30 1.30 1.60Bobwhite Quail 0.30 1.40 1.90Roadrunner 0.50 Z.00 3.50Figure 1. Positioning and pinningof a Formax preserved specimen.Note position of S-shaped support inmouth and throat.MARTIND. FLOYDand GARYA.HEIDT, Dept. ofBiology,University ofArkansas atLittle Rock, LittleRock, AR 72204.TRICHOMES OF SOME MEMBERS OF THE LOASACEAE, A SCANNING ELECTRON MICROSCOPE STUDY.The plant familyLoasaceae contains herbs, shrubs, and some woody vines. The leaves are alternate or opposite, entire or variously divided,and generally covered with rough bristly or barbed hairs. Fifteen genera of about 250 species are native to the Americas, and one species toSouthwest Africa.Metcalfe and Chaulk (1950) have presented a summary of the known trichome types; however, most of this seems tohave been from the workof Solereder (1908). Thurston and Lersten (1969) and Thurston (1969, 1974) provided data on the stinging emergences of Loasa tricolor. Thomp-son (1963) and his co-workers (Davis and Thompson, 1967; Ernst and Thompson, 1963; Thompson and Roberts, 1971;and Thompson and Zavor-tink, 1968) have supplied taxonomic data on species ofLoasa of the United States; they have done only limited work with trichome morphology.They worked with trichomes onall parts of the plant, and provided an illustration of the dendritic or candelabra type as described by Payne(1978). Hill(1975, 1976, 1977) worked with the seeds, and Jensen et al(1978) have recently worked with protein chemistry and chromosome num-bers of the group.This report provides some new morphological data concerning the types of trichomes, employing the scanning electron microscope.tOnespecies of Mentzelia was examined from live material, four species of Mentzelia and one of Loasa were rehydrated from herbariumrial, and all other specimens were frompressed herbarium materials. The livingleaf samples were fixedinCRAF V solution, dehydrated withfied DMP,and critical point dried from CO2. The rehydrated specimens were prepared byplacing them inwater which was then heated untilample dropped to the bottom of the container. After soaking there forone week, they were dehydrated withDMP and critical point driedCOi. Allspecimens were then mounted onto stubs and coated with approximately 50 A of carbon and 50 A of 40/60 gold palladium byurnevaporation. They were then examined and photographed in a Cambridge S-600 using Polaroid 665 P/N film.IThe trichomes observed during this study which appear to be different from those described in the literature are numerically listed belowg withthe genus from which they were observed:1. Ananchor trichome withelongated, downwardlydirected barbs alternately arranged on the stalk. Eucnide. Fig. 1.2. Very elongated, pointed trichome covered withsmall, oppositely or whorled upwardly directed spines. Mentzelia. Fig. 2.3. Short conical-shaped with straight barbs inwhorls, and anchor-like cap, and a raised, apparently multicellular base. Mentzelia. Fig. 3.4. Long tapering trichome with spines pointing straight out and seemingly randomly arranged. Petalonyx. Fig. 4. There may be two differ-ent sizes within this group.5. Long tapering trichome withelongated horizontal protuberances randomly arranged. Loasa. Fig. 5.6. Thin tapering trichome withoutward pointing spines which do not extend to the apical end. This type is attached to a large, flat basal oraccessory cell. Mentzelia. Fig.6.These types give onlyapartial presentation of the data gathered. Figures 7-10illustrate apicalportions of trichomes seen onotherwise similartrichomes. Further investigation must be completed before a clear understanding of these observations can be achieved.1Although Loasaceae trichome morphology has been organized into groups ofsix unicellular forms and one multicellular form, this examina-using the scanning electron microscope has revealed trichome morphology that does not satisfy previous criterion.Several other modifications not presented here also have been observed, but it is not yet known whether these are developmental stages ordistinct forms. A study utilizing livingspecimens is inprogress from which it is anticipated that the developmental stages of some of these tri-chomes can be determined. From current morphological data, it appears that a classification system dividingLoasaceae trichomes into at leastthe followingfour (4) majorcategories wouldbe more appropriate:1. Stinging emergences2. Simple acerate to attenuate, of variable length, withor without tuberous swelling3. Branching or candelabra type4. Attenuate anchor hairs, with categorization based upon:a. Apex formb. Barb type and arrangement on shaftc. Length of shaftd. Base type77Journal of the Arkansas Academy of Science, Vol. 33 [1979], Art. 28http://scholarworks.uark.edu/jaas/vol33/iss1/28

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Modifications and Improvements in the Formax Method of Preparing Small Avian Study Specimens

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