Unmounted Specimens Research Articles

Unmounted Doesn’t Mean Unprocessed: A Case Study of Exporting Australian Herbarium Specimens

In 2017, two incoming international herbarium loans were confiscated and destroyed by the Commonwealth Department of Agriculture and Water Resources (DAWR). Following these regrettable incidents, communication has improved between Managers of Australasian Herbarium Collections (MAHC) and Plant Import Operations Branch of DAWR. The outcome is that new protocols now exist for shipment of plant material between herbaria based on the recognition by DAWR that all herbarium specimens are in fact processed (pressed, dried and frozen) even if they are not fully mounted (Brown et al, this conference). Simultaneously, the United States Department of Agriculture (USDA) had been reported to be destroying incoming unmounted specimens, thus making it difficult to send unmounted exchange material overseas, and particularly to the USA. Using the documentation developed for incoming overseas loans by MAHC and, encouraging the receiving institution in the USA to acquire a USDA 588 Importing Permit for unprocessed plant specimens (non-loan, unmounted herbarium specimens), it was possible to export unmounted herbarium specimens and dried DNA samples into the USA. Excluding the USA specific requirements, we were also able to export exchange material and dried DNA samples to Singapore and Sweden.

Rethinking the links between systematic studies and ex situ living collections as a contribution to the Global Strategy for Plant Conservation

The Museum national d’histoire naturelle (MNHN) in Paris holds ca. 70 million specimens. The collections were in need of a strategy to ensure their long-term conservation. We discuss how the Department of Botanical and Zoological Gardens (DJBZ; tropical living collections), and the Department of Systematics and Evolution (DSE; herbarium) contribute to achieving GSPC’s Target 1 (‘a widely accessible working list of known plant species as a step towards a complete world flora’). The DJBZ started encouraging better management of the collections, evolving towards focused reference collections, where all specimens have well-documented collection data. The objective is to link all collections to a scientific referee. This has already been achieved for a number of taxa. The herbarium of the DSE (acronym P) is among the world’s largest (11 million specimens, including 400,000 types). The collection’s heterogeneity impedes access to its data, since P is a mix of recent well-documented collections and historical collections at various curational levels. P is currently under renovation, which started by mounting all ca. 2 million unmounted specimens. The project also includes databasing and imaging of every specimen. The database now holds around 1,000,000 records. For taxonomic studies, living collections are crucial, especially for plants that are not easily preserved as herbarium specimens. Living collections also enable studies impossible to forecast at the time of collecting. Herbaria and living collections should therefore be conceived as interoperable entities requiring common scientific curation. Through a combination of its assets and the expertise of its researchers, the MNHN is well prepared to tackle the new objectives of the GSPC beyond 2010.

Rapid decline of PCR amplification from genomic extracts of DESS-preserved, slide-mounted nematodes

Abstract Many studies use integrative methods to study both morphology and gene sequences of nematode species, yet there is little evidence to indicate the optimum criteria for merging taxonomic and molecular protocols. Preliminary evidence suggests that standard methods of desiccation and slide mounting nematode specimens in glycerin can sporadically result in degradation of genomic DNA. A time series experiment was constructed in order to assess whether this degradation of genomic DNA could be recorded and quantified. Two groups of nematode specimens were desiccated, mounted on slides, and stored at either 4°C or at room temperature for specified time intervals. Genomic DNA was extracted and PCR was conducted to amplify a section of the 18S rRNA gene at each time point. The resulting gel photographs were used to record the outcome of PCR and quantify the strength of amplification if successful. Results from slide-mounted specimens were compared to PCR products derived from unmounted nematodes extracted directly from the preservative solution at specified time intervals. The desiccation and slide mounting process appears to reduce overall band intensity after 1 day in slide mounts. Unmounted specimens consistently exhibit high success rates of PCR and a high overall DNA content per band at all time points, whereas slide-mounted nematodes show a decrease in the number of successful PCRs and weakening band intensity as time progresses. Results clearly indicate a steady degradation of genomic DNA in nematodes stored in slide mounts for more than 2 weeks, whereas unmounted specimens extracted from preservative solution showed no decline in PCR success or quality. We suggest a maximum storage period of 2 weeks on slides if mounted nematodes are to be used for molecular analyses.

Thomas F. Lucy's Plant Collections, Donated to the Buffalo Society of Natural Sciences in the Early 20Th Century

The botanical collection of Thomas Francis Lucy (1844–1906), which includes voucher specimens for The Flora of the Upper Susquehanna and its Tributaries (Clute 1898), was rediscovered at Elmira College, Elmira, New York in 1984 (Kelloff et al. 1990). These specimens were originally part of the ‘‘Elmira Academy of Sciences, The Upper Susquehanna Flora’’ herbarium, as identified by Lucy’s herbarium labels, and they were transferred to Elmira College around 1881 (Kelloff et al. 1990). This rediscovered herbarium formed the basis for the Elmira College Herbarium (ECH), established in 1985 (Holmgren et al. 1990). Lucy’s Upper Susquehanna flora specimens at ECH-Lucy were believed to be the key to another set of his specimens donated to the herbarium of the Buffalo Society of Natural Sciences (BUF), Buffalo, New York following Lucy’s death in 1906. Much of the history of the Lucy collection has been documented by Kelloff et al. (1990) and Kelloff and Kass (1993). Since Lucy’s field notebooks and catalogue did not accompany the collection that BUF acquired in the early 1920s, the BUF curators had no way to identify the species of most of the unmounted specimens, which were coded only with numbers written on the newspapers that contained them (Kelloff and Kass 1993). Learning that ECH had Lucy’s fully identified and labeled original collection, RHODORA, Vol. 110, No. 944, pp. 484–491, 2008 E Copyright 2008 by the New England Botanical Club

Life Cycle Details of Solenopotes capillatus (Anoplura: Linognathidae)

The duration of immature life stages and the preoviposition period of Solenopotes capillatus Enderlein (Anoplura: Linognathidae) were determined by daily observation in vivo of individual specimens. The duration of the egg and nymphal stages was obtained by daily observation of eggs deposited by females transferred to normal predilection sites of uninfested cattle. The preoviposition period was determined by transferring third instars to uninfested cattle and then recording the period from adult eclosion to first oviposition. The egg stage required 11–13 d. Both first and second instars required 3–4 d, and third instars, based on observations of females, required 5–6 d. The preoviposition period was 2 d. Between 27 and 29 d was required for completion of the cycle (egg to egg). The size (length) of unmounted specimens, in millimeters, was egg 0.70; first instar 0.69; second instar, 0.82; third instar, 1.06; adult female, 1.50; and adult male, 1.08.

Life Cycle Details of <I>Solenopotes capillatus</I> (Anoplura: Linognathidae)

The duration of immature life stages and the preoviposition period of Solenopotes capillatus Enderlein (Anoplura: Linognathidae) were determined by daily observation in vivo of individual specimens. The duration of the egg and nymphal stages was obtained by daily observation of eggs deposited by females transferred to normal predilection sites of uninfested cattle. The preoviposition period was determined by transferring third instars to uninfested cattle and then recording the period from adult eclosion to first oviposition. The egg stage required 11-13 d. Both first and second instars required 3-4 d, and third instars, based on observations of females, required 5-6 d. The preoviposition period was 2 d. Between 27 and 29 d was required for completion of the cycle (egg to egg). The size (length) of unmounted specimens, in millimeters, was egg 0.70; first instar 0.69; second instar, 0.82; third instar, 1.06; adult female, 1.50; and adult male, 1.08.

Volume changes during preparation of mouse embryonic tissue for scanning electron microscopy

AbstractThis paper describes the results of experiments in which the volume changes in mouse embryo limb samples were followed more or less continuously after fixation through dehydration and critical point drying, with in some instances data relating to post critical point drying shrinkage.14 and 15 day p. c. mouse embryos were fixed in 3 % glutaraldehyde in cacodylate buffer and stored in this fixative until use. Single specimens were studied using a Quantimet image analysing computer to record the changes in projected area of the unmounted specimens as they were passed through the usual series of reagents according to various commonly used dehydration schedules. The area changes were converted to volume changes for the purposes of presentation in this paper. The Quantimet system could not be used to follow volume changes in the CPD bomb so that most experiments detail the volume in the intermediate fluid before CPD and the size of the specimen immediately after it was removed from the CPD bomb. A few experiments were conducted in which the specimens were measured whilst they were in the CPD bomb. The measurements relating to dehydration and CPD procedures were compared with measurements of air dried and freeze dried specimens.All three drying methods cause considerable shrinkage: freeze drying to 85 % of the glutaraldehyde fixed tissue volume; critical point drying to 41% (after 24 h); and air drying from a volatile solvent to about 18% of the fixed tissue volume. Air drying from water caused a shrinkage to about 12% of the original volume. There was no significant difference between the various commonly used CPD schedules or between GA only and GA + Os O4 fixed tissue. CPD via cellosolve and CO2 caused substantially more shrinkage than other methods.Dimensional changes during specimen preparation are probably associated with changes in shape and in relative relationships between organelles, cells and tissues having different compositions. This should be borne in mind by all those interpreting scanning electron micrographs of dried animal soft tissue specimens.

Amblyomma imitator, a New Species of Tick from Texas and Mexico, and Remarks on the Synonymy of A. cajennense (Fabricius) (Acarina-Ixodidae)

Holotype: Female, from goat, Brownsville, Texas, February 8, 1934 (RML 10021) L. Demieville. In the Rocky Mountain Laboratory. Allotype: Male, data as for holotype. In the Rocky Mountain Laboratory. Paratypes: 3 males, 5 females, 7 nymphs with same data as holotype. 14 males, 12 females, from cow, Brownsville, Texas, May 18, 1935 (RML 11087) L. Demieville. 3 males, 7 females, 1 nymph, from man, El Baiito, near Valles, San Luis Potosi, Mexico, 1939 (RML 17459) R. Traub. Paratypes are deposited in the U. S. National Museum, Chicago Natural History Museum, Laboratorio de Entomologia de Instituto de Salubridad y Enfermedades Tropicales, Mexico, D.F., and the Rocky Mountain Laboratory. Over 700 other specimens examined, designated by the accession numbers assigned to them at the Rocky Mountain Laboratory, are as follows: 10036, 10048, 11049, 11060, 11063, 32241, 34576, 34585, 32165, 32172, 32175, 32180, 32178, 32169, 25328, 25329, 15492, 15493, 15494, 17470, 17467, 17458, 17461, 17465, 17468, 29974, 24010, 24011, 24015, 32160, 32176. These are all from Texas and Mexico, and from these hosts: man, dogs, horses, burros, cattle, goats, deer, peccary, opossum, Sciurus aureogaster, Meleagris gallopavo, Geococcyx californianus; some were from vegetation or the ground. Female: Resembles A. cajennense but tends to be smaller; differs particularly from it in lacking chitinous tubercles at the postero-internal angle of the festoons, and in details of the external genitalia. Length of holotype, excluding capitulum, 3.18; width 2.56. Length and width of largest and smallest unfed specimens seen, 3.49 by 2.67 and 2.87 by 2.15, respectively. Contour oval, broadest at about middle. Color of unfed specimens paler than usual in Amblyomma. Largest fed specimen seen is 9.74 long by 6.66 wide and both ends are equally broad. Capitulum: Length (holotype) 0.98; width of basis, 0.63. Details as in A. cajennense. Scutum: Triangular, 1.57 long by 1.70 wide (holotype); size ranges from about 1.34 by 1.52 to 1.84 by 1.90. Shape, grooves and punctations as in A. cajennense. Ornamentation variable in distinctness and extent; pattern, when well developed, as in Figure 1B, similar to that of A. cajennense, Figure 1D. Legs and Spiracular Plate: As in A. cajennense. Genital aperture: Located between coxae II as in A. cajennense. Genital apron overlaid posterolaterally on either side by a blunt flattened projection darker in color than the apron and adjacent integument, as illustrated in Figure 2A. In A. cajennense these projections are long and narrow as shown in Figure 2B. This differentiating character is easily observed in clean, unfed or fed, unmounted specimens properly lighted under 36x magnification. Male: Closely resembles A. cajennense and not always distinguishable from it with certainty but usually paler and tending to be narrower and smaller. Ornamentation variable in extent and definition; pattern, when well developed, as in Figure 1A and similar to that of A. cajennense as shown in Figure 1C. Length of allotype, excluding capitulum, 2.97; width 2.36. Largest and smallest specimens seen are 3.48 by 2.66 and 2.26 by 1.77, respectively. Nymph: Similar to A. cajennense except scutum shagreened instead of smooth and punctations somewhat larger, deeper, and more distinct.

Collections of Ectoparasitic Mites from Alaska

This report on ectoparasitic mites from Alaska is presented to supplement the few published accounts of mites from the territory (see references). All species concerned are members of the family LAELAPTIDAE. Specimens reported were obtained from three sources: Rocky Mountain Laboratory of the United States Public Health Service, through Mr. Glen M. Kohls; United States National Museum, through Dr. E. W. Baker; and Arctic Aeromedical Laboratory of the United States Air Force, through Captain John M. Geary. Among the specimens sent by Mr. Kohls were three collections made on Nunivak Island by personnel of the Arctic Health Research Center of the United States Public Health Service at Anchorage. Unmounted specimens were prepared for study by enlisted personnel of the survey section, 498th Preventive Medicine Company, Fort Sam Houston, Texas. Specimens reported in this paper were taken from rodents and birds at several widely separated localities from Nunivak Island off the coast of southwestern Alaska to Umiat and Barter Island far above the Arctic circle. Eight species are represented in these collections. In the list which follows, Rocky Mountain Laboratory is abbreviated as RML, and Arctic Health Research Center is abbreviated as AHRC. Identified specimens have been returned to the institutions from which they were obtained.

A Modification of Alizarin Red S Technic for Demonstrating Bone Formation: Methods of Mounting, Photographing and Demonstrating the Specimens

This paper shows that by using solutions heated in the incubator during certain stages, the alizarin red S method of staining the ossified centers in embryos has been shortened, with a consequent saving in time.New methods of mounting the specimens have been evolved and are described in detail.The technic of photographing mounted and unmounted specimens is outlined and illustrated by diagrams.Diagrammatic illustrations are provided of the various types of apparatus used, including a plan of the cabinet for demonstrating clearly the smaller embryos mounted between watch glasses. Photographic examples of the results achieved are also shown.

A revision of the species of the genus Trichobius

This paper deals with the species of the commonest genus of the American Streblidae. The descriptions of the species were made from specimens mounted on slides, in Canada balsam. Here I wish to point out again that spirit preserved specimens are unsuitable for the study of these Diptera. These specimens do not show clearly the details of the structure of some sclerotized parts, and especially the arrangement of the small setae on the thorax, which in some cases, are the only reliable characters for the separation of the species. The study of unmounted specimens of these Diptera may easily lead to incorrect identification, or to the increase of undesirable synonyms.