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Pollen from the Exoskeletons of Stable Flies, Stomoxys Calcitrans (Linnaeus 1758), in Gainesville, Florida, U.S.A

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Pollen from the Exoskeletons of Stable Flies, Stomoxys Calcitrans (Linnaeus 1758), in Gainesville, Florida, U.S.A University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln U.S. Department of Agriculture: Agricultural Publications from USDA-ARS / UNL Faculty Research Service, Lincoln, Nebraska 2008 Pollen From The Exoskeletons Of Stable Flies, Stomoxys Calcitrans (Linnaeus 1758), In Gainesville, Florida, U.S.A. David M. Jarzen University of Florida, [email protected] Jerome Hogsette United States Department of Agriculture-ARS-Center for Medical, [email protected] Follow this and additional works at: https://digitalcommons.unl.edu/usdaarsfacpub Part of the Agricultural Science Commons Jarzen, David M. and Hogsette, Jerome, "Pollen From The Exoskeletons Of Stable Flies, Stomoxys Calcitrans (Linnaeus 1758), In Gainesville, Florida, U.S.A." (2008). Publications from USDA-ARS / UNL Faculty. 1031. https://digitalcommons.unl.edu/usdaarsfacpub/1031 This Article is brought to you for free and open access by the U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Publications from USDA-ARS / UNL Faculty by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. POLLEND.M. Jarzen and J.A. FROM Hogsette: Pollen THE from the EXOSKELETONS exoskeletons of stable flies, Stomoxyx OFcalcitrans in Gainesville, Florida, U.S.A. 77 STABLE FLIES, STOMOXYS CALCITRANS (LINNAEUS 1758), IN GAINESVILLE, FLORIDA, U.S.A. DAVID M. JARZEN Paleobotany and Palynology Laboratory Florida Museum of Natural History University of Florida Gainesville, Florida 32611-7800 U.S.A. e-mail: [email protected] JEROME A. HOGSETTE U.S. Department of Agriculture Agricultural Research Service Center for Medical, Agricultural, and Veterinary Entomology Gainesville, Florida 32608 U.S.A. e-mail: [email protected] Abstract Stable flies are an important pest of humans and livestock. Despite being blood feeders, they also visit flowers to eat nectar. Stable flies with pollen adhering to their exoskeletons were collected at the University of Florida Horse Teaching Unit and taken to the Paleobotany and Palynology Laboratory at the Florida Museum of Natural History for processing and identification. The pollen on all the specimens was identified as Carolina willow, Salix caroliniana Michaux 1803. This small shrub or tree is found throughout Florida in wetland areas. This study demonstrates the utility of pollen analysis in interdisciplinary studies. Key words: Stable flies; pollen; Salix caroliniana; Stomoxys calcitrans; Florida. INTRODUCTION foraging and nectar-feeding habits of insects is important for several reasons. Pollen identification may be useful in Stable flies, Stomoxys calcitrans (Linnaeus 1758), are the determination of feeding habits, migration patterns, hematophagous, pestiferous flies that are a major pest of migration routes, and origins (Jones and Jones, 2001). humans and livestock in the United States and many other Migration patterns and routes are important because corri- regions (Hogsette and Farkas, 2000; Broce et al., 2005). dors of movement may be predicted, and origins of dis- Although both sexes feed on blood, they are known to visit persal can be identified. flowers and eat nectar (Jones et al., 1985). A list of plants visited by stable flies in Florida has been compiled by MATERIAL AND METHODS observation and identification of pollen adherent to the flies (Tseng et al., 1983). If the source of the pollen could be During a March, 2008 study of stable flies at the Horse identified, the habits of the flies would be better defined, Teaching Unit (HTU), University of Florida, Gainesville thereby adding to the information on stable fly dispersal (Alachua County), Florida, stable flies with lateral and and feeding habits (Hogsette and Ruff, 1985; Hogsette et ventral surfaces covered with a bright yellow pollen-like al., 1987). Stable flies are not pollen feeders because their substance were captured on sticky traps. Several of these mouth parts are unsuitable (Jones et al., 1985). However, flies were brought to the Paleobotany and Palynology these flies visit plants for a nectar source and, in doing so, Laboratory at the Florida Museum of Natural History for become contaminated with pollen. An understanding of the investigation and possible identification of the pollen. Palynology, 32 (2008): 77–81 © 2008 by AASP Foundation ISSN 0191-6122 78 PALYNOLOGY, VOLUME 32 — 2008 Initially, a single fly was used as a test to determine are stored at the Paleobotany and Palynology Laboratory, suitable processing methods that would produce satisfac- Florida Museum of Natural History, Gainesville, Florida, tory recovery of the pollen adhering to the body of the U.S.A. Photographs of the pollen grains were made using flies. A single fly, bearing pollen grains, was placed into a Nikon Coolpix 4500™ camera mounted on a Leitz a 4 ml vial. A small amount of 100% ethanol was added Dialux 20™ transmitted light research microscope. The to the vial and gently shaken. This effectively freed the specimens are located by stage coordinates marking an x pollen grains from the fly body and hairs. The liquid was and y axis location for the Leitz Dialux 20 microscope. then poured off into a 15 ml centrifuge tube and processed Pollen used for comparison with the pollen adhering to the using standard palynological procedures for modern pol- fly bodies was collected from the herbarium at the Florida len (Traverse, 2007). Twelve additional samples of pollen Museum of Natural History (FLAS), and processed using washed from the bodies of flies with 100% ethanol were the techniques outlined by Traverse (2007) for modern then processed. An optical check, using a dissecting pollen. Species of Salix were used as a comparison with microscope, of the flies made before the ethanol rinse, the pollen attached to the bodies of the flies. Measure- indicated very little pollen was present in these twelve ments were made on 10–15 grains at x400 magnification. samples. Because of this, the samples were transferred to Table 1 gives the herbarium sheet data for the specimens 1.5 ml caped, conical, plastic centrifuge tubes examined, and representative specimens are illustrated in (Eppendorf™ microcentrifuge tubes), and washed once Plate 1. with distilled water to remove most of the ethanol. A single wash in glacial acetic acid removed the water in SYSTEMATIC SECTION preparation for acetolysis. The samples were then treated with about 1.0 ml of acetolysis mixture (acetic anhydride Ten of the thirteen samples contained pollen in varying and concentrated sulfuric acid in a 9:1 ratio), and heated amounts. All the pollen recovered from the flies was in a water bath for 10–12 minutes at about 95˚C. Follow- identified as Carolina willow, Salix caroliniana Michaux ing acetolysis, the samples were centrifuged, decanted, 1803 (Salicaceae), and is described here. washed once in glacial acetic acid, and washed using distilled water. The residues were retained in the Eppendorf Family SALICACEAE Mirbel 1815 tubes for slide preparation. Extreme care was taken at this Salix Linnaeus 1753 stage, because the residues appeared clear, suggesting Salix caroliniana Michaux 1803 that little or no pollen was recovered. Plate 1, figs. 1–6 The microscope slides were prepared using glycerin jelly as the mounting medium. The coverslips were sealed Description. Pollen grains single, tricolporate or with clear nail polish. All slides and remaining residues tricolporoidate, prolate to mostly subprolate; colpi Table 1. Data on Salix specimens examined for comparison with the pollen attached to the stable flies. The habit and habitat data are provided as recorded from the herbarium sheet. D.M. Jarzen and J.A. Hogsette: Pollen from the exoskeletons of stable flies, Stomoxyx calcitrans in Gainesville, Florida, U.S.A. 79 meridionally arranged, long, narrow, reaching to the poles; shape to accommodate variations in the volume of the pores indistinct, appearing as a thinned area within the cytoplasm caused by hydration,” (Punt et al., 1994). copal margins; surface reticulate, heterobrochate, reticu- lum finer at poles and along colpi margins, lumina maxi- RESULTS AND DISCUSSION mum size 1.5 µm diameter, muri about 0.5 µm wide; exine 1.0–1.5 µm thick, two-layered, nexine about as thick as All the pollen recovered from the fly bodies is of Salix sexine; dimensions 21(22.8)24 x 17(19)21 µm; p/e = 1.2 caroliniana. Of the 400 species of Salix (Mabberley, 1997), subprolate (10 grains measured). six species occur in Florida (Argus, 1986). These include Remarks. It is of interest to note that cursory examina- Salix babylonica C. Linneaus 1753 (weeping willow), tion of the pollen, using a dissecting microscope, while Salix caroliniana A. Michaux 1803 (Carolina willow), still attached to the body of the flies appeared more prolate Salix eriocephala A. Michaux 1803 (Missouri river wil- to perprolate in general shape. Once the pollen was low), Salix floridana Chapman 1860 (Florida willow), transferred to centrifuge tubes and washed in water, the Salix humilis H. Marshall 1785 (dwarf willow), and Salix shape changed dramatically to subprolate. The take-up of nigra H. Marshall 1785 (black willow). Salix caroliniana, water demonstrated the principle of harmoegathy, i.e. Salix floridana, and Salix humilis occur in Alachua County, “…the process by which pollen grains and spores change Florida (Wunderlin and Hansen, 2004). PLATE 1 Salix pollen recovered from stable fly body hairs, showing polar and equatorial views. Scale bar = 20 µm. 1–3 Salix caroliniana, polar view, PY01A 26.0x99.5, x 4–6 Salix caroliniana, equatorial view, PY01A 29.4x101.5, 1500. Figure 2 illustrates the reticulate surface. x 1500. Figure 6 shows the nature and length of the colpus. 80 PALYNOLOGY, VOLUME 32 — 2008 Measurements for the three Alachua County species of Salix pollen has previously been found on adult stable Salix, based on a count of 10 grains for each species are: flies in west Florida (Tseng et al., 1983). The fact that stable flies from the HTU have also been visiting Salix flowers Salix caroliniana 21(21.5)24 x 17(19)21 µm; p/e ratio = further confirms their tendency to utilize nectar even when 1.13.
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