University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln USDA National Wildlife Research Center - Staff U.S. Department of Agriculture: Animal and Plant Publications Health Inspection Service 2017 Clinostomum album n. sp. and Clinostomum marginatum (Rudolphi, 1819), parasites of the great egret Ardea alba L. from Mississippi, USA Thomas G. Rosser Mississippi State University Neely R. Alberson Mississippi State University Ethan T. Woodyard Mississippi State University Fred L. Cunningham USDA/APHIS/WS National Wildlife Research Center, [email protected] Linda M. Pote Mississippi State University See next page for additional authors Follow this and additional works at: https://digitalcommons.unl.edu/icwdm_usdanwrc Part of the Life Sciences Commons Rosser, Thomas G.; Alberson, Neely R.; Woodyard, Ethan T.; Cunningham, Fred L.; Pote, Linda M.; and Griffin,a M tt .,J "Clinostomum album n. sp. and Clinostomum marginatum (Rudolphi, 1819), parasites of the great egret Ardea alba L. from Mississippi, USA" (2017). USDA National Wildlife Research Center - Staff Publications. 1930. https://digitalcommons.unl.edu/icwdm_usdanwrc/1930 This Article is brought to you for free and open access by the U.S. Department of Agriculture: Animal and Plant Health Inspection Service at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in USDA National Wildlife Research Center - Staff ubP lications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors Thomas G. Rosser, Neely R. Alberson, Ethan T. Woodyard, Fred L. Cunningham, Linda M. Pote, and Matt .J Griffin This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/icwdm_usdanwrc/ 1930 Syst Parasitol (2017) 94:35–49 DOI 10.1007/s11230-016-9686-0 Clinostomum album n. sp. and Clinostomum marginatum (Rudolphi, 1819), parasites of the great egret Ardea alba L. from Mississippi, USA Thomas G. Rosser . Neely R. Alberson . Ethan T. Woodyard . Fred L. Cunningham . Linda M. Pote . Matt J. Griffin Received: 13 May 2016 / Accepted: 21 November 2016 Ó Springer Science+Business Media Dordrecht 2016 Abstract Members of the genus Clinostomum great egrets preying on catfish ponds remains mostly Leidy, 1856, colloquially known as yellow grubs, are understudied. Thirteen great egrets were captured cosmopolitan parasites of piscivorous birds, freshwa- from commercial catfish ponds in northeast Missis- ter snails, fish and amphibians. In the southeastern sippi, and examined for trematode infections. Two United States, piscivorous birds present a continuous morphologically distinct Clinostomum spp. were challenge for producers of farm-raised catfish. Ciconi- observed in the great egrets sampled, one morpholog- iform birds are common hosts of Clinostomum spp. in ically consistent with Clinostomum marginatum (Ru- North America and are endemic on most commercial dolphi, 1819) and one morphologically unique catfish operations. The great egret Ardea alba L. is an species. These morphological descriptions were sup- avian predator often found foraging on commercial plemented with molecular sequence data (c.4,800 bp catfish operations, but to date the trematode fauna of of ribosomal DNA and c.600 bp of mitochondrial DNA). Gene sequences confirmed the identification of C. marginatum. However, the second species differed Electronic supplementary material The online version of significantly from its congeners in both morphology this article (doi:10.1007/s11230-016-9686-0) contains supple- and DNA sequence. Given these distinct morpholog- mentary material, which is available to authorized users. ical and molecular characters we propose this second species as Clinostomum album n. sp. T. G. Rosser Á N. R. Alberson Á E. T. Woodyard Á L. M. Pote Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Introduction MS 39762, USA F. L. Cunningham Digenetic trematodes of the genus Clinostomum Mississippi Field Station, National Wildlife Research Leidy, 1856 are widely distributed parasites of Center, Wildlife Services, United States Department of piscivorous birds, molluscs, fishes and amphibians Agriculture, Mississippi State, MS 39762, USA (Kanev et al., 2002). Adults are found in the oral cavity M. J. Griffin (&) or oesophagus of the bird hosts, the cercariae develop Thad Cochran National Warmwater Aquaculture Center, in freshwater snails and metacercariae encyst in Aquatic Research & Diagnostic Laboratory, Delta freshwater fish or amphibians (Dias et al., 2003). Research and Extension Center, College of Veterinary The debate on the number of species is ongoing, but Medicine, Mississippi State University, Stoneville, MS 38776, USA contemporary studies investigating the genetic varia- e-mail: matt.griffi[email protected] tion between and among species have revealed new 123 36 Syst Parasitol (2017) 94:35–49 insights into the taxonomy of the genus (Locke et al., Pinacho, Garcı´a-Varela & Pe´rez-Ponce de Leo´n, 2015). As with other groups of digeneans, discrimi- 2013 from great egret, great blue heron, and bare- natory morphological characters that differentiate throated tiger heron Tigrisoma mexicanum Swainson. closely related species are lacking. This has led to an Furthermore, Caffara et al. (2011) combined morpho- underestimation of true species richness, which was logical and molecular data to differentiate adult and revealed primarily through molecular DNA sequenc- metacercaria stages of C. complanatum and C. ing (Caffara et al., 2011; Locke et al., 2015; Rosser marginatum and concluded that C. complanatum is et al., 2016a). Supplemental molecular data coupled the ‘‘European’’ species and is not present in the with detailed morphological descriptions have facil- Americas. itated the identification of new species and offer more A recent collection of great egrets from catfish sound support of amended descriptions of established production operations in the northeastern area of species. Genetic markers typically used to distinguish Mississippi was evaluated for Clinostomum spp. between species include ribosomal (e.g. internal infection and a novel species is described herein. transcribed spacer regions) and mitochondrial (e.g. cytochrome c oxidase subunit 1) genes (Caffara et al., 2011; Gustinelli et al., 2010; Sereno-Uribe et al., 2013; Materials and methods Locke et al., 2015). The great egret Ardea alba L. (Pelecaniformes: Trematode collection and morphological characteri- Ardeidae) is a species of piscivorous bird ranging sation throughout the USA, southern Canada, Central Amer- Thirteen great egrets were collected from commercial ica and South America. Given the frequent occurrence catfish operations in Noxubee County, Mississippi of the great egret on commercial catfish operations in using soft catch leg hold traps and euthanized using the southeastern USA, the great egret is widely CO2. Immediately following euthanasia, the oral considered a nuisance species in catfish aquaculture cavity and sublingual area were inspected for adult (Glahn & King, 2004). While the effect of predatory Clinostomum spp. These were removed manually with foraging by great egrets on catfish aquaculture has sterile featherweight forceps (BioQuip Products, been documented (Glahn et al., 1999; Werner et al., Rancho Dominguez, California) and placed in 0.09% 2001), the impact of the trematodes they introduce to sterile saline. The oesophagus and trachea were catfish production systems is largely understudied. separated and opened longitudinally, the contents In North America the number of Clinostomum emptied into a 38-lm aperture brass sieve and washed species continues to expand. Clinostomum heluans with dechlorinated water. The intestinal lining was Braun, 1899 was reported in the great blue heron Area scraped manually and the contents rinsed onto the herodias L. and Clinostomum intermedialis Lamont, screen. The entire screen contents were then examined 1920 in the Brandt’s cormorant Phalacrocorax peni- in a lined Petri dish under a dissecting stereomicro- cillatus Brandt from Mexico (Bravo-Hollis, 1947). In scope (Olympus SZ60, Olympus Optical Co. Ltd., a survey of the helminth parasites of great egrets in Tokyo, Japan). Remaining intestinal contents were Florida, USA, two species of Clinostomum were removed and examined for additional trematodes. All reported, Clinostomum attenuatum Cort, 1913 and Clinostomum spp. adults were washed into containers Clinostomum complanatum (Rudolphi, 1814) (see with 0.09% saline. Sepu´lveda et al., 1999). Overstreet & Curran (2004) Adult trematodes were relaxed in slightly boiling reported Clinostomum marginatum (Rudolphi, 1819) saline and fixed in 70% ethanol. A subsample of each from herons, egrets and catfish obtained from produc- suspected species was stained with acetocarmine for at tion ponds in Louisiana and Mississippi, USA. In least 5 h, destained in 1% acidic ethanol, and rinsed in Mexico, C. complanatum has been found in great increasing concentrations of ethanol (70–100%) for at egrets (Violante-Gonza´lez et al., 2012); however least 1 h each. Specimens were cleared in Hemo-De recently Sereno-Uribe et al. (2013) suggested that (Fisher Scientific, Pittsburgh, Pennsylvania, USA) and previous records of C. complanatum in Mexico are mounted on clean glass microscope slides using likely C. marginatum or the more recently recognized PermountTM Mounting Medium (Fisher Scientific, Clinostomum tataxumui Sereno-Uribe, Pinacho- Pittsburgh,