Southeastern Naturalist 365 A.D. Boggs, C.S. DePerno, and J.R. Flowers 22001188 SOUTHEASTERN NATURALIST 1V7o(2l.) :1376,5 N–3o7. 02 Range Expansion of Fascioloides magna in North Carolina April D. Boggs1,*, Christopher S. DePerno1, and James R. Flowers2 Abstract - In North Carolina, Odocoileus virginianus (White-tailed Deer) are the definitive natural hosts of Fascioloides magna (Giant Liver Fluke). Previous research identified the enzootic range of Giant Liver Fluke in North Carolina to be within the Tar River and Roanoke River Basins in Halifax and neighboring counties. Recent Giant Liver Fluke infections of Ovis aries (Domestic Sheep), Capra hircus (Domestic Goat), Lama glama (Llama), Vicugna pacos (Alpaca), and Bos taurus (Cattle) outside the historic enzootic range prompted us to investigate the current range of Giant Liver Fluke in North Carolina. From September 2014 to January 2015, we examined livers from hunter-harvested Whitetailed Deer within 16 North Carolina counties. We detected Giant Liver Fluke in livers from 5 counties, with an overall prevalence of 10.3%. Besides reporting the first Giant Liver Fluke infections of livestock in North Carolina, we documented new geographic localities (Cabarrus, Franklin, Mecklenburg, Union, Wake, and Washington counties) for Giant Liver Fluke. An increased impact on North Carolina livestock is likely with the possible range expansion of Giant Liver Fluke, which may be related to the increase in populations of White-tailed Deer. Introduction Fascioloides magna (Bassi) (Giant Liver Fluke) is a parasite of North American wild and domestic ruminants. Odocoileus virginianus (Zimmermann) (White-tailed Deer) is the definitive host of Giant Liver Fluke (Pybus 2001), but little pathology occurs in this host species. However, infections in domestic livestock cause death or condemnation of livers (Davidson 2006, Králová-Hromadová et al. 2016, Pybus 2001, Swales 1935). Only 3 studies (Flowers 1996, Harlow and Jones 1965, Pursglove et al. 1977) have reported Giant Liver Fluke from North Carolina. In 2013, a fatal case of Giant Liver Fluke was diagnosed in a Lama glama L. (Llama) from Cabarrus County, NC (NCDA Griffin Diagnostic Laboratory, Monroe, NC). In 2014, two fatal cases of Giant Liver Fluke in Vicugna pacos (L.) (Alpaca) were diagnosed from Franklin County, NC, and in 2016, an Ovis aries L. (Domestic Sheep) from Wake County, NC died from a Giant Liver Fluke infection (NCDA Rollins Diagnostic Laboratory). In addition, a Capra hircus L. (Domestic Goat) infection and numerous Bos taurus L. (Cattle) infections have been detected in Washington County (NCDA Rollins Diagnostic Laboratory). Livestock may experience morbidity and mortality from liver fluke disease; thus, the range expansion of Giant Liver Fluke is of concern, especially where wild hosts share habitat 1North Carolina State University, Department of Forestry and Environmental Resources, Fisheries, Wildlife, and Conservation Biology, 110 Brooks Avenue, Raleigh, NC 27607. 2North Carolina State University, College of Veterinary Medicine, Department of Population Health and Pathobiology, 1060 William Moore Drive, Raleigh, NC 27607. *Corresponding author - adboggs@ncsu.edu. Manuscript Editor: Roger Applegate Southeastern Naturalist A.D. Boggs, C.S. DePerno, and J.R. Flowers 2018 Vol. 17, No. 2 366 with livestock. Significant financial losses can affect livestock producers; therefore, our objective was to document and report an increase in the historic distribution of White-tailed Deer infected with Giant Liver Fluke in North Carolina. Methods From September 2014 to January 2015, we collected 165 livers from hunter-harvested White-tailed Deer within 16 counties in North Carolina (Fig. 1). Collections were opportunistic, based on hunter participation, and included contributions from 4 hunt clubs (Washington, Halifax, and Stanly counties), 3 nature preserves (Mecklenburg County), 1 research station (Washington County), 1 custom processing center (various counties), and donations from private citizens (various counties). Our research team received White-tailed Deer livers from hunters that were either freshly harvested or that had been stored on ice or in freezers. Livers from the custom processor were donated directly to our research team, placed in a cooler for transport back to our North Carolina State University (NCSU) laboratory, and necropsied within 24 h. Livers collected in Mecklenburg County were placed on ice, transported back to NCSU, and necropsied weekly. Other livers were collected by hunters or hunt clubs, stored in freezers, transported to NCSU, thawed and necropsied monthly or after the end of the hunting season. At the NCSU laboratory, we examined the livers for signs of Giant Liver Fluke or other parasites. Besides the number of flukes detected in each liver, we also noted pathologic signs characteristic of Giant Liver Fluke infections, such as melanoid and fibrous tracks in the liver tissue, liver damage, and the number of fluke pockets (Králová-Hromadová et al. 2016, Pursglove 1977, Pybus 2001, Pybus et al. 2015, Swales 1935) . We deposited specimens (HWML 99829–99844) in the Harold W. Manter Laboratory of Parasitology. Our parasite–host statistical analyses (prevalence and intensity) followed Bush et al. (1997). Results We collected 165 livers from 16 counties and detected Giant Liver Fluke in 17 livers (10.3%) from 5 of the counties sampled (Table 1, Fig. 1). The average number of liver flukes per infected liver was 5.4; the highest number of liver flukes was 29 in a single liver from Mecklenburg County (Table 1). Discussion Harlow and Jones (1965) reported a prevalence of 11.5% out of 87 North Carolina White-tailed Deer infected with Giant Liver Fluke; however, they did not report the counties of collection. In the 1960s and 1970s, parasitologists from North Carolina State University conducted various surveys of helminth parasites of wildlife in North Carolina. Giant Liver Fluke had been detected in White-tailed Deer from hunting clubs near the towns of Roanoke Rapids and Scotland Neck, in Halifax County (Fig. 1), but had not been detected at Fort Bragg hunting clubs in Cumberland County (G. Miller, Emeritus, North Carolina State University, Raleigh, NC, Southeastern Naturalist 367 A.D. Boggs, C.S. DePerno, and J.R. Flowers 2018 Vol. 17, No. 2 pers. comm.). Within the Roanoke River basin of North Carolina, Pursglove et al. (1977) reported enzootic areas with prevalence of 56% in Halifax County, 63% in Bertie County, and 80% in Northampton County, and a focus of infection in Bladen County (29%) within the Cape Fear River Basin (Fig. 1). Later, Flowers (1996) reported 73% prevalence in White-tailed Deer from a hunting club in Halifax County, near Fishing Creek, a tributary of the Tar River in North Carolina (Fig. 1). The prevalence of Giant Liver Fluke from our study (10.3%) was similar to the prevalence (12.8%) detected by Pursglove et al. (1977) in their study of 13 southeastern states. However, in North Carolina, Pursglove et al. (1977) reported Giant Liver Fluke from only 4 of 22 locations, with no Giant Liver Fluke detected east of Bertie County or west of Bladen County (Fig.1). Our research identified Giant Liver Fluke infections in Washington County, ~84 km (~52 mi) east of the Halifax enzootic area; in Wake and Franklin counties, ~101 km (~63 mi) west of the Halifax enzootic area; and in Cabarrus, Union, and Mecklenburg counties, ~322 km (~200 mi) southwest of the Halifax enzootic area (Table 1, Fig. 1). Of special interest is the continued enzootic status of Halifax County, in North Carolina’s Upper Coastal Plain. The prevalence of Giant Liver Fluke in Whitetailed Deer from Halifax County has been reported as 56% (Pursglove et al. 1977), 73% (Flowers 1996), and 70% (present study). Halifax County is bordered to the north by the Roanoke River and only 12.5 miles to the south by Fishing Creek, a major tributary of the Tar River. This physiography results in lowland to swampy habitat throughout much of the county, which is excellent for the endemicity of Giant Liver Fluke, providing browse and cover for the definitive host (i.e., Whitetailed Deer), and slow-moving to stagnant aquatic habitat for its lymnaeid snail host, Pseudosuccinea columnella (Say) (Mimic Lymnaea) (Flowers and Miller 1993, Walter 1954). Similar habitats, including marshlands, moist lowlands, and swamps associated with major river drainages, have been reported as enzootic habitats for Giant Liver Fluke (Pursglove et al. 1977, Pybus 2001, Pybus et al. 2015, Vanderwaal et al. 2014, Vannatta 2016). Pybus et al. (2015) described the Giant Figure 1. Collection sites of White-tailed Deer livers from September 2014 to January 2015, and locations of previously reported Fascioloides magna (Giant Liver Fluke) infections in North Carolina. Southeastern Naturalist A.D. Boggs, C.S. DePerno, and J.R. Flowers 2018 Vol. 17, No. 2 368 Liver Fluke ’s habitat tolerances to be relatively broad and characterized by standing water, ubiquitous aquatic snails, and emergent vegetation. Pursglove et al. (1977) reported Giant Liver Fluke from the Halifax enzootic region but did not detect this fluke from collection sites outside this region (Fig. 1). The present study reports new geographic locales, which may indicate a range expansion of this fluke. Our White-tailed Deer and livestock reports of Giant Liver Fluke from Franklin and Wake counties, as well as Washington County suggests expansion west and east, respectively, from the Halifax enzootic region. Our reports of this fluke from Cabarrus, Mecklenburg, and Union counties suggest the Giant Liver Fluke may have dispersed into North Carolina from populations reported by Pursglove et al (1977) in South Carolina’s Chester and Union counties. Range expansions of the Giant Liver Fluke in North Carolina would unlikely be due to recent expanded distributions of snail hosts, as the Mimic Lymnaea, likely the fluke’s main snail host in North Carolina, has historically been common, ubiquitous, and widespread (Adams et al. 1990, Flowers 1996, Flowers and Miller 1993, Turgeon et al. 1998, Walter 1954). Although marshland habitats are important for Giant Liver Fluke enzootic regions, Peterson et al. (2013) concluded the occurrence of Giant Liver Fluke in Alces alces (L.) (Moose) populations was more correlated to the population density of White-tailed Deer than the proportion of wetland habitats. In North Carolina, White-tailed Deer populations have increased from 1976 to 2015 (NCWRC 2015), and have the potential to spread parasites and diseases, such as the Giant Liver Fluke and tick-borne pathogens (Paddock and Yabsley 2007). Future research is necessary to determine if the Giant Liver Fluke has expanded to other locations in North Carolina and to understand financial impacts that infections may have on livestock producers. Table 1. Fascioloides magna (Giant Liver Fluke) from 17 of 165 Odocoileus virginianus (Whitetailed Deer) collected from September 2014 to January 2015 in North Carolina. Intensity County Livers examined Prevalence (%) Mean Range Beaufort 1 0.0 - - Chatham 2 0.0 - - Columbus 5 0.0 - - Franklin 13 0.0 - - Granville 5 0.0 - - Halifax 10 70.0 3.3 1–9 Lee 1 0.0 - - Mecklenburg 78 6.4 8.8 1–29 Moore 1 0.0 - - Nash 1 0.0 - - Stanly 22 0.0 - - Union 1 100.0 6.0 - Vance 1 0.0 - - Wake 7 14.3 13.0 - Warren 1 0.0 - - Washington 15 20.0 2.0 1–4 Southeastern Naturalist 369 A.D. Boggs, C.S. DePerno, and J.R. Flowers 2018 Vol. 17, No. 2 Acknowledgments We thank J. Tetterton, J. Bland, J. Giles, G. Marshall, J. 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