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Infection in the Virginia Opossum (Didelphis Virginiand)

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Infection in the Virginia Opossum (Didelphis Virginiand) RESEARCH NOTES 253 •—, and J. Sarmiento. 1985. Observations on the Parenti, L. R. 1984. A taxonomic revision of the fishes of the Bolivian part of Lake Titicaca. II. Andean killifish genus Orestias, Cyprinodonti- Orestias agassii, Pisces, Cyprinodontidae. Revue formes, Cyprinodontidae. Bulletin of the Ameri- d'Hydrobiologie Tropicale 18:159-170. can Museum of Natural History 178:110-214. J. Helminthol. Soc. Wash. 62(2), 1995, pp. 253-256 Research Note Histopathology of Oligacanthorhynchus tortuosa (Oligacanthorhynchidae) Infection in the Virginia Opossum (Didelphis virginiand) DENNIS J. RICHARDSON AND EARL B. BARN A WELL School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 68588-0118 ABSTRACT: Oligacanthorhynchus tortuosa, a common globular proboscis bears 6 spiral rows of 6 hooks acanthocephalan of the Virginia opossum (Didelphis each and has a length of 0.22-0.23 mm and width virginiana} in North America, has been reported to be associated with large, nodule-like lesions at points of of 0.23-0.29 mm (Van Cleave, 1953). Leidy attachment of the proboscides. Three lesions resulting (1850) reported a specimen of O. tortuosa as hav- from the attachment of individuals of O. tortuosa, 1 ing the anterior 3 lines of its length buried in an each from 3 infected opossums, were prepared for his- oval tumor, 4 lines in diameter, in the mesentery tological examination to further characterize histo- of an opossum. Based on this statement, Van pathologic changes elicited by this parasite. Histolog- ically, lesions involved the mucosa, submucosa, and Cleave (1924) concluded that the worm had pen- muscularis. The proboscides were contained within ab- etrated the intestinal wall, entered the body cav- scesses characterized by necrotic debris interspersed ity, and attached to the mesentery. Feldman et with many pycnotic nuclei. The abscesses were ap- al. (1972) reported severe ulcerative lesions proximately 1.4 mm in diameter and were surrounded by regions of dense connective tissue (collagen), ap- evoked at points of attachment of unidentified proximately 142 /um wide. The bands of dense con- acanthocephalans from opossums. Brief descrip- nective tissue were surrounded by regions of active tion and a photomicrograph (Feldman et al., fibroblast and fibrocyte proliferation, approximately 1972) suggest that these specimens were O. tor- 169 jum wide, in which evidence of collagen synthesis tuosa. Richardson et al. (1992) reported 2 poorly was observed. Both longitudinal and smooth muscle layers of the muscularis had been completely destroyed developed individuals of O. tortuosa from the in the area of the lesion. Absence of polymorphonu- small intestine of a raccoon (Procyon lotor) that clear leukocytes were indicative of chronic lesions. His- caused "severe lesions" at points of attachment; topathologic changes elicited by O. tortuosa include however, histological examination was not con- chronic inflammatory response to mechanical trauma resulting from injury caused by the proboscis with sub- ducted. The only histological examination of le- sequent fibrosis and nodule formation. sions caused by O. tortuosa was conducted by KEY WORDS: histopathology, Oligacanthorhynchus Babero (1957), who reported elevated nodules tortuosa, Didelphis virginiana, opossum, Acanthoce- over the serosal surface of the small intestines of phala. 2 Illinois opossums having a base diameter of 2-7 mm. He reported the nodules to have a bright Oligacanthorhynchus tortuosa, a common red appearance due to congestion of intestinal acanthocephalan of the Virginia opossum (Di- blood vessels. Histologically, lesions reported by delphis virginiand) in North America, has been Babero (1957) resulted in complete mechanical reported to be associated with large, nodule-like destruction of the mucosal and submucosal lay- lesions at points of attachment of proboscides. ers with some focal atrophy and necrosis of the Oligacanthorhynchus tortuosa is represented by muscularis. He further noted limited leukocytic large worms with females achieving lengths of infiltration and some pigment deposition. Ba- up to 350 mm (Richardson, unpubl. data). The bero (1960) examined opossums from Georgia Copyright © 2011, The Helminthological Society of Washington 254 JOURNAL OF THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON, 62(2), JUL 1995 infected with O. tortuosa in which no such hem- orrhagic lesions were observed. The purpose of this investigation was to fur- ther characterize lesions resulting from O. tor- tuosa infections in the Virginia opossum. Three lesions resulting from the attachment of individuals of O. tortuosa, 1 each from 3 opos- sums, were prepared for histological examina- tion. Material was obtained from opossums col- lected in the course of a survey of Acanthoce- phala of opossums from Arkansas, results of which were reported by Richardson (1993). All lesions examined were caused by mature worms. '%''*'' ^^^&^3^£^& '*' Immediately after sacrificing the opossum, the '^ '"•':fcSr^te"^"'/.^s-^sS^' ' ^ small intestine was examined for nodules on the serosal surface, then removed and longitudinally i-^^P^^M ;£ v\-vr>^;:"- ~r$g. --;m dissected. Mature worms were severed so as to leave the proboscis intact and undisturbed in the lesion. The lesion along with normal tissue im- mediately surrounding the lesion was excised and placed directly into Bouin's fixative. After fixing in Bouin's solution for 24 hr, tis- sues were stored in 70% ethanol. Tissues were dehydrated by treating in a graded series of eth- anol. Tissues were cleared in toluene, infiltrated and embedded in paraffin blocks, and sectioned at a thickness of 5-7 ^m using a rotary micro- Figure 1. Photomicrograph of a cross-section of a tome. Ribbons were attached to slides with al- lesion elicited by Oligacanthorhynchus tortuosa in the bumin and stained folio wing Masson's trichrome intestine of a Virginia opossum (Didelphis virginiand) showing proboscis (arrow), necrotic abscess (asterisk), technique as described by Luna (1968). Two per- ring of collagen (C), region of active fibrocyte prolif- cent light green was substituted in place of aniline eration (arrowhead), muscularis (M), normal submu- blue to enhance staining of connective tissue. cosa (Sm), serosal side (S), and luminal side (L). Scale Stained sections were mounted in Canada bal- bar = 200 pm. Figure 1 appeared in Foundations of Parasitology, 5th edition, Wm. C. Brown, Publishers, sam and examined using light microscopy. and is used here with permission of the company. Worms were restricted to the anterior ¥2 of the small intestine, with most occurring in the an- terior '/3. Grossly, hard, white nodules were ap- mately 1.14 mm in diameter, were surrounded parent on the serosal surface of the intestine cor- by regions of dense connective tissue (collagen), responding to points of worm attachment. Pro- approximately 142 ^m wide. These regions of boscides were firmly embedded in the intestinal dense connective tissue, which appeared to have wall resulting in nodule formation; however, there effectively contained the abscesses, were inter- was no apparent evidence of hemorrhage as ob- spersed with small numbers of fibrocytes and served grossly by Babero (1957). At the base, fibroblasts. Spaces were noted between strands nodules ranged from approximately 1 to 5 mm of collagen, many of which appeared to be lym- in diameter. phatics. Bands of dense connective tissue were Lesions involved the mucosa, submucosa, and surrounded by regions of active fibroblast and muscularis; however, the serosa was intact and fibrocyte proliferation, approximately 169 ^m apparently unaffected (Fig. 1). Proboscides were wide, in which evidence of collagen synthesis contained within abscesses characterized by ne- could be observed. Within these regions, occa- crotic debris interspersed with many pycnotic sional plasma cells, lymphocytes, and mast cells nuclei, particularly abundant around the periph- were observed (approximately 1/0.133 mm2 [x 40 ery of abscesses. Small aggregations of collagen field]). These areas were infiltrated with many were present. No evidence of recent hemorrhage blood vessels and lymphatics. Lumina of arte- was observed. Abscesses, which were approxi- rioles were occluded by contraction of smooth Copyright © 2011, The Helminthological Society of Washington RESEARCH NOTES 255 muscle in the arteriole wall. Abscesses along with ment of the worm does not occur after it has bands of connective tissue resulted in drastic en- established and embedded its proboscis into the largement of the submucosa to over 7 times its intestinal wall, unlike Moniliformis moniliformis normal width, resulting in formation of the gross- in rats, which was found to attach superficially, ly observable nodule. Width of the true sub- penetrating only the mucosa and tunica propria, mucosal region of unaffected tissue was approx- with no fibrosis (Taraschewski et al., 1989). Tar- imately 240 Aim, whereas that of affected regions aschewski et al. (1989) interpreted these lesions was 1.87 mm. In the regions of the lesions, both as evidence that worms frequently changed their longitudinal and smooth muscle layers of the sites of attachment. muscularis had been completely eroded; how- Pathologic changes induced by O. tortuosa in ever, the serosa remained intact and appeared to the opossum were similar, with notable excep- be unaffected. No evidence of hypertrophy of any tions, to those elicited by M. hirudinaceus and of the muscular layers in the vicinity of the le- M. ingens, which
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