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Transmural Ileal Ganglioneuromatosis in a Young Boer Goat (Capra hircus)
J. Comp. Path. 2014, Vol. 151, 190e194
Available online at www.sciencedirect.com
ScienceDirect www.elsevier.com/locate/jcpa
NEOPLASTIC DISEASE
Transmural Ileal Ganglioneuromatosis in a Young Boer Goat (Capra hircus) M. F. Sheley*, R. J. Higgins† and A. Mete* * California Animal Health and Food Safety Laboratory and † Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
Summary A diagnosis of transmural ileal ganglioneuromatosis was made in a 15-day-old goat that was found dead following a period of diarrhoea and inappetence. Grossly, the entire length of the wall of the ileum was pale and firm with a variably segmental to transmural thickening. Microscopically, the ileal transmural thickening was due to a diffuse proliferation of both ganglionic and glial cells forming cell nests or packets that infiltrated the wall and into the mesentery surrounding a mesenteric lymph node. The neoplastic ganglionic cells were immunoreactive for S100, synaptophysin and triple neurofilament, while the glial spindle cells were immunoreactive with glial fibrillary acidic protein, S100 and laminin confirming their Schwann cell identity. Nerve fibres expressing neurofilament protein 200 and phosphorylated neurofilament (SMI-31) were observed rarely. Ganglioneuromatosis is defined as diffuse exuberant proliferation of all components of the intestinal ganglionic plexuses. In man, the transmural form has more grave clinical consequences than a focal pattern and is commonly associated with germline mutations in the RET proto-oncogene. Whether there is any comparable molecular genetic abnormality in animals remains unknown; however, ganglioneuromatosis needs to be included in the differential diagnosis of tumours of the autonomic enteric nervous system. Published by Elsevier Ltd. Keywords: ganglioneuromatosis; goat; immunohistochemistry; small intestine
Intestinal ganglioneuromatosis (GN) is a rare disorder of the myenteric plexus of the autonomic enteric nervous system and defined as diffuse exuberant proliferation of ganglionic cells and their processes admixed with a spindle cell stroma of Schwann cells (Smith et al., 1999; Thway and Fisher, 2009). In man, GN can affect the urinary bladder, gall bladder or any portion of the gastrointestinal tract, but most commonly involves the ileum, colon and appendix (Thway and Fisher, 2009). It can occur in two patterns described as either mucosal or the clinically more severe transmural form. In animals, to date, only the mucosal pattern of GN has been reported in the colon in two dogs (Fairley and McEntee, 1990; Bemelmans et al., 2011), one steer (Cole et al., 1990) and incidentally in a horse (Porter et al., 2007), and in two dogs with either ileal
*Correspondence to: A. Mete (e-mail: [email protected]). 0021-9975/$ - see front matter http://dx.doi.org/10.1016/j.jcpa.2014.04.012
or jejunal localization (Hazell et al., 2011; Paris et al., 2012). This report describes for the first time the transmural form of ileal GN in a neonatal goat. A female Boer goat that was unremarkable at birth according to the owner underwent horn debudding and vaccination against Clostridium perfringens types C and D and tetanus (Clostridium tetani) with a CDT vaccine at 5 days of age. The health of the kid then appeared to decline progressively with an onset of inappetence and diarrhoea. Despite bottle feeding and symptomatic treatment with electrolytes (ResorbÒ, Zoetis, Kalamazoo, Michigan, USA), bismuth subsalicylate (Pepto-BismolÒ, Procter & Gamble, Cincinnati, Ohio, USA), a non-steroidal anti-inflammatory agent (PrevailÒ, VetOne, Meridian, Idaho, USA) and antibiotics (penicillin) there was no apparent clinical response and the kid was found dead at 15 days old and was submitted for necropsy examination to the California Animal Health and Food Safety laboratory, Davis, California. Published by Elsevier Ltd.
Ileal Ganglioneuromatosis in a Goat
At necropsy examination 10 ml of brown turbid fluid containing flecks of milk curd was present within the abdominal cavity. The abomasum was filled with a similar content and had a 1 cm diameter antemortem perforation of the greater curvature; the mucosal and serosal surfaces of the abomasum were affected by extensive dark red to brown coalescing petechial haemorrhages. In the caudal small intestines, beyond the normal jejunum, was a poorly defined transition zone leading to a circumferentially thickened (1e5 mm thick), pale, slightly firm and corrugated ileal wall. On transverse sections the thickened wall was segmentally transmural and asymmetrical, with regions of concentric thickening causing mild stenosis of the lumen. The small intestine overall contained scant yellow-tinged liquid. Additional gross findings were epicardial petechiations and redness within both of the ocular anterior chambers. Samples of brain, trachea, oesophagus, heart, lung, thymus, liver, kidney, adrenal gland, mesenteric lymph node, spleen, eye, umbilical stump, diaphragm, rumen, reticulum, abomasum, small and large intestines and urinary bladder were fixed in 10% neutral buffered formalin. Tissues were processed routinely and embedded in paraffin wax. Sections (5 mm) were stained with haematoxylin and eosin (HE). Microscopically, within the ileal wall there was confluent proliferation of a densely-packed heterogeneous population of spindle and polygonal cells, which asymmetrically expanded the wall (Fig. 1)
Fig. 1. Asymmetrical transmural expansion of the ileal wall by dense, confluent cellular proliferations. HE.
191
and infiltrated multifocally into the mesentery and the adipose tissue abutting a mesenteric lymph node. The majority of the ileum was affected transmurally, with the cells infiltrating the lamina propria and expanding the villi (Fig. 2), partially to diffusely obliterating the submucosa and dissecting into and partly effacing the tunica muscularis. The neoplastic cells were in bundles and frequently formed nests and packets interspersed with large neuronal ganglionic cell bodies that occasionally formed clusters of 2e5 cells. Serial sections were subjected to immunohistochemistry (IHC) and the immunohistochemical markers and immunoreactivity profiles of the tumour cells are summarized in Table 1. Neural components were confirmed by diffuse cytoplasmic immunoreactivity of the neoplastic cells to S100 and synaptophysin antibodies (Fig. 3) and the majority of cells expressed triple neurofilament protein (TNF) (Fig. 4), which highlighted scattered large neuronal cell bodies. The proliferating cells did not express neurofilament protein 200 (NF200) or phosphorylated neurofilament SMI-31; however, these markers revealed sparse fragments of nerve fibres that were scattered randomly throughout the affected segments. Nerve fibres and processes of ganglia in the unaffected regions of the ileal wall were discernible with SMI-31, but not with NF200. Few to a majority of the spindle stromal cells were immunoreactive for glial fibrillary
Fig. 2. Mucosal and submucosal infiltration of the cells with obliteration of the lamina propria and marked villus expansion. HE.
M.F. Sheley et al.
192
Table 1 Summary of immunohistochemical markers and pattern of labelling Antibody
Immunoreactivity
Specificity
Type*
Dilution
Source
Glial cells
Neurons
S100 Synaptophysin Triple neurofilament
RP MM MM
1 in 400 1 in 80 1 in 50
+ (90%) + (90%)
+ (90%) + (90%) + (80%)
Neurofilament 200 Phosphorylated neurofilament (SMI-31) Glial fibrillary acidic protein Laminin
MM MM
1 in 50 1 in 2,000
RP RP
1 in 500 1 in 30
+ (80%) + (90%)
Ki67 (MIB-1) Chromogranin A
MM RP
1 in 75 1 in 300
Neuronal nucleus
MM
1 in 500
Novocastra Laboratories, Newcastle, UK Dako, Carpinteria, California, USA Novocastra Laboratories, Richmond, Virginia, USA Novocastra Laboratories Covance Laboratories, Folsom, California, USA Dako Biogenex Laboratories, Fremont, California, USA Dako Novus Biologicals, Littleton, Colorado, USA Chemicon International, Temecula, California, USA
2%†
(%) is the % of subpopulations of cells that are immunoreactive. * Type of antibody: RP, rabbit polyclonal; MM, mouse monoclonal. † MIB-1 proliferation index is determined as % of immunoreactive cells in a total of 2,000 cell counted.
acidic protein (GFAP). Laminin outlined the cellular membranes of cell nests and packets, confirming the glial (Schwann cell) component. In the abomasum there was severe multifocal to transmural neutrophilic abomasitis with abundant mucosal fibrin deposition. Focally extensive and transmural dissecting fibrin with degenerate neutrophils and oedema characterized the site of the perforation. Additional microscopical findings included an acute mild cranioventral suppurative bronchopneumonia (presumed to be the result of inhalation of foreign material), fibrin effusion in the anterior chambers of the eyes and mild multifocal acute tubular necrosis in both kidneys attributed to septicaemia.
Fig. 3. Diffuse expression of synaptophysin by proliferating ganglionic cells. IHC.
Anaerobic and aerobic bacterial cultures of the peritoneal contents isolated Clostridium tertium and an unidentified gram-negative rod, while cultures of swabs taken from the liver, lung and meninges did not yield bacterial growth. Screening the liver for lead, mercury, arsenic, copper, manganese, zinc, cadmium and iron did not reveal any accumulation of these metals. Hepatic selenium levels were also within normal limits and no parasites were detected by faecal flotation. The gross and histological findings suggest that the acute cause of death of this neonatal goat was sepsis, stemming from an abomasal rupture. Bacterial overgrowth leading to abomasal bloat and perforation of the wall may occur due to overeating, decreased immunity and disorders inhibiting gastrointestinal motility (Songer and Miskiminis, 2005). The primary outcome of GN in man is loss of gastrointestinal function in the affected segment leading to constipation or diarrhoea if enterocolitis is involved or often frank obstruction in infants (Smith et al., 1999). In the present case, the extensive nature of the GN affecting the complete ileal segment with transmural involvement and infiltration into the mesentery may have led to motility dysfunction and the development of the abomasitis. In addition, debudding and vaccination at 5 days of age may have had adverse stressful effects on innate immunity, possibly facilitating the abomasitis. The only bacterium identified was C. tertium from the peritoneal contents, which is an opportunistic pathogen that has been previously associated with sepsis in a Moluccan Cockatoo (Hess et al., 1998) and shown
193
Ileal Ganglioneuromatosis in a Goat
Fig. 4. TNF immunoreactivity of polygonal ganglionic cells in packets and nests. IHC.
to cause sepsis in immunocompromised people as well as primary necrotizing fasciitis (Vanderhofstadt et al., 2010). However, the prior administration of antibiotics likely contributed to the negative culture results. GN must be distinguished from other autonomic nervous system tumours including ganglioneuromas, neuroblastomas, ganglioneuroblastomas and neurofibromas. Ganglioneuromas are relatively benign tumours of the enteric nervous tissues like GN, but manifest as single or multiple well-demarcated masses (Cole et al., 1990; Smith et al., 1999; Porter et al., 2007). Neuroblastomas or variants such as ganglioneuroblastoma are malignant tumours with nests of poorly-differentiated neuroblastic cells and minimal stroma (Ulrich et al., 2008). Neurofibromas are a subtype of peripheral nerve sheath tumour, are nodular and are a mixture of Schwann cells, perineurial cells and fibroblasts, with patchy distribution of S100 expression (Sch€oniger and Summers, 2009). The morphology and immunolabelling characteristics of the proliferating cells, as well as the distribution and the pattern of ileal involvement, were most compatible with a diagnosis of transmural ileal GN. Most other reports of GN have utilized IHC minimally since the diagnosis is fundamentally histological (Thway and Fisher, 2009). Here, the selected antibodies were applied to confirm histologically both the neuronal and Schwann cell components. The classification of human GN as either a transmural or mucosal variant, depending on whether the lesion diffusely invades all layers of the intestinal wall (transmural form) or is isolated in the mucosae (mucosal form), is an important distinction for both treatment and prognosis (d’Amore et al., 1991; Thway and Fisher, 2009). Although few isolated spontaneously arising cases are reported (Carter and Laurini, 2008), transmural ileal GN is most commonly clinically linked to diarrhoea, has a poorer prognosis
and is associated with the development of forms of multiple endocrine neoplasia type 2B due to germline mutations in the RET proto-oncogene (Smith et al., 1999; Lee and Norton, 2000; Camacho et al., 2008). Focal mucosal human GN is rare, colonic in localization, clinically results in constipation and is sporadic or associated mainly with NF1 gene mutations. The molecular pathogenesis of GN beyond the RET proto-oncogene mutation is not well understood, but it has been shown that proliferation of multiple nerve fibre types, including those with peptidergic, cholinergic and adrenergic function is involved (Smith et al., 1999). Based on these data, it is possible that this goat had a spontaneous mutation in the RET proto-oncogene, since there was no historical evidence of this problem following three similar prior matings of the dam.
Acknowledgements We wish to thank Mr. M. Manzer in the VMTH immunohistochemical laboratory for technical support.
Conflict of Interest Statement The author(s) declare no potential conflicts of interest with respect to the research, authorship and/or publication of this article. The author(s) declare no financial support for the research, authorship, and/or publication of this article.
References Bemelmans I, K€ ury S, Albaric O, Hordeaux J, Bertrand L et al. (2011) Colorectal hamartomatous polyposis and ganglioneuromatosis in a dog. Veterinary Pathology, 48, 1012e1015. Camacho CP, Hoff AO, Lindsey SC, Signorini PS, Valente FO et al. (2008) Early diagnosis of multiple endocrine neoplasia type 2B: a challenge for physicians. Arquivos Brasileiros Endocrinologia e Metabologia, 52, 1393e1398. Carter JE, Laurini JA (2008) Isolated intestinal neurofibromatous proliferations in the absence of associated systemic syndromes. World Journal of Gastroenterology, 14, 6569e6571. Cole DE, Migaki G, Leipold HW (1990) Colonic ganglioneuromatosis in a steer. Veterinary Pathology, 27, 461e462. d’Amore ES, Manivel JC, Pettinato G, Niehans GA, Snover DC (1991) Intestinal ganglioneuromatosis: mucosal and transmural types, a clinicopathologic and immunohistochemical study of six cases. Human Pathology, 22, 276e286. Fairley RA, McEntee MF (1990) Colorectal ganglioneuromatosis in a young female dog (Lhasa Apso). Veterinary Pathology, 27, 206e207.
194
M.F. Sheley et al.
Hazell KLA, Reeves MP, Swift IM (2011) Small intestinal ganglioneuromatosis in a dog. Australian Veterinary Journal, 89, 15e18. Hess L, Bartick T, Hoefer H (1998) Clostridium tertium infection in a Moluccan Cockatoo (Cacatua moluccensis) with megacolon. Journal of Avian Medicine and Surgery, 12, 30e35. Lee NC, Norton JA (2000) Multiple endocrine neoplasia type 2B e genetic basis and clinical expression. Surgical Oncology, 9, 111e118. Paris JK, McCandlish IAP, Schwarz T, Simpson JW, Smith SH (2012) Small intestinal ganglioneuromatosis in a dog. Journal of Comparative Pathology, 148, 323e328. Porter BF, Storts RW, Payne HR, Edwards JF (2007) Colonic ganglioneuromatosis in a horse. Veterinary Pathology, 44, 207e210. Sch€oniger S, Summers BA (2009) Localized, plexiform, diffuse, and other variants of neurofibroma in 12 dogs, 2 horses, and a chicken. Veterinary Pathology, 46, 904e915. Smith VV, Eng C, Milla PJ (1999) Intestinal ganglioneuromatosis and multiple endocrine neoplasia type 2B: implications for treatment. Gut, 45, 143e146.
Songer GJ, Miskiminis DW (2005) Clostridial abomasitis in calves: case report and review of the literature. Anaerobe, 11, 290e294. Thway K, Fisher C (2009) Diffuse ganglioneuromatosis in small intestine associated with neurofibromatosis type 1. Annals of Diagnostic Pathology, 13, 50e54. Ulrich R, Stan AC, Koch A, Beineke A (2008) Expression of brain-derived neurotrophic factor and tropomyosinrelated kinase-B in a bovine jejunal nodular ganglioneuroblastoma. Veterinary Pathology, 45, 355e360. Vanderhofstadt M, Andre M, Lonchay C, Levecque P, Holemans X et al. (2010) Clostridium tertium bacteremia: contamination or true pathogen? A report of two cases and a review of the literature. International Journal of Infectious Diseases, 14, e335ee337.
January 12th, 2014 ½ Received, Accepted, April 21st, 2014
Available online at www.sciencedirect.com
ScienceDirect www.elsevier.com/locate/jcpa
NEOPLASTIC DISEASE
Transmural Ileal Ganglioneuromatosis in a Young Boer Goat (Capra hircus) M. F. Sheley*, R. J. Higgins† and A. Mete* * California Animal Health and Food Safety Laboratory and † Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
Summary A diagnosis of transmural ileal ganglioneuromatosis was made in a 15-day-old goat that was found dead following a period of diarrhoea and inappetence. Grossly, the entire length of the wall of the ileum was pale and firm with a variably segmental to transmural thickening. Microscopically, the ileal transmural thickening was due to a diffuse proliferation of both ganglionic and glial cells forming cell nests or packets that infiltrated the wall and into the mesentery surrounding a mesenteric lymph node. The neoplastic ganglionic cells were immunoreactive for S100, synaptophysin and triple neurofilament, while the glial spindle cells were immunoreactive with glial fibrillary acidic protein, S100 and laminin confirming their Schwann cell identity. Nerve fibres expressing neurofilament protein 200 and phosphorylated neurofilament (SMI-31) were observed rarely. Ganglioneuromatosis is defined as diffuse exuberant proliferation of all components of the intestinal ganglionic plexuses. In man, the transmural form has more grave clinical consequences than a focal pattern and is commonly associated with germline mutations in the RET proto-oncogene. Whether there is any comparable molecular genetic abnormality in animals remains unknown; however, ganglioneuromatosis needs to be included in the differential diagnosis of tumours of the autonomic enteric nervous system. Published by Elsevier Ltd. Keywords: ganglioneuromatosis; goat; immunohistochemistry; small intestine
Intestinal ganglioneuromatosis (GN) is a rare disorder of the myenteric plexus of the autonomic enteric nervous system and defined as diffuse exuberant proliferation of ganglionic cells and their processes admixed with a spindle cell stroma of Schwann cells (Smith et al., 1999; Thway and Fisher, 2009). In man, GN can affect the urinary bladder, gall bladder or any portion of the gastrointestinal tract, but most commonly involves the ileum, colon and appendix (Thway and Fisher, 2009). It can occur in two patterns described as either mucosal or the clinically more severe transmural form. In animals, to date, only the mucosal pattern of GN has been reported in the colon in two dogs (Fairley and McEntee, 1990; Bemelmans et al., 2011), one steer (Cole et al., 1990) and incidentally in a horse (Porter et al., 2007), and in two dogs with either ileal
*Correspondence to: A. Mete (e-mail: [email protected]). 0021-9975/$ - see front matter http://dx.doi.org/10.1016/j.jcpa.2014.04.012
or jejunal localization (Hazell et al., 2011; Paris et al., 2012). This report describes for the first time the transmural form of ileal GN in a neonatal goat. A female Boer goat that was unremarkable at birth according to the owner underwent horn debudding and vaccination against Clostridium perfringens types C and D and tetanus (Clostridium tetani) with a CDT vaccine at 5 days of age. The health of the kid then appeared to decline progressively with an onset of inappetence and diarrhoea. Despite bottle feeding and symptomatic treatment with electrolytes (ResorbÒ, Zoetis, Kalamazoo, Michigan, USA), bismuth subsalicylate (Pepto-BismolÒ, Procter & Gamble, Cincinnati, Ohio, USA), a non-steroidal anti-inflammatory agent (PrevailÒ, VetOne, Meridian, Idaho, USA) and antibiotics (penicillin) there was no apparent clinical response and the kid was found dead at 15 days old and was submitted for necropsy examination to the California Animal Health and Food Safety laboratory, Davis, California. Published by Elsevier Ltd.
Ileal Ganglioneuromatosis in a Goat
At necropsy examination 10 ml of brown turbid fluid containing flecks of milk curd was present within the abdominal cavity. The abomasum was filled with a similar content and had a 1 cm diameter antemortem perforation of the greater curvature; the mucosal and serosal surfaces of the abomasum were affected by extensive dark red to brown coalescing petechial haemorrhages. In the caudal small intestines, beyond the normal jejunum, was a poorly defined transition zone leading to a circumferentially thickened (1e5 mm thick), pale, slightly firm and corrugated ileal wall. On transverse sections the thickened wall was segmentally transmural and asymmetrical, with regions of concentric thickening causing mild stenosis of the lumen. The small intestine overall contained scant yellow-tinged liquid. Additional gross findings were epicardial petechiations and redness within both of the ocular anterior chambers. Samples of brain, trachea, oesophagus, heart, lung, thymus, liver, kidney, adrenal gland, mesenteric lymph node, spleen, eye, umbilical stump, diaphragm, rumen, reticulum, abomasum, small and large intestines and urinary bladder were fixed in 10% neutral buffered formalin. Tissues were processed routinely and embedded in paraffin wax. Sections (5 mm) were stained with haematoxylin and eosin (HE). Microscopically, within the ileal wall there was confluent proliferation of a densely-packed heterogeneous population of spindle and polygonal cells, which asymmetrically expanded the wall (Fig. 1)
Fig. 1. Asymmetrical transmural expansion of the ileal wall by dense, confluent cellular proliferations. HE.
191
and infiltrated multifocally into the mesentery and the adipose tissue abutting a mesenteric lymph node. The majority of the ileum was affected transmurally, with the cells infiltrating the lamina propria and expanding the villi (Fig. 2), partially to diffusely obliterating the submucosa and dissecting into and partly effacing the tunica muscularis. The neoplastic cells were in bundles and frequently formed nests and packets interspersed with large neuronal ganglionic cell bodies that occasionally formed clusters of 2e5 cells. Serial sections were subjected to immunohistochemistry (IHC) and the immunohistochemical markers and immunoreactivity profiles of the tumour cells are summarized in Table 1. Neural components were confirmed by diffuse cytoplasmic immunoreactivity of the neoplastic cells to S100 and synaptophysin antibodies (Fig. 3) and the majority of cells expressed triple neurofilament protein (TNF) (Fig. 4), which highlighted scattered large neuronal cell bodies. The proliferating cells did not express neurofilament protein 200 (NF200) or phosphorylated neurofilament SMI-31; however, these markers revealed sparse fragments of nerve fibres that were scattered randomly throughout the affected segments. Nerve fibres and processes of ganglia in the unaffected regions of the ileal wall were discernible with SMI-31, but not with NF200. Few to a majority of the spindle stromal cells were immunoreactive for glial fibrillary
Fig. 2. Mucosal and submucosal infiltration of the cells with obliteration of the lamina propria and marked villus expansion. HE.
M.F. Sheley et al.
192
Table 1 Summary of immunohistochemical markers and pattern of labelling Antibody
Immunoreactivity
Specificity
Type*
Dilution
Source
Glial cells
Neurons
S100 Synaptophysin Triple neurofilament
RP MM MM
1 in 400 1 in 80 1 in 50
+ (90%) + (90%)
+ (90%) + (90%) + (80%)
Neurofilament 200 Phosphorylated neurofilament (SMI-31) Glial fibrillary acidic protein Laminin
MM MM
1 in 50 1 in 2,000
RP RP
1 in 500 1 in 30
+ (80%) + (90%)
Ki67 (MIB-1) Chromogranin A
MM RP
1 in 75 1 in 300
Neuronal nucleus
MM
1 in 500
Novocastra Laboratories, Newcastle, UK Dako, Carpinteria, California, USA Novocastra Laboratories, Richmond, Virginia, USA Novocastra Laboratories Covance Laboratories, Folsom, California, USA Dako Biogenex Laboratories, Fremont, California, USA Dako Novus Biologicals, Littleton, Colorado, USA Chemicon International, Temecula, California, USA
2%†
(%) is the % of subpopulations of cells that are immunoreactive. * Type of antibody: RP, rabbit polyclonal; MM, mouse monoclonal. † MIB-1 proliferation index is determined as % of immunoreactive cells in a total of 2,000 cell counted.
acidic protein (GFAP). Laminin outlined the cellular membranes of cell nests and packets, confirming the glial (Schwann cell) component. In the abomasum there was severe multifocal to transmural neutrophilic abomasitis with abundant mucosal fibrin deposition. Focally extensive and transmural dissecting fibrin with degenerate neutrophils and oedema characterized the site of the perforation. Additional microscopical findings included an acute mild cranioventral suppurative bronchopneumonia (presumed to be the result of inhalation of foreign material), fibrin effusion in the anterior chambers of the eyes and mild multifocal acute tubular necrosis in both kidneys attributed to septicaemia.
Fig. 3. Diffuse expression of synaptophysin by proliferating ganglionic cells. IHC.
Anaerobic and aerobic bacterial cultures of the peritoneal contents isolated Clostridium tertium and an unidentified gram-negative rod, while cultures of swabs taken from the liver, lung and meninges did not yield bacterial growth. Screening the liver for lead, mercury, arsenic, copper, manganese, zinc, cadmium and iron did not reveal any accumulation of these metals. Hepatic selenium levels were also within normal limits and no parasites were detected by faecal flotation. The gross and histological findings suggest that the acute cause of death of this neonatal goat was sepsis, stemming from an abomasal rupture. Bacterial overgrowth leading to abomasal bloat and perforation of the wall may occur due to overeating, decreased immunity and disorders inhibiting gastrointestinal motility (Songer and Miskiminis, 2005). The primary outcome of GN in man is loss of gastrointestinal function in the affected segment leading to constipation or diarrhoea if enterocolitis is involved or often frank obstruction in infants (Smith et al., 1999). In the present case, the extensive nature of the GN affecting the complete ileal segment with transmural involvement and infiltration into the mesentery may have led to motility dysfunction and the development of the abomasitis. In addition, debudding and vaccination at 5 days of age may have had adverse stressful effects on innate immunity, possibly facilitating the abomasitis. The only bacterium identified was C. tertium from the peritoneal contents, which is an opportunistic pathogen that has been previously associated with sepsis in a Moluccan Cockatoo (Hess et al., 1998) and shown
193
Ileal Ganglioneuromatosis in a Goat
Fig. 4. TNF immunoreactivity of polygonal ganglionic cells in packets and nests. IHC.
to cause sepsis in immunocompromised people as well as primary necrotizing fasciitis (Vanderhofstadt et al., 2010). However, the prior administration of antibiotics likely contributed to the negative culture results. GN must be distinguished from other autonomic nervous system tumours including ganglioneuromas, neuroblastomas, ganglioneuroblastomas and neurofibromas. Ganglioneuromas are relatively benign tumours of the enteric nervous tissues like GN, but manifest as single or multiple well-demarcated masses (Cole et al., 1990; Smith et al., 1999; Porter et al., 2007). Neuroblastomas or variants such as ganglioneuroblastoma are malignant tumours with nests of poorly-differentiated neuroblastic cells and minimal stroma (Ulrich et al., 2008). Neurofibromas are a subtype of peripheral nerve sheath tumour, are nodular and are a mixture of Schwann cells, perineurial cells and fibroblasts, with patchy distribution of S100 expression (Sch€oniger and Summers, 2009). The morphology and immunolabelling characteristics of the proliferating cells, as well as the distribution and the pattern of ileal involvement, were most compatible with a diagnosis of transmural ileal GN. Most other reports of GN have utilized IHC minimally since the diagnosis is fundamentally histological (Thway and Fisher, 2009). Here, the selected antibodies were applied to confirm histologically both the neuronal and Schwann cell components. The classification of human GN as either a transmural or mucosal variant, depending on whether the lesion diffusely invades all layers of the intestinal wall (transmural form) or is isolated in the mucosae (mucosal form), is an important distinction for both treatment and prognosis (d’Amore et al., 1991; Thway and Fisher, 2009). Although few isolated spontaneously arising cases are reported (Carter and Laurini, 2008), transmural ileal GN is most commonly clinically linked to diarrhoea, has a poorer prognosis
and is associated with the development of forms of multiple endocrine neoplasia type 2B due to germline mutations in the RET proto-oncogene (Smith et al., 1999; Lee and Norton, 2000; Camacho et al., 2008). Focal mucosal human GN is rare, colonic in localization, clinically results in constipation and is sporadic or associated mainly with NF1 gene mutations. The molecular pathogenesis of GN beyond the RET proto-oncogene mutation is not well understood, but it has been shown that proliferation of multiple nerve fibre types, including those with peptidergic, cholinergic and adrenergic function is involved (Smith et al., 1999). Based on these data, it is possible that this goat had a spontaneous mutation in the RET proto-oncogene, since there was no historical evidence of this problem following three similar prior matings of the dam.
Acknowledgements We wish to thank Mr. M. Manzer in the VMTH immunohistochemical laboratory for technical support.
Conflict of Interest Statement The author(s) declare no potential conflicts of interest with respect to the research, authorship and/or publication of this article. The author(s) declare no financial support for the research, authorship, and/or publication of this article.
References Bemelmans I, K€ ury S, Albaric O, Hordeaux J, Bertrand L et al. (2011) Colorectal hamartomatous polyposis and ganglioneuromatosis in a dog. Veterinary Pathology, 48, 1012e1015. Camacho CP, Hoff AO, Lindsey SC, Signorini PS, Valente FO et al. (2008) Early diagnosis of multiple endocrine neoplasia type 2B: a challenge for physicians. Arquivos Brasileiros Endocrinologia e Metabologia, 52, 1393e1398. Carter JE, Laurini JA (2008) Isolated intestinal neurofibromatous proliferations in the absence of associated systemic syndromes. World Journal of Gastroenterology, 14, 6569e6571. Cole DE, Migaki G, Leipold HW (1990) Colonic ganglioneuromatosis in a steer. Veterinary Pathology, 27, 461e462. d’Amore ES, Manivel JC, Pettinato G, Niehans GA, Snover DC (1991) Intestinal ganglioneuromatosis: mucosal and transmural types, a clinicopathologic and immunohistochemical study of six cases. Human Pathology, 22, 276e286. Fairley RA, McEntee MF (1990) Colorectal ganglioneuromatosis in a young female dog (Lhasa Apso). Veterinary Pathology, 27, 206e207.
194
M.F. Sheley et al.
Hazell KLA, Reeves MP, Swift IM (2011) Small intestinal ganglioneuromatosis in a dog. Australian Veterinary Journal, 89, 15e18. Hess L, Bartick T, Hoefer H (1998) Clostridium tertium infection in a Moluccan Cockatoo (Cacatua moluccensis) with megacolon. Journal of Avian Medicine and Surgery, 12, 30e35. Lee NC, Norton JA (2000) Multiple endocrine neoplasia type 2B e genetic basis and clinical expression. Surgical Oncology, 9, 111e118. Paris JK, McCandlish IAP, Schwarz T, Simpson JW, Smith SH (2012) Small intestinal ganglioneuromatosis in a dog. Journal of Comparative Pathology, 148, 323e328. Porter BF, Storts RW, Payne HR, Edwards JF (2007) Colonic ganglioneuromatosis in a horse. Veterinary Pathology, 44, 207e210. Sch€oniger S, Summers BA (2009) Localized, plexiform, diffuse, and other variants of neurofibroma in 12 dogs, 2 horses, and a chicken. Veterinary Pathology, 46, 904e915. Smith VV, Eng C, Milla PJ (1999) Intestinal ganglioneuromatosis and multiple endocrine neoplasia type 2B: implications for treatment. Gut, 45, 143e146.
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January 12th, 2014 ½ Received, Accepted, April 21st, 2014