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Short Communications
Pyogranulomatous meningoencephalitis in a goat due to Corynebacterium ulcerans
W. E. Morris, F. A. Uzal, A. L. Cipolla
PYOGRANULOMAS or cerebral abscesses caused by Staphylococcus aureus, Fusobacterium necrophorum, Arcanobacterium pyogenes or Corynebacterium species infections have been reported in goats and other animal species (Moriwaki and others 1972, Altman and Bogokovsky 1973, Amand and others 1973, Glass and others 1993, Smith and Sherman 1994). Corynebacterium ulcerans, a microorganism closely related to Corynebacterium pseudotuberculosis, has been isolated from clinical samples from various species (Carter 1995, Radostits and others 2000, Tejedor and others 2000), including human beings (Barret 1986) but, to the authors knowledge, there are no reports of infections by this microorganism in goats, nor has it been associated with brain lesions in any other animal species. This short communication describes a pyogranulomatous meningoencephalitis in a goat due to C ulcerans. A six-year-old female Creole goat was submitted to the animal health unit of the National Institute of Agricultural Technology in Bariloche, Patagonia, Argentina. Its clinical signs consisted of circling, vertical nystagmus of the left eye and tilting of the head to the left. The animal was able to drink and eat but it was in poor body condition. Its body temperature was 389C. The goat was anaesthetised using sodium pentobarbitone and then euthanased by intravenous injection of an oversaturated solution of magnesium sulphate, and a full postmortem examination was carried out. There was pasty, yellowish green, thick pus inside the right acoustic meatus, surrounding the acoustic nerve. A mass, approximately 25 cm in diameter, composed of caseous material and surrounded by a thin, fibrous capsule, was observed under the dura mater, infiltrating the cerebellum, cerebellar peduncles, brainstem and occipital cerebral cortex on the right side (Fig 1). This mass extended into the fourth ventricle. No other gross lesions were observed. Impression smears from the brain lesion were stained with Gram stain, and caseous material from the lesion and cerebrospinal fluid were collected asceptically and inoculated on to 7 per cent sheep blood agar and MacConkeys agar plates. The plates were then incubated at 37C for 48 hours under aerobic, microaerophilic and anaerobic conditions. Gramstained smears of the brain lesion demonstrated groups of pleomorphic Gram-positive, non-sporulated rods, often with clubbed ends, occurring singly or in pairs. A pure culture of small, greyish-white, beta-haemolytic colonies grew on the blood agar plates inoculated with caseous material or cerebrospinal fluid and incubated under microaerophilic and aerobic conditions. Gram-stained smears of these colonies revealed microorganisms morphologically identical to those observed on impression smears of the brain lesion. These colonies were submitted to conventional biochemical tests. The microorganism isolated was nitrate negative, catalase and urease positive and fermented trehalose. Based on these and other conventional biochemical tests, the microorganism was identified as C ulcerans. No growth was observed on MacConkeys agar or on blood agar plates incubated under anaerobic conditions.

FIG 1: Pyogranuloma occupying part of the fourth ventricle of a goat and involving the cerebellum, cerebellar peduncles and pons

The brain was fixed by immersion in 10 per cent buffered formalin for seven days, and haematoxylin and eosin-stained sections were prepared by routine methods from the internal capsule, thalamus, parietal cortex, midbrain at the level of the superior coliculi, cerebellum, cerebellar peduncles, pons and obex. Histological analysis revealed multifocal to coalescent pyogranulomas in the cerebral cortex, cerebellum, cerebral peduncles and brainstem. These lesions consisted of a centre of caseous necrosis surrounded by a layer of neutrophils, macrophages, lymphocytes, plasma cells and multinucleated giant cells. A thin capsule of fibrous connective tissue separated the lesions from the surrounding parenchyma, in which there was mild to moderate haemorrhage and perivascular cuffing of lymphocytes, plasma cells and a few neutrophils. The meninges in the area were severely infiltrated by lymphocytes, plasma cells and macrophages. A diagnosis of chronic active, multifocal to coalescent pyogranulomatous encephalitis caused by C ulcerans was established based on pathological and microbiological findings. The microorganism isolated was urease positive, a characteristic which is used to distinguish C ulcerans and C pseudotuberculosis (both urease positive) from Corynebacterium diphtheriae, which is urease negative (Funke and Bernard 1999). Some strains of C pseudotuberculosis are phenotypically similar to those of C ulcerans, and it has been suggested that carbohydrate fermentation is often an unreliable method for differentiating between these two species (Sutherland and others 1996). However, it has been shown that fermentation of trehalose reliably differentiates C pseudotuberculosis from C ulcerans (Thompson and others 1983, Centicaya and others 2002). In a recent study, 93 strains of C pseudotuberculosis isolated from the lymph nodes of goats and sheep with a caseous lymphadenitis, and which had been identified by a PCR technique using a primer pair specific for the 16S rRNA of C pseudotuberculosis, were subjected to the trehalose test, and all of the strains were negative in this test (Centicaya and others 2002). However, C ulcerans is always positive for trehalose fermentation (Thompson and others
The Veterinary Record, March 5, 2005

Veterinary Record (2005) 156, 317-318 W. E. Morris, DVM, MSc, National Institute of Agricultural Technology, Bariloche, Argentina F. A. Uzal, DVM, MSc, PhD, California Animal Health and Food Safety Laboratory System, School of Veterinary Medicine, University of California, Davis, 105 West Central Avenue, San Bernardino, CA 92408, USA A. L. Cipolla, DVM, National Institute of Agricultural Technology, Balcarce, Argentina Correspondence to Dr Uzal

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Short Communications

1983). Although the differentiation between C ulcerans and C pseudotuberculosis was based on a single biochemical reaction, the trehalose test, the present results are in agreement with previous studies (Centicaya and others 2002) and indicate that the microorganism isolated in the present study was C ulcerans. Trehalose fermentation also differentiates C ulcerans from A pyogenes (Thompson and others 1983), another microorganism phenotypically similar to C ulcerans and a common pathogen of goats (Smith and Sherman 1994). The presence of pus in the right acoustic meatus of the goat in the present study suggests that the infection in the central nervous system originated in the middle ear and then progressed to the inner ear, meninges and brain. In many animal species, middle ear infections often progress to the brain, and in some animals, such as lambs and pigs, the infection often ascends from the pharynx via the auditory tube to the middle ear (Wilcock 1992, Summers and others 1995, Radostits and others 2000). In other species, such as dogs and cats, chronic otitis externa is an important predisposing factor for brain infections (Wilcock 1992). In the present study, there was no sign of otitis externa, suggesting that the bacteria had reached the middle ear through the auditory tube from the pharynx. Fibrous proliferation in the central nervous system is usually rudimentary unless the inflammatory process is close to the meningeal surface (Glass and others 1993, Summers and others 1995). In the present case, the connection of the abscess to the meninges might have been responsible for the fibrous capsule that was found. To the authors knowledge the present study is the first report of an infection by C ulcerans in a goat, as well as the first reported case of pyogranulomatous meningoencephalitis due to the same agent reported in any species. This microorganism is also a human pathogen, and the zoonotic potential of the infection needs to be emphasised (Kisley and others 1994). ACKNOWLEDGEMENTS The authors thank Ms E. Vidal for excellent technical assistance, Mrs S. J. Uzal for reviewing and Mrs S. Fitisemanu for typing the manuscript.

References
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The Veterinary Record, March 5, 2005

Downloaded from veterinaryrecord.bmj.com on March 28, 2012 - Published by group.bmj.com

Pyogranulomatous meningoencephalitis in a goat due to Corynebacterium ulcerans

W. E. Morris, F. A. Uzal and A. L. Cipolla Veterinary Record 2005 156: 317-318

doi: 10.1136/vr.156.10.317

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