N. J. Davison

Meningoencephalitis associated with Brucella species in a live-stranded striped dolphin (Stenella coeruleoalba) in south-west England

THE first recovery of a Brucella species from marine mammals was reported in Scotland in 1994 ([Ross and others 1994][1]). Since then, there have been many reports of both serological evidence and isolations of Brucella species from many marine mammal species around the world ([Dawson 2005][2], [

Association of a lukM-positive clone of Staphylococcus aureus with fatal exudative dermatitis in red squirrels (Sciurus vulgaris).

Fatal exudative dermatitis (FED) is a recently described condition affecting red squirrels (Sciurus vulgaris) on the Isle of Wight and Jersey (Simpson et al., 2010a). Staphylococcus aureus strains isolated from skin lesions in cases of FED were characterised by molecular and phenotypic approaches. The strains were found to belong to a single MLST clonal complex (CC49) representing either ST49 or a novel single locus variant thereof (ST1957), were closely related by other molecular typing approaches, and all possessed the leukotoxin M encoding gene (lukM). In contrast S. aureus was either not isolated from none-FED cases or belonged to distinct and diverse molecular types that, with one exception, did not encode lukM. All isolates from FED cases were susceptible to all antimicrobials tested, including penicillin, and all proved negative for mecA and mecC as well as 14 other staphylococcal toxin genes. As all squirrels affected by FED were infected with S. aureus of the same lineage and encoded the lukM gene, it is possible that strains of this lineage may be involved in the pathogenesis of the dermatitis.

Postmortem evidence of interactions of bottlenose dolphins (Tursiops truncatus) with other dolphin species in south-west England

Reports of violent interactions between bottlenose dolphins (Tursiops truncatus) and harbour porpoises (Phocoena phocoena) in the coastal waters of the UK are well documented. Examination of stranded cetaceans by the Cornwall Wildlife Trust Marine Strandings Network and the UK Cetacean Strandings Investigation Programme has indicated that seven animals, of four other species, found stranded in south-west England, had pathology consistent with bottlenose dolphin interaction, including two juvenile and two adult common dolphins (Delphinus delphis), one juvenile pilot whale (Globicephala melas), one juvenile Risso’s dolphin (Grampus griseus) and one adult striped dolphin (Stenella coeruleoalba). Although recorded traumatic lesions were often not as severe as those found in harbour porpoises, it is probable that the interactions did contribute to stranding and/or death in all four of the juvenile animals examined. Furthermore, analysis of photographs taken before establishment of the Marine Strandings Network revealed rake (teeth) marks consistent with bottlenose dolphin interaction on one stranded common dolphin in 1992. A number of causes have been suggested for these interactions in harbour porpoises stranded in the UK and it is possible that any combination of these factors may also be implicated in the cases described in this report.

Isolation of Brucella species from a bottlenosed dolphin (Tursiops truncatus)

Brucella was first recovered from marine mammals in 1994 (Ross and others 1994). Since then, there have been numerous further accounts of serological evidence and isolations of Brucella species originating from marine mammals inhabiting many of the world’s oceans. This short communication describes the first confirmed case of Brucella species infection in a wild bottlenosed dolphin (Tursiops truncatus) in the UK. In December 2004, a male bottlenosed dolphin was found dead on a beach at Gwithian Hayle, Cornwall. It was taken to the Veterinary Laboratories Agency (VLA) – Truro for postmortem examination. The animal, considered to be in moderate condition with moderate autolysis, was 258 cm long and weighed 160 kg. There were numerous deep rake or bite marks present over the entire body (Fig 1), possibly caused by other bottlenosed dolphins. There had been some scavenger damage to the left eye and left pectoral flipper. Several longstanding cestode (Phyllobothrium delphini) cysts were present in the blubber around the vent; abscesses associated with these cysts were similar to the abscess from which Brucella species was later isolated. Postmortem examination revealed numerous nematodes in the bronchi and bronchioles, which were associated with a parasitic pneumonia. All three sections of the stomach were empty, and chyle was not present in the mesentery, indicating that the animal had not fed recently. The rake or bite marks were not associated with any additional pathology and the cause of death was not established. The exact age of the animal had yet to be determined. Samples of heart blood, lung, liver, kidney and intestine were prepared for routine bacteriological culture on blood agar and MacConkey’s agar. A mixed growth of Edwardsiella tarda and a non-haemolytic Staphylococcus species was recovered from the heart blood, lungs and intestine. Analysis of pericardial fluid by the rose bengal plate test revealed antibodies to Brucella species. Tissue samples of lung, liver, kidney, spleen, testes and brainstem, together with samples of pericardial and abdominal fluid, were therefore submitted to VLA – Weybridge to examine for the presence of Brucella species. Nematode worms recovered from the cardiac section of the stomach and from the lung were also subjected to examination by culture. Brucella was isolated from a purulent abscess within blubber taken from around the vent, and a fine bacterial growth was produced after seven days of incubation in enrichment broth and two days after subculture on Farrell’s media (Farrell 1974). The isolate was initially confirmed as Brucella species by classical biotyping methods (Alton and others 1988). It did not require additional carbon dioxide for growth and produced lysis by phages BK2 and Wb (Table 1). Amplification by PCR of an IS711 element downstream of the bp26 gene (Cloeckaert and others 2000) confirmed that the isolate possessed this unique feature specific to marine mammal strains of Brucella species. Molecular characterisation of the outer membrane protein of the strain using a selection of restriction enzymes (Cloeckaert and others 2001) revealed the type to be N-(K), as is found in common dolphins (Delphinus delphis) and striped dolphins (Stenella coeruleoalba). A competitive ELISA (cELISA) and two indirect ELISAs (iELISAs) were used to screen the pericardial and abdominal fluids for the presence of Brucella antibodies. The cELISA (MacMillan 1990) uses a lipopolysaccharide (LPS) Brucella melitensis antigen and a monoclonal antibody conjugate. For the two iELISAs, B melitensis 16M antigen was used for the first, as described for the cELISA, and for the other a Brucella abortus LPS antigen was used. The iELISAs require an antiglobulin conjugate with specificity for the immunoglobulin isotypes of the species under test; however, protein A has been shown to bind to the immunoglobulin G of a range of marine mammals (Eliasson and others 1989, Sikkema 1989). Positive/negative thresholds for these assays were set with some uncertainty, but were based on those used for testing a wide range of terrestrial mammals from Britain for brucellosis (Table 2). Samples of both pericardial and abdominal fluids were found to be positive for Brucella antibodies by all three assays. A previous report described the characterisation of Brucella species isolated from an aborted fetus of a captive FIG 1: Rake or bite marks on the body of a bottlenosed dolphin (Tursiops truncatus) found dead on a beach in Cornwall

Hepatozoon species infection in wild red squirrels (Sciurus vulgaris) on the Isle of Wight

Postmortem examinations of 49 red squirrels (Sciurus vulgaris) found dead on the Isle of Wight revealed the presence of a Hepatozoon species in 18 of them (37 per cent). The prevalence of infection was highest in subadult animals and no juveniles were infected. The prevalence was higher in the squirrels dying from natural causes (nine of 12) than in squirrels killed in road accidents (seven of 27). The weight of infection varied, and there were heavy infections in squirrels dying from toxoplasmosis and bacterial pneumonia. A PCR-based assay was used to identify the presence of Hepatozoon species dna in the lungs, and immunoperoxidase staining was used to confirm the identity of schizonts observed in histological sections. The nucleotide base sequence of the pcr products indicated that the organism was a novel species closely related to, but distinct from, Hepatozoon erhardovae of bank voles (Clethrionomys glareolus).

Mycobacterium bovis infection in a grey seal pup (Halichoerus grypus)

Mycobacterial disease has been reported in both free-ranging and captive southern hemisphere pinnipeds in Australasia, South America and Europe. Although isolates in cases published prior to 2001 had biochemical characteristics which suggested Mycobacterium bovis , genetic analyses revealed differences and Cousins and others (2003) demonstrated that these isolates were phenotypically and genetically distinct from M bovis and belonged to a unique member of the Mycobacterium tuberculosis complex, given the name Mycobacterium pinnipedii . M pinnipedii has since been isolated from other southern hemisphere pinnipeds, for example, Kriz and others (2011). This paper describes the isolation and identification of what is believed to be the first confirmed case of M bovis in a pinniped. It was isolated from a grey seal pup ( Halichoerus grypus ) undergoing rehabilitation. The seal was found on the south coast of Cornwall in January 2012. It was estimated to be four months of age and weighed 26 kg. The seal was hospitalised in a facility which rehabilitated up to 50 grey seal pups a year and also held a variety of indigenous and non-indigenous pinniped species, otters, penguins, ponies, goats and sheep. On examination, the most significant findings were one 5 cm long open infected wound on the perineum, puncture wounds over both hocks and a tear in the webbing of the right hind flipper. A two-week course of amoxycillin/clavulanic acid was given (Noroclav, initially at 8.75 mg/kg intramuscular, then at 12.5 mg/kg orally, Norbrook, UK). The wounds healed, but due to persistent neutrophilia (initially 22.95 × 109/l, increasing to 72.25 × 109/l; reference range: 2–12 × 109/l; Barnett and Robinson 2003) and failure to gain weight consistently, a two-week course of enrofloxacin (Enrox, 5 mg/kg orally, Virbac, UK) and one injection of long-acting oxytetracycline (Alamycin LA, 20 mg/kg intramuscular, Norbrook, UK) were then given. Over the next month, the seal developed granulomas …

Infection with Brucella ceti and high levels of polychlorinated biphenyls in bottlenose dolphins ( Tursiops truncatus ) stranded in south-west England

Eight bottlenose dolphins (Tursiops truncatus) that stranded in Cornwall, south-west England, between June 2004 and December 2007 were examined using standardised postmortem examination and bacteriological methods. Evidence of Brucella species infection was found in four of these dolphins on culture. In addition, of the eight dolphins, four were positive and two were weakly positive for antibodies to Brucella species on serological analyses of pericardial and other fluids using a competitive ELISA and two indirect ELISAs. High or very high levels of the sum of 25 individual chlorobiphenyl congeners (∑25CBs) were also determined in blubber samples from two of the dolphins (45.5 and 446.6 mg/kg lipid weight).

Staphylococcus aureus ST49 infection in red squirrels.

A PAPER in this issue of Veterinary Record describes dermatitis in red squirrels ( Sciurus vulgaris ) (Simpson and others, [pp 59-62][1]). We wish to report some additional observations. In 2009, a red squirrel on the Isle of Wight died after showing respiratory distress. On postmortem examination

Fatal candidiasis in a wild red squirrel (Sciurus vulgaris)

Candida species are commonly present as commensal organisms in the alimentary tract of mammals and birds, but they are also opportunistic pathogens. Many species may cause disease, but the great majority of cases in both human beings and animals are due to Candida albicans . This is a dimorphic

Isolation of Bisgaardia hudsonensis from a seal bite

We read with interest the recent case report of Bisgaardia hudsonensis from a patient with an infected seal bite. We notice with some concern that there was no mention of whether testing for Mycoplasma was attempted as Mycoplasma phocicerebrale is the most commonly isolated organism from “seal finger” or “speck finger”, an infection that can occur from handling seals or from infected bite wounds. Studies by our group on UK common and grey seals recovered M. phocicerebrale from various sites, including the teeth. Furthermore our studies also found M. phocicerebrale to be the most common organism isolated from infected bite wounds following aggressive interactions between seals. The GP in this case selected to use amoxicillin/clavulanic acid treatment and not tetracycline, the treatment indicated for seal finger cases. The eventual response of the patient to amoxicillin/clavulanic acid would suggest that Mycoplasma infection was not the cause on this occasion. We feel that it is of major importance however, to emphasise the association of M. phocicerebrale with seal finger, the importance of testing for Mycoplasma infections in all suspect cases and the use of prompt and appropriate antibiotic treatment for this serious but little known infection, which can often have significant sequelae. We are also aware of two cases of similar finger infections over the last two decades involving injuries from working with harbour porpoise carcases in the UK. In both cases initial treatment did not improve the condition of the patients and it was only following change of antibiotic regime that the infection subsided, however in one case this had been supported by surgical debridement of the infected digit. In both casesMycoplasma culture was not attempted, however we have recently reported the isolation ofM. phocicerebrale from harbour porpoises increasing our suspicions of Mycoplasma involvement in these cases. We wish to raise the awareness of the medical fraternity to the possibility of seal finger-like infections following injury while working with cetaceans and the importance of prompt treatment with tetracycline in such patients. With respect to the incriminated organism in the case reported by Sundeep and colleagues, we would wish to offer