Geoffrey Foster

A review of Brucella sp. infection of sea mammals with particular emphasis on isolates from Scotland

Brucellae recovered from sea mammals were first reported in 1994. In the years since both culture and serological analysis have demonstrated that the infection occurs in a wide range of species of marine mammals inhabiting a vast amount of the worlds oceans. Molecular studies have demonstrated that the isolates differ from those found amongst terrestrial animals and also distinguish between strains which have seals and cetaceans as their preferred hosts. At the phenotypic level seal and cetacean strains can also be differed with respect to their CO(2) requirement, primary growth on Farrells medium and metabolic activity on galactose. Two new species B. cetaceae and B. pinnipediae have been proposed as a result. This paper provides a review of Brucella in sea mammals and updates findings from the study of sea mammals from around the coast of Scotland.

ISOLATION OF BRUCELLA SPECIES FROM CETACEANS, SEALS AND AN OTTER

Brucella organisms which differed from the recognised species of the genus, were isolated from nine seals, eight cetaceans and one otter. A method is described for the isolation of Brucella species from sea mammals and the first isolations of Brucella species are recorded from an Atlantic white-sided dolphin (Lagenorhynchus acutus), two striped dolphins (Stenella coeuleoalba), a hooded seal (Cystophora cristata), a grey seal (Halichoerus grypus) and a European otter (Lutra lutra). There were differences in the culture media required for the primary isolation of the organisms and in their dependency on carbon dioxide. Subcutaneous lesions, when present, always yielded a confluent growth. The organisms were isolated from seven of 14 spleen samples and also from the mammary glands, uterus, testes and blood and the mandibular, gastric, iliac, sub-lumbar and colorectal lymph nodes.

The newly described mecA homologue, mecALGA251, is present in methicillin-resistant Staphylococcus aureus isolates from a diverse range of host species

OBJECTIVES A previously unidentified mecA homologue, mecA(LGA251), has recently been described in methicillin-resistant Staphylococcus aureus (MRSA) from humans and dairy cattle. The origin and epidemiology of this novel homologue are unclear. The objective of this study was to provide basic descriptive information of MRSA isolates harbouring mecA(LGA251) from a range of host animal species. METHODS A number of S. aureus isolates from historical animal isolate collections were chosen for investigation based on their similarity to known mecA(LGA251) MRSA isolates. The presence of mecA(LGA251) was determined using a multiplex PCR and antimicrobial susceptibility testing performed by disc diffusion. RESULTS MRSA harbouring mecA(LGA251) were found in isolates from a domestic dog, brown rats, a rabbit, a common seal, sheep and a chaffinch. All of the isolates were phenotypically MRSA, although this depended on which test was used; some isolates would be considered susceptible with certain assays. All isolates were susceptible to linezolid, rifampicin, kanamycin, norfloxacin, erythromycin, clindamycin, fusidic acid, tetracycline, trimethoprim/sulfamethoxazole and mupirocin. Five multilocus sequence types were represented (2273, 130, 425, 1764 and 1245) and six spa types (t208, t6293, t742, t6594, t7914 and t843). CONCLUSIONS The discovery of MRSA isolates possessing mecA(LGA251) from a diverse range of host species, including different taxonomic classes, has important implications for the diagnosis of MRSA in these species and our understanding of the epidemiology of this novel mecA homologue.

Brucella species infection in North Sea seal and cetacean populations.

albumin to 1:750 and 1:1500. Overnight incubation at 4°C was the best condition for the different primary antibodies employed. Swine anti-rabbit immunoglobulins 1:20 for use as a secondary antibody and PAP complex 1:100 were both made up in phosphate buffered saline with I per cent bovine serum albumin and incubated for 30 minutes at room temperature. To detect the immune complexes, the sections were incubated in a solution of 60 mg of 3,3-diamino-benzidine tetrahydrochloride, 100 ,ul hydrogen peroxide and 100 ml of phosphate buffered saline for eight minutes and counterstained with Mayers haematoxylin. Ganglia from all horses with either mal seco or EGS showed

MLVA-16 typing of 295 marine mammal Brucella isolates from different animal and geographic origins identifies 7 major groups within Brucella ceti and Brucella pinnipedialis

BackgroundSince 1994, Brucella strains have been isolated from a wide range of marine mammals. They are currently recognized as two new Brucella species, B. pinnipedialis for the pinniped isolates and B. ceti for the cetacean isolates in agreement with host preference and specific phenotypic and molecular markers. In order to investigate the genetic relationships within the marine mammal Brucella isolates and with reference to terrestrial mammal Brucella isolates, we applied in this study the Multiple Loci VNTR (Variable Number of Tandem Repeats) Analysis (MLVA) approach. A previously published assay comprising 16 loci (MLVA-16) that has been shown to be highly relevant and efficient for typing and clustering Brucella strains from animal and human origin was used.Results294 marine mammal Brucella strains collected in European waters from 173 animals and a human isolate from New Zealand presumably from marine origin were investigated by MLVA-16. Marine mammal Brucella isolates were shown to be different from the recognized terrestrial mammal Brucella species and biovars and corresponded to 3 major related groups, one specific of the B. ceti strains, one of the B. pinnipedialis strains and the last composed of the human isolate. In the B. ceti group, 3 subclusters were identified, distinguishing a cluster of dolphin, minke whale and porpoise isolates and two clusters mostly composed of dolphin isolates. These results were in accordance with published analyses using other phenotypic or molecular approaches, or different panels of VNTR loci. The B. pinnipedialis group could be similarly subdivided in 3 subclusters, one composed exclusively of isolates from hooded seals (Cystophora cristata) and the two others comprising other seal species isolates.ConclusionThe clustering analysis of a large collection of marine mammal Brucella isolates from European waters significantly strengthens the current view of the population structure of these two species, and their relative position with respect to the rest of the Brucella genus. MLVA-16 is confirmed as being a rapid, highly discriminatory and reproducible method to classify Brucella strains including the marine mammal isolates. The Brucella2009 MLVA-16 genotyping database available at http://mlva.u-psud.fr/ is providing a detailed coverage of all 9 currently recognized Brucella species.

Causes of death of wild birds of the family Fringillidae in Britain.

The provision of supplementary food for wild birds in gardens during the winter months is common in the UK, but it is possible that it may precipitate infectious diseases in the birds. This paper describes the results of postmortem examinations of 116 wild finches carried out over a period of four years. The two commonest causes of death in areas where high mortality had been reported were infections with the bacteria Salmonella typhimurium DT4o and Escherichia coli O86. Coccidia of the genera Atoxoplasma or Isospora were found in several of the birds but were considered to be incidental. Megabacteria were also identified in some of the birds, for the first time in flocks of wild birds in the UK, but they were not considered to be significant.

Detection of Escherichia coli serogroups O26, O103, O111 and O145 from bovine faeces using immunomagnetic separation and PCR/DNA probe techniques.

Aims: The aim of this study was to isolate Escherichia coli O26, O103, O111 and O145 from 745 samples of bovine faeces using (i) immunomagnetic separation (IMS) beads coated with antibodies to lipopolysaccharide, and slide agglutination (SA) tests and (ii) PCR and DNA probes for the detection of the Verocytotoxin (VT) genes.

Distribution of Escherichia coli O157 in Bovine Fecal Pats and Its Impact on Estimates of the Prevalence of Fecal Shedding

ABSTRACT The distribution of Escherichia coli O157 in bovine feces was examined by testing multiple samples from fecal pats and determining the density of E. coli O157 in immunomagnetic separation (IMS)-positive fecal samples. The density of E. coli O157 in bovine feces was highly variable, differing by as much as 76,800 CFU g−1 between samples from the same fecal pat. The density in most positive samples was <100 CFU g−1, the limit of reliable detection by IMS. Testing only one 1-g sample of feces per pat with IMS may result in a sensitivity of detection as low as 20 to 50%. It is therefore probable that most surveys have greatly underestimated the prevalence of E. coli O157 shedding in cattle and the proportion of farms with shedding cattle. The sensitivity of the detection of E. coli O157 in bovine feces can be as much as doubled by testing two 1-g samples per pat rather than one 1-g sample.

Prophage Sequences Defining Hot Spots of Genome Variation in Salmonella enterica Serovar Typhimurium Can Be Used To Discriminate between Field Isolates

ABSTRACT Sixty-one Salmonella enterica serovar Typhimurium isolates of animal and human origin, matched by phage type, antimicrobial resistance pattern, and place of isolation, were analyzed by microbiological and molecular techniques, including pulsed-field gel electrophoresis (PFGE) and plasmid profiling. PFGE identified 10 profiles that clustered by phage type and antibiotic resistance pattern with human and animal isolates distributed among different PFGE profiles. Genomic DNA was purified from 23 representative strains and hybridized to the composite Salmonella DNA microarray, and specific genomic regions that exhibited significant variation between isolates were identified. Bioinformatic analysis showed that variable regions of DNA were associated with prophage-like elements. Subsequently, simple multiplex PCR assays were designed on the basis of these variable regions that could be used to discriminate between S. enterica serovar Typhimurium isolates from the same geographical region. These multiplex PCR assays, based on prophage-like elements and Salmonella genomic island 1, provide a simple method for identifying new variants of S. enterica serovar Typhimurium in the field.

Salmonella enterica subspecies enterica serotype Typhimurium and Escherichia coli 086 in wild birds at two garden sites in south-west Scotland

Salmonella enterica subspecies enterica serotype Typhimurium and Escherichia coli 086:K61 :NM are two bacteria that can cause outbreaks of mortality in garden birds visiting bird tables and other feeding stations. Two sites in south-west Scotland were monitored for the two organisms for 12 months. At site A, large numbers of birds fed throughout the year, and at site B smaller numbers of birds fed only in the winter months. Samples of composite faeces were collected from the feeding stations and screened for the organisms, and any dead birds were also screened. S Typhimurium definitive type (DT) 56 (variant) was found to be endemic at site A, and was recovered from 48 per cent of samples of composite faeces collected from the bird table, from 42 per cent of composite faeces from underneath a hanging feeder, and from 33 per cent of composite faeces from below a roost used by house sparrows; the organism was also isolated from the carcases of six wild birds found dead at the site. In contrast, S Typhimurium (DT41) was recovered only once at site B, from 2 per cent of the composite faeces from below a hanging feeder, and no dead birds were recovered from the site. E coli 086 was not recovered from the faeces collected from either site, but was isolated from a bird that died from trauma at site A.