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Tryland M.,Section for Arctic Veterinary Medicine | Nesbakken T.,Section for Food Safety | Robertson L.,Section for Microbiology | Grahek-Ogden D.,Norwegian Scientific Committee for Food Safety | Lunestad B.T.,National Institute of Nutrition And Seafood Research
Zoonoses and Public Health | Year: 2014

Only a few countries worldwide hunt seals and whales commercially. In Norway, hooded and harp seals and minke whales are commercially harvested, and coastal seals (harbour and grey seals) are hunted as game. Marine mammal meat is sold to the public and thus included in general microbiological meat control regulations. Slaughtering and dressing of marine mammals are performed in the open air on deck, and many factors on board sealing or whaling vessels may affect meat quality, such as the ice used for cooling whale meat and the seawater used for cleaning, storage of whale meat in the open air until ambient temperature is reached, and the hygienic conditions of equipment, decks, and other surfaces. Based on existing reports, it appears that meat of seal and whale does not usually represent a microbiological hazard to consumers in Norway, because human disease has not been associated with consumption of such foods. However, as hygienic control on marine mammal meat is ad hoc, mainly based on spot-testing, and addresses very few human pathogens, this conclusion may be premature. Additionally, few data from surveys or systematic quality control screenings have been published. This review examines the occurrence of potential human pathogens in marine mammals, as well as critical points for contamination of meat during the slaughter, dressing, cooling, storage and processing of meat. Some zoonotic agents are of particular relevance as foodborne pathogens, such as Trichinella spp., Toxoplasma gondii, Salmonella and Leptospira spp. In addition, Mycoplasma spp. parapoxvirus and Mycobacterium spp. constitute occupational risks during handling of marine mammals and marine mammal products. Adequate training in hygienic procedures is necessary to minimize the risk of contamination on board, and acquiring further data is essential for obtaining a realistic assessment of the microbiological risk to humans from consuming marine mammal meat. © 2013 Blackwell Verlag GmbH.


Asmare K.,Hawassa University | Sibhat B.,Haramaya University | Ayelet G.,National Veterinary Institute | Shiferaw J.,National Veterinary Institute | Godfroid J.,Section for Arctic Veterinary Medicine
Acta Tropica | Year: 2013

A cross-sectional study was conducted to investigate the seroprevalence of brucellosis and identify risk factors in exotic and cross bred cattle in Ethiopia. A total of 2334 cattle from 273 farms were tested serially for Brucella antibodies using the Rose Bengal Plate Test (RBPT) and the Compliment Fixation Test (CFT). The overall animal level seroprevalence was 1.9% (95% CI: 1.2, 2.6), with urban and peri-urban dairy 2.4% (95% CI: 1.4, 3.4), commercial 1.5% (95% CI: 0.5, 2.5) and breeding farms 1.5% (95% CI: 0.2, 3.2). The overall farm level prevalence was 10.6% (95% CI: 6.9, 14.3), with 8.6% (95% CI: 4.8, 12.4) in urban and peri-urban dairy followed by 16.9% (95% CI: 7.3, 26.6) in commercial and 20.0% (95% CI: 0.0, 59.4) in breeding farms. At individual animal level, purchased cows and adult age groups were observed to associate with Brucella seropositivity while presence of small ruminants on the farm was the only factor associated with increased risk of herd level Brucella infection. The lack of association between reproductive disorders and Brucella seroprevalence suggest that other causes largely outweigh as causes of the aforesaid disorder in studied production systems and demands an investigation. Finally, the need for isolation and characterization of circulating Brucella spp. and institution of regulatory measures to reinforce farm biosecurity was suggested. © 2013.


Larsen A.K.,Section for Arctic Veterinary Medicine | Larsen A.K.,Member of the Fram Center | Nymo I.H.,Section for Arctic Veterinary Medicine | Nymo I.H.,Member of the Fram Center | And 5 more authors.
PLoS ONE | Year: 2013

A high prevalence of Brucella pinnipedialis serology and bacteriology positive animals has been found in the Northeast Atlantic stock of hooded seal (Cystophora cristata); however no associated gross pathological changes have been identified. Marine mammal brucellae have previously displayed different infection patterns in human and murine macrophages. To investigate if marine mammal Brucella spp. are able to invade and multiply in cells originating from a presumed host species, we infected alveolar macrophages from hooded seal with a B. pinnipedialis hooded seal isolate. Hooded seal alveolar macrophages were also challenged with B. pinnipedialis reference strain (NCTC 12890) from harbor seal (Phoca vitulina), B. ceti reference strain (NCTC 12891) from harbor porpoise (Phocoena phocoena) and a B. ceti Atlantic white-sided dolphin (Lagenorhynchus acutus) isolate (M83/07/1), to evaluate possible species-specific differences. Brucella suis 1330 was included as a positive control. Alveolar macrophages were obtained by post mortem bronchoalveolar lavage of euthanized hooded seals. Phenotyping of cells in the lavage fluid was executed by flow cytometry using the surface markers CD14 and CD18. Cultured lavage cells were identified as alveolar macrophages based on morphology, expression of surface markers and phagocytic ability. Alveolar macrophages were challenged with Brucella spp. in a gentamicin protection assay. Following infection, cell lysates from different time points were plated and evaluated quantitatively for colony forming units. Intracellular presence of B. pinnipedialis hooded seal isolate was verified by immunocytochemistry. Our results show that the marine mammal brucellae were able to enter hooded seal alveolar macrophages; however, they did not multiply intracellularly and were eliminated within 48 hours, to the contrary of B. suis that showed the classical pattern of a pathogenic strain. In conclusion, none of the four marine mammal strains tested were able to establish a persistent infection in primary alveolar macrophages from hooded seal. © 2013 Larsen et al.


Davidson R.K.,Norwegian Veterinary Institute | Romig T.,University of Hohenheim | Jenkins E.,University of Saskatchewan | Tryland M.,Section for Arctic Veterinary Medicine | Robertson L.J.,Parasitology Laboratory
Trends in Parasitology | Year: 2012

In the past three decades, . Echinococcus multilocularis, the cause of human alveolar echinococcosis, has been reported in several new countries both in definitive hosts (canids) as well as in people. Unless treated, infection with this cestode in people is fatal. In previously endemic countries throughout the Northern Hemisphere, geographic ranges and human and animal prevalence levels seem to be increasing. Anthropogenic influences, including increased globalisation of animals and animal products, and altered human/animal interfaces are thought to play a vital role in the global emergence of this pathogenic cestode. Molecular epidemiological techniques are a useful tool for detecting and tracing introductions, and differentiating these from range expansions. © 2012 Elsevier Ltd.


Bhagwat S.S.,University of Tromsø | Larsen A.K.,Section for Arctic Veterinary Medicine | Winberg J.-O.,University of Tromsø | Seternes O.-M.,University of Tromsø | Bang B.E.,University of Tromsø
Food and Chemical Toxicology | Year: 2014

Occupational skin symptoms are prevalent among the workers of the seafood processing industry. In this study we investigate the role of salmon ( Salmo salar) and king crab trypsin ( Paralithodes camtschaticus) as inducers of inflammation in skin via secretion of inflammatory mediators. Human skin keratinocytes (HaCaT cells) were exposed to purified salmon and king crab trypsin. We observed that salmon trypsin enhanced the secretion of IL-8 and MMP-2 and crab trypsin enhanced the secretion of IL-8, MMP-2 and MMP-9 in a dose dependent manner. As protease activated receptors (PAR)-2 in skin are known to play an important role in physiology and pathology, we explored the involvement of these receptors in mediating the release of interleukin (IL)-8 and matrix metalloproteinase (MMP)-2 and -9 subsequent to exposure of skin keratinocytes to salmon and crab trypsin. In addition we observed that salmon and crab trypsin exhibit individual differences in stimulating the release of these inflammatory mediators. Finally, using specific small interfering RNA (siRNA) against PAR-2, we confirmed that the increase in secretion of IL-8, MMP-2 and MMP-9 in skin keratinocytes following exposure to salmon and crab trypsin was mediated via activation of PAR-2. These results suggest that exposure to proteases from the seafood may lead to inflammatory reactions in skin. © 2014 Elsevier Ltd.


Larsen A.K.,Section for Arctic Veterinary Medicine | Larsen A.K.,Fram Center | Nymo I.H.,Section for Arctic Veterinary Medicine | Nymo I.H.,Fram Center | And 4 more authors.
PLoS ONE | Year: 2013

Marine mammal Brucella spp. have been isolated from pinnipeds (B. pinnipedialis) and cetaceans (B. ceti) from around the world. Although the zoonotic potential of marine mammal brucellae is largely unknown, reports of human disease exist. There are few studies of the mechanisms of bacterial intracellular invasion and multiplication involving the marine mammal Brucella spp. We examined the infective capacity of two genetically different B. pinnipedialis strains (reference strain; NTCT 12890 and a hooded seal isolate; B17) by measuring the ability of the bacteria to enter and replicate in cultured phagocytes and epithelial cells. Human macrophage-like cells (THP-1), two murine macrophage cell lines (RAW264.7 and J774A.1), and a human malignant epithelial cell line (HeLa S3) were challenged with bacteria in a gentamicin protection assay. Our results show that B. pinnipedialis is internalized, but is then gradually eliminated during the next 72-96 hours. Confocal microscopy revealed that intracellular B. pinnipedialis hooded seal strain colocalized with lysosomal compartments at 1.5 and 24 hours after infection. Intracellular presence of B. pinnipedialis hooded seal strain was verified by transmission electron microscopy. By using a cholesterol-scavenging lipid inhibitor, entrance of B. pinnipedialis hooded seal strain in human macrophages was significantly reduced by 65.8% (± 17.3), suggesting involvement of lipid-rafts in intracellular entry. Murine macrophages invaded by B. pinnipedialis do not release nitric oxide (NO) and intracellular bacterial presence does not induce cell death. In summary, B. pinnipedialis hooded seal strain can enter human and murine macrophages, as well as human epithelial cells. Intracellular entry of B. pinnipedialis hooded seal strain involves, but seems not to be limited to, lipid-rafts in human macrophages. Brucella pinnipedialis does not multiply or survive for prolonged periods intracellulary. © 2013 Larsen et al.


Jensen S.-K.,Section for Arctic Veterinary Medicine | Jensen S.-K.,University of St. Andrews | Nymo I.H.,Section for Arctic Veterinary Medicine | Forcada J.,Natural Environment Research Council | And 2 more authors.
Diseases of Aquatic Organisms | Year: 2013

Brucellosis is a worldwide infectious zoonotic disease caused by Gram-negative bacteria of the genus Brucella, and Brucella infections in marine mammals were first reported in 1994. A serosurvey investigating the presence of anti-Brucella antibodies in 3 Antarctic pinniped species was undertaken with a protein A/G indirect enzyme-linked immunosorbent assay (iELISA) and the Rose Bengal test (RBT). Serum samples from 33 Weddell seals Leptonychotes weddelli were analysed, and antibodies were detected in 8 individuals (24.2%) with the iELISA and in 21 (65.6%) with the RBT. We tested 48 southern elephant seal Mirounga leonina sera and detected antibodies in 2 animals (4.7%) with both the iELISA and the RBT. None of the 21 Antarctic fur seals Arctocephalus gazella was found positive. This is the first report of anti-Brucella antibodies in southern elephant seals. The potential impact of Brucella infection in pinnipeds in Antarctica is not known, but Brucella spp. are known to cause abortion in terrestrial species and cetaceans. Our findings suggest that Brucella infection in pinnipeds is present in the Antarctic, but to date B. pinnipedialis has not been isolated from any Antarctic pinniped species, leaving the confirmation of infection pending. © Inter-Research 2013.


Biffa D.,Center for Epidemiology and Biostatistics | Biffa D.,University of Arizona | Johansen T.B.,National Veterinary Institute | Godfroid J.,Section for Arctic Veterinary Medicine | And 3 more authors.
Infection, Genetics and Evolution | Year: 2014

Bovine tuberculosis (BTB) remains a major threat to animal and human health, and obstructs international and inter-regional livestock trade in Ethiopia. Many aspects of epidemiology of BTB and its causative agent, Mycobacterium bovis (M. bovis) are not well known. Aims of the study were to elucidate molecular characteristics of M. bovis strains using MLVA typing method. Further aim was to determine infection pressure associated with occurrence of multiple genotypes in individual infected cattle. Data and samples were collected in the period July 2006-January 2007 in cattle slaughtered at five representative abattoirs across the country. Molecular investigation of the isolates was carried out using multilocus variable-number tandem-repeat analysis (MLVA) of 28 variable numbers of tandem repeat (VNTR) loci, and the results were compared to spoligotyping. This study is believed to contribute to the knowledge of molecular genetics and epidemiology of M. bovis in Ethiopia and elsewhere with similar BTB epidemic situation and livestock production settings. Four-hundred and six tissue samples from 337 carcasses revealing gross pathologic lesions compatible with BTB were collected from five abattoirs. Fifty-eight isolates obtained from cultured samples were subjected to region of difference (RD) analysis and MLVA typing. RD confirmed all isolates as being M. bovis. MLVA revealed a high heterogeneity of M. bovis (19 genotypes) and the discriminatory power of MLVA was higher than for spoligotyping (Hunter-Gaston Diversity Index (HGDI) 0.92 vs. 0.82). Adoption of the nine VNTR loci with ≥3 alleles provided good differentiation between the isolates. However, differentiation was optimized when MLVA was combined with spoligotyping (HGDI=0.99). MLVA confirmed infections with multiple genotypes in 38.5% (10/26) of individual animals. In conclusion, the usefulness of MLVA for genotyping M. bovis in high prevalence settings was demonstrated. BTB in Ethiopia is caused by heterogeneous populations of M. bovis and individual carcasses were often infected with different genotypes, indicating a high infection pressure perhaps related to the absence of protective immunity conferred by infection. © 2014 Elsevier B.V.


Biffa D.,Center for Epidemiology and Biostatistics | Bogale A.,Tuskegee University | Godfroid J.,Section for Arctic Veterinary Medicine | Skjerve E.,Center for Epidemiology and Biostatistics
Tropical Animal Health and Production | Year: 2012

Bovine TB is a disease of high economic and public health importance particularly in resource poor countries. Many aspects of pathogenesis of bovine TB in cattle have not been well understood. We carried out an investigation on 337 Ethiopian cattle with characteristic TB-like lesions to describe severity of pathology and factors associated with it. Severity of pathology was determined based upon gross lesion characteristics, distribution and presence/absence of viable mycobacteria. Molecular speciation of mycobacteria was performed using Gene-Probe's Accu-Probe method. Mycobacterium bovis was identified by genomic deletion analysis and spoligotyping. Data were analysed using descriptive statistics and regression model. The results showed that TB-like lesions and M. bovis were more frequently observed in lungs and respiratory lymph nodes. Mammary lesions yielded significant proportion of M. bovis upon culturing. Intestinal lesions were the second most frequently encountered pathology; upon culturing, however, the tissue specimens yielded the lowest proportion of M. bovis isolates. Sex, breed and management system were found to significantly affect TB manifestation. Female (β ± SE = 4.1 ± 1.0; P = 0.00) and exotic breed (β ± SE = 1.7 ± 0.9; P = 0.045) were at a relatively higher risk of developing severe tuberculosis. TB pathology was more severe in cattle raised under large-scale farming (β ± SE = 2.3 ± 0.5; P = 0.00). The fact that severe tuberculosis is linked to high degree of disease transmission potential warrants implementation of proper disease surveillance programs in large-scale farms. Isolation of M. bovis from mammary and muscle tissues implies a potential threat of zoonotic transmission, where raw milk and raw beef constitute a customary dietary regimen in Ethiopia. © 2011 Springer Science+Business Media B.V.


PubMed | Section for Arctic Veterinary Medicine
Type: Journal Article | Journal: PloS one | Year: 2013

A high prevalence of Brucellapinnipedialis serology and bacteriology positive animals has been found in the Northeast Atlantic stock of hooded seal (Cystophoracristata); however no associated gross pathological changes have been identified. Marine mammal brucellae have previously displayed different infection patterns in human and murine macrophages. To investigate if marine mammal Brucella spp. are able to invade and multiply in cells originating from a presumed host species, we infected alveolar macrophages from hooded seal with a B. pinnipedialis hooded seal isolate. Hooded seal alveolar macrophages were also challenged with B. pinnipedialis reference strain (NCTC 12890) from harbor seal (Phocavitulina), B. ceti reference strain (NCTC 12891) from harbor porpoise (Phocoenaphocoena) and a B. ceti Atlantic white-sided dolphin (Lagenorhynchusacutus) isolate (M83/07/1), to evaluate possible species-specific differences. Brucella suis 1330 was included as a positive control. Alveolar macrophages were obtained by post mortem bronchoalveolar lavage of euthanized hooded seals. Phenotyping of cells in the lavage fluid was executed by flow cytometry using the surface markers CD14 and CD18. Cultured lavage cells were identified as alveolar macrophages based on morphology, expression of surface markers and phagocytic ability. Alveolar macrophages were challenged with Brucella spp. in a gentamicin protection assay. Following infection, cell lysates from different time points were plated and evaluated quantitatively for colony forming units. Intracellular presence of B. pinnipedialis hooded seal isolate was verified by immunocytochemistry. Our results show that the marine mammal brucellae were able to enter hooded seal alveolar macrophages; however, they did not multiply intracellularly and were eliminated within 48 hours, to the contrary of B. suis that showed the classical pattern of a pathogenic strain. In conclusion, none of the four marine mammal strains tested were able to establish a persistent infection in primary alveolar macrophages from hooded seal.

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