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This study aimed to comprehensively characterize the gut microbiota in diarrheal mink. We conducted Shotgun metagenomic sequencing on samples from five groups of diarrheal mink and five groups of healthy mink. The microbiota α-diversity and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology did not show significant differences between the groups. However, significant differences were observed in microbiota β-diversity and the function of carbohydrate-active enzymes (CAZymes) between diarrheal and healthy mink. Specifically, The relative abundance of Firmicutes was lower, whereas that of Bacteroidetes was higher in diarrheal mink. Fusobacteria were enriched as invasive bacteria in the gut of diarrheal mink compared with healthy mink. In addition, Escherichia albertii was identified as a new bacterium in diarrheal mink. Regarding functions, nicotinate and nicotinamide metabolism and glycoside hydrolases 2 (GH2) family were the enhanced KEGG orthology and CAZymes in diarrheal mink. Furthermore, the diversity and number of antibiotic-resistant genes were significantly higher in the diarrheal mink group than in the healthy group. These findings enhance our understanding of the gut microbiota of adult mink and may lead to new approaches to the diagnosis and treatment of mink diarrhea., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Klebsiella pneumoniae is a zoonotic opportunistic pathogen, and also one of the common pathogenic bacteria causing mink pneumonia. The aim of this study was to get a better understanding of the whole-genome of multi-drug resistant Klebsiella pneumoniae with K2 serotype in China. This study for the first time to analyze Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, resistance and virulence genes of Klebsiella pneumoniae in mink., Results: The isolate was Klebsiella pneumoniae with serotype K2 and ST6189 by PCR method. The string test was positive and showed high mucus phenotype. There was one plasmid with IncFIB replicons in the genome. The virulence factors including capsule, lipopolysaccharide, adhesin, iron uptake system, urease, secretory system, regulatory gene (rcsA, rcsB), determinants of pili adhesion, enolase and magnesium ion absorption related genes. The strain was multi-drug resistant. A total of 26  resistance genes, including beta-lactam, aminoglycosides, tetracycline, fluoroquinolones, sulfonamides, amide alcohols, macrolides, rifampicin, fosfomycin, vancomycin, diaminopyrimidines and polymyxin. Multidrug-resistant efflux protein AcrA, AcrB, TolC, were predicted in the strain., Conclusion: It was the first to identify that serotype K2 K. pneumonia with ST6189 isolated from mink in China. The finding indicated that hypervirulent and multi-drug resistant K. pneumoniae was exist in Chinese mink. The whole-genome of K. pneumoniae isolates have importance in mink farming practice., (© 2024. The Author(s).)

Bovine tuberculosis (bTB) is a chronic infectious-contagious disease with worldwide distribution, caused by the zoonotic pathogen Mycobacterium bovis. It is believed that the existence of wild cycles may hamper the success of bTB control strategies worldwide, where wildlife species could be reservoirs of this bacterial agent across their native (e.g., European badgers, wild boars) or non-indigenous (e.g., brushtail possum in New Zealand) ranges. However, further studies are required to understand the potential risk posed by non-native wildlife in becoming carriers of M. bovis in other neglected latitudes, such as the Southern Cone of South America. In this study, we performed a specific M. bovis-RD4 real-time PCR (qPCR) assay to detect bacterial DNA in tissues from the invasive American mink (Neogale vison) in Los Ríos region, Chile. We detected M. bovis DNA in blood samples collected from 13 out of 186 (7 %) minks with known sex and age. We did not find any significant differences in bacterial DNA detection according to mink sex and age. We found that 92 % (12/13) of specimens were positive in lung, 39 % (5/13) in mediastinal lymph node, and 15 % (2/13) in mesenteric lymph node, which suggest that both respiratory and digestive pathways as possible routes of transmission between infected hosts and minks. Our study is the first report on M. bovis molecular detection in invasive minks in an area where the largest cattle population in the country is located. Furthermore, this area is characterized by a low within-herd prevalence of M. bovis infection in cattle, with a relatively low number of infected herds, and so far, no attempts at eradicating the disease have been successful., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Objectives: Klebsiella pneumoniae is a major opportunistic pathogen that is a member of the Enterobacteriaceae. Klebsiella pneumoniae causes pneumonia in mink and has become the primary infectious disease that limits mink farming. In this study, we report the draft genome sequence of a multidrug-resistant (MDR) strain of K. pneumoniae that harbours the mcr-1 gene isolated from a mink in China., Methods: The agar microdilution method was used to determine the minimum inhibitory concentration of the strain. The entire genomic DNA was sequenced using an Illumina MiSeq platform. A multilocus sequence type (MLST) and a core genome SNP phylogenetic tree analysis with a heatmap of the resistance genes and virulence genes were performed., Results: The size of the genome was 5451.826 kb, and it included one chromosome and one plasmid. The draft genome of K. pneumoniae indicated that the isolate was a member of MLST 661. Four types of virulence genes were detected. The results of antimicrobial susceptibility testing showed multiple drug resistance, and 17 resistance genes were identified., Conclusion: The genome sequence reported in this study will help to reveal the key role of antibiotic resistance and pathogenic mechanisms. It will provide useful information for the role of mobile genetic elements in the adaptive translocation and spread of antimicrobial resistance., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Enterocytozoon bieneusi, a zoonotic pathogen prevalent in both humans and animals, is the most frequently diagnosed microsporidian species in humans and presents significant public health risks. However, data on the prevalence and genotypes of E. bieneusi in farmed minks (Neovison vison) and raccoon dogs (Nyctereutes procyonoides) in China are limited. Therefore, 275 minks (89 from Hebei Province, 57 from Heilongjiang Province, 109 from Liaoning Province, 20 from Shandong Province) and 235 raccoon dogs (114 from Hebei Province, 27 from Heilongjiang Province, 61 from Liaoning Province, 33 from Jilin Province) were examined for the prevalence and genotypes of E. bieneusi through sequence analysis of the internal transcribed spacer (ITS) region of the rRNA gene. The overall prevalence of E. bieneusi was 18.6% (95/510), with 10.5% (29/275) in farmed minks and 28.1% (66/235) in raccoon dogs. Ten genotypes (CHN-F1, genotype D, Type IV, EbpC, NCF2, NCF5, NCF6, Peru8, Henan V, and MJ5) were identified in minks and raccoon dogs. This study is the first to detect the CHN-F1, NCF2, NCF6, Peru8, and Henan V genotypes in minks and the NCF5, NCF6, and MJ5 genotypes in raccoon dogs. Additionally, the D, Type IV, and Peru8 genotypes, previously identified in humans, were also found in minks and raccoon dogs, suggesting that these animals could be potential sources of human microsporidiosis. These findings expand the understanding of E. bieneusi's host distribution in China and contribute to the prevention of zoonotic E. bieneusi infections among farmed animals., (© N.-Y. Xue et al., published by EDP Sciences, 2024.)

Streptococcus phocae infection has been described in salmon, sea otters, and several families of pinnipeds. The pathology of the infected animals has mainly been located in the respiratory tract and reproductive system, and with indications of septicemia. In this study, we report the finding of S. phocae in diagnostic material from three unrelated cases of farmed mink. Since S. phocae initially has been described in pinnipeds, two isolates from wild harbor seals were included. All isolates originated from Denmark. To our knowledge, this is the first report of S. phocae infection in mink. The animals (three mink, two seals) were necropsied, and samples were collected for bacteriology, virology, and histopathology. Additionally, the S. phocae isolates were whole genome sequenced and compared to sequences of previously reported isolates from other host species. S. phocae was isolated from the lungs of one mink and one seal with bacteremia, and from one seal with pneumonia. The two remaining mink had dermal infections on the paws and S. phocae was isolated from the lesions. The analysis of the sequence data showed that the three mink isolates and one seal isolate were closely related. Further investigation is needed to conclude whether S. phocae is establishing as commensal in farmed mink and to uncover the infection related pathology in mink. Streptococcus phocae has been described as an emerging pathogen in other species, therefore future awareness and surveillance of this pathogen is crucial., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Antibiotic-resistant infections caused by extended-spectrum β-lactamases (ESBLs) and carbapenemases are increasing worldwide. Bacteria resistant to extended-spectrum cephalosporins and last resort carbapenems have been reported from food animals and their environments. Other concentrated nonfood-producing animals such as mink farming can be a reservoir of bacteria resistant to these critically important antibiotics. The objective of this study was to determine the prevalence of ESBL-producing bacteria and carbapenem-resistant (CR) bacteria from mink fecal ( n  = 42) and feed ( n  = 8) samples obtained from a commercial mink farm in the United States. The most prevalent ESBL-producing bacteria identified from the fecal samples were Escherichia coli (93%), Klebsiella pneumoniae (76%), and Proteus species (88%). E. coli (100%) and K. pneumoniae (75%) were also the most prevalent ESBL-producing bacteria identified from feed samples. All ESBL E. coli isolates were resistant to penicillin and most cephem beta-lactam antibiotics. Among the ESBL E. coli isolates, co-resistance was observed to ciprofloxacin (33%) and gentamicin (28%) indicating multidrug resistance. ESBL E. coli isolates predominantly carried bla CTX-M-14 and bla CTX-M-15 genes. Although all feed K. pneumoniae isolates carried bla CTX-M-9 , all fecal K. pneumoniae isolates carried bla SHV . CR Pseudomonas species (7%), Hafnia alvei (24%), and Myroides odoratimimus (9.5%) were detected from fecal samples. H. alvei (37.5%) was the only CR bacteria detected from the feed samples. All CR isolates were polymerase chain reaction negative for the tested carbapenemases that are commonly reported, which may indicate intrinsic rather than acquired resistance. This study indicates that mink production can be a reservoir for bacteria resistant to the highest priority critically important antibiotics for human health.

American mink ( Neovison vison ) is a significant source of global fur production. Except for a few studies from Denmark and Canada reporting antimicrobial resistance in bacteria isolated from clinical cases, studies from the general mink population are scarce and absent in the United States. Mink feces ( n  = 42) and feed ( n  = 8) samples obtained from a mink farm were cultured for the enumeration and detection of tetracycline-resistant (TET r )- and third-generation cephalosporin-resistant (TGC r )- Escherichia coli . Isolates were characterized phenotypically for their resistance to other antibiotics and genotypically for resistance genes. TET r E. coli were detected from 98% of feces samples (mean concentration = 6 log 10 ) and from 100% of feed samples (mean concentration = 3.2 logs). Among TET r E. coli isolates, 44% ( n  = 41) of fecal- and 50% ( n  = 8) of feed isolates were multidrug resistant (MDR; resistance to ≥3 antimicrobial classes), and 96% ( n  = 49) of TET r isolates were positive for tet (A) and/or tet (B). TGC r E. coli were detected from 95% of feces and 75% of feed samples with 78% ( n  = 40) of fecal isolates, and all six of the feed isolates were MDR. Nearly two-thirds (65%) of the TGC r E. coli isolates ( n  = 46) were positive for bla CMY-2 ; the remaining 35% were positive for bla CTX-M, with the bla CTX-M-14 being the predominant (75%, n  = 16) variant detected. Metagenomic DNA was extracted directly from feces and feed samples, and it was tested for 84 antimicrobial resistance genes by using quantitative polymerase chain reaction (PCR) array; selected genes were also quantified by droplet digital PCR. The genes detected from the fecal samples belonged mainly to five antimicrobial classes: macrolide-lincosamide-streptogramin B (MLS B ; 100% prevalence), TETs (88.1%), β-lactams (71.4%), aminoglycosides (66.7%), and fluoroquinolones (47.6%). β-Lactam, MLS B , and TET resistance genes were also detected from feed samples. Our study serves as a baseline for further studies and to streamline antimicrobial use in mink production in accordance with current regulations as in food animals.

On many mink farms, antibiotics are used extensively during the lactation period to reduce the prevalence and severity of pre-weaning diarrhoea (PWD) in mink kits (also referred to as greasy kit syndrome). Concerns have been raised, that routine treatment of PWD with antibiotics could affect the natural successional development of the gut microbiota, which may have long lasting consequences. Here we investigated the effects of early life antibiotic treatment administered for 1 week (postnatal days 13-20). Two routes of antibiotic administration were compared to a non-treated control group (CTR, n = 24). Routes of administration included indirect treatment, through the milk from dams receiving antibiotics by intramuscular administration (ABX_D, n = 24) and direct treatment by intramuscular administration to the kits (ABX_K, n = 24). A tendency for slightly increased weight at termination (Day 205) was observed in the ABX_K group. The gut microbiota composition was profiled by 16S rRNA gene sequencing at eight time points between Day 7 and Day 205. A clear successional development of the gut microbiota composition was observed and both treatment regimens caused detectable changes in the gut microbiota until at least eight days after treatment ceased. At termination, a significant positive correlation was identified between microbial diversity and animal weight.

In recent decades, the number of studies on the occurrence of resistant strains in wildlife animals has increased significantly, but data are still fragmentary. The aim of this study was to evaluate drug resistance of Escherichia coli strains isolated from wild carnivorous mammals, common in Poland. Selective media with antimicrobials (tetracycline, kanamycin, chloramphenicol, and cefotaxime) were used for isolation. Of 53 isolates shown to be distinct by the amplification of DNA fragments surrounding rare restriction site-fingerprinting method, 77.8% were multidrug-resistant (multidrug-resistant). All strains were resistant to ampicillin and many of them also exhibited resistance to tetracycline (76.2%), sulfamethoxazole (57.1%), streptomycin and kanamycin (49.2%), chloramphenicol (30.1%), and nalidixic acid (46%). In most cases, the phenotypic resistance profile was confirmed by detection of relevant genes mostly occurring in strains isolated from livestock animals and humans. Extended-spectrum β-lactamase-producing strains were detected in one mink and three martens. The strains were carriers of bla TEM-1, bla TEM-135 , and bla CTX-M-15 genes. Our research confirmed a high carrier rate of MDR E. coli , even more than one MDR strain in a single individual; therefore, wider monitoring in this group of animals should be considered.

Mortality of mink kits represents a significant loss to production. However, causes of post-weaning mortality in mink kits in modern Danish mink production systems are still relatively poorly documented. We performed a cross-sectional mortality study on eight Danish mink farms including 1893 post mortem examinations of mink kits found dead or euthanized. We assessed the prevalence of cystitis and urolithiasis leading to mortality. Gross pathological findings as well as animal characteristics were recorded and associations with post mortem microbiology (using culture and MaldiTof-MS Vitek MS system) were investigated. Cystitis and/or urolithiasis were associated with death in 33 % (n = 476) and 37 % (n = 166) of the examined mink kits in 2015 and 2017. On farm level, the prevalence of cystitis and/or urolithiasis leading to mortality varied from 0.25 % to 1.27 % with a low overall mortality of 0.9-4.5 %. The bacterial agent most frequently isolated in post mortem bladder swabs from mink with a post mortem diagnosis of urolithiasis and cystitis was Staphylococcus delphini group A (51/283) with a significant (p < 0.0001, CI = [19.5;4745.7]) association to gross pathological findings in the urinary tract. Staphylococcus delphini group A was cultured from 70 % of the skin swabs obtained from apparently healthy mink euthanized at pelting (n = 222). In conclusion urinary tract disease (cystitis and urolithiasis) was the most prevalent post mortem diagnosis during the growth period and was associated with Staphylococcus delphini group A., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

More than 55 million mink skins were produced globally in 2017. As a consequence, a large number of people are employed in mink production worldwide. In Denmark, farmed mink were found to constitute a reservoir of methicillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC) 398 and 6000 mink farm workers in Denmark are potentially exposed to LA-MRSA CC398. The study aim was to elucidate the source of LA-MRSA CC398 in mink farms and to investigate possible transmission to humans. In total, 161 LA-MRSA CC398 isolates from mink (n = 65), mink feed (n = 16) and humans (n = 80) with reported contact to mink, were whole-genome sequenced and compared to 183 LA-MRSA CC398 isolates from Danish pigs and an international collection of 89 S. aureus CC398 isolates. Most of the mink-associated isolates clustered within the predominant LA-MRSA CC398 lineages circulating in the Danish pig production, supporting that pigs are a source of LA-MRSA CC398 in mink feed, mink, and mink farmers., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The present study was designed to identify nine Arcanobacterium phocae strains isolated from cases of mink dermatitis of a single farm in Finland and characterize the strains for epidemiological relationships. All nine strains and previously described A. phocae used for comparative purposes were identified and further characterized phenotypically, by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), by Fourier Transform Infrared Spectroscopy (FT-IR) and genotypically by detection of phocaelysin encoding gene phl with a previously developed loop-mediated isothermal amplification (LAMP) assay and by sequencing 16S rRNA gene and gene phl, the elongation factor tu encoding gene tuf and the β subunit of bacterial RNA polymerase encoding gene rpoB. Genetic relatedness among isolates was determined using whole-genome single nucleotide polymorphism (wgSNP) analysis. The wgSNP results, partly the MALDI-TOF MS and FT-IR analyses and sequencing of the genes, revealed that the nine A. phocae strains recovered from a single farm showed close sequence similarities among each other and differed from previously investigated A. phocae strains isolated from other farms and animals in Finland and from the A. phocae type strain. This indicated a close epidemiological relationship of the A. phocae strains isolated from a single farm and that the nine A. phocae strains of the present study might have developed from a common ancestor., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this study, 20 P. aeruginosa strains were isolated from 112 farmed mink exhibiting hemorrhagic pneumonia in mideastern Shandong province, China. Serotype G (18/20) was the dominant serotype among the isolates with prevalence in mink, followed by serotype B (1/20), serotype C (1/20). The 9 virulence-associated genes of P. aeruginosa were tested using PCR. The prevalence of the virulence genes for the isolates were algD 95% (19/20), plcH 85% (17/20), exoY 80% (16/20), aprA 75% (15/20), lasB 70% (14/20), exoS 65% (13/20), exoT 60% (12/20) and toxA 60% (12/20), respectively. The 20 isolates were negative for exoU gene. The isolates exhibited multidrug resistance and cross resistance, using antimicrobial disc susceptibility assays. The animal experiments demonstrated that LD50% of the P.aeruginosa-CS-2 in the intratracheally challenged mink was 2.2 × 10 7.0  CFU, and 6.8 × 10 4.0  CFU in the intraperitoneally challenged mink. It implied that both the inoculation doses and the routes of inoculation could have influences on the pathogenicity of P. aeruginosa in mink. Therefore, the evolutionary and epidemiological surveillance of P. aeruginosa in mink should be further strengthened for public health., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Influenza A virus (IAV) and bacteria co-infection can influence the host clinical conditions. Both H9N2 IAV and Pseudomonas aeruginosa (P. aeruginosa) are potential pathogens of respiratory diseases in mink. In this study, to clarify the effects of H9N2 IAV and P. aeruginosa co-infections on hemorrhagic pneumonia in mink, we carried out to establish the mink models of the two-pathogen co-infections in different orders. Compared with the single infections with H9N2 IAV or P. aeruginosa, the mink co-infected with H9N2 IAV and P. aeruginosa showed severe respiratory diseases, and exacerbated histopathological lesions and more obvious apoptosis in the lung tissues. H9N2 IAV shedding and viral loads in the lungs of the mink co-infected with H9N2 IAV and P. aeruginosa were higher than those in the mink with single H9N2 IAV infection. Furthermore, the clearance of P. aeruginosa in the co-infected mink lungs was delayed. In addition, the anti-H9N2 antibody titers in mink with P. aeruginosa co-infection following H9N2 IAV infection were significantly higher than those of the other groups. This implied that H9N2 IAV and P. aeruginosa co-infection contributed to the development of hemorrhagic pneumonia in mink, and that P. aeruginosa should play a major role in the disease. The exact interaction mechanism among H9N2 IAV, P. aeruginosa and the host needs to be further investigated., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, the prevalence, phenotypes, and clonal relationships of Escherichia coli ( E. coli ) strains isolated from minks were investigated. In July 2017, a total of 62 fresh faecal swab samples were randomly collected from one large-scale mink farm in Zhucheng, Shandong Province, China. In all the samples, 50 E. coli strains were isolated and then assigned to serotyping, antimicrobial susceptibility test, detection of antimicrobial resistance genes and the Class 1 integrons, and multilocus sequence typing (MLST). Four pathogenic serotypes were identified among all the isolates, while the most common serotype was enterohemorrhagic E. coli O104:H4 (6.0 %). Antimicrobial sensitivity testing revealed that most isolates were susceptible to cefoxitin (96.0 %) and amikacin (82.0 %), while most isolates were resistant to ampicillin (92.0 %) and tetracycline (90.0 %). An analysis of the nucleotide sequences revealed that 7 isolates (14.0%) carried 4 types of Class 1 integron cassette, including dfrA27+aadA2+qnrA (57.1%), dfrA17+aadA5 (14.3%), dfrA12+aadA2 (14.3%), and dfrA1+aadA1 (14.3%). PCR screening showed that 14 antibiotic resistance genes were presented in 50 isolates, while the most prevalent resistance gene was qnrS , which was detected in 60.0 % of isolates, followed by sul2 (40.0%) and oqxA (38.0%). MLST analysis showed that 32 sequence types (STs) were identified, while ST46 was the predominant genotype among all isolates. Clonal complex 3 (CC3) was dominant. Compared with 340 human E. coli STs reported in China, the ST10 clonal complex, known as the largest human clonal complex, was also found in the 50 mink E. coli isolates. Meanwhile, mink-derived strain ST206 formed a new clonal complex, CC206, which was different from human ST strains. Our results showed that farmed minks could be reservoirs of antimicrobial-resistant E. coli with Class 1 integron cassettes and resistance genes, which were likely to pose a threat to public health. Therefore, continuous inspections and monitoring of E. coli in minks are essential for detecting and controlling emerging E. coli with different serovars as well as antibiotic resistance.

Hemorrhagic pneumonia in mink is a fatal disease caused by Pseudomonas aeruginosa. Very little is known about P. aeruginosa in relation to genotype and the mechanisms underlying antimicrobial resistance in mink. A total of 110 P. aeruginosa samples were collected from mink from Chinese mink farms between 2007 and 2015. Samples underwent molecular genotyping using pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST), antimicrobial susceptibility and its mechanism were investigated at the molecular level. The PFGE identified 73 unique types and 15 clusters, while MLST identified 43 (7 new) sequence types (ST) and 12 sequence type clonal complexes (STCC). Sequence types and PFGE showed persistence of endemic clones in cities Wendeng (Shandong, China) and Dalian (Liaoning, China), even in different timelines. The MLST also revealed the gene correlation of the mink P. aeruginosa across different time and place. The ST1058 ( n = 14), ST882 ( n = 11), and ST2442 ( n = 10) were the predominant types, among which ST1058 was the only one found both in Shandong province and Dalian (Liaoning, China). The MLST for P. aeruginosa infection in mink was highly associated with that in humans and other animals, implying possible transmission events. A small proportion of mink exhibited drug resistance to P. aeruginosa (9/69, 13%) with resistance predominantly to fluoroquinolone, aminoglycoside, and β-lactamase. Eight strains had mutations in the quinolone-resistance determining regions (QRDR). High proportions (65%; 72/110) of the fosA gene and 2 types of glpt deletion for fosmycin were detected. Furthermore, in the whole genome sequence of one multidrug resistant strain, we identified 27 genes that conferred resistance to 14 types of drugs.

The aim of the present study was to identify the animal prevalence and environmental reservoir of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in mink farms. LA-MRSA on mink constitutes a human health hazard to farmers and farm workers, who handle the animals and are at risk of bites and scratches from colonized sites. The primary route of LA-MRSA colonization of mink is suspected to be by ingestion of contaminated pig by-products. We performed a cross-sectional study with repeated measurements during May-July 2017. A total of 644 mink carcasses (542 mink kits and 102 breeding animals) from five Danish farms were sampled. From each carcass, pharynx was swabbed and the right forepaw dissected. In addition, environmental samples covering feed, air, glove, cages (top and between) and nest boxes were collected on the farms. MRSA was selectively cultured from each sample and suspect colonies were assessed using matrix-assisted laser desorption ionisation (MALDI-TOF) for species confirmation. Further, from each farm, three isolates from mink and one isolate per positive environmental site were sent for whole genome sequencing. We isolated LA-MRSA from mink in four out of the five farms, but LA-MRSA bacterium was detected on all farms. On farms with LA-MRSA positive animals, the overall apparent animal prevalence ranged from 20% [13;29] CI95% to 29% [22;38] CI95% . LA-MRSA was isolated from kits before weaning, most likely due to a contaminated environment or transfer from the dam. Further, the apparent prevalence of LA-MRSA in kits tended to increase during the first months of age. On farms where LA-MRSA was isolated from mink, LA-MRSA was also isolated from the environment. LA-MRSA was isolated from all environmental sites tested (i.e. glove, on top of and between the cages and in the nest boxes), apart from air. The negative air samples contrast with the high concentrations of LA-MRSA in air found in the pig production. Hence, the risk of human exposure to LA-MRSA on mink-farms tends to be associated mainly with direct contact with contaminated environmental sites and the handling of colonized mink. All sequenced isolates were confirmed as LA-MRSA CC398 and genetically similar to clones previously isolated from the Danish pig production, supporting the hypothesis of LA-MRSA being transmitted by contaminated pig by-products., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is widely distributed in the Danish pig production. Spillover to the mink production is hypothesized to occur via contaminated pig by-products used in the production of mink feed. The aim of the present longitudinal experimental cohort study was to confirm the potential of LA-MRSA being transmitted to naïve mink after exposure to contaminated feed, and to study the persistence of the bacterium on the animals after ceased exposure to contaminated feed. LA-MRSA-negative mink (n = 28) were housed in pairs in 14 mesh cages. Twenty-four mink (12 cages) received around 5.1 × 10 8 cfu/mink in the feed for five days, while four mink (two cages) were kept as negative controls and fed with LA-MRSA negative feed. Twenty-four hours after initiation of spike, all 28 mink were tested LA-MRSA-positive by paw swabs. After cease of the spiking period, one mink per cage were moved to a clean housing facility to study the potential effect of environmental contamination in persistence of the LA-MRSA. All mink were re-tested three times per week for the subsequent 26 days to study whether the mink cleared off the bacterium. The results showed that LA-MRSA can be transmitted to paws and pharynx on mink after exposure to contaminated feed and that LA-MRSA may spread indirectly through contaminated environmental sites. Mink tend to clear off LA-MRSA, however, the bacterium may persist on mink for more than 26 days., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Chlamydia (C.) abortus , a globally distributed obligate intracellular bacterium, has attracted increasing interest according to its veterinary importance and zoonotic nature. C. abortus can infect a variety of animals and cause foetal loss in livestock resulting in economic loss. In this study, the samples collected from two farms of foxes (n=20), raccoon dogs (n=15) and minks (n=20), were investigated by Chlamydiaceae - and Chlamydia species-specific real-time PCR. The results showed that all the tested foxes (20/20) and raccoon dogs (15/15) harbored Chlamydia spp., while 5% of minks (1/20) were positive for Chlamydia spp. C. abortus was identified in all positive samples as the dominant Chlamydia species, with C. pecorum DNA coexistence in some of the rectal samples (7/20) taken from foxes. Phylogenetic analysis based on specific gene fragments of 16S rRNA, IGS-23S rRNA, and ompA revealed that all sequences obtained in this study were assigned to the Chlamydiaceae family with high similarity to C. abortus S26/3 and B577 previously identified in ruminants. This is the first report confirming that farmed foxes, raccoon dogs, and minks carry C. abortus . Further studies are needed to fully elucidate the epidemiology and pathogenicity of this pathogen in farmed fur animals as well as the potential risks to public health.

The gastrointestinal microbiome is known to play a critical role in animal health but has been relatively poorly characterized in commercial mink, an obligate carnivore. Whether the microbiota can be manipulated in mink to improve pelt quality, health, and well-being is unknown. The objectives of this study were to characterize the fecal microbiota of commercial mink, and to evaluate potential changes due to year (2014 vs 2015), life stage (adult female vs weaned kit), season (summer vs winter), and between Canadian farms. Pooled fecal samples were collected from adult females and weaned kits in the summers of 2014 (n = 173) and 2015 (n = 168), and from females in the winter of 2016 (n = 39), a time when females undergo marked calorie restriction, from 49 mink farms in Ontario. Bacterial DNA was extracted and the V4 region of the 16S rRNA gene was amplified. Approximately 22 million sequences were identified following quality control filtering. A total of 31 bacterial phyla were identified; however, only 3 comprised >1% of the total sequences identified, with Firmicutes and Proteobacteria together comprising 95% of the total sequences. Comparisons were made by life stage, season and year; no differences were found in the relative abundance of any taxa between samples collected from adult females and weaned kits from the same year and the greatest number of differences at each taxonomic level were noted between 2014 and 2015. Significantly more operational taxonomic units (OTUs) were found in 2014 than 2015 or 2016 (p<0.05) and samples from 2014 were more even, but less diverse than in 2015 (p = 0.002 and 0.001, respectively). There were significant differences in community population and structure by year and season (all p-values <0.001). The predominant phyla and genera at the farm level were similar from year to year. Together, these indicate that mink environment, season, and time are important factors in the stability of gastrointestinal microbiota, once mink reach maturity., Competing Interests: The authors have declared that no competing interests exist.

Pseudomonas aeruginosa is an important animal pathogen and contributes to hemorrhagic pneumonia in mink. Between April 2011 and December 2016, samples of lung, liver, and spleen were collected from mink with this disease on 11 mink farms in 5 Chinese provinces. From these samples, we obtained 98 isolates of P. aeruginosa that belonged to 5 serotypes: G ( n = 58), I ( n = 15), C ( n = 8), M ( n = 5), and B ( n = 2); 10 isolates were not typeable (10/98). More than 90% of the isolates formed biofilms, and 85% produced slime. All 98 isolates were resistant to 10 antibiotics (oxacillin, ampicillin, penicillin G, amoxicillin, ceftriaxone, cefazolin, cefaclor, tilmicosin, tildipirosin, and sulfonamide). However, almost all were susceptible to gentamicin, polymyxin B, and amikacin. We identified 56 unique genotypes by pulsed-field gel electrophoresis. These findings have revealed genetic diversity and high antimicrobial resistance in P. aeruginosa isolated from mink with hemorrhagic pneumonia and will facilitate the prevention and control of this disease.

MRSA is an increasing problem in humans as well as livestock. The bacterial co-colonization of the skin in MRSA carriers has been poorly investigated and moreover, there have been no methods for high resolution investigations of the Staphylococcus genus apart from tediously culturing or doing multiple PCRs. On 120 samples from pig ear, skin and nose, we generated amplicons from the V1-V2 region of the 16S rRNA gene to gather an overview of the genus-level microbiome, along with using MRSA specific plates to count MRSA. In parallel with this, amplicons of the tuf gene were generated, targeting only a region of the tuf gene found only in the Staphylococcus genus. Using these methods, we determined a core microbiota across the healthy pig and determined the Staphylococcus genus to be dominated by S. equorum. Moreover, we found Streptococcus to be inversely associated with Staphylococcus and MRSA, suggesting a role for this genus in combating MRSA. In this work, we have thoroughly investigated the skin and nose microbiome of the pig and developed a high throughput method for profiling the Staphylococcus genus which we believe will be useful for further investigations.

Wildlife species are involved in the transmission of diverse pathogens. This study aimed to monitor raccoons (Procyon lotor), American minks (Neovison vison), and red foxes (Vulpes vulpes) as potential reservoirs in central Spain. Specifically, 200 spleen and fecal samples (from 194 raccoons, 3 minks, and 3 foxes) were analyzed molecularly by PCR/qPCR and sequencing for the presence of piroplasmids, Hepatozoon spp., Toxoplasma gondii, and Ehrlichia canis infections in the Community of Madrid (Spain). Biological samples were obtained in the years 2014, 2015, and 2016. No pathogen DNA was found in fecal samples. In contrast, analysis of raccoon spleen samples revealed that Toxoplasma was the most prevalent pathogen (prevalence 3.6 ± 2.6%), followed by Hepatozoon canis and E. canis (each with a prevalence of 2.57 ± 2.2%). Hepatozoon canis was also diagnosed in all three of the analyzed foxes. Analysis of yearly prevalence showed that tick-borne pathogens were less frequent in raccoon in 2015, a dry and warm year compared both to 2014 and 2016. These data suggest that fecal PCR assays are unsuitable for detection of DNA of non-erythrocytic pathogens. Furthermore, they demonstrate that the raccoon (an invasive species often living in proximity to domestic areas) and the red fox are putative reservoirs for pathogenic organisms in the Community of Madrid.

In the present study 12 Arcanobacterium phocae strains isolated from fur animals in Finland, including foxes, minks and Finnraccoons, could successfully be identified phenotypically, by matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) and genotypically by sequencing 16S rDNA and phocaelysin (PHL) encoding gene phl. The PHL of all 12 A. phocae strains in the present study and reference strains A. phocae DSM 10002 T and A. phocae DSM 10003 displayed, as typical members of the cholesterol dependent cytolysin-group of toxins, the variant undecapeptide sequence EATGLAWDPWW which appeared to be most closely related to arcanolysin of Arcanobacterium haemolyticum and pyolysin of Trueperella pyogenes. In addition, gene phl could be determined with a newly designed loop-mediated isothermal amplification (LAMP) assay. The detection of mass spectra by MALDI-TOF MS and the LAMP assay based on gene phl might help to reliably identify A. phocae in future and also elucidate the role this species plays in infections of fur animals., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Microsporidiosis is an emerging and opportunistic disease, and Enterocytozoon bieneusi is the main cause of this disease in humans. Little information is available on prevalence and genotyping of E. bieneusi in minks. We collected 559 feces samples of minks from Heilongjiang and Jilin provinces in 2017, and studied E. bieneusi prevalence by nested PCR. A total of 23 out of 559 minks (4.1%) were detected as E. bieneusi-positive, and were raised in five of the seven investigated farms. Age was the only risk factor associated with E. bieneusi prevalence in investigated minks through logistic regression analysis. Sequence analysis of the ITS gene revealed that five E. bieneusi ITS genotypes, including Peru11, EbpC, and three novel genotypes (HLJM-1, HLJM-2 and JLM-1) were present, suggesting minks may be a potential source of human microsporidiosis., (© W. Cong et al., published by EDP Sciences, 2018.)

Background: Basic pathologic characteristics for farmed minks were previously reported worldwide. However, its status in the wild has not been studied in detail., Objective: Serology and electrophoresis were carried out for evidence of exposure to 12 mink pathogens on two different locations., Animals and Methods: Serology was done in 87 wild minks by reference techniques against Toxoplasma gondii, Encephalitozoon cuniculi, Neospora caninum, Brucella abortus, Mycobacterium bovis, Leptospira interrogans, canine distemper virus (CDV), canine adenovirus (CAV), canine parvovirus (CPV), rabies virus (RV), Influenza A virus (FLUAV) and Aleutian disease virus (ADV). Hypergammaglobulinemia, the ADV main clinical feature, was determined by conventional electrophoresis., Results: Seventy-one percent of the 87 sera had antibodies against one or more pathogens. ADV accounted for the highest seroprevalence (29%), followed by T. gondii (26%), L. interrogans (14%), M. bovis (12%), B. abortus (9%), N. caninum (3%), CPV (3%) and CDV (2%). Seroprevalence was influenced by location but not sex or age. Additionally, 16% of the seropositive samples for ADV had gammaglobulin levels >40.0 g/L. Antibody titers for CDV and CPV were low and difficult to interpret as almost all these cases had borderline concentrations., Conclusion: A cautious interpretation of the results is urged as the epidemiological role of the wild mink is largely unexplored for most of these agents. Nevertheless, the information may be clinically relevant..

Background: Infectious skin disorders are not uncommon in mink. Such disorders are important as they have a negative impact on animal health and welfare as well as on the quality and value of the fur. This study presents the isolation of Arcanobacterium phocae from mink with severe skin lesions and other pathological conditions, and from wild seals and otters., Results: In 2015, A. phocae was isolated for the first time in Denmark from outbreaks of dermatitis in mink farms. The outbreaks affected at least 12 farms. Originating from these 12 farms, 23 animals cultured positive for A. phocae. The main clinical findings were necrotizing pododermatitis or dermatitis located to other body sites, such as the lumbar and cervical regions. A. phocae could be isolated from skin lesions and in nine animals also from liver, spleen and lung, indicating a systemic spread. The bacterium was also, for the first time in Denmark, detected in dead seals (n = 9) (lungs, throat or wounds) and otters (n = 2) (throat and foot)., Conclusions: An infectious skin disorder in mink associated with A. phocae has started to occur in Danish farmed mink. The origin of the infection has not been identified and it is still not clear what the pathogenesis or the port of entry for A. phocae infections are.

It is well documented that the microbial interactions and biodiversity play an important role in health and disease in mammalian species. There is a rare study about gut microbiota of Mustelidae family. In this study, 40 male and female minks from Northeast China were divided into three groups and fed until they reached maturity. The V3 region of 16S rRNA genes was amplified and sequenced using NGS. There were 526 OTUs principally concentrated among five bacterial phyla. Two points about mink's body weight gaining were observed: (1) the weight of male individuals increased more rapidly than female individuals; (2) the weight of individuals whose feed was supplemented with Chinese herb additives increased more rapidly than individuals without giving food additives. The differences of microorganism abundance were shown in two points: (1) two genera which had ≥2-fold change difference were found between male and female minks. (2) Ten genera which had a ≥2-fold change difference were found among minks with and without Chinese herb additive. Findings from this study provide new and fundamental knowledge on the gut microbiota composition of minks farmed in Northeast China, which can contribute to the general well-being of minks worldwide.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) clonal complex (CC) 398 is widespread in the Danish pig production with around 90% of herds being positive. Since 2009, cases of CC398 LA-MRSA infections in Danish mink farmers have been observed. The objective of the study was to examine the presence of LA-MRSA in farmed mink. The investigation comprised three different sample types 1) clinical samples from carcasses submitted to the laboratory for diagnostic examination, 2) paws and pharyngeal swabs from healthy animals collected at pelting, and 3) feed samples from mink feed producers. In clinical samples, LA-MRSA was found in 34% of submissions and was most prevalent in samples from paws (33%) and pharynx (17%), followed by nasal and intestinal samples (each 13%), whereas it was never detected in perineal samples. LA-MRSA was found in healthy animals on 40% of the investigated farms, including paw samples (29%) and pharyngeal samples (16%). Twenty out of the 108 feed samples from feed producers were positive for LA-MRSA. The dominant spa-types were t034 and t011 associated to CC398, corresponding to the dominant spa-types detected in the Danish pig production, from which slaughter offal is used for mink feed. The spa-types, the high prevalence of LA-MRSA on paws and in pharynx, and its detection in feed samples, suggest feed as a possible source of LA-MRSA in mink., (Copyright © 2017 Elsevier B.V. All rights reserved.)

It is well documented that antibiotics have pronounced modulatory effects on the intestinal bacterial community of both humans and animals, with potential health consequences. The gut microbiota of mink has however attracted little attention due to low bacterial load and fast gastrointestinal transit time, questioning its relevance. In this study, we hypothesise that oral amoxicillin treatment affects the gut microbiota in mink. This was investigated in a controlled trial including 24 animals of which 12 were treated with amoxicillin for 7 days. By applying 16S rRNA gene sequencing, we found that the faecal microbiota was markedly altered already after 2 days of treatment, with a surprising increase in diversity to resemble the feed. The diversity within the mucosa at termination was however reduced, which indicates this compartment as an important colonisation site in mink. No impact on blood biochemistry, lipid metabolism, serum amyloid A, vitamins A and E and histomorphology of the gut and liver was found; however, a slight decrease in fat digestibility was observed. We suggest that early-life use of amoxicillin in mink production may be counteractive as dysbiosis of the microbiota during infancy is increasingly being recognised as a risk factor for future health., (© FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Disease in mink clinically characterized by abortion and increased mortality among pregnant female mink on 28 Danish farms was observed during April and May 2015. Most of these farms suffered extensive disease problems, including a significant increase in the number of mated females without litters. Pathological, microbiological and molecular biological methods were applied to investigate the cause of disease. Necropsies of animals found dead revealed fragile and partially dissolved (liquefying) uterine tissue, with the presence of Gram positive rod-shaped bacteria. These slow growing bacteria were isolated by anaerobic culturing and identified as Clostridium limosum by both MALDI-TOF mass spectrometry analysis and 16S rRNA gene sequencing. All the performed tests for relevant differential diagnoses were negative. Foodborne disease was indicated because all the affected farms were served by the same feed factory. A specific PCR-based analysis was developed for positive identification of C. limosum and used to screen archived feed samples from the implicated feed factory. Both C. limosum 16S rRNA genes and C. limosum collagenase genes were identified in both mixed feed and more specifically in raw chicken carcass used as one of the components in the mixed feed, which was therefore identified as the most likely source of contamination. Based on the results of this investigation it is concluded that C. limosum can be associated with abortion and increased mortality in pregnant mink females and it is consequently recommended that raw materials contaminated with C. limosum should be avoided in mink feed, in particular during the whelping season., (Copyright © 2017. Published by Elsevier B.V.)

Toll-like receptors (TLRs) can recognize conserved molecular patterns and initiate a wide range of innate and adaptive immune responses against invading infectious agents. The aim of this study was to assess the transcript profile of mink TLRs (mTLRs) in mink peripheral blood mononuclear cells (PBMCs) and a range of tissues, and to explore the potential role of mTLRs in the antiviral immune response process. The results indicated that the mTLR partial nucleotide sequences had a high degree of nucleotide identity with ferret sequences (95-98%). Phylogenetic analysis showed that mammalian TLRs grouped into five TLR families, with a closer relationship of the mTLRs with those of ferret than the other mammalian sequences. Moreover, all the mTLRs were ubiquitously expressed in lymphoid organs (spleen and lymph nodes) and PBMCs. Interestingly, the mTLR expression patterns in lung, uterus, and heart showed quite a lot of similarity. Another remarkable observation was the wide expression of mTLR1-3 mRNAs in all tissues. Among the analyzed tissues, skeletal muscle was revealed to being the lowest repertoire of mTLR expression. Additionally, mink PBMCs exposed to the canine distemper virus revealed significant upregulation of mTLR2, mTLR4, mTLR7, and mTLR8 mRNAs, indicating that mTLRs have a role in innate immunity in the mink. Collectively, our results are the first to establish the basic expression patterns of mTLRs and the relationship between mTLRs and a virus, which will contribute to better understanding of the evolution and the functions of mTLRs in the innate immune system in minks.

Fur Animal Epidemic Necrotic Pyoderma (FENP) is a severe, often lethal infectious disease affecting all three fur animal species: mink (Neovision vision), foxes (Vulpes lagopus) and finnraccoons (Nyctereutes procyonoides). Previous studies showed an association between Arcanobacterium phocae and FENP. An experimental infection was conducted to confirm the ability of A. phocae to infect mink either alone or concurrently with a novel Streptococcus sp. found together with A. phocae in many cases of FENP. Different inoculation methods were tested to study possible routes of transmission. Typical signs, and gross- and histopathological findings for FENP were detected when naïve mink were infected with the tissue extract of mink with FENP, using a subcutaneous/ intradermal infection route. Edema, hemorrhage, necrosis and pus formation were detected in the infection site. A pure culture preparation of A. phocae alone or concurrently with the novel Streptococcus sp. caused severe acute signs of lethargy, apathy and anorexia and even mortality. The histopathological findings were similar to those found in naturally occurring cases of FENP. In contrast, the perorally infected mink presented no clinical signs nor any gross- or histopathological lesions. This study showed that A. phocae is able to cause FENP. The study also indicated that predisposing factors such as the environment, the general condition of the animals, temperature and skin trauma contribute to the development of the disease., Competing Interests: We have the following interests: This study was funded in part by the Finnish Fur Breeders association. HN was affiliated to the Finnish Fur Breeders association and to FinFur Lab during the course of the study. HN is currently affiliated to University of Helsinki, Finland. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

We investigated the evolution and epidemiology of a novel livestock-associated methicillin-resistant Staphylococcus aureus strain, which colonizes and infects urban-dwelling Danes even without a Danish animal reservoir. Genetic evidence suggests both poultry and human adaptation, with poultry meat implicated as a probable source., (© The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America.)

Pododermatitis is a disease of concern for mink breeders in Canada and worldwide, as it causes discomfort and lowers the breeding rates on farms affected by the disease. Unfortunately, the etiology and pathogenesis of pododermatitis are still unknown. In this study, we compared Staphylococcus spp. and Streptococcus canis isolates from healthy mink with isolates from animals with pododermatitis on 2 farms in Ontario. Almost all hemolytic Staphylococcus spp. isolated were shown to be Staphylococcus delphini Group A by 16S ribosomal ribonucleic acid (rRNA) sequence analysis and polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) did not reveal any S. delphini or S. canis clonal lineages specifically associated with pododermatitis, which suggests that these bacteria do not act as primary pathogens, but does not dismiss their potential roles as opportunistic pathogens. While S. delphini and S. canis were the most prevalent bacterial pathogens in mink pododermatitis, they were also present in samples from healthy mink. Arcanobacterium phocae is occasionally isolated from pododermatitis cases, but is difficult to recover with conventional culture methods due to its slow growth. A quantitative real-time PCR was developed for the detection of A. phocae and was tested on 138 samples of footpad tissues from 14 farms. The bacterium was detected only in pododermatitis-endemic farms in Canada and was at higher concentrations in tissues from infected footpads than in healthy tissues. This finding suggests that A. phocae is involved in the pathogenesis of pododermatitis.

The objective of this study was to investigate the potential of using phages as a therapy against hemorrhagic pneumonia in mink both in vitro and in vivo. Five Pseudomonas aeruginosa (P. aeruginosa) strains were isolated from lungs of mink with suspected hemorrhagic pneumonia and their identity was confirmed by morphological observation and 16S rDNA sequence analysis. Compared to P. aeruginosa strains isolated from mink with hemorrhagic pneumonia in 2002, these isolates were more resistant to antibiotics selected. A lytic phage vB&#95;PaeP&#95;PPA-ABTNL (PPA-ABTNL) of the Podoviridae family was isolated from hospital sewage using a P. aeruginosa isolate as host, showing broad host range against P. aeruginosa. A one-step growth curve analysis of PPA-ABTNL revealed eclipse and latent periods of 20 and 35 min, respectively, with a burst size of about 110 PFU per infected cell. Phage PPA-ABTNL significantly reduced the growth of P. aeruginosa isolates in vitro. The genome of PPA-ABTNL was 43,227 bp (62.4% G+C) containing 54 open reading frames and lacked regions encoding known virulence factors, integration-related proteins and antibiotic resistance determinants. Genome architecture analysis showed that PPA-ABTNL belonged to the "phiKMV-like Viruses" group. A repeated dose inhalational toxicity study using PPA-ABTNL crude preparation was conducted in mice and no significantly abnormal histological changes, morbidity or mortality were observed. There was no indication of any potential risk associated with using PPA-ABTNL as a therapeutic agent. The results of a curative treatment experiment demonstrated that atomization by ultrasonic treatment could efficiently deliver phage to the lungs of mink and a dose of 10 multiplicity of infection was optimal for treating mink hemorrhagic pneumonia. Our work demonstrated the potential for phage to fight P. aeruginosa involved in mink lung infections when administered by means of ultrasonic nebulization.

The biological characteristics and molecular epidemiology of Pseudomonas aeruginosa associated with mink hemorrhagic pneumonia from Shandong province of eastern China were determined in this study. From 2010 to 2011, 30 mink P. aeruginosa isolates were identified from lung, fecal and feed samples of clinical cases and subjected to serotyping, antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE) using SpeI. The P. aeruginosa isolates belonged to four serotypes-21 of type G, four of type I, three of type M, one of type B, and one non-typable strain. The strains were divided into four large groups as determined by PFGE. Isolates from the group 2 were highly homologous and were obtained from the same region as an epidemic. All of the isolates were sensitive to piperacillin, piperacillin/tazobactam, ceftazidime, cefepime, imipenem, amikacin, gentamicin and tobramycin and resistant to ampicillin, cefuroxime and cefuroxime axetil. A high frequency of resistance was found to ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, nitrofurantoin, and trimethoprim/sulfamethoxazole (96.7%). Resistance to ticarcillin/clavulanic acid, ciprofloxacin and levofloxacin was less common (13.3%). There was no relationship between antibiotic resistance and serotype distribution of the isolates. The epidemic serotype of P. aeruginosa from the mink hemorrhagic pneumonia in Shandong province was type G, which was a clone of commonly found in this province. These findings reveal the genetic similarities and antimicrobial susceptibility profiles of P. aeruginosa from clinical cases of mink hemorrhagic pneumonia and will facilitate the prevention and control of the disease in Shandong province of China., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This study evaluated the cholesterol-lowering property of lactic acid bacteria (LAB) isolated from mink. Two strains, Enterococcus faecium MDF1104 and Lactobacillus plantarum MDL1118, were shown to remove cholesterol from broths of natural hen egg yolk and skimmed milk. The cholesterol in hen egg yolk was reduced by 58.15% and 38% by L. plantarum and E. faecium, respectively. When the bacteria were used in combination, 48.95% (p < 0.01) of cholesterol was removed from skimmed milk. Experimental mice remained healthy when fed different doses of the LAB, and the total serum cholesterol concentration was the lowest (0.90 mmol/L) (p < 0.01) when a combination of L. plantarum and E. faecium was used. Based on our results, we suggest that L. plantarum MDL1118, E. faecium MDF1104, or a combination of the 2 strains could be considered as promising cholesterol-lowering probiotics.

Hemorrhagic pneumonia can be a major cause of mortality in farmed mink in the fall. In its classic form, hemorrhagic pneumonia is caused by the bacterium Pseudomonas aeruginosa. In recent years, however, outbreaks of this type of pneumonia that are associated with hemolytic Escherichia coli have also occurred in farmed mink. The purpose of this study was to compare histological lesions of acute hemorrhagic pneumonia associated with both P. aeruginosa and E. coli in mink, including a description of tissue distribution of pathogens, in an attempt to differentiate between the 2 disease entities based on histopathology. The study included material submitted for diagnostic investigation to the National Veterinary Institute in Denmark from 2006 to 2009. Altogether, 19 cases of hemorrhagic pneumonia with a pure lung culture of P. aeruginosa and 18 cases of hemorrhagic pneumonia with a pure lung culture of E. coli were examined. Formalin-fixed paraffin-embedded lung tissue obtained from the mink was examined by histology and fluorescence in-situ hybridization (FISH). It was possible to detect a slight histological difference between hemorrhagic pneumonia caused by P. aeruginosa and by E. coli, as P. aeruginosa was most often found surrounding blood vessels and lining the alveoli, while E. coli showed a more diffuse distribution in the lung tissue. Furthermore, P. aeruginosa often elicited a very hemorrhagic response in the lung, while infection with E. coli was associated with a higher frequency of alveolar edema and mild lymphoid cuffing in the lungs.

Hemorrhagic pneumonia is an acute and fatal disease of farmed mink caused by Pseudomonas aeruginosa. The pathogenesis of this disease has not yet been resolved. Mink are the only animals known to be susceptible to acute, contagious, and fatal lung infections caused by P. aeruginosa. The purpose of this study was to investigate the correlation between dose-response and season of infection and to clarify whether Danish mink are carriers of P. aeruginosa on their nasal mucosa during the season for hemorrhagic pneumonia. To elucidate the pathogenesis of the disease, an infectious dose-response trial was carried out on adult mink and mink kits, both in the season for hemorrhagic pneumonia (November) as well as out of season (July). It proved difficult to infect mink via the intra-nasal route. Only 4 out of 60 infected mink developed clinical disease and were euthanized, all of them in November, illustrating that predisposing factors in the mink itself and not infectious dose might be crucial for disease development. We were able to culture P. aeruginosa from the nasal cavity of the clinically healthy experimental mink 8 d after inoculation. This indicated that the mink can carry P. aeruginosa on their nasal mucosa without developing the disease. It was not possible, however, to culture P. aeruginosa from the nasal cavity of clinically healthy mink obtained from farms in November, which indicates that the organism is not a normal part of the nasal mucosal flora of mink.

Hemorrhagic pneumonia in mink (Neovison vison) is caused by Pseudomonas aeruginosa and is an acute and fatal disease in farmed mink. Earlier work has demonstrated that some outbreaks of hemorrhagic pneumonia are caused by pathogenic strains while most outbreaks are caused by local strains. The objective of this study was to determine the genetic and geographical relationship among outbreaks of hemorrhagic pneumonia by pulsed-field gel electrophoresis typing of P. aeruginosa isolates. Furthermore, chosen isolates were typed by a commercial genotyping method based on single nucleotide polymorphisms (SNPs) and compared to a larger dataset of human and environmental origin. The bacterial isolates were obtained from diagnostic samples from 2002 to 2009 and contained 164 isolates from 95 outbreaks on 90 farms. Our results show that most outbreaks of hemorrhagic pneumonia in mink are caused by distinct strains of P. aeruginosa. We also identified related P. aeruginosa strains which, together with two prevalent but unrelated clones, caused one third of the outbreaks of hemorrhagic pneumonia supporting the sparse literature on this subject. None of the SNP typed strains were identified in a large dataset of human and environmental origin., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Granulomatous myelitis due to infection with Mycobacterium avium was diagnosed in a 4-year-old male neutered European mink (Mustela lutreola). The causative agent was detected by an acid-fast stain and further characterized by polymerase chain reaction and DNA sequencing of the PCR product. A thorough histological evaluation of the remaining organs revealed no granulomatous lesions or detectable acid-fast organisms. Although minks are generally highly susceptible for mycobacteria, localised infections, especially of the central nervous system, are unusual and may represent an atypical chronic form of the disease.

To study the possible role of disease in the decline of endangered European mink (Mustela lutreola), we conducted a survey of antibody prevalence and renal carriage of pathogenic leptospira (Leptospira interrogans sensu lato) using serum and kidney samples collected from 1990 to 2007 from several free-ranging small carnivores and farmed American mink (Mustela vison) in southwestern France. An indirect microscopic agglutination test using a panel of 16 serovars belonging to 6 serogroups (Australis, Autumnalis, Icterohæmorrhagiæ, Grippotyphosa, Panama, Sejroe) revealed antibodies in all species, with significant differences in antibody prevalences: 74% in European mink (n=99), 65.4% in European polecats (Mustela putorius, n=133), 86% in American mink (n=74), 89% in stone martens (Martes foina, n=19), 74% in pine martens (Martes martes, n=19), 35% in common genets (Genetta genetta, n=79), and 31% in farmed American mink (n=51). Serogroups Australis and Icterohæmorragiæ were dominant in most free-ranging species; serogroup Grippotyphosa had high prevalences in European mink. Such high antibody prevalences have never been reported. They are probably related to the large number of known reservoirs, rats (Rattus spp.), muskrat (Ondatra zibethicus), and coypu (Myocastor coypu), in the study area. The polymerase chain reaction test specific for pathogenic leptospiral DNA detected renal carriage in 23% of 34 European mink, 22% of 18 polecats, and 15% of 33 free-ranging American mink, with no significant differences. Renal carriage shows that mustelids may shed leptospira for short periods, but their epidemiologic role is probably limited. High antibody prevalences suggest that the disease is unlikely to be highly pathogenic for these species; however, chronic forms of the disease (abortions, renal lesions) could reduce the reproductive success or life span of infected animals. Further studies on the pathogenicity of leptospirosis in these populations are needed to measure its impact on the population dynamics of these rodent predators.

An outbreak of diarrhea on a large commercial mink farm affected 5,000 of 36,000 neonatal mink kits, with 2,000 dying within a 2-week period. Affected kits were severely dehydrated, and their furcoats and paws were covered with yellow- to green-tinged mucoid feces. On necropsy, the small intestines of examined animals were markedly distended by serous to mucoid fluid. Microscopically, there was prominent colonization of the intestinal villar epithelium by gram-positive bacterial cocci in the absence of inflammation and morphologic changes in villous enterocytes. The colonizing bacteria were phenotypically identified as belonging to the Staphylococcus intermedius group of bacteria. This was confirmed by nucleic acid sequence analysis of the 16S ribosomal RNA gene. Further nucleic acid sequencing of polymerase chain reaction (PCR) amplicons from the superoxide dismutase gene and the heat shock protein 60 gene differentiated the isolate as Staphylococcus delphini. Production of staphylococcal enterotoxins A and E was demonstrated with a commercial ELISA-based immunoassay. Sequencing of PCR amplicons confirmed the presence of the enterotoxin E gene, but PCR amplification of the enterotoxin A, B, C, or D genes was not successful. Although direct causation was not confirmed in this study, the authors postulate that the observed hypersecretory diarrhea in these mink kits was the result of colonization of the small intestine by S delphini and subsequent production of enterotoxin.

The usage of antimicrobials for treatment of mink and the occurrence of antimicrobial resistance among the most important bacterial pathogens in mink was investigated. The aim of the study was to provide data, which may serve as a basis for the formulation of recommendations for prudent use of antimicrobials for mink. A total of 164 haemolytic staphylococci, 49 haemolytic streptococci, 39 Pseudomonas aeruginosa, 13 Pasteurella multocida, and 1093 Escherichia coli isolates from Danish mink were included in the study. A high frequency of resistance among S. intermedius was found for tetracyclines (54.7%), followed by penicillin (21.7%), lincosamides (20.4%), macrolides (19.1%), and spectinomycin (18.5%). Very low frequencies of resistance were recorded for other antimicrobials. The highest frequency among the E. coli isolates was recorded for ampicillin, streptomycin, sulphonamides, and tetracyclines, whereas resistance to other antimicrobials was rare. All P. aeruginosa were sensitive to gentamicin and colistin and sensitive or intermediate to enrofloxacin, whereas most isolates were resistant to all other antimicrobials. All P. multocida and haemolytic streptococci were sensitive to penicillin. There was a steady increase in the use of antimicrobials during the period 2001-2006, the majority of the prescribed amount being extended spectrum penicillins followed by aminoglycosides, sulphonamides with trimethoprim, and macrolides.

Colibacillosis is responsible for significant losses to the mink and cattle industries. Previous work in our laboratory and by others has suggested that possession of cnf1, the gene encoding cytotoxic necrotizing factor (CNF1), may contribute to the virulence of isolates of E. coli from mink and cattle. The cnf1 gene from E. coli isolated from a mink with colisepticaemia and a bovid with scours was amplified and cloned as a 3.5 kb fragment, and the fragment was sequenced. The cnf1 sequences from the mink and bovine isolates of E. coli were compared to each other and to cnf1 sequences of E. coli from urinary tract and diarrhoea-associated infections of humans. The difference was only 7 nucleotides between the cnf1 sequences of the mink and bovine isolates of E. coli, which translated into 7 differences in amino acids. The cnf1 sequence of the mink isolate of E. coli had 15 nucleotide differences from the cnf1 sequences of the human isolate of E. coli (GenBank X70670), which translated into 11 differences in amino acids between these proteins. The cnf1 sequence of the bovine isolate of E. coli had 14 nucleotide differences from the cnf1 sequence of the human isolate of E. coli (GenBank X70670), which translated into 10 differences in amino acids between these proteins. The highly conserved sequences of the amino acids of CNF1 proteins make them a promising target for detection and control of the CNF1-producing E. coli involved in disease among various host species.

Isolates of Pseudomonas aeruginosa from clinical infections in mink were subjected to serotyping and pulsed-field gel electrophoresis (PFGE) using SpeI. A total of 212 isolates of P. aeruginosa from the year 1998 to 2001 were included in this study: 168 isolates from mink obtained from 74 farm outbreaks of haemorrhagic pneumonia. Isolates from mink were separated into 34 distinct clones by PFGE subtyping. All isolates from mink infected during the same farm outbreak were identical, except in one case where two different strains were isolated from mink obtained from the same farm outbreak. P. aeruginosa of specific PFGE types were found to cause clusters of outbreaks on several farms within a few weeks of each other. However, PFGE types of strains causing clusters of farm outbreaks changed from year to year. These results suggest that some outbreaks of haemorrhagic pneumonia are caused by pathogenic strains of P. aeruginosa spread between farms and animals either mechanically, or through feed or water from a common source, rather than by random nosocomial infections with strains from the farm environment.