Complete Genome Sequences of Mycobacterium bovis Strains Affiliated with Bovine Tuberculosis Outbreaks in Canada in 2016 and 2018.

Mycobacterium bovis is the primary causative agent of bovine tuberculosis, a zoonotic infectious disease that presents a risk to public health, livestock, and wildlife. Here, we report complete genome sequences of two Mycobacterium bovis strains affiliated with bovine tuberculosis outbreaks in Canadian cattle farms in 2016 and 2018.

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario, Canada.

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog's condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal's disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.

Unfavorable polysomnographic sleep patterns predict poor sleep and poor psychological functioning 3 years later in patients with restless legs syndrome.

BACKGROUND: Amongst the variety of disorders affecting sleep, restless legs syndrome (RLS) merits particular attention. Little is known about long-term outcomes for sleep or psychological functioning following a diagnosis of RLS. The aim of the present study was thus to evaluate sleep and psychological functioning at a 3-year follow-up and based on polysomnographic measurements. METHOD: Thirty-eight patients (18 female and 20 male patients; mean age: 56.06, SD = 12.07) with RLS and sleep electroencephalographic recordings were followed-up 33 months later. Participants completed a series of self-rating questionnaires related to sleep and psychological functioning. Additionally, they completed a sleep log for 7 consecutive days. RESULTS: Age, male gender, increased light sleep (S1, S2) and sleep onset latency, along with low sleep efficiency, predicted psychological functioning and sleep 33 months later. Specifically, sleep fragmentation predicted poor psychological functioning, and both sleep fragmentation and light sleep predicted poor sleep. CONCLUSIONS: In patients with RLS, irrespective of medication or duration of treatment, poor objective sleep patterns at diagnosis predicted both poor psychological functioning and poor sleep about 3 years after diagnosis. The pattern of results suggests the need for more thorough medical and psychotherapeutic treatment and monitoring of patients with RLS.

Mobility of ß-lactam resistance under ampicillin treatment in gut microbiota suffering from pre-disturbance.

Ingestion of food- or waterborne antibiotic-resistant bacteria may lead to dissemination of antibiotic resistance genes (ARGs) in the gut microbiota. The gut microbiota often suffers from various disturbances. It is not clear whether and how disturbed microbiota may affect ARG mobility under antibiotic treatments. For proof of concept, in the presence or absence of streptomycin pre-treatment, mice were inoculated orally with a ß-lactam-susceptible Salmonella enterica serovar Heidelberg clinical isolate (recipient) and a ß-lactam resistant Escherichia coli O80:H26 isolate (donor) carrying a blaCMY-2 gene on an IncI2 plasmid. Immediately following inoculation, mice were treated with or without ampicillin in drinking water for 7 days. Faeces were sampled, donor, recipient and transconjugant were enumerated, blaCMY-2 abundance was determined by quantitative PCR, faecal microbial community composition was determined by 16S rRNA amplicon sequencing and cecal samples were observed histologically for evidence of inflammation. In faeces of mice that received streptomycin pre-treatment, the donor abundance remained high, and the abundance of S. Heidelberg transconjugant and the relative abundance of Enterobacteriaceae increased significantly during the ampicillin treatment. Co-blooming of the donor, transconjugant and commensal Enterobacteriaceae in the inflamed intestine promoted significantly (P<0.05) higher and possibly wider dissemination of the blaCMY-2 gene in the gut microbiota of mice that received the combination of streptomycin pre-treatment and ampicillin treatment (Str-Amp) compared to the other mice. Following cessation of the ampicillin treatment, faecal shedding of S. Heidelberg transconjugant persisted much longer from mice in the Str-Amp group compared to the other mice. In addition, only mice in the Str-Amp group shed a commensal E. coli O2:H6 transconjugant, which carries three copies of the blaCMY-2 gene, one on the IncI2 plasmid and two on the chromosome. The findings highlight the significance of pre-existing gut microbiota for ARG dissemination and persistence during and following antibiotic treatments of infectious diseases.

Mobility of ß-Lactam Resistance Under Bacterial Co-infection and Ampicillin Treatment in a Mouse Model.

Ingestion of food- or waterborne antibiotic-resistant bacteria may lead to the dissemination of antibiotic-resistance genes in the gut microbiota and the development of antibiotic-resistant bacterial infection, a significant threat to animal and public health. Food or water may be contaminated with multiple resistant bacteria, but animal models on gene transfer were mainly based on single-strain infections. In this study, we investigated the mobility of ß-lactam resistance following infection with single- versus multi-strain of resistant bacteria under ampicillin treatment. We characterized three bacterial strains isolated from food-animal production systems, Escherichia coli O80:H26 and Salmonella enterica serovars Bredeney and Heidelberg. Each strain carries at least one conjugative plasmid that encodes a ß-lactamase. We orally infected mice with each or all three bacterial strain(s) in the presence or absence of ampicillin treatment. We assessed plasmid transfer from the three donor bacteria to an introduced E. coli CV601gfp recipient in the mouse gut, and evaluated the impacts of the bacterial infection on gut microbiota and gut health. In the absence of ampicillin treatment, none of the donor or recipient bacteria established in the normal gut microbiota and plasmid transfer was not detected. In contrast, the ampicillin treatment disrupted the gut microbiota and enabled S. Bredeney and Heidelberg to colonize and transfer their plasmids to the E. coli CV601gfp recipient. E. coli O80:H26 on its own failed to colonize the mouse gut. However, during co-infection with the two Salmonella strains, E. coli O80:H26 colonized and transferred its plasmid to the E. coli CV601gfp recipient and a residential E. coli O2:H6 strain. The co-infection significantly increased plasmid transfer frequency, enhanced Proteobacteria expansion and resulted in inflammation in the mouse gut. Our findings suggest that single-strain infection models for evaluating in vivo gene transfer may underrepresent the consequences of multi-strain infections following the consumption of heavily contaminated food or water.

HEALTH STATUS OF REINTRODUCED WOOD BISON ( BISON BISON ATHABASCAE): ASSESSING THE CONSERVATION VALUE OF AN ISOLATED POPULATION IN NORTHWESTERN CANADA.

A central goal for reintroduced populations of threatened wood bison ( Bison bison athabascae) is to maintain them free of diseases of concern, particularly bovine tuberculosis (caused by Mycobacterium bovis) and brucellosis (caused by Brucella abortus). A wood bison population in southwestern Yukon, Canada was reintroduced into the wild in 1988, but no health assessment has been done since then. To provide an initial assessment of the health status and, hence, the conservation value of this population, we serologically tested 31 wood bison (approximately 3% of the population) for pathogens of interest and obtained histopathology results for select tissues. We found no evidence of exposure to M. bovis or Brucella spp., but antibodies were present to bovine parainfluenza virus 3, bovine coronavirus, Leptospira interrogans, and Neospora caninum, with seroprevalences of 87, 7, 61, and 7% of the tested animals, respectively. Reintroduced wood bison in southwestern Yukon may be of high value for wood bison recovery because it is a large and geographically isolated population with no bacteriologic, histopathologic, or serologic evidence of exposure to Brucella spp. or M. bovis.

Draft Genome Sequences of Two Strains of Salmonella enterica Serovar Typhimurium Displaying Different Virulence in an Experimental Chicken Model.

Salmonella enterica serovar Typhimurium strains 22495 and 22792, obtained from wild birds, were found to display different virulence attributes in an experimental chicken model. Closed genome sequences were assembled after sequencing with the Roche 454 and Illumina MiSeq platforms. An additional plasmid was present in the more virulent strain 22495.

FUNCTIONAL STATUS OF PATIENTS AFTER STROKE.

INTRODUCTION: Patients who have suffered from stroke become disabled and have specific problems due to the physical and mental disability that requires the implementation of rehabilitation and the creation of conditions for independent living, economic and social reintegration. The aim of this study was to evaluate the functional recovery of patients after ischemic stroke, during the subacute phase of medical rehabilitation. MATERIAL AND METHODS: The study was organized as a prospective study, which included 74 patients (44 men, 30 women) treated after stroke at the Department of Medical Rehabilitation, Clinical Center of Vojvodina during 2013. Motor recovery was assessed by Signe-Brunnstrom scale, and Barthel Index, Rivermead Mobility Index, and modified Rankin scale were applied to assess the function. RESULTS: The average age of patients after stroke was 66.59 ± 9.607 years. The mean hospital stay was 34.35 days. The majority of patients in this study had right-hand hemiparesis 47 (63.5%), and 27 (36.5%) had left-hand hemiparesis. By analyzing the average valueof motor recovery of the affected limb by S.Brunnstrom's scale during rehabilitation at the Department of Medical Rehabilitation, it was found that the value at the end of subacute rehabilitation phase was significantly increased (p<0.01). The results of this testing showed a statistically significant improvement (p<0.0l) in the average values of Barthel Index, Rivermead Mobility Index and modified Rankin scale during the rehabilitation treatment of stroke patients. CONCLUSION: The obtained results showed that the rehabilitation treatment resulted in better functional and motor recovery in the patients who had had ischemic stroke.

Sensitive diagnosis of bovine tuberculosis in a farmed cervid herd with use of an MPB70 protein fluorescence polarization assay.

After histopathological examination of a lesion found in a herd member returned a diagnosis of mycobacteriosis, a farmed herd (n = 47) of elk (Cervus elaphus nelsoni) and red deer (C. elaphus elaphus) was investigated for bovine tuberculosis with a battery of antemortem and postmortem diagnostic tests. Every animal was tested with the mid-cervical tuberculin skin test; all 47 had negative results. All of the 16 adult animals and 15 of the 31 calves (approximately 2-years-old) were blood-tested with a lymphocyte stimulation test (LST) and a fluorescence polarization assay (FPA), which detects antibody to the MPB70 protein antigen. At necropsy of the 31 blood-tested animals, tissues were harvested for histopathological examination and culture of mycobacteria. Mycobacterium bovis was isolated from 16 of the 31 animals, and a scotochromogen was also isolated from 1 of the 16 whose tissues yielded M. bovis. Each of these 16 animals, 15 of which were calves, also received a histopathological diagnosis of mycobacteriosis. Other species of mycobacteria, including those belonging to the M. avium and M. terrae complexes, were isolated from an additional 7 animals. The FPA was scored "positive" or "suspect" for 16 animals, 13 (81%) of which were culture-positive for M. bovis. The other 3 animals that were culture-positive for M. bovis had negative FPA results. Of the 3 FPA-positive or FPA-suspect animals that were culture-negative, 2 were suspected to have mycobacteriosis on the basis of the histopathological examination. The 7 animals from which Mycobacterium species other than M. bovis were cultured were all FPA-negative. The only animal with positive LST results was also FPA-positive and culture-positive for M. bovis. The M. bovis isolates had an identical spoligotype pattern, with an octal code of 664073777777600. This is the first report of the isolation and identification of this strain type in Canada.