The First Report of Mycobacterium celatum Isolation from Domestic Pig (Sus scrofa domestica) and Roe Deer (Capreolus capreolus) and an Overview of Human Infections in Slovenia

Mycobacterium celatum, a slowly growing potentially pathogenic mycobacterium first described in humans, is regarded as an uncommon cause of human infection, though capable of inducing invasive disease in immunocompromised hosts. According to some reports, a serious disease due to M. celatum may also occur in individuals with no apparent immunodeficiency. In animals, an M. celatum-related disease has been described in three cases only: twice in a domestic ferret (Mustela putorius furo) and once in a white-tailed trogon (Trogon viridis). In this paper, we report the first detection of M. celatum in a domestic pig (Sus scrofa domestica) and roe deer (Capreolus capreolus). A nation-wide overview of human M. celatum infections recorded in Slovenia between 2000 and 2010 is also given. Pulmonary disease due to M. celatum was recognized in one patient with a history of a preexisting lung disease

Assessment of tuberculosis biomarkers in paratuberculosis-infected cattle

Introduction: Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis, respectively the causative agents of bovine tuberculosis (bTB) and bovine paratuberculosis (PTB), share a high number of antigenic proteins. This characteristics makes the differential diagnosis of the diseases difficult. The interferon gamma (IFN-γ), C-X-C motif chemokine ligand 10 (CXCL10), matrix metallopeptidase 9 (MMP9), interleukin 22 (IL-22) and thrombospondin 1 (THBS1) bovine genes have already been shown to be accurate transcriptional biomarkers of bTB. In order to improve the diagnosis of bTB and PTB, in the present study we evaluated the risk of false positivity of these bTB biomarkers in cattle with PTB. Material and methods: The transcription of these genes was studied in 13 PTB-infected cattle, using Mycobacterium avium subsp. paratuberculosis (MAP)-stimulated peripheral blood mononuclear cells (PBMC). Results: Overall, the levels of IFN-γ, CXCL10, MMP9 and IL-22 transcripts in MAP-stimulated PBMC failed to differentiate animals with PTB from healthy animals. However, as bTB-afflicted cattle do, the MAP-infected group also displayed a lower level of THBS1 transcription than the non-infected animals. Conclusion: The results of this study add new specificity attributes to the levels of transcription of IFN-γ, CXCL10, MMP9 and IL-22 as biomarkers for bTB.Instituto de BiotecnologíaFil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Klepp, Laura Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Colombatti Olivieri, Maria Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Colombatti Olivieri, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moyano, Roberto Damian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Moyano, Roberto Damian. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romano, Maria Isabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Malovrh, Tadej. University of Ljubljana. Veterinary Faculty. Institute for Microbiology and Parasitology; EsloveniaFil: Ocepek, Matjaž. University of Ljubljana. Veterinary Faculty. Institute for Microbiology and Parasitology; EsloveniaFil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Bigi, Fabiana. Consejo Nacional de investigaciones Científicas y Tecnológicas; Argentin

Molekularna epidemiologija infekcije vrstom Mycobacterium tuberculosis u goveda i ljudi - prikaz slučaja.

We describe a case of transmission of Mycobacterium (M.) tuberculosis infection from a man to cattle. M. tuberculosis was isolated from the bronchial lymph nodes of a heifer that reacted positively to bovine tuberculin but showed no gross pathological changes at slaughter. The cattle owner died of tuberculosis the same year the heifer was diagnosed with M. tuberculosis infection. M. tuberculosis strains isolated from the heifer and its owner were genotyped by mycobacterial interspersed repetitive units - variable number tandem repeat (MIRUVNTR) typing, which revealed identical MIRU profiles for both isolates. This is the first described case of M. tuberculosis infection in cattle and the first case of human-to-animal transmission of M. tuberculosis in Croatia.Opisan je slučaj prijenosa zaraze vrstom Mycobacterium (M.) tuberculosis s čovjeka na govedo. M. tuberculosis je izdvojen iz bronhalnih limfnih čvorova junice koja je pozitivno reagirala na tuberkulin, a prilikom klanja nisu utvrđene patomorfološke promjene karakteristične za tuberkulozu. Iste godine vlasnik goveda je preminuo od posljedica tuberkuloze. Izolati M. tuberculosis iz goveda i čovjeka bili su genotipizirani pomoću metode određivanja promjenjivog broja opetovanih sljedova nukleotida (engl. mycobacterial interspersed repetitive units - variable-number tandem repeat [MIRU-VNTR]) i u oba je slučaja bio utvrđen identičan rezultat genotipizacije. Ovo je prvi opisani slučaj zaraze vrstom M. tuberculosis u goveda i prvi slučaj prijenosa ove bolesti s čovjeka na govedo u Hrvatskoj

Effect of Linseed Supplementation on Carcass, Meat Quality and Fatty Acid Composition in Pigs

The effect of linseed supplementation on carcass, meat quality and fatty acid profile of fat tissue was studied. No differences in carcass and meat quality traits were observed, the exception being drip loss that was lower in pigs supplemented with linseed. As regards fatty acids, linseed supplementation led to the increased content of unsaturated, polyunsaturated and n-3 fatty acids and decreased content of saturated fatty acids and n-6/n-3 ratio in the subcutaneous and intramuscular fat of pigs

Mycobacteria in aquarium fish

Mycobacteria potentially pathogenic for humans are commonly present in aquarium fish sold in Slovenian pet shops. Mycobacterium marinum is the most important causative agent of skin mycobacteriosis in humans. The purpose of the present study was to evaluate, by means of a questionnaire, aquarium hobbyists’ and pet shop salespersons’ awareness of the health risks related to aquarium fish handling. A total of 198 participants took part in the study, 76.3% of these were aquarium hobbyists, and 23.7% were pet shop salespersons with up to 15 years of fish handling experience. About one third (35.8%) of all participants recognized that fish may be a source of infection for humans and that fish may contract tuberculosis. Fewer (24.4%) were aware of the fact that fish tuberculosis is a zoonotic disease. The vast majority of respondents were unfamiliar with the clinical manifestations of either fish tuberculosis or cutaneous mycobacteriosis in humans. Despite the generally acknowledged belief that aquarium water may pose a risk to human health, the respondents’ aquarium handling practices were surprising as the majority never used waterproof gloves. Several differences were revealed between the two groups of respondents. Pet shop salespersons were better educated on fish handling, and more were aware of the health risks linked to aquarium fish than aquarium hobbyists. The latter often perceived pet shop salespersons as relevant sources of information. However, overall awareness of the zoonotic potential of fish mycobacteria proved to be fairly low. Therefore, more effort should be made to increase the awareness of the role of mycobacteria in infections associated with exposure to aquarium fish.Za ljudi potencialno patogene vrste mikobakterij so v Sloveniji splošno prisotne pri akvarijskih ribah, ki so naprodaj v trgovinah z domačimi ljubljenčki, med drugimi tudi vrsta Mycobacterium marinum, ki je najpomembnejša povzročiteljica kožne mikobakterioze pri ljudeh. Namen raziskave je bil, s pomočjo vprašalnika, oceniti zavedanje ljubiteljskih akvaristov in prodajalcev v trgovinah z akvarističnim programom o zdravstvenih tveganjih, povezanih z rokovanjem z ribami. V raziskavi je sodelovalo 198 oseb, 76,3 % ljubiteljskih akvaristov in 23,7 % prodajalcev v trgovinah, ki so v povprečju imeli do 15 let izkušenj pri rokovanju z akvarijskimi ribami. Približno tretjina (35,8 %) sodelujočih se je strinjala s trditvijo, da so ribe lahko vir okužbe za ljudi in da ribe lahko zbolijo za tuberkulozo. V manjšem deležu (24,4 %) so se sodelujoči strinjali s trditvijo, da je ribja tuberkuloza zoonoza. Velika večina anketirancev ni poznala kliničnih znakov ribje tuberkuloze in kožne mikobakterioze pri ljudeh. Kljub splošno izraženemu prepričanju, da lahko voda iz akvarija predstavlja tveganje za zdravje ljudi, je bila splošna praksa rokovanja z ribami presenetljiva, saj velika večina sodelujočih pri rokovanju z ribami nikoli ne uporablja vodoodpornih rokavic. Med obema skupinama anketirancev smo odkrili več razlik. Prodajalci v trgovinah so bolj poučeni o rokovanju z ribami in bolj ozaveščeni o zdravstvenih tveganjih povezanih z akvarijskimi ribami, kot ljubiteljski akvaristi. Slednji prodajalce dojemajo kot zaupanja vreden vir informacij. V splošnem je ozaveščenost o zoonotskem potencialu mikobakterij pri akvarijskih ribah precej slaba, zato bi morali nameniti več pozornosti ozaveščanju o vlogi mikobakterij pri okužbah, povezanih z akvarijskimi ribami

Implications of cattle trade for the spread and control of infectious diseases in Slovenia

The objectives of this study were to gain insight into the structure of the cattle trade network in Slovenia and to evaluate the potential for infectious disease spread through movements. The study considered cattle movements between different types of premises that occurred between August 1, 2011 and July 31, 2016 with the exclusion of the movements to the end nodes (e.g., slaughterhouses). In the first part, we performed a static network analysis on monthly and yearly snapshots of the network. These time scales reflect our interest in slowly spreading pathogensnamely Mycobacterium avium subsp. paratuberculosis (MAP), which causes paratuberculosis, a worldwide economically important disease. The results showed consistency in the network measures over timenevertheless, it was evident that year to year contacts between premises were changing. The importance of individual premises for the network connectedness was highly heterogeneous and the most influential premises in the network were collection centers, mountain pastures, and pastures. Compared to random node removal, targeted removal informed by ranking based on local network measures from previous years was substantially more effective in network disassociation. Inclusion of the latest movement data improved the results. In the second part, we simulated disease spread using a Susceptible-Infectious (SI) model on the temporal network. The SI model was based on the empirically estimated true prevalence of paratuberculosis in Slovenia and four scenarios for probabilities of transmission. Different probabilities were realized by the generation of new networks with the corresponding proportion of contacts which were randomly selected from the original network. These diluted networks served as substrates for simulation of MAP spread. The probability of transmission had a significant influence on the velocity of disease spread through the network. The peaks in daily incidence rates of infected herds were observed at the end of the grazing period. Our results suggest that network analysis may provide support in the optimization of paratuberculosis surveillance and intervention in Slovenia. The approach of simulating disease spread on a diluted network may also be used to model other transmission pathways between herds

Management of a Coxiella burnetii-infected sheep flock after an outbreak of Q fever in humans

Following an outbreak of Q fever in a group of students who contracted the infection during a training course on a sheep farm, a detailed investigation of the sheep flock involved was conducted. Of 478 flock animals, 60 Coxiella burnetii ELISA-positive and 60 ELISA-negative ewes were selected for the trial and divided into four groups. A month after the initial ELISA screening, all ewes in the flock (except the control group) were vaccinated. Sequentially collected blood samples were tested with ELISA and PCRfeces, milk, manure, bedding, and soil were tested with PCR. The immune response to the vaccination was 92.7%, while the overall C. burnetiiseroprevalence in the flock after the human outbreak was 64.9%. PCR was positive for 0.2% of milk samples and 34.4% of fecal samples of animals from all four trial groups. C. burnetii DNA was not detected in any of the blood samples. Manure was PCR-positive for about 35 monthsbedding from the stable was also positive while samples of pasture soil were negative. It appears that extensive cleaning and disinfection combined with vaccination could be regarded as an appropriate approach to control/prevent Q fever in farm settings even in the short ter

Transmission of Mycobacterium tuberculosis from Human to Cattle

We describe the first transmission of Mycobacterium tuberculosis from human to cattle confirmed by molecular typing of isolates involved in the transmission. IS6110-based restriction fragment length polymorphism analysis showed that the isolates from the cattle and farm worker who suffered from pulmonary tuberculosis 1 year prior to this case were the same strains

Phylogeography of Brucella suis biovar 2 with focus on Slovenian wildlife

Brucella suis commonly infects swine but occasionally also other animal species and humans. Wild boars are the most important reservoir of B. suis biovar 2, continually infecting susceptible hosts through close contact. Nevertheless, the genetic diversity of B. suis in wildlife remains understudied. Here, we typed 17 Slovenian B. suis biovar 2 isolates obtained in 2017–2019 from wild boars (n = 16) and a hare (n = 1) using whole-genome sequencing (WGS). To assess the global phylogenetic diversity of B. suis, we compared them to 126 publicly available B. suis genomes. All Slovenian isolates fell within the biovar 2 lineage, confirming the previous multiplex PCR typing results. According to MLST-21, the wild boar isolates were of sequence types (STs) ST16 (n = 8) and ST153 (n = 8)the maximum genetic distance between isolates of the same ST was 28 wgMLST alleles. The ST153 isolates were restricted to the Slovenian-Croatian border and clustered together with the Croatian ST153 isolates from swine, indicating cross-border transmission of B. suis ST153 strain. The hare isolate was of ST40 and was genetically distant (≥ 489 alleles) from the wild boar isolates. The genome-wide phylogeny clearly separated different B. suis biovars. The present study is the first report on the population structure of B. suis in wildlife in Slovenia and shows that the Slovenian B. suis population is genetically heterogeneous. At the species level, B. suis biovars are clearly separated in the WGS-based phylogenetic tree and can therefore be reliably predicted using WGS

Identification of risk factors influencing Clostridium difficile prevalence in middle-size dairy farms

Farm animals have been suggested to play an important role in the epidemiology of Clostridium difficile infection (CDI) in the community. The purpose of this study was to evaluate risk factors associated with C. difficile dissemination in family dairy farms, which are the most common farming model in the European Union. Environmental samples and fecal samples from cows and calves were collected repeatedly over a 1 year period on 20 mid-size family dairy farms. Clostridium difficile was detected in cattle feces on all farms using qPCR. The average prevalence between farms was 10% (0-44.4%) and 35.7% (3.7-66.7%) in cows and calves, respectively. Bacterial culture yielded 103 C. difficile isolates from cattle and 61 from the environment. Most C. difficile isolates were PCR-ribotype 033. A univariate mixed effect model analysis of risk factors associated dietary changes with increasing C. difficile prevalence in cows (P = 0.0004)and dietary changes (P = 0.004), breeding Simmental cattle (P = 0.001), mastitis (P = 0.003) and antibiotic treatment (P = 0.003) in calves. Multivariate analysis of risk factors found that dietary changes in cows (P = 0.0001) and calves (P = 0.002) increase C. difficile prevalencemastitis was identified as a risk factor in calves (P = 0.001). This study shows that C. difficile is common on dairy farms and that shedding is more influenced by farm management than environmental factors. Based on molecular typing of C. difficile isolates, it could also be concluded that family dairy farms are currently not contributing to increased CDI incidence