Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. ivniGhielmetti Giovanni Potential Zoonotic and Situation Southern Epidemiological from Improvements, scrofa) Diagnostic (Sus Switzerland: Boars Wild in Infections Mycobacterial idiei h aneac ftecua gnso T.Ti eut ncniuu neseistransmissions interspecies continuous in by results played This role bTB. fundamental of the agents causal shown the have domestic of investigations different maintenance numerous epidemiological the from the Recent in isolated of wildlife been species. have members animal MTBC by wild 2014). caused and al., disease et chronic (Rodriguez-Campos a (MTBC) plex is (bTB) tuberculosis Bovine Introduction (3.6%; method. possible reference not based present was sequence the identification the to Overall, where isolates) species 100/111 111 changes. (90.1%; Mycobacterium The macroscopic rate undescribed without concordance area. and of high study examined a hominissuis with the exception the had subsp. from nodes the MALDI-TOF on avium originating lymph 4/111), with M. based boars in that, addition, CI95% species. wild found Mycobacterium show In (2.8%; the was undescribed findings of boars potentially and methods. 64.5) three wild (22.5%) molecular – and 176 predominated by 50.0 species of thereby known microti (CI95% out 24 Mycobacterium 57.4% to five in be belonged a in detected to isolates of detected were confirmed analysis were NTM all sequence MTBC nodes, were with lymph the and was compared of desorption 6.5) Histology was laser Mycobacteria - matrix-assisted members. identification 1.2 of MTBC genes. species accuracy for housekeeping Moreover, spectrometry specific inspection, of lesions. mass PCR gross visible combination Following real-time (MALDI-TOF) gross based flight presenting collected. IS6110 nodes of were an of lymph ionization–time Switzerland, to of number of and Ticino, selection decades. nodes increasing culture a Canton lymph last of to an of Mandibular performed the subjected region diagnostics, aspects. was over pre-Alpine laboratory possible node observed the the improved lymph been from of each some one: boars has are wild than infections damage hunted assumed more NTM lung 176 are with probably in NTM individuals are trend and the augmentation patients increasing of immunocompromised such an majority large of individuals, the reasons most over Although (MTBC), The to complex described. non-pathogenic - been tuberculosis be have Mycobacterium the to remain species of pathogens (NTM) members opportunistic mycobacteria pathogenic or well-known nontuberculous obligate the as 180 to implication addition their In and hosts unclear. different mainly in infections mycobacterial of occurrence The Abstract 2020 28, April 6 5 4 3 2 1 Stephan Roger ainlRfrneCne o neoahgncBcei n Listeria and Bacteria Enteropathogenic for Center Hygiene Reference and National Safety Food for Institute Office Veterinary Cantonal available not Affiliation Zurich of University Faculty Vetsuisse Zurich of University 5 n ud Bloemberg Guido and , 1 oiaHilbe Monika , 2 t Friedel Ute , 6 1 3 haaMenegatti Chiara , yoatru tuberculosis Mycobacterium 4 uaBacciarini Luca , com- 4 , Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. rd ewe euneaayi n AD-O.Ti td ie i odtrieteocrec and consensus occurrence the the verified determine and to source (i) common used aimed a research from study present isolated This The species MALDI-TOF. of and evaluated. the this range analysis be to of wide sequence should a superior limitations between origin showing grade remains and veterinary NTM approaches accuracy from of genetic the panel samples a et ordinary with Therefore, Murugaiyan on present 2015, 2014). based at workflow al., (Tortoli, method achievable the et identification accuracy in Mediavilla-Gradolph Because species the integrated 2018, MALDI-TOF-based However, been al., diagnostic. nowadays et routine 2018). has (Alcaide in ionization–time al., technology laboratories personal desorption MS) diagnostic laser laboratory numerous (MALDI-TOF matrix-assisted for of spectrometry the characterization challenge throughput mass The a high flight mycobacteria. and often of cost-effectiveness isolated is rapidness, the samples It its of clinical of comparison 2014). from direct (Tortoli, mycobac- a NTM decades in impeding of past progress levels, describe the different the the publications, of at over and mentioned result dissection formed improvements techniques the latter the molecular diagnostic exclusively that the in enormous noteworthy, be advance that, not is to the may note led but differences to characterization Such NTM, significant terial of Pate isolated. is 2011, distribution species It al., of geographic et spectrum of Gortazar 2006). the 2015, al., in al., of differences et et spectrum Trcka significant (Garcia-Jimenez the 2016, methods evaluated molecular comprehensively al., using second Slovenia et pigs Ticino, and black of Brazil in Canton Republic, species the Czech NTM in Spain, mammalians from or to hunted studies sex most order Recent without the In September of in deer. measurements. one year red popu- each control is to animals the boar without the only consequently Wild months hunt to 100%-180%, 12 restrictions. allowed bag at in the are / hunting hunters triplicate estimated Territorio in licensed (Dipartimento even is observed growth, valley or this increase density Maggia regulate double population animal the theoretically territory The of highest it can part the are the that 2010). lation lower boars in with 1981 the Economia, wild presence since land, and The e boar Lugano only hilly Finanze 2010). Mendrisio, is wild and Economia, of it The e flat- districts and Finanze along southern territory the disappeared / region. exclusively Territorio in The it most alpine (Dipartimento almost concentrated Thereafter, the the again is Italy. distributed century. sighted is of area with officially XVI Ticino urban third shared Naranjo been the Canton the one has 2015, is during of about 2006). documented al., majority border cover al., been the et his prevalence forests where has et Michelet of the kilometre and Vicente 2012, on proportion square land 2009, al., infections 2‘812 large flat al., et of NTM a et Marco area of and Santos Di an impact Switzerland 2010, encompasses 2016, species the of al., al., this (ii) Canton et et concerning and Richomme southern Chiari literature MTBC 2008, 2014, the of al., al., 1993), presence et et al., most the (Boniotti the et (i) among MTBC (Oliver are on of world boars focused the wild although mainly in Moreover, on is mammals slow information available. extremely is large restricted wildlife only or distributed in infections, infections fastidious widely diversity mycobacterial their of NTM in and detection relevance opportunistic occurrence increasing accurate 2007, the to the more al., Despite susceptible et enabled species. have Griffith now highly growing diagnostics 2014, are are Boschiroli, laboratory pathogens individuals and improved emerging (Biet Immunocompromised and these countries with industrialized infections 2014). namely in and Tortoli, humans, tuberculosis mammals, for than other pathogens common and true more Ghielmetti humans as 2015, to recognized (Falkinham, pathogenic sources, are tunistic animals of species variety or two a ulcerans protozoa these, from invertebrates, Of 2018). M. isolated al., aerosol, been 2018). et have dust, al., (Gupta they soil, et and described et feed, environment been Martinelli the water, have 2008, in including (NTM) and encountered al., mycobacteria commonly MTBC et nontuberculous are the (Gortazar of NTM in- of E species several ongoing members 180 hepatitis of The over the and monitoring disease, Britain. bTB Beside the Great like regarding including 2015). Badger concerns plagues Europe, raised al., zoonotic 2018). have in including al., bTB populations diseases, of et boar fectious reservoir Nigsch wild a of programs 2015, as expansion al., eradication acting geographic et international animals Maria and wild 2016, known national relevant al., hindering et ee meles Meles Jhne ta. 00.Nvrhls,mr hn6 pce fNMaekont eoppor- be to known are NTM of species 60 than more Nevertheless, 2020). al., et (Johansen ,fe–agn e er( deer red free–ranging ), evseahselaphus elaphus Cervus .avium M. ope MC ntemnindpbiain a per- was publications mentioned the in (MAC) complex Mycobacterium 2 n idba ( boar wild and ) ere h gn htcue Hansen’s causes that agent the leprae, u scrofa Sus .marinum M. r h most the are ) and Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. Tlnie l,19)hueepn ee a efre ndpiae olwdb eehmlg analy- homology gene by followed duplicates in performed was 2018), of al., genes combination et housekeeping a (Scherrer of 1993) rRNA analysis al., 16S sequence et of MTBC by (Telenti sequencing negative characterized (MALDI- Sanger and spectrometry further MS). mass staining time-of-flight and TOF (ZN) desorption/ionization laser NTM Ziehl-Neelsen matrix-assisted as and by genes classified housekeeping (AFBs) were bacilli results acid-fast 200 qPCR presented 13000 into that at materials reactions. cultures centrifuged cell downstream Pure in and of used Switzerland) loop-full was Cressier, a supernatant The Matrix, inoculating minutes. (InstaGene by extracted resin 2006). double- was ion-exchange assay 7500 al., was mycobacteria a chelating qPCR et on probe cultured (Hoffmann the performed IS6110 Switzerland) from Moreover, and Zug, MTB colleagues respectively. DNA Biosystems, and the 3IABkFQ, Hoffmann (Applied Briefly, by and system described 2016). PCR ZEN as real-time al., of eGFP Fast by et instead substituted (Reed was BHQ1 control modifications and internal sequence slight iQ500 insertion with targeting with assay al. quenched qPCR et the Reinach, using Scientific, Reed ng/ detection Fisher by 100 MTBC (Thermo direct of reading Spectrophotometer for concentration by 2000c used measured maximal was NanoDrop was a eluate a to final using diluted the nm QIAamp Switzerland), in 260 the concentration overnight. with at DNA (Qiagen) accordance absorbance (Qiagen). in K the protocol instrument Proteinase Kit QIAcube Mini a with the Pathogen using using digestion cador performed lysis enzymatic was cell and purification mechanical Germany) DNA 400 through Automated 37 Hilden, Simultaneously, extracted (Qiagen, was at days. DNA II weeks ten Genomic TissueLyser eight Germany). to slants Eschwege, to agar three Biomedicals, Stonebrink up (MP of on incubated and 100 intervals and agar-plates in at 7H10 suspended inoculum on performed same subcultures were cultures, the mycobacterial (BD) with pure L obtain one inoculated to addition, were order In (BD) specimen. slants homogenized agar and decontaminated mixture antibiotic of 50 al., Azlocillin) ml and Trimethoprim, BBL Switzerland) et Allschwil, supplement, acid, growth BD, (Ghielmetti Nalidixic 960 Dickinson, B, elsewhere MGIT (Becton, Bactec Amphotericin described with B, supplemented (Polymyxin as tubes PANTA media MGIT performed liquid MGIT were BBL Briefly, cultures mycobacterial 2018). and preparation sterile Sample using composed transported by years and nodes two containers Procedures lymph sterile than Microbiological conditions. into excised older packed cooled Office animals immediately under Veterinary was adult laboratory Cantonal sample the and Each the to yearlings, classi- of scalpels. Age as and personnel knives age field. Trained dissection the of on group. years directly last two recorded animals the and of were patterns: months weight animal animals, eruption six each the 2500-3000 tooth of between and on of position study based geographical size was the exact period. population fication of two-year the estimated a and periods over age an hunting analyzed Sex, lymph two Assuming was Mandibular the population respectively. determined. the during 2018, of was collected total, and 5% were boar 2017 approximatively In animals in wild independently. Ticino 90 sampled Canton Offices and every in Veterinary 86 hunted Cantonal were of the boars node by wild regulated 1588 are and 1436 Switzerland in seasons Hunting Samples of Collection Methods and Materials identification ge- diagnostic the different identify two to compare (ii) to Ticino, genus (iii) Canton the and in for species, hunted approaches mycobacterial boar of wild healthy distribution among ographical species mycobacterial of diversity μ T ue Qae,Hle,Gray n rnfre noaLsn arxEtubes E Matrix Lysing a onto transferred and Germany) Hilden, (Qiagen, buffer ATL l Mycobacterium . 3 μ < /lsdu-yuaewr ahiouae ih0.5 with inoculated each were sodium-pyruvate g/ml otso g eercre sjvnls those juveniles, as recorded were age of months 6 μ n trda -20 at stored and l rpoB wnti-esnadoeStonebrink one and ¨ owenstein-Jensen A´ kmie l,20)and 2003) al., (Ad´ekambi et μ ° fclueiouu was inoculum culture of l ni s.PrfidDNA Purified use. until C IS6110 sdescribed as g ° o 10 for hsp65 .In C. μ of l Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. itlgcleaiain ape eefie n1%bffrdfrai n meddi aan w to Two paraffin. for Additionally, in submitted (HE). were embedded eosin PCR and and hematoxylin real-time formalin with by buffered stained MTBC and 10% obtained for in were positive sections fixed tested tissue were three-micron-thick that Samples and nodes examination. macroscopically lymph histological nodes all prevalent most lymph addition, criteria seven In Selection the the histology. lesions. for with inspected submitted infections staff additionally on was qualified focusing samples were of tissue, subset connective A changes. and pathological recorded fat of removal After Examination Histological MBT the and against matched Macroscopic profiles was spectrum MSP main Each specific software. 4.0. in 4.1 exported Explorer Library were Compass Mycobacteria isolate Biotyper one MALDI of the spectra using measured (MSP) the isolated, species each For Hz 60 system of operating benchtop frequency LT laser Microflex maximum the using a Da at 20,000 mode to F 2,000 ion of GmbH, positive (m/z) Daltonik linear ratio 1 (Bruker charge a Thereafter to in mass conditions. acquired a 2 across were level plates spectra biosafety target to mass the (HCCA, taken Peptide point, the spot then this each at and At to centrifuged temperature duplicates. added were room in was tubes at USA) matrix the dry MA, conclusion, Billerica, to 25–50 Daltonics, allowed In and Bruker were steel; pellets. min ground the 1 1 target of and for 96 volume above speed as the maximal conditions on same at depending vortexing added, by were disrupted acid were cells Mycobacterial ossetwe h aeseiswsteol n ugse ytesfwr ihaLVaoe18 Lower 1.8. above considered LSV and a with caution software with the treated by were suggested between 2.0 one (LSV) only and values the scores 1.8 spectrum was Log between ( main control. species LSV acceptable, quality same 880 as for the containing and considered when calibrator were consistent 4.0, a 3.0 Library as and run Mycobacteria each 2.0 MBT in highest An used The the species. was GmbH). mycobacterial manufacturer with Daltonik 159 (Bruker compared representing software (MSP), was 3.3 profiles value FlexAnalysis the score by log analysed and method FlexControl Bartlesville, BioSpec, (Zirconia/Silica; suspension bead pellets of the full enabling open spatula-tip left A 10-50 were tubes 13000 and temperature. The at USA) room discharged. min at was 2 dry supernatant the medium of to 300 and solid step in repeated from centrifugation was re-suspended culture a trifugation was of After pellet loopful conditions. the A for 300 discharged 3 medium 2010). with culture level tube al., as Eppendorf biosafety et chosen ml Lotz solid under were 1.5 on 2016, agar-plates a grown al., into 7H10 isolates transferred optical et My- media, from was liquid enable (Kodana obtained the from spectra analysis to using the obtained MS order of those performed MALDI-TOF In quality than was the better manufacturer. because analysis is the and media MS colonies with tested accordance MALDI-TOF the in of for evaluation (MycoEX) isolates Method the Extraction of cobacteria preparation and Inactivation (Ghielmetti described Spectrometry previously Mass as MALDI-TOF nodes spoli- lymph using of described determination extracts 2017). species the DNA al., and of using et (MIRU-VNTR) any interspersed performed repeats mycobacterial from therefore by tandem cultivated were characterization number gotyping Molecular be variable incubation. not and of unit could months repetitive 12 mycobacteria after PCR, even real-time media, culture MTBC direct by performed positive was alignment were sequence multiple included 25291, for Genomics strains searching CLC Control similarity using assembled BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi). were and sequences complete (Qiagen) Resulting 7.5.1 the Switzerland). Workbench MAC, (Balgach, Microsynth the at of performed members as Turenne identified by isolates For ses. < .avium M. .)wr nepee sicretadrcre s“oietfiainpsil”(aebe l,2011). al., et (Saleeb possible” identification “no as recorded and incorrect as interpreted were 1.8) tal. et μ sn rmr Ahp5 n Ahp5 Trnee l,20) N eunigwas sequencing DNA 2006). al., et (Turenne MAChsp65R and MAChsp65F primers using subsp L μ faeoirl eeaddt h elt,dpnigo h oueo h pellets. the of volume the on depending pellets, the to added were acetonitrile of l hominissuis . ladn wteln) ahso a esrdtieuigteMBT the using twice measured was spot Each Switzerland). ¨ allanden, μ ftespraatwssotdo h AD-O agtpae (MSP plates target MALDI-TOF the on spotted was supernatant the of l TC700898, ATCC α cao4hdoyinmcacid). -cyano-4-hydroxycinnamic μ fHL-ae n 900 and HPLC-water of L 4 .peregrinum M. μ fHL-ae n nciae o 0mna 99 at min 30 for inactivated and HPLC-water of L Mycobacterium shrci coli Escherichia hsp65 TC708.Frsmlsresulted samples For 700686. ATCC pce n rsnigmacroscopic presenting and species eewssqecda proposed as sequenced was gene .avium M. eeec tanpoie ythe by provided strain reference μ fehnl hratr cen- Thereafter, ethanol. of L g h uentn was supernatant the , subsp μ f7%formic 70% of l avium . FC.par ATCC μ of l ° C Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. fmcbcei.Ol 49 n 58 ftejvnlsadyalnspeetdval T respectively NTM viable presented of or yearlings isolation animals and the juvenile juveniles between growth to the showed association comparison animals of adult in 45.8% from mycobacteria and originating nodes ( with 54.9% lymph infected Only analysed mycobacteria. the be of of to 74.4% Overall, prone 2). more (Fig. were yearlings boars with wild infected transmission five Adult boars a of wild or 1A. out the infection Figure of of four in localization source on displayed geographical common is performed The a S1). suggesting was material code, loci (Supplementary identical chain MIRU-VNTR an showed 24 and on samples containing based characterization Molecular . ( NTM three with is one ( and comprising NTM group, two 539 and with ST (n=2) co-infections as juveniles nonchromogenicum presented and SpolDB4 animals database (n=7) Seven spoligotyping adults node. (www.Mbovis.org). international from 37-38 the originating spacers in (5.1%), of known presence also for the is presented indicative by samples signature characterized five All PCR. identification same SB0118 performed. real-time species was signature The achieved, MTBC nodes be spoligotype by lymph could the same positive from period were the DNA incubation extracted boars months using 12 wild spoligotyping a direct adult over by two mycobacteria of and growth juvenile no three Since from derived samples by Five followed isolates, species. the new of represent 22.5% may with and rRNA, species (16S mycobacterial genes described sequenced (57.4%; 24 The 101 observed. to from were nodes lymph staining mandibular ZN 2018, by and 2017 during collected boars CI wild investigated 176 the Of Results animal Swiss for permit with or accordance approval in ethical An boars 455). wild SR hunted Act applicable. legally Welfare not Animal from was and originated experimentation 922 study SR this Law (Hunting in legislation used was samples significance animals Statistical All test. same the Statement the with investigated Ethics of was isolated of age NTM groups isolation viable and age the of between three to presence sex Diego, set association the the regarding and San possible and groups a Software, data species, age Moreover (GraphPad isolated collected different nodes. evaluate 8.2.1 the lymph to Prism their used on GraphPad was from investigated Based test using were exact 3.6.1. performed territory Fisher’s the was USA). Desktop on analysis QGIS NTM statistical different software the animals, free of circulation the the using and mycobacteria of growth Analysis. showing Statistical and additional an Distribution staining, Geographical HE after observed performed. were was infections mycobacterial staining with ZN consistent lesions where cases in h egahcldt olce.Tehne idpgpplto a iie nosvndsrcso h study on the of based districts investigated seven was into divided animals was analyzed population the pig wild among hunted circulating The collected. NTM data different geographical the the of distribution The adults versus juveniles P 95% -value P 00–6.)aiassoe yia rwhof growth typical showed animals 64.5) – 50.0 .scrofulaceum M. -value < .5.Acreainbtenifce nml n hi e a o on,nrasignificant a nor found, not was sex their and animals infected between correlation A 0.05). .microti M. < Mycobacterium 0.05. .fortuitum M. , .phlei M. P / .florentinum M. Bue ta. 06.Mcbceilc-netoswr eetdi iesamples nine in detected were co-infections Mycobacterial 2006). al., et (Brudey .0;yalnsvru adults versus yearlings 0.804; = / complex/ .nonchromogenicum M. pce Tbe1.Tresnl sltscudntb lsie oayknown any to classified be not could isolates single Three 1). (Table species Mah .nonchromogenicum M. .egakiM vaccae engbaekii/M. M. , .vaccae M. n h he g rus(ueie essyearlings versus (juveniles groups age three the and / 5 Mycobacterium Mycobacterium , .avium M. .nonchromogenicum M. P h egahcldsrbto ftelmhnodes lymph the of distribution geographical The 0.384). = .avium M. ih2.%o h isolates. the of 21.6% with rpoB n n animal one and ) subsp. p.(1 sltsi oa)adAFBs and total) in isolates (111 spp. p)oeaia rsne co-infection presented animal one sp.) , hsp65 / .vaccae M. hominissuis sind18o 1 isolates 111 of 108 assigned ) / .ciar intracelullare chimaera M. .microti M. Mah .mcoiM neoaurum microti/M. M. twice , ( Mah h otprevalent most the , predominated ) .avium M. oiielymph positive P .microti M. .microti M. 0.151; = / M. Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. h oa idba ouainoe tlatadcd.Tesoioyigpol B18 hrdb all by shared showing SB0118, and profile species host spoligotyping the one The from prevalence of independently higher far, decade. presence slightly so a persistent a Switzerland least in a period, 2014). detected at this suggests al., profile over Over only et This population the (Boniotti 2009. is boar shown Italy. and isolates, wild is colleagues 2007 in and Como local between described Boniotti analyzed of the by was isolates Province observed 5.8% 26 the was of loci) of along presence (ETRs out region transboundary profile five of border a MIRU-VNTR chain in literature, and Italian transmission from spoligotyping the a information identical available and or The al., the Swiss boar infection and et the of wild findings (Schoning source in these in based 3.6% common on characterization study was molecular Based a and previous prevalence revealing spoligotyping material). a observed by loci, analyzed with the further 24 were accordance where on samples five cantons, in present Swiss is The different prevalence 2013). observed three CI from The (2.8%; RT-PCR. originating boars by wild detected analyzed 176 was of out five In Discussion MALDI-TOF by possible” identification as “no analysis monacense as sequence classified 1). by isolates, (Table identified 9 were assigned remaining MS, correctly as et The were misidentified (Alcaide S2-S4). species and material species 111 mentary 19 species mycobacterial of same new for out the a proposed isolates from representing previously 80 (90.1%) one as identify isolates isolates, 1.8 correctly 100 Two to 2015), [?] gene, able Pranada, at rRNA was 2018, identification (16s MS al., species analysis MALDI-TOF for sequence 2.0, using cut-off species the [?] 19 at to cut-off appertaining (72.1%) LSV a setting observed By were bacilli Acid-fast Spectrometry 1998). lesions Mass al., visible et MALDI-TOF 3B). Soolingen presented (Fig. (van that macrophages species within Samples this and scales. with extracellular associated grading for commonly positive WHO is revealed tested and which staining and IUATLD Ziehl-Neelsen tuberculosis macrosco- entity. the with observed moderate to to compatible lesions mild and the according of Overall, 3A AFBs and present. (Fig. scanty circumscribed markedly eosinophils were were histologically and calcifications and cells dystrophic infiltration pically giant cases inflammatory three Langhans mild In multinucleated and 3C). infections neutrophils, mycobacterial necrosis macrophages, with epithelioid focal-extensive compatible nodules. of by by lesions calcified infected characterized macroscopic and animals lymphadenitis presenting of necrotic 14) granulomatous nodes caseous, = Lymph as histologically. (n such examined nodes moderate lesions were lymph to visible of mild with subset contrary, associated a A the always visible. and On were was changes. hyperplasia infections macroscopic necrosis reactive (n=3) without focal-extensive severe and moderate with to nodes to moderate lymph giant mild a in scarce a lymphadenitis. additionally Histologically, with of showed eosinophilic lymphadenitis 26). adjacent granulomatous nodes = or moderate lymph surrounding (n a The type recorded lesions, nodular Langhans were nodular described of the presenting induration necrotic cells to samples and addition caseous, the In multiple discoloration of (n=14). or enlargement, observed 70% single were node mm) as (1-30 lymph such sizes of lesions lesions, different growth of visible showing nodules Macroscopic boars calcified 2). and wild (Table 101 lesions the pathological 1B). from copic obtained; (Fig nodes were districts lymph samples four the of remaining Of number the low in Examination a shown Histological is where and cultured districts Macroscopic NTM northern the three of the distribution of homogeneous exception a the with and, area and .terrae M. .vulneris M. ope oeioaeec) n w diinlundescribed additional two and each), isolate (one complex Mah iietfidas misidentified h rdmnn pce dnie ntepeetsuy a on in found was study, present the in identified species predominant the , .colombiense M. 95% .colombiense M. .arupense M. . .)DAaprann oamme fteMTBC the of member a to appertaining DNA 6.5) - 1.2 .microti M. 6 33isolates) (3/3 LV18)ldt iceatrsls(Supple- results discrepant to led 1.81) (LSV Mycobacterium LV20)adascn n,presumably one, second a and 2.08) (LSV hwd”risn-ie rSsae AFBs, S-shaped or ”croissant-like” showed .microti M. .microti M. .scrofulaceum M. rpoB and p. 57 hwdmacros- showed 25.7% spp., .microti M. .microti M. n5 and (n=5) .florentinum M. and Mycobacterium hsp65 22isolates), (2/2 (Supplementary tanaffecting strain .florentinum M. .B lowering By ). hwdall showed spp.. M. Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. o eetd ugsigadvriyo T pce tgorpia ee.Mmeso h A eealso were MAC the al., of Members et Trcka level. 2016, geographical at al., species et NTM Pate of detected: 2011, diversity irregularly al., a suggesting et detected, detect Gortazar presence not not 2015, the did investigated al., colleagues human extensively et and for publications (Garcia-Jimenez suitable few boar condition Only wild healthy 2006). overall in concern. an NTM health in of public the be veterinary Since to a probabilities. judged area represent contact were study animal-to-human the study the in the present as population consumption, the well the including of hunters, Cantons, as by Swiss animals Gethoffer animal-to-animal reduction several hunted 2012, the in population annual al., law augmenting annual by et an the rising, prohibited (Ebert and is enable is 2015). order conditions ungulates ungulates, this al., weather wild Despite and of et among feeding Ticino. food (Massei capacity Supplemental favorable Asia reproductive 2007). under South-east al., highest 250% et and of the Japan rate with growth Russia, Western associated population Eastern from ( mobility extends to boars range high basin natural Wild Their The Mediterranean all analysis. worldwide. be the were mammals sequence to and large study gene has distributed Europe present MALDI-TOF widely additional most the with require the level in might among subspecies tested are and MAC isolates carefulness at MAC infection differentiation with However, 25 of performed The source (80%). correct the 2017). generally however, al., was cases, et be most Ricchi to In and 2017, confirmed human al., 2006). between 2008, et al., relatedness al., (Ravva et genetic of et studies Mobius discrimination close Falkinham The 2007, a 2012, al., unclear. reported al., environmental remained et studies et included (Johansen certain (Eisenberg with These Noteworthy, isolates tubs co-infected swine hot analyses. 2008). being and al., molecular bathrooms animal et through water, Hilborn animals one drinking and and humans e.g. animals observed surveillances, for cumulative be positive sources a could MTBC infection infection be MTBC and of may and individuals number isolation unknown. NTM older remains which small from and one unclear in effect the mostly second co-infection no is a to Overall of it from due number exposure. cases, infection and such increased first time In the this two with of 2017). for effect or effect al., explanation Hlokwe, protective et MTBC/NTM and possible host Yilmaz with Gcebe the 2019, A 2015, Co-infections and al., al., et first, et Stepanyan infected species. (Garcia-Jimenez 2011, agent humans multiple co- al., and et that by animals Lim observation in infected 2017, the described was been by have group corroborated NTM prolonged juvenile different is a the This of from results by presence. the infected boars an transient probably animal with are a one correlated mycobacteria including than infection isolated 2015), infections, more mycobacterial the al., was colonization that of animals et or suggesting increase the (Garcia-Jimenez 2), infection significant of implications (Figure age study a clinical and age Overall, Spanish MAC of and between survey. a increase correlation present host-adaptation from no the subadults, observations distribution, from in the often its observed more to understand isolated Contrary was better MAC where 2006). to members al., host-specific necessary and et is environmental (Turenne between MAC differentiation clear the a of nevertheless, 2006). MAC, al., the of et members (Turenne subspecies namely these subspecies, between four comprises (van and species mycobacterium mycobacterial growing important slowly avium clinically thermophilic a several belonged is comprises study complex this (n=3) 2018). in This colombiense identified al., species 1). et and predominant (Table molecular Ingen The by wild MAC negative different tested the preparation). all South Interestingly, were in to cats, MTBC domestic (manuscript results). for from examined (unpublished methods areas isolated spots Switzerland cultural been hot has in from originating spologotype gibbons species this captive rodent boars, and wild to camelids Additionally American range. host large a subsp. .neoaurum M. paratuberculosis .chelonae M. ( Maa ) .avium M. , Mah Mah nteps eae,vrosrsac rilsatmtdt ecieMCreservoirs MAC describe to attempted articles research various decades, past the In . a h otfeun T pce sltdb Garcia-Jimenez by isolated species NTM frequent most the was , .chimaera/intracelullare M. .intracellulare M. .aim(n=25) avium M. ysqec nlss n h rtsbpce ugsinpoie yMALDI-TOF by provided suggestion subspecies first the and analysis, sequence by ( neto aeo 4 bevdi h adblrlmhnd sntwrh n may and noteworthy is node lymph mandibular the in observed 14% of rate infection Map subsp. .Sqecn fte3 eino the of region 3’ the of Sequencing ). .chelonae M. silvaticum , Map Maa a h otpeaetmme ftecmlx olwdby followed complex, the of member prevalent most the was n14wl or etd ntepeetstudy present the In tested. boars wild 124 in .microti M. ihteMLITFtcnqehsbe sesdi previous in assessed been has technique MALDI-TOF the with ( Mas , Mah group Mah ), 7 and .avium M. (n=2) a h raeths ag oprdt h other the to compared range host broadest the has eemsl nautaiasadnn ftewild the of none and animals adult in mostly were , .colombiense M. and hsp65 subsp. .vulneris(n=1) M. eecnuabgosydistinguish unambiguously can gene hominissuis r oaascasfida MAC as classified nowadays are tal et ( Mah Tbe1). (Table ,wis Gortazar whilst ., .avium M. .chelonae M. and ) u scrofa Sus .microti M. .avium M. .avium M. subsp. was M. ) Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. ytetoaoemnindseiswr nitnusal.Teefidnshglgtteiprac of importance the from members highlight MTBC caused findings of differentiation lesions These reliable the and indistinguishable. histologically rapid a were and allows Macroscopically species that methods mentioned characterization 2005). To above molecular al., two 2007). et al., the by (Tortoli et by caused pulmonary Emmanuel various lymphadenitis lesions with growing 1998, patients and similar slow immunocompromised al., disorders and a that et immunocompetent from as time Soolingen isolated Classified van pathogen first human 2000, the et medicine. al., (Frank is veterinary et individuals this in Niemann immunocompetent knowledge, 2005, in authors’ al., members The the et MTBC 2007). Geiss other al., 2009, by et al., caused relevant Martin-Hernando those a 2007, in from al., and indistinguishable et animals (Dondo adult by affected in 2014). caused MTBC organ al., lesions by only visible caused the lesion of is mentioned visible occurrence this The for cases organ systems. This feed and of likely organ infected nodes most other 2019). proportion and throughout lymph the al., disseminate contaminations are mandibular eventually et environmental in nodes or (Queiros of concentrate lymph persist intake boars often eliminated, have are microorganisms the wild there nodes viable allowing in from lymph infection, was habits mycobacteria Mandibular oro-nasal it eating for After carcass. study, its point boar present sources. entry to wild the preferred due the of the probably in design lesions is be lesions the further to the of of of described presence Because nature the been paucibacillary assess process. the to chronic and possible and not calcifications circumscribed which These a in calcifications. however, samples dystrophic suggests, particular, cases with In compatible some al., lesions analyzed. in et granulomatous and nodes of (Acevedo-Whitehouse lymph presence of species the florentinum subset was wild mycobacterial M. finding a of other interesting in and resistance An observed possibly enable genetic generalization 2007). tuberculosis suggested can and cause al., previously carrion agents environmental et to the Dondo other with causing seem of 2005, and contact not bTB because rodents does in presumably against individuals, this small often boars all However, dead are in of mycobacteria. boars disease consumption clinical pathogenic wild Ticino. of occasional habits, of Canton transmission and the eating one across animal direct Moreover be isolated and of also species spread behavior may NTM species. mobility the the rooting high km range NTM in of This their 50 role another spread 1992). homogenous crucial to to of Spitz, the a move up 2019, Because for playing al., and range explanations basis, et possible can territory daily Schulz the 2015, that their at of al., territory territory of et entire single (Nugent portion the pathogens a small across zoonotic has a move 2004). to male only (Leuenberger, able adult use packs are the neighboring to of intervals, tend ranges regular which home km at with packs, of 1-3 overlap to may Jura-region area: territory contrast the core the its In in dependent of defends rest km2 to strongly the tends 8-30 while animals areas, covered 1994) of Packs geographical group (Briedermann, single between influence. A animals human widely km 10 1994). and 2-40 vary and water 1993), 5 may (Baettig, of between packs Switzerland presence vary by can landscape, covered and size the piglets pack territory from newborn average The of The consists a maturity. group around 2009). the sexual grouped of reach packs rest they social The time in sows. organized breeding ( generally mature al., individuals isolated are sexually et juvenile species boars three (Pate remaining or Wild Slovenia two time, the animals. of 2014), Among first affected nucleus al., three the 2011). et the al., for in (Boniotti et study observed Italy Gortazar present 2015, 2006), al., the al., et in et (Trcka (Garcia-Jimenez Spain Republic (Pate and Czech 18.2% 2016) 2011), to from al., 2015) varying NTM, et al., pigs mentioned (Lara et above wild (Garcia-Jimenez the in engbaeki 16.8% to 2011), M. rates addition nonchromogenicum, al., In isolation et potential NTM 2016). (Gortazar as al., described 16.1% relevance et 2006), their reports al., estimate Previous et and (Trcka implement involved to 8.9% consumers. species fundamental therefore for exact at is the MAC It source identify the infection 2006). to of al., order dissection et (Turenne in The trivial approaches always medicine. advanced not veterinary is level and subspecies human or in species pathogens relevant as implicated members .microti M. eedtce,soe rnlmtu ypaeii hrceie yfcletnienecrosis focal-extensive by characterized lymphadenitis granulomatous showed detected, were < lhuhblee ob esptoei,hsbe ecie ocueetnielesions, extensive cause to described been has pathogenic, less be to believed although , 0mnh fae rmtepeiu itr ae r xeldfo h akb the by pack the from expelled are Males litter. previous the from age) of months 20 and 2 .septicum M. nsuhr rne(pt,19)ad14km 1-4 and 1992) (Spitz, France southern in .microti M. .florentinum M. eeioae rmwl ori eetsuisfo Brazil from studies recent in boar wild from isolated were nwl or a endsrbdpeiul Bnot et (Boniotti previously described been has boars wild in 8 Mycobacterium sntwrh eadn h rnlmtu lesions granulomatous the regarding noteworthy is .prgiu,M erses,M etflvm M. lentiflavum, M. nebraskense, M. peregrinum, M. 2 , Siz 1992), (Spitz, .florentinum M. .florentinum M. 2 nIay(otn tal., et (Boitani Italy in < km 1 2 hs individuals These . sa opportunistic an is 2 .microti M. Biaie al., et (Boitani r described are and Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. eeac.Rgrigtetoseiso T hthv enms rqetyioae ntepeetstudy present the great in of isolated therefore frequently is most sources been have infection potential that NTM local of of identification species ( Latshang two and countries. 2012, the 2003), different al., Regarding al., et between (Kuznetcova compare et humans relevance. and in Taillard to incidence infections 2011, difficult NTM the al., with or regarding country, lacking et other data be any are countries, NTM as not agents most faced, Because these could is in by Switzerland samples. sequence authorities caused analysed submitted health diseases the a public of of where to IP141170001 prevalence misidentification notifiable case, a strain not rare to from are a leading derived infections potentially is sequence search, this shotgun similarity experience, by genome for our found whole search In nucleotide namely PRJNA354248. NCBI sequence, a BioProject identical Hence, suggested an strains. analysis of three MS finding present all MALDI-TOF for the The 2.0 during [?] 97%. noticed of aspect identified score the unambiguously interesting identity of be An analysis BLAST to by assignment. challenge diernhoferi classification accurate most a erroneous for represent the The was still analysis study, 4.0. species sequence library the Library related require in closely Mycobacteria present and of profiles) that MBT isolate spectrum demonstrating the (main one MSP Moreover, 2.08, in of number reference. present restricted as although the this used is identified In this be for misidentification. explanation not as probable that MALDI- or is could the identification rRNA, isolate) of any 16S drawback without (one Mediavilla- e.g. species: important either like 2014, NTM an undetected, genes three opinion, housekeeping go al., study, of authors’ will et analysis the species sequence Buchan In undescribed with 2019, yet 2011). comparison in al., al., technology et MS et Saleeb values TOF cut-off (Alcolea-Medina 2015, lower the al., results evaluated by et different for authors satisfactory recommended Gradolph reliable different from threshold with 72.1%, be mycobacteria the only to mycobacteria Although showed of viable MS for identification specimens. of MALDI-TOF correct veterinary Overall, number the from enable boars. high derived manufacturer wild adopted of NTM alarming hunted standards of from growth an an hygiene identification meat showing that though the ordinary raw in Even indicates the nodes present above This are lymph days. were species different all collection processing. samples on in meat for processed for histology used animals instruments with from RT-PCR and proven and specific technique not MTBC lesions mandibular the was visible the excluded. of presenting of sensitivity infection be disinfection high nodes can samples, the the lymph samples the on of in of Based the environmental size elimination of with possible. small degree the cross-contamination not the contamination was in adopted, of certain area Because results a superficial probably excluded. hand, nodes be other lymph lymph cases mycobacteria to the most on viable not infection, On in of is the focused 2014). mycobacteria and presence of al., studies stage The identifiable et two early morphologically (Mann 2012). an by microorganism yet al., represent described not et might Muwonge lesions recently is granulomatous 2014, et which were histopathological al., Tortoli findings of et 2010, evidence Similar (Mann al., without pigs et slaughter 2). Syed from (Table 2014, nodes al., general et of in (Nukui proportion finding US significant the a and Interestingly, florentinum Japan Finland, 2005). Italy, al., in described of been distribution have geographical The NTM. other h ag aoiy(=8 a endsrbda ua ahgn.I re oass h osberole possible has the including MS assess investigations MALDI-TOF that, to epidemiological show order further findings present In (n=24), mycobacteria, the Moreover, species of emphasized pathogens. required. isolated spread be human are the the must sources as different among into it described from and animals isolates unclear, been consumption wild remains has human by NTM (n=18) for viable played isolated intended presented majority the boars were large unknown. wild of animals the remains from risk hunted infection collected the zoonotic of nodes route that, the lymph and affected mandibular Although source localization of the classical number mycobacteria. et cases, The remarkable (Nishiuchi most a 2015). Japan in al., conclusion, 2013), and et In al., system (Boyle et America respiratory North (Hoefsloot the and is Europe 2018) MAC Northern al., by in et human Korea(Ko in 2017), diseases al., NTM of cause common .avium M. subsp. sltstse.Acrigt CIBATtecoetseiswas species closest the BLAST NCBI to According tested. isolates eeioae rmtsu ape htddntpeetgauoaoslsoso pathological or lesions granulomatous present not did that samples tissue from isolated were , hominissuis .colombiense M. and .nonchromogenicum M. .vulneris M. 33isolates) (3/3 .florentinum M. 9 Mycobacterium a iietfidas misidentified was ,MCmmeshv endsrbda h most the as described been have members MAC ), .scrofulaceum M. per ob iepedsnehmncases human since widespread be to appears .microti M. .denoeirpoB diernhoferi M. rpoB .diernhoferi M. p.ohrthan other spp. eesqecsdrvdfo three from derived sequences gene 22ioae)and isolates) (2/2 N a dnie exclusively identified was DNA .colombiense M. .aurum M. iha nqioa LSV unequivocal an with Mycobacterium eune loe the allowed sequences .microti M. ihapercentage a with .monacense M. ihaLVof LSV a with p,the sp., and M. M. Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. oiti .B,A aui .Glet,e l,21:Dtcinadmlclrcaatrzto of characterization Tuscany, molecular in and Detection (2014). boars 2014: wild al., of et patterns Gelmetti, microti D. activity Gaffuri, and A. B., Spatial M. Boniotti, 1994: Corsi, F. and Nonis relevance. D. veterinary Italy. Mattei, of L. infections the L., mycobacterial Boitani, of Non-tuberculous relevance 2014: Boschiroli, Britain: Sci., L. in Vet. M. badgers and F. and Biet, tuberculosis Bovine 1993: M., 2013: the Baettig, Robinson, for method A. P. simple Mass improved and Ionization past. An J. Desorption- P. 2019: Laser Atkins, al., (Matrix-Assisted et MS Montiel, MALDI-TOF N. by Spectrometry). Fernandez, Mycobacteria C. of T. identification M. A., non-tuberculous Alcolea-Medina, identify to: How Diseases, Infectious (2018). and 2018: Microbiology late- Clinical al., of and et Society European spectrometry. nonpigmented Bou, mass of G. MALDI-TOF Amlerova, identification using J. rpoB-based F., 2003: Alcaide, Drancourt, M. mycobacteria. in growing and tuberculosis rapidly Colson pigmenting bovine P. to resistance T., Genetic Adekambi, 2005: al., boar. et wild Gortazar, Iberian C. the Vicente, J. K., Acevedo-Whitehouse, References reasonable upon author corresponding the from available are study this of findings request. the support authors that data the The from request on available Data interest. of conflict no have they that declare authors The Statement Interest of Hubschke Conflict Ella acknowledge their gratefully without authors Ticino, The Canton assistance. from technical possible. wardens skilled game been for the have Rademacher and not Fenja hunter would and local study the this thank to cooperation, like would authors The high and medicine. veterinary cost-effectiveness Acknowledgments in rapidness, species his NTM of of because identification for and tool method diagnostic reference valid the a to represent throughput, rate concordance high a pdmooyadinfection, and Epidemiology Mammal, J sltsi idba rmnrhr Italy. northern from boar wild in isolates 7Spl 6-7 o:011/.vc21.807(2014). doi:10.1016/j.rvsc.2014.08.007 S69-77, Suppl, 97 cetfi reports, Scientific 5 0-1,diDi1.371857(1994). 10.2307/1382507 doi:Doi 600-612, 75, a Schwarzwild Das oeua ecology, Molecular 4,13-44 o:011/05281027 (2013). doi:10.1017/S095026881200297X 1437-1444, 141, ,226 o:013/458095647(2019). doi:10.1038/s41598-019-56604-7 20216, 9, eihedrS.alshnNtrisncatihnGesellschaft. 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Mycobacterium Sus-scrofa lnMicro- Clin J International ,e54253, 8, non- a , infection ,4189, 9, ,32, 5, L.). 48, Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. mogenicum nonchro- Mycobacterium avium Mycobacterium and rRNA, (16S no i) analysis as sequence intended by results identified Discrepant Species sequence MALDI- to shown. on compared are species based discrepant offs identifications a of cut of highlighted. assignment value are ii) Results or analysis scores analysis method. log MALDI-TOF by standard different identification gold two species as applying used MS was human TOF analysis and Sequence boars wild nodes. from undescribed potentially isolated three mycobacteria nontuberculous relevance. of clinical Identification 1. Mycobacteria Nontuberculous Table and Tuberculosis Mycobacterium Lungs. 2017: the Saglam, of L. and Coinfection Ucar Y. E. 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(Sus V. boar D. the wild Semret, of European M. 2005. in Y., to infections C. 2002 Mycobacterial Turenne, years 2006: the al., during Republic et Suchy, Czech R. the Lamka, J. I., Trcka, microbiology, 2005: evolutionary al., and et (2005). systematic Goh, of S. genus journal K. the International Rindi, of L. species E., new Tortoli, of relevance clinical and features . Microbiological 2014: E., Tortoli, lnclmcoilg reviews, microbiology Clinical hsp 8 6637,di1.09ism0032 (2018). doi:10.1099/ijsem.0.003026 3666-3677, 68, yoatru avium Mycobacterium 65) rpoB oa f11nnueclu yoatraioae eogn o2 nw pce and species known 24 to belonging isolates mycobacteria nontuberculous 111 of total A 41 4+(aa tal., et (Sawai + et Ingen (van + 24 25 15 24 LSV 24 25 isolates of No. ukToa J, Thorac Turk nentoa ora fifciu iess:II ffiilpbiaino the of publication official : IJID : diseases infectious of journal International complex. 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J. 8 32,di1.12TrToaJ21.63 (2017). doi:10.5152/TurkThoracJ.2017.16034 23-26, 18, nentoa ora fsseai n vltoaymicrobiol- evolutionary and systematic of journal International 7 56,di1.06jii.071.0 (2018). doi:10.1016/j.ijid.2017.12.001 65-69, 67, AD-O MS MALDI-TOF by Identification pce eeclue rm16wl ormniua lymph mandibular boar wild 176 from cultured were species yoatru florentinum Mycobacterium > e Med-Czech, Vet 16 yoatru neoaurum Mycobacterium LSV 2 7 0-1,di1.01vte:054 (2006). doi:10.1051/vetres:2005044 107-119, 37, netosdiseases, Infectious lnMicrobiol, Clin J yoatru scrofulaceum Mycobacterium 4 3-4,doi:10.1128/JCM.44.2.433-440.2006 433-440, 44, IDCases, 5 1110,doi:10.1099/ijs.0.63485-0 1101-1106, 55, 1 2-3 (2006). 320-332 51, AD-O MS MALDI-TOF by Identification 1 89,doi:10.1016/j.idcr.2018.01.004 88-90, 11, hsp65 yoatru avium Mycobacterium > p o. sltdfo humans. from isolated nov., sp. 1 0-0,doi:10.1080/23744235.2019.1634281 602-609, 51, 6 8014 (1998). 1840-1845 36, 1.8 itnuse mn subsets among distinguishes ierltdbceei with bacteremia line-related yoatru lentiflavum Mycobacterium yoatru microti Mycobacterium ies:experience disease: Mycobacterium 2006) 2018) al., relevance clinical Human ope for complex +) Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. vulneris Mycobacterium terrae Mycobacterium septicum Mycobacterium monacense Mycobacterium interjectum Mycobacterium holsaticum Mycobacterium complex fortuitum Mycobacterium phlei Mycobacterium scrofulaceum Mycobacterium intermedium Mycobacterium group intracelullare chimaera- Mycobacterium celatum Mycobacterium bourgelatii Mycobacterium peregrinum Mycobacterium florentinum Mycobacterium diernhoferi Mycobacterium colombiense Mycobacterium lentiflavum Mycobacterium nebraskense Mycobacterium neoaurum Mycobacterium engbaekii Mycobacterium vaccae Mycobacterium and rRNA, (16S analysis sequence by identified Species hsp complex 65) rpoB Troi 2014) (Tortoli, + / et (Schinsky + 2014) (Tortoli, + (Dholakia, + 0 al., et (Richter + 0 et (Hoefsloot + 1 al., et (Tanaka + 0 al., et (Wilson + 1 0 - 1 0 1 1 et Ingen (van + 2 0 et (Chavarria + 0 1 1 - 2 1 1 et (Hoefsloot + 1 1 al., et (Tortoli + 2 1 1 - 2 0 1 2006) (Tortoli, + 2 2 1 al., et (Yagi + 3 1 2006) (Tortoli, + 3 2 2 al., et (Walayat + 3 2 2 - 0 2 2 - 4 3 4 2 2 6 3 2 6 0 2 8 4 3 1 3 4 3 3 3 8 4 4 6 6 8 AD-O MS MALDI-TOF by Identification 17 AD-O MS MALDI-TOF by Identification l,2000) al., 2017) 2002) 2013) al., 2019) 2019) 2018) al., 2018) al., 2013) al., 2005) 2018) 2018) relevance clinical Human +) Posted on Authorea 30 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158559989.96244770 — This a preprint and has not been peer reviewed. Data may be preliminary. 5–n dAlseislisted species All nd nd nd +) done. not nd, findings; histological or macroscopic no – present; finding nd histological or macroscopic +, – + 75 or Single 12 4 100 or Single Macroscopic 10 80 n nodes lymph in of infection Examination mycobacterial with for compatible positive boars findings wild histological hunted of and nodes unknown Macroscopic lymph +; 111 pathogen: 2. (opportunistic Table cased reported -) previously saprophytes: to or according pathogenicity relevance clinical Human +) Total described) sp. Mycobacterium and rRNA, (16S analysis sequence by identified Species (not hsp 65) rpoB utpenodules multiple nodules multiple findings nodes lymph of Examination / 0 0 3 +) – – + + findings Histological nodes lymph of Examination AD-O MS MALDI-TOF by Identification ++) 18 Mycobacterium findings Histological nodes lymph of Examination ++) AD-O MS MALDI-TOF by Identification p.aeresumed. are spp. identified mycobacteria of Species .neoaurum M. 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