is present in the normal intestinal flora but some strains can cause intestinal and extraintestinal diseases, and research on its presence in food of animal origin is in the interests of public ...health. This study was designed to characterise
strains according to their origin, their carriage of virulence genes specific for certain pathogroups, and phylogenetic group affiliation.
The study was carried out on 100
strains isolated from food samples of various animal origin as well as pig and cattle carcass swabs. Isolation of the strains was performed using two methods. One method included colony count and the other an overnight enrichment of the samples. Isolation was followed by DNA extraction and detection of virulence genes and phylogenetic group with conventional and multiplex PCRs.
In this study, the most prevalent gene was
(20%) and strains which carried it were identified as enteroadherent
. Other pathogroups were represented in lower incidences. Phylogenetic group analysis revealed the prevalence of the A and B1 groups, with B1 mainly present in game and cattle strains, while the majority of pig and poultry strains were assigned to group A.
This study provides an overview of the presence of potentially pathogenic strains and
phylogenetic groups in Croatia, for which the data are limited. Further microbiological and molecular research is required to examine the epidemiological situation in the country.
In order to detect thermotolerant
spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase ...chain reaction/restriction enzyme analysis for exact species determination.
spp. prevalence was 73.86%.
and
were isolated from 53.53 and 15.35% of the samples, respectively. In 4.98% of isolates thermotolerant
spp. were not determined. The multilocus sequence typing method was used to evaluate genetic diversity of eight
and four
isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs) and clonal complexes (CCs) of
and
.
isolates in Croatia. By applying the multilocus sequence typing, a new allele of
gene locus was discovered and marked
508. The
.
ST 6182 and
.
ST 6183 genotypes were described for the first time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of
and
in Croatia.
Campylobacteriosis is the most common gastrointestinal bacterial disease in the European Union (EU). Wild birds are one of the natural reservoirs of these pathogens. In this study we tested cloacal ...swabs of 643 gulls captured on rubbish tip in Zagreb, Croatia for the presence of Campylobacter spp. and found 168 Campylobacter positive samples. We used multilocus sequence typing (MLST) to genotype 62 random C. jejuni isolates from gulls, 24 isolates from broiler caeca, 27 isolates from broiler neck skins and 23 human isolates. Altogether, we identified 44 different STs, from which 19 were newly described. Most of the new STs (14) originate from gulls. Although humans and broilers share the majority of STs and isolates from gulls are separated from these, there was one ST present in all three hosts: 45. Additionally antimicrobial susceptibility to six antimicrobials was performed on 123 C. jejuni strains isolated from broiler caeca (n = 22), neck skins of broilers (n = 20), gulls cloacal swabs (n = 50) and human faeces (n = 31) by the broth microdilution method. Results show lower resistance of gull isolates to NAL and CIP, while resistance to TET was as high as in human and broiler isolates.
Brucellosis in swine was surveyed from 2011 to 2015 in 13 counties in Croatia. A total of 3230 breeding males were tested serologically, and positive reactions were confirmed in 42 (1.3%) males from ...17 farms. A total of 641 sows with abortion or reproductive problems were tested, and positive reactions were confirmed in 34 (5.3%). Organs from 68 swine were tested for bacteria, and Brucella spp. was isolated from 47 (69.1%). B. suis was identified in 45 isolates from domestic swine and 2 isolates from wild boar in six counties in Croatia, and all isolates were found to be B. suis biovar 2 based on Bru-up/Bru-low, Bruce-ladder, Suis-ladder and RFLP-based PCR assays. These results indicate that brucellosis is difficult to eradicate in free-range and semi-free-range swine farming, particularly in areas where contact with wild boar is possible. Further disease control measures are required.Key words: Brucella suis biovar 2; swine; abortion; prevalence; CroatiaBRUCELOZA PRAŠIČEV, POVZROČENA Z BAKTERIJO Brucella suis BIOVAR 2 NA HRVAŠKEMBrucelozo pri prašičih smo spremljali od leta 2011 do leta 2015 v 13 hrvaških občinah. S serološko analizo smo preverili prisotnost bruceloze pri 3230 samcih v razplodu. Pozitivne reakcije smo ugotovili pri skupno 42 merjascih s 17 farm, kar predstavlja 1,3 % živali. S serološkimi testi smo preverili prisotnost protiteles proti bruceli tudi pri 641 plemenskih svinjah, ki so zvrgle ali imele težave z zabrejitvijo. Pozitivna reakcija je bila ugotovljena pri 34 svinjah, kar predstavlja 5,3 % vseh testiranih živali. Notranje organe 68 svinj iz šestih občin smo uporabili za osamitev bakterij Brucella spp. Bakterije smo ugotovili pri 47 vzorcih (69,1 %). Bakterijo Brucella suis smo odkrili v vseh 47 vzorcih, izmed katerih jih je bilo 45 od domačih plemenskih svinj, dva vzorca pa sta bila od divjih svinj. Vse izolirane bakterije so pripadale sevu B. suis biovar 2, kot so pokazale dodatne analize z uporabo metod Bruup/Bru-low, Bruce-ladder, Suis-ladder in RFLP. Ti rezultati kažejo, da je popolno izkoreninjenje bruceloze težavno, še posebej v prostih rejah prašičev, kjer obstajajo možnosti stika z divjimi prašiči. Zato bi bilo v prihodnje potrebno razmisliti o dodatnih načinih nadzora nad to nevarno boleznijo prašičev.Ključne besede: Brucella suis biovar 2; prašiči; zvrg; pojavnost; Hrvaška
Patogene mikobakterije imaju svoje domaćine, a većina njih i prirodne rezervoare u prirodi. Tuberkuloza prouzročena vrstom M. tuberculosis i dalje ostaje glavni uzročnik tuberkuloze u svijetu. M. ...bovis ima najrašireniji raspon prirodnih rezervoara od svih poznatih patogena. Različite vrste divljih životinja predstavljaju trajan rezervoar infekcije vrstom M. bovis za domaće životinje i ljude. U novije vrijeme otkrivene su i nove vrste mikobakterija (M. caprae, M. pinnipedii, M. mungi, M. orygis i M. suricattae) koje po svojim karakteristikama pripadaju u kompleks M. tuberculosis. Sve navedene vrste mikobakterija imaju svoje rezervoare u prirodi i kao zoonoze predstavljaju stalnu prijetnju za infekciju ljudi. M. leprae prouzroči lepru u ljudi. Dugo vremena se mislilo da je samo čovjek domaćin i rezervoar uzročnika. Danas je poznato da su prirodni rezervoari lepre devetopojasni pasanci (Dasypus novemcinctus) u SAD i Južnoj Americi, crvene vjeverice (Sciurus vulgaris) u Velikoj Britaniji i različite vrste majmuna u Africi i Aziji.
Bakterije
mikroorganizmi koji za svoj rast u okolišu, kao i u laboratorijskim uvjetima trebaju mikroaerofilne uvjete. Za rutinsku analitiku klasičnim metodama primjerena je primjena priznatih ...standardiziranih protokola i propisanih visoko selektivnih tekućih i/ili krutih hranjivih podloga. Nakon porasta karakterističnih kolonija radi određivanja vrste kampilobaktera primjenjuju se mikroskopski, fenotipski, biokemijski, imunozimski testovi, rezistotipizacija i/ili metoda masene spektrometrije. Za razlikovanje sojeva kampilobaktera postoje dvije klasične metode serotipizacije. Do sada je razvijen znatan broj PCR protokola usmjerenih prema otkrivanju Campylobacter roda, vrste ili podvrste, bilo iz uzoraka hrane, okoliša ili izmeta. Glavne prednosti PCR metoda su u njihovoj brzini, osjetljivosti i specifičnosti, a nedostatak PCR metodologije je u tome što ne razlikuje žive od mrtvih stanica, zbog čega se razvijaju novi protokoli bazirani na RT-PCR, NASBA i QRT- PCR molekularnim metodama.
Različite metode genotipizacije omogućuju razlikovanje bakterijskih sojeva ispod razine vrste ili podvrste. Gel elektroforeza u pulsirajućem polju (PFGE) neprestano se usavršava od 1980-tih godina i ...do danas je upotrebljavaju mnogi javnozdravstveni laboratoriji. Od ostalih metoda koriste se: metoda analize duljine polimorfizma restrikcijskih odsječaka (RFLP), metoda nasumično umnoženih poliformnih odsječaka (RAPD) i metoda ribotipizacije. Određivanje redoslijeda nukleotidnih sljedova unutar DNK naziva se sekvenciranje. Na osnovi sekvenciranja bazira se rad novih metoda tipizacije određivanjem sljedova nukleotida na više genskih lokusa poput MLST, rMLST i cgMLST. One istražuju odnose između bakterija roda Campylobacter, posebice C. jejuni i C. coli. Korisne su u istraživanju mogućih izvora i rezervoara uzročnika bolesti, njihovih odnosa te molekularne epidemiologije bolesti. Metoda sekvenciranja cijelog genoma (WGS) u mikrobiologiji revolucionarna je metoda za koju se smatra da će postupno zamijeniti ostale metode genotipizacije.
Unutar roda Mycobacterium (M.) identificirano je više od 170različitih vrsta mikobakterija. U odnosu prema čovjeku podijeljene su u tri skupine.M. tuberculosis kompleks u koji spada deset vrsta koje ...uzrokuju tuberkulozu usisavaca. Vrsta M. tuberculosis, unutar M. tuberculosis kompleksa i dalje ostaje glavniuzročnik tuberkuloze u ljudi u svijetu. Osim već do sada dugo poznatih vrsta (M.bovis, M. africanum, M. microti, M. canetti, M. caprae), u novije vrijeme otkriveni sui novi pripadnici te skupine. M. pinnipedii u tuljana, M. munghi u mungosa, M. orygisu arapske oriks antilope i M. suricattae u merkata. Isti uzročnici dokazani su u ljudikoji žive u njihovoj blizini. M. leprae je uzročnik lepre u ljudi. Bolest je dokazana uoklopnika (aramadila) (Dasypus novemcinctus), koji predstavljaju prirodni rezervoarM. leprae. Najveći broj mikobakterija (više od 160 vrsta) pripada netuberkuloznimmikobakterijama. Nalaze se u okolišu, vrlo su otporne i prilagođene rastu i razvoju,a njima su često izloženi ljudi i životinje. Neke su važne potencijalno patogene vrstei ponekad uzrokuju teške patološke procese u ljudi i životinja.
Within the genus Mycobacterium (M.) more than 170 different types ofmycobacteria have been identified. In relation to humans, they are divided into threegroups. M. tuberculosis complex, which ...includes ten species that cause tuberculosisin mammals. The type of M. tuberculosis within M. tuberculosis complex remainsthe major cause of tuberculosis in humans in the world. Except for a long time knownspecies (M. bovis, M. africanum, M. microti, M. canetti, M. caprae), new membersof the group have recently been discovered. M. pinnipedii in seals, M. munghi inmongoose, M. orygis in arabian oryx and M. suricattae in the meerkat. The samecauses have been proven in people living nearby. M. leprae is the cause of theleprosy in humans. The disease has been proven in nine banded armadillos (Dasypusnovemcinctus) and represents the natural reservoir of M. leprae. The largest numberof mycobacteria (more than 160 species) belongs to nontuberculous mycobacteria.They are found in the environment, they are very resistant and adapted to growthand development, and to humans and animals are often exposed to them. Someare important potentially pathogenic species and they sometimes cause severepathological processes in humans and animals.
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