Prevalencija Coxiella burnetii protutijela kod ovaca i koza na području Istarske županije u Hrvatskoj

A seroepidemiological study was conducted on the presence of Coxiella burnetii (C. burnetii) antibodies in sheep and goats in Istria, the largest peninsula in Croatia. Random blood samples were taken from 634 sheep and goats at different localities throughout the region. The aim of the study was to assess the prevalence of C. burnetii infection in sheep and goats, which represent the most important reservoir of infection in humans. C. burnetii antibody detection was performed by ELISA (LSIVet Ruminant Q Fever-Serum/Milk). Seroprevalence of C. burnetii was proven in 6.2% of sheep and 3.5% of goats. Larger herds, poor hygienic conditions on farms, a higher presence of goats in a restricted area, and the northern part of Istria proved to be significant risk factors for the seropositivity of animals.The southern part of Istria is known to have endemic Q fever, but no studies have been carried out so far to explore this issue. To gain a more complete epidemiological picture of Q fever in Istria, studies in humans, especially those professionally exposed to C. burnetii infection, should be performed.U radu prikazani su rezultati seroepidemiološkog istraživanja prisutnosti protutijela protiv bakterije Coxiella burnetii (C.burnetii) kod ovaca i koza. Uzeto je slučajnim odabirom 634 uzoraka krvi ovaca i koza s različitih područja Istarske županije, najzapadnije regije u Hrvatskoj. Cilj istraživanja bio je procijeniti proširenost infekcije koksijelom kod tih životinja koje predstavljaju glavni rezervoar infekcije za ljude. Prisutnost protutijela protiv C. burnetii utvrđena je serološkom metodom ELISA (LSIVet Ruminant Q Fever-Serum/Milk). Prevalencija C. burnetii bila je 6,2% u ovaca i 3,5% u koza. Veća stada, lošiji higijenski uvjeti na farmi, veća prisutnost koza na jednom ograničenom području te sjeverna regija Istre dokazani su kao statistički znakoviti čimbenici rizika za seropozitivnost životinja. Poznato je da je južni dio Istre endemski za Q-groznicu, ali do sada nije provedeno istraživanje koje bi obuhvatilo cijelo područje Istarske županije. Da bi se dobila još kompletnija epidemiološka slika Q-groznice na ovom području, trebalo bi uključiti u istraživanje i ljude, osobito one koji su profesionalno eksponirani ovoj bolesti

Tipizacija izolata Brucella melitensis izdvojenih u Republici Hrvatskoj analizom broja uzastopnih ponavljanja na više lokusa (MLVA).

In the period from 2009 to 2013, bacteriological testing was conducted on 336 sheep, goat and cattle samples. Using classical bacteriological and molecular procedures, B. melitensis was confirmed in 14 (4.2 %) samples. Brucella was isolated in the Karlovac, Lika-Senj and Split-Dalmatia Counties. Brucella isolates were genotyped using the MLVA method and compared with isolates from neighbouring Bosnia and Herzegovina (BH). A total of 14 isolates (strains) originating from Croatia and 25 from BH were analysed. Complete matches between Croatian and BH isolates were found in two genotype groups. Overall, the MLVA analysis indicated that the Croatian and BH genotypes of B. melitensis from animals were highly homogenous. The Hunter Gaston diversity index (HGDI) showed that diversity was found among the 16 tested loci for 5 loci of panel 2B (Bruce 04, 07, 09, 16, 30). The discriminating loci are optimal for the use in epidemiological investigation of B. melitensis infections in these two countries.U razdoblju od 2009. do 2013. u Hrvatskoj je izvršena bakteriološka pretraga uzoraka ukupno 336 ovaca, koza i goveda. Klasičnim bakteriološkim i molekularnim postupcima B. melitensis je potvrđena u 14 (4,2%) izolata. Brucele su izdvojene u Karlovačkoj, Ličko-senjskoj i Splitsko-dalmatinskoj županiji. Analiza broja uzastopnih ponavljanja na više lokusa (engl. multiple locus variable number of tandem repeat analysis, MLVA) primijenjena je radi genotipizacije i usporedbe s izolatima iz susjedne Bosne i Hercegovine. Ukupno je analizirano 14 izolata podrijetlom iz Hrvatske i 25 iz susjedne Bosne i Hercegovine. U dva slučaja genotipovi izolata iz Hrvatske u potpunosti su se poklapali s genotipovima u Bosni i Hercegovini. Korištenjem metode MLVA utvrđen je visok stupanj homogenosti izolata u ove dvije zemlje. Od korištenih 16 MLVA lokusa raznolikost je utvrđena samo na 5 lokusa panela 2B (Bruce 04, 07, 09, 16, 30) što je potvrđeno računanjem HGDI indeksa. Korištenje ovih 5 razlikovnih lokusa optimalno je za provođenje epidemioloških istraživanja u Hrvatskoj i Bosni i Hercegovini

Mycobacterium avium subsp. hominissuis izdvojen iz divlje svinje (Sus scrofa) u Republici Hrvatskoj.

During the research period (2001-2005), the lymph nodes of 123 wild boars from seven locations in the Republic of Croatia were tested for the presence of mycobacteria. Mycobacteria were isolated from the lymph nodes of 15 (12.2%) wild boars from fifi ve locations. Specific hybridization showed that 11 (8.9%) isolates belonged to M. avium, 2 (1.6%) isolates were typed as M. fortuitum, and 2 (1.6%) isolates belonged to M. gordonae. All 11 isolates belonging to M. avium sp. were tested by means of the specififi c primers P1 FR300 and P2 FR300. Product size in all isolates was 300 bp, which confirms that M. avium subsp. hominissuis was present in all of them. As in swine from intensive farming, M. avium subsp. hominissuis is a dominant mycrobacteria type in wild boar in Croatia. The results lead to the conclusion that the most important source for wild boar is an environment suitable for the development and survival of M. avium subsp. hominissuis, as confirmed in earlier research studies by other authors.Tijekom istraživanog razdoblja (2001. do 2005.) radi pretrage na prisutnost mikobakterija obrađeni su limfni čvorovi 123 divlje svinje iz sedam lokaliteta u Republici Hrvatskoj. Mikobakterije su izdvojene iz limfnih čvorova 15 (12,2%) divljih svinja iz pet lokaliteta. Specifičnom hibridizacijom dokazano je da 11 (8,9%) izolata pripada vrsti M. avium sp., 2 (1,6%) izolata tipizirana su kao M. fortuitum, a 2 (1,6%) izolata pripadali su vrsti M. gordonae. Svih 11 izolata koji su pripadali M. avium sp. testirani su pomoću specifičnih početnica P1 FR300 i P2 FR300. U svih izolata veličina proizvoda iznosila je 300 parova baza što potvrđuje da se u svih radi o vrsti M. avium subsp. hominissuis. Kao i u svinja iz intenzivnog uzgoja, M. avium subsp. hominissuis predstavlja dominantnu vrstu mikobakterija u divljih svinja u Hrvatskoj. Na temelju dobivenih rezultata može se zaključiti da je po svemu sudeći najvažniji izvor za divlje svinje okoliš koji pogoduje razvoju i opstanku M. Avium subsp. hominissuis, što potvrđuju i ranija istraživanja drugih autora

Spontaneous Mycobacterium avium serovar 2 infection in a Muscovy duck (Cairina moschata)

Avian tuberculosis is an avian disease of worldwide importance. It is typically caused by serovars 1, 2 and 3 of Mycobacterium avium subsp. avium. This paper describes spontaneous, subclinical tuberculosis in a 5-year-old male Muscovy duck. Necropsy revealed a characteristic distribution pattern of granulomatous lesions involving the liver and spleen. Histopathological analysis confirmed the formation of tubercles containing a large number of giant and epitheloid cells, while PCR analysis (according to insertion sequence IS901) and an agglutination test detected M. avium serovar 2 as the causative agent of this condition

Serological, Bacteriological, and Molecular Diagnosis of Brucellosis in Domestic Animals in Croatia

Aim To present the surveillance data on Brucella melitensis, B. suis, and B. ovis infection in cattle, sheep, goats, and swine in Croatia obtained in 2008 by serological, bacteriological, and molecular methods for diagnostics of brucellosis in domestic animals. Methods We serologically tested 42 785 cattle serums, 22 686 sheep and goat serums, and 28 520 swine serums using the Rose Bengal test, complement fixation test, and various immunosorbent assays. We also tested 10 173 ram blood samples for B. ovis infection using the complement fixation test. Bacteriological examination was conducted on 214 samples collected from 34 serologically positive animals. Different molecular methods were employed in the identification and typing of 20 isolates from the samples. Results B. melitensis biovar (bv.) 3 was confirmed with different identification methods in 2 flocks in 2 Croatian counties and B. suis bv. 2 in 3 flocks in 3 counties. B. melitensis in cows was confirmed for the first time in Croatia. Infection with B. ovis was serologically confirmed in 202 rams in 12 counties. Conclusions In 2008, the size of the brucellosis-affected area in Croatia and the efficiency of detection and prevention of brucellosis in sheep, goats, and swine were satisfactory. Infection with B. melitensis in cattle was confirmed for the first time and possible links for infection in humans were detected. More efficient measures for suppression and control of ovine epididymitis are required and a new strategy may be necessary for complete eradication of this disease

Dokazivanje gena za enterotoksine koagulaza-negativnih stafilokoka iz autohtonih hrvatskih fermentiranih kobasica.

The aim of this study was to determine the presence of enterotoxin genes in coagulase - negative staphylococci from traditional Croatian fermented sausages. A total of 39 isolates were collected from the dry sausage “kulen” and Slavonian homemade sausages as presumptive CNS, and were subjected to PCR identification to the genus level. All isolates belonged to the Staphylococcus genus. Strains were tested for the presence of 13 enterotoxin genes; seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN and seO by the PCR method. All strains were negative to all enterotoxin genes. The observed results indicate the absence of risk related to enterotoxogenic potential of coagulase - negative staphylococci as indigenous microbiota from autochthonous Croatian traditional sausages.Cilj ovog rada bio je utvrditi prisutnost gena za enterotoksine u koagulaza - negativnih stafilokoka iz tradicionalnih hrvatskih fermentiranih kobasica. Ukupno je prikupljeno 39 izolata iz kulena i domaće slavonske kobasice te identificirano PCR - om do razine roda. Svi izolati pripadali su rodu Staphylococcus. PCR - om je određivana prisutnost 13 gena za enterotoksine: seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN i seO. Ni u jednom izolatu nije utvrđena prisutnost gena za enterotoksine. Dobiveni rezultati upućuju na odsutnost rizika povezanog s toksogenim potencijalom koagulaza-negativnih stafilokoka kao dijela autohtone mikroflore hrvatskih fermentiranih kobasica

Bruceloza u uzgoju turopoljskih svinja u Hrvatskoj.

In the period 2008-2011, a program of diagnostics and control of swine brucellosis was conducted in the remaining two farms of the Turopolje pig breed, including a total of 729 samples of porcine blood. Serological diagnostics were carried out by means of the Rose Bengal test (RBT), complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA). Agreement between serological methods was determined by the Kappa test. Almost perfect agreement was found between the results obtained by the CFT and RBT methods (0.764-0.936), while the agreement between the results obtained by RBT and CFT in comparison with the ELISA test results was substantial (0.547-0.767; 0.647-0.843). In addition, in 2009 and 2011, samples from 50 pigs were subjected to bacteriological tests, and a total of 15 isolates were obtained. Bacteriology was conducted with various time delays from the determination of positive serological reactions, and the method efficacy was determined considering the duration of the infection in pigs. The average efficacy of the isolation of Brucella (B.) suis ranged between 10-43.3 % (P = 0.01) and was significantly higher in acute brucellosis. Isolation was successful for all age categories, regardless of their sexual maturity. Molecular identification confirmed B. suis bv. 2 for the first time in this breed. The extensive breeding of Turopolje pigs and daily contact with live reservoirs of the disease agents are the reasons why the disease cannot be completely eradicated according to the test-and-remove principle. A condition for the successful control and eradication of brucellosis in a Turopolje pig herd is to ensure the conditions for individual and permanent supervision of each breeding pig.U razdoblju od 2008. do 2011. godine proveden je program dijagnostike i kontrole bruceloze u preostala dva uzgoja turopoljskih svinja na ukupno 729 uzoraka krvi. Serološka dijagnostika provedena je rose bengal testom (RBT), reakcijom vezanja komplementa (RVK) i imunoenzimskim testom (ELISA). Međusobna podudarnost metoda određena je kappa testom. Između rezultata dobivenih primjenom RVK i RBT utvrđena je vrlo visoka podudarnost (0,764-0,936). Nešto niža, ali još uvijek visoka podudarnost utvrđena je između rezultata dobivenih RBT i RVK u usporedbi s rezultatima dobivenim ELISA testom (0,547-0,767; 0,647-0,843). Također, tijekom 2009. i 2011. godine bakteriološki je pretražen materijal podrijetlom od 50 svinja te je ukupno izdvojeno 15 izolata. S različitim vremenskim odmakom od utvrđenih pozitivnih seroloških reakcija učinjena je i bakteriološka pretraga te određena uspješnost metode s obzirom na starost infekcije u uzgoju. Prosječna uspješnost izdvajanja vrste Brucella (B.) suis varirala je od 10 - 43,3 % (P = 0,01), te je bila značajno veća pri akutnoj brucelozi. Izdvajanje uzročnika bilo je uspješno u svih dobnih kategorija, bez obzira na spolnu zrelost svinja. Molekularnom identifikacijom prvi je put u ove pasmine potvrđena infekcija vrstom B. suis bv. 2. Ekstenzivni način uzgoja turopoljske svinje i svakodnevni kontakt sa živim rezervoarima uzročnika određuju da se bolest ne može u potpunosti iskorijeniti prema principu pretraži i ukloni. Preduvjet uspješne kontrole i eradikacije bruceloze u stadu turopoljskih svinja jest osiguravanje uvjeta individualnog i trajnog nadzora nad svakom rasplodnom svinjom