ORIGINAL SCIENTIFIC ARTICLE / IZVORNI ZNANSTVENI ČLANAK Research of Chlamydiosis presence in dogs population in Bosnia and Herzegovina

B. Čengić, A. Ćutuk, E. Šatrović, N. Varatanović, R. Lindtner Knifc, B. Slavec L. Velić and A. Dovč*

Abstract Our research describe epidemiological study was conducted in twelve cities in Bosnia presence of Chlamydiosis in diferent and Herzegovina with cooperation between categories of dogs in Bosnia and Herzegovina. two departments for contagious disease in Problem of stray dogs, inordinately examined Veterinary faculty of Sarajevo and Veterinary and not vaccinated dogs is one of the faculty of Ljubljana. The aim of the research most complex problems among citizens, was to determine presence of Chlamydial nongovernment organisations and institutions infections in diferent categories of dogs, using in Bosnia and Herzegovina. Chlamydiosis is modern serological and molecular diagnostic zoonotic disease caused by Gram-negative, methods. Blood serum samples were taken intracellulare bacteria, which include strains: during 2012/2013. In total, 294 samples were Chlamydophila felis, Chlamydophila abortus, assessed for presence of specifc Chlamydial Chlamydophila psitaci and Chlamydophila antibodies using method of indirect caviae. Disease have endemic characteristics immunofuorescence, while method of RT- and there is litle information about natural PCR was used for determination of antigen. infections in dogs, which were mostly related After assessing 294 blood serum samples, to conjuctivitis, encephalitis and symptoms 2.04% (6 samples) were positive for Cp. psitaci. characteristic for pneumonia. In Europe, Most of the positive samples originated from research of clamidiosis in dogs has been stray dogs. From serology positive animals, conducted in a small number of countries nose swabs were taken and assessed using which include Germany, Slovakia, Sweden RT-PCR. The presence of nucleic acid from Cp. and Lithuania. This was a frst of its kind psitaci was not confrmed in any of them. study of Clamidiosis in dog population, Key words: chlamydiosis; antigen; antibody; carried out in Bosnia and Herzegovina. The dogs; Sarajevo

Introduction Pets and working dogs (Shepherds, and Herzegovina. Unfortunately, because Hounds and Watchdogs) had always their of unfavorable economic conditions in traditional place among citizens in Bosnia country during last several years, there

Benjamin ČENGIĆ, DVM, PhD, Amel ĆUTUK, DVM, PhD, Edin ŠATROVIĆ, DVM, PhD, Nazif VARATANOVIĆ, DVM, PhD, Lejla VELIĆ, DVM, PhD, Veterinary Faculty University in Sarajevo, Bosnia and Herzegovina; Renata LINDTNER KNIFIC, DVM, Brigita SLAVEC, DVM, PhD, Alenka DOVČ*, DVM, PhD, (Corresponding author, e-mail: [email protected]), Veterinary Faculty University of Ljubljana, Slovenia

VETERINARSKA STANICA 50 (6), 2019. 541 B. ČENGIĆ, A. ĆUTUK, E. ŠATROVIĆ, N. VARATANOVIĆ, R. LINDTNER KNIFIC, B. SLAVEC L. VELIĆ and A. DOVČ was a notable increase in number of Chlamydophila psitaci and Chlamydophila abandoned dogs, regardless of their caviae (Pantchev et al., 2010). Chlamydias breed. However, many citizens still kept are group of bacterias resistent to envi- their dogs, but could not perform regular ronmental factors such as cooling and de- health inspections or vaccinations, where hydration (Gylstrof and Grimm, 1987). together with abandoned dogs as a result, Chlamydias with their biological unfavorable epizootiological situation and morphological characteristics looks has developed. Anadolu Agency quotes like bacterias and viruses. However that a problem of stray dogs is one of the only characteristic that chlamydias the most complex problems among share with viruses is that they are forced citizens, nongovernment organisations obligate intracellular “parasites” and (NGOs) and institutions in Bosnia and use the hosts ATP, as well as guanosine Herzegovina (BiH). There is no ofcial triphosphate (GTP) (Acha and Szyfres, information about number of stray dogs 2003). Unlike viruses, chlamydias have in the cities, but estimates suggests it to cellular membrane, similar to gram- be around 7000 of them just in Sarajevo. negative bacterias, contain DNA and (htp://www.oslobodjenje.ba/vijesti/bih/ RNA, have own ribosomes, synthetize psi-su-strah-i-trepet-sarajeva-lutalice-do- their own proteins, nuclein acids and fats sada-ugrizle vise-od-dvije-hiljade-ljudi). and are responsive to tetracyclins as well Stray dogs has become a serious threat as certain antibiotics that inhibit their for public health, because their number multiplication (Moulder, 1984). incerase all the time and during last Chlamydias were found and several years and fear among citizens described, for the frst time in dogs is justifed by the notion that a large more then 50 years ago in Germany proportion of the stray dogs are potential (Sprague et al., 2009). Disease have carriers of contagious diseases (CD), endemic characteristics and there is litle which may jeopardise health in humans information about natural infections and animals. in dogs, which were mostly related to Chlamydiosis in dogs is disease conjuctivitis, encephalitis and symptoms caused by gram-negative, intracellulare characteristic for pneumonia (Gresham et bacteria, which include strains: Chla- al., 1996). mydophilia felis, Chlamydiophila abortus, Clinical cases of chlamydiosis are manifested via increased body temperature, apathy, vomiting, difcult breathing, uncoordinated movements, conjuctivitis, enteritis, joint pains and neurological disorders (Greene and Sykes, 2006; Liutkeviciene et al., 2009). Researches has found that pathogenicity is dependant of toxin potency (Chahota et al., 2006; Corsaro and Greub, 2006). Importance of chlamydiosis is also zoonotical. Several types of Chlamydiales can be transmited from animals to humans. Most often it is: Cp. psitaci, Cp. abortus and Cp. felis. Humans infected with Cp. psitaci may result in symptoms similar Figure 1. Sampling locations in territory of like atypical pneumonia, sometimes Bosnia and Herzegovina. with complications like endocarditis,

542 VETERINARSKA STANICA 50 (6), 541-547, 2019. Research of Chlamydiosis presence in dogs population in Bosnia and Herzegovina Istraživanje klamidioze u populaciji pasa lutalica u Bosni i Hercegovini miocarditis, hepatitis, encefalitis, artritis, Zenica. Dogs were divided in three keratoconjunctivitis (Chau et al., 2015), categories: 1. stray dogs; 2. pets; 3. while infection with Cp. abortus in women working dogs. The overall study was may cause miscarriage (Smith et al., conducted between two departments for 2005). The greatest zoonotic potential in contagious disease in Veterinary faculty dogs develop Cp. psitaci (Public Health of Sarajevo and Veterinary faculty of England, 2014), but zoonotic potential Ljubljana. Method applied for verifying also have Cp. felis. specifc clamidial antibodies was Indirect Transmission is most likely through Immunofuorescence (IIF), while Real air, as well as by eye or nose secretions Time-Polimerase Chain Reaction (RT- from diseased cats and dogs. Clinical PCR) was used for proving antigens. In cases of Chlamydiosis in humans caused addition, in the current study, clamidial by Cp. felis is mostly manifested with antigen were verifed using nose swab conjunctivitis. Sometimes, there are cases samples in cases of serology positive with complicated additional clinical dogs. symptoms (Browning, 2004; Wu et al., Blood samples were taken in 2013). vacutainer tubes, containing etilen Because of potential for spreading diamin tetraacetate (EDTA) (BD disease from animal to humans Vacutaner Systems, Belliver Industrial (zoonosis), presence of Chlamydiosis Estate, Plymouth, UK). Blood samples both in animal and public, increased were transported in temperature of +4 °C number of stray dogs, reduced number to laboratory. In the laboratory, blood of vaccinated pets and working dogs was immidiately centrifugated (4000 rpm and insufcient data about widespread for 15 minutes, LC 320) to separate the of this disease, were the main reasons for serum. Separated serum was transferred the research study about the presence of with pipete to sterile eppendorf tubes Chlamydiosis in the stray dog population and stored in a freezer (-20 °C), where in Sarajevo. The need for further research it was held until transportation to comes from the fact that Bosnia and Veterinary faculty of Ljubljana. Before Herzegovina is a country in transition samples of nasal epithelial cells from with the intention of joining the European dogs were taken, nasal surroundings Union (EU). Implementation of measures were cleaned with sterile gauze to for monitoring of contagious diseases, remove dirt. Sampling swab was then especially zoonosis, their prevention and dipped in 2-sacharose-phosphate therapy is one of the preconditions for medium for purposes of transport (2SP) joining EU. that lasted 10 to 20 seconds. 2SP contain

sacharose (74.6 g/L), KH2PO4 (0.512 g/L),

K2HPO4 (1.237 g/L) and L-glutamin acid Materials and methods (0.721 g/L). Following this, a swab was The study into the presence of used (Clearview Chlamydia, Dacton- Chlamydiosis in dogs population was Tipped swabs, UK) to “wipe” dog’s nasal conducted in period from September mucous membrane. During wiping, 2012 to August 2013 and it involved in special care was taken not to touch skin total 294 dogs of diferent breeds, 197 or hairs arround nostrils, which could male and 97 female, age 6 months to potentially contaminate the sample. 10 years. Our study included twelve Every swab with nasal epithelial cells cities: Busovača, Goražde, Jablanica, was dipped in eppendorf tube, flled Ključ, Konjic, Lukavac, Prijedor, Sanski with transport medium for clamidia. Most, Sarajevo, Travnik, Tuzla and These samples were again transported to

VETERINARSKA STANICA 50 (6), 541-547, 2019. 543 B. ČENGIĆ, A. ĆUTUK, E. ŠATROVIĆ, N. VARATANOVIĆ, R. LINDTNER KNIFIC, B. SLAVEC L. VELIĆ and A. DOVČ laboratory of Department for Contagious charge, urinary, digestive, respiratory Diseases with Epizootiology department and reproductive problems and other for in Veterinary faculty of Sarajevo, minor health disorders. where they were frozen (-20 °C), until Presence of specifc antibodies IgG transportation and PCR processing for Cp. psitaci were conducted using in laboratory of Institute for Poultry, method of IIF. Out of 294 tested serums, Birds, Small Mammals and Reptiles in 2.04% (6/294) were positive. None of the Veterinary faculty of Ljubljana, Slovenia. six serologicaly positive animals, had any The sampling methology of the clinical manifested signs of disease. biological material, was performed in Titar of proven antibodies for Cp. accordance with the law for animal psitaci had range from 1:80 to 1:640. welfare and protection (Ofcial Gazete of In three dogs, determined titar was 1:80, the Bosnia and Herzegovina no. 316/09). in two dogs was 1:160, while in one dog determined titar was 1:640. In the seropositive dogs, chlamydial Results infection could not been confrmed using Most of the animals had relative- method of nasal swabing and molecular ly good general health status, but in detection. 17.3% certain pathological conditions RT-PCR was used for direct were observed, like eye and nose dis- measurement of chlamydia, from nasal

Table 1. Representation of seropositive and seronegative dogs according to categories. Categories Stray dogs (n=107) Pets (n=98) Working dogs (n=89) Positive dogs 3.73% (n=4) 1.02% (n=1) 1.12% (n=1) Negative dogs 96.27% (n=103) 98.98% (n=97) 98.88% (n=88)

Table 2. Number and percentage of seropositive animals to Chlamydophila psittaci according to dilutions and cities Titar IgG antibodies of Chlamydophila psittaci CITY 1:40 1:80 1:160 1:320 1:640 % positive Busovača (n=9) 0 0 0 0 1 11.1 Goražde (n=17) 0 0 0 0 0 0 Jablanica (n=8) 0 0 1 0 0 12.5 Ključ (n=11) 0 0 0 0 0 0 Konjic (n=20) 0 0 0 0 0 0 Lukavac (n=32) 0 0 0 0 0 0 Prijedor (n=11) 0 0 0 0 0 0 Sanski Most (n=23) 0 0 0 0 0 0 Sarajevo (n=59) 0 3 1 0 0 6.8 Travnik (n=48) 0 0 0 0 0 0 Tuzla (n=31) 0 0 0 0 0 0 Zenica (n=25) 0 0 0 0 0 0 TOTAL (n=294) 0 3 2 0 1 2.04

544 VETERINARSKA STANICA 50 (6), 541-547, 2019. Research of Chlamydiosis presence in dogs population in Bosnia and Herzegovina Istraživanje klamidioze u populaciji pasa lutalica u Bosni i Hercegovini

Figure 2. Photo of negative result after Figure 3. Photo of positive result after assessment of blood serums samples, using assessment of blood serums samples, using method of IIF and tied antigen of Cp. psittaci on method of IIF and tied antigen of Cp. psittaci on glass slide (microscop enlargement 400 x). glass slide (microscop enlargement 400 x). swabs, taken from animals, that were Chlamydiosis in dogs could serologicaly positive. In none of the six be described as one of the best serologicaly positive animals, nucleic „impersinator“ among infective acid of Cp. psitaci could be confrmed, diseases, given that it can look very using this method. similar to other contagious diseases, Figures 2. and 3. shows negative and which with their clinical condition positive serology results with IIF method, „mask“ noncharacteristic clinical signs using tied Cp. psitaci, antigen on slide of chlamydiosis (Liutkeviciene et al., and magnifcation 400 x. 2009). In the same manner, chlamydiosis is followed by polymorphism of clinical signs, which can result in confusion in Discussion terms of diagnostics and therapy. Because All animals as well as humans are of mentioned facts, chlamydiosis in dogs susceptible to chlamidial infections. is diferentially difcult to separate from Chlamydiosis in dogs has been rhinitis and keratoconjunctivitis. Another insufciently investigated in terms of reason for the need of fast and accurate its: pathogenesis, clinical condition, diagnosis of chlamydiosis in dogs is diagnostics, as well as contamination the lack of standardized diagnostic rates and prevalence between animal procedures, diagnostics for routine species. testing, as well as source of adequate In Europe, research of chlamydiosis information. in dogs has been conducted in a small Indirect immunofuorescence (IIF) number of countries which include is one of the serological methods for Germany, Slovakia, Sweden and diagnosing Chlamydiosis in dogs. This Lithuania (Kocianova et al., 1992; method is based on marking specifc Liutkeviciene et al., 2001; Liutkeviciene antibodies with fuorescent colour – et al., 2009; Holst et al., 2010; Takačova fuorohromom, which is covalently tied et al., 2010). Their prevalence rates were with antibodies specifc for chlamydia. As in ranges from 0% in Sweden to 38.1% in such this technique allows for recording Lithuania Research into the epizootical of the disease, without changes in its appearance of chlamydiosis have been functionality and immune specifcity conducted in Asia and North America (Owen et al., 2003). Molecular methods of with seroprevalence range from 9.4% to chlamydial diagnostics are based on use 12.1% (Fukushi et al., 1985; Arizmendi et of PCR, the most common used method al., 1992; Wu et al., 2013). for amplifcation of nucleic acids. Such

VETERINARSKA STANICA 50 (6), 541-547, 2019. 545 B. ČENGIĆ, A. ĆUTUK, E. ŠATROVIĆ, N. VARATANOVIĆ, R. LINDTNER KNIFIC, B. SLAVEC L. VELIĆ and A. DOVČ procedure is sensitive and specifc, while of this infection in wider territory of based on segmental DNA replication. Bosnia and Herzegovina. As prophylactic PCR procedure allows exact verifcation measures, increased zootechnic of nucleic acids from causative agent in procedures could be used for the purpose biological materials, taken from animals. of breeding and lodging (birds, cats, In addition to classic PCR, real-time PCR dogs), as well as routine monitoring of (RT-PCR) is also often applied. their health condition and development Considering there is an absence of of vaccine to build up immunity in mandatory control for chlamydiosis in animals. dogs, the presence of this disease had not been researched previously in Bosnia References and Herzegovina and this is the frst of its 1. ACHA, P. N. and B. SZYFRES (2003): Zoonoses kind study performed with the main aim and communicable diseases common to man and to determine the presence of chlamdial animals : Volume II. Chlamydioses, Ricketsioses, infections in the population of stray dogs and Viroses. Pan American Health Organization, Pan American Sanitary Bureau, Regional Ofce of in larger cities. The serology assessment the World Health Organization. Washington, DC. (IIF) of blood serum in dogs (n=294), 2. ARIZMENDI, F., J. E. GRIMES and R. L. RELFORD found seroprevalence of chlamydiosis (1992): Isolation of Chlamydia psitaci from to be 2.04% (6/294). Serology positive pleural efusion in a dog. Journal of veterinary diagnostic investigation : ofcial publication of the dogs however did not have clinically American Association of Veterinary Laboratory manifested signs of the disease, neither Diagnosticians, Inc. 4, 460-463. did they have signs of other contagious 3. BROWNING, G. F. (2004): Is Chlamydophila felis a signifcant zoonotic pathogen? Austral. Vet. J. 82, disease or disorders. By using molecular 695-596. diagnostics, we could not prove the 4. CHAHOTA, R., H. OGAWA, Y. MITSUHASHI, presence of chlamydia in serologically K. OHYA, T. YAMAGUCHI, T. and H. FUKUSHI positive animals. Given the negative (2006): Genetic diversity and epizootiology of Chlamydophila psitaci prevalent among the results in terms of the presence of captive and feral avian species based on VD2 region antigen, as well as the absence of clinical of ompA gene. Microbiol. Immunol. 50, 663-678. signs of the disease, we can assum that 5. CHAU, S., E. Y. TSO, W. S. LEUNG, K. S. FUNG (2015): Three cases of atypical pneumonia caused these dogs do not secrete antigen in by Chlamydophila psitaci. Hong Kong Med. J. 21, their surroundings, as they are not their 272-275. immediate source of infection. Presence of 6. CORSARO, D. and G. GREUB (2006): Pathogenic antibodies, without confrmed presence potential of novel Chlamydiae and diagnostic approaches to infections due to these obligate of antigen, leads us to assume that dogs intracellular bacteria. Clin. Microbiol. Rev. 19, 283-297. who had contact with antigen, have 7. FUKUSHI, H., H. OGAWA, N. MINAMOTO, developed immunological reaction, but A. HASHIMOTO, K. YAGAMI, H. TAMURA, S. SHIMAKURA and K. HIRAI (1985): they do not inevitably have to become ill Seroepidemiological surveillance of Chlamydia psitaci or be a threat to other animals and public in cats and dogs in Japan. Vet. Rec. 117, 503-504. health. 8. GREENE, C. E. and J. E. SYKES (2006): Chlamydial Research of chlamydial infections in Infections. In: C. E. Greene (Ed.): Infectious diseases of the dog and cat, Saunders Elsevier, St. Louis, dogs, conducted in other countries so Mo., London, pp. 246-252. far (Kocianova et al., 1992; Liutkeviciene 9. GRESHAM, A. C., C. E. DIXON and B. J. BEVAN et al., 2001; Liutkeviciene et al., 2009; (1996): Domiciliary outbreak of psitacosis in dogs: potential for zoonotic infection. Vet. Rec. 138, 622-623. Holst et al., 2010; Takačova et al., 2010) 10. GYLSTROFF, I. and F. GRIMM (1987): have covered bigger regions and larger Chlamydiales, Vogelkrankheiten, Verlag Ulmer, samples, it is therefore difcult to draw Stutgart, pp. 317-322. a parallel between our and other research 11. HOLST, B. S., S. HANAS, G. BOLSKE and C. L. FORSBERG (2010): An investigation on the across the world. However, it does leaves presence of Chlamydiaceae in Swedish dogs. Acta us enough space to continue research Vet. Scand. 16, 52-63.

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12. KOCIANOVA, E., V. LISAK and M. KOPCOK 18. PANTCHEV, A., R. STING, R. BAUERFEIND, J. (1992): Coxiella burnetii and Chlamydia psitaci TYCZKA and K. SACHSE (2010): Detection of all infection in dogs. Vet. Med. Czech 37, 177-183. Chlamydophila and Chlamydia spp. of veterinary 13. LIUTKEVICIENE, V., J. BAGDONAS, M. interest using species-specifc real-time PCR assays. STANKEVICIENE, V. BALIUKONIENE, L. Comp. Immunol. Microbiol Infect. Dis. 33, 473-484. GANUSAUSKAITE, G. GERULIS, K. MASILIONIS, A. 19. PUBLIC HEALTH ENGLAND (2014): Chlamydial STANKEVICIUS and O. AKUNYTE (2001): Serological zoonotic infections. NHS. London. investigations of chlamydiosis in Lithuanian dog’s 20. SMITH, K. A., K. K. BRADLEY, M. G. STOBIERSKI population (in Lithuanian). Vet. Zootechn. 13, 20-25. and L. A. TENGELSEN (2005): Compendium 14. LIUTKEVICIENE, V., J. MOCKELIUNIENE, J. of measures to control Chlamydophila psitaci SENGAUT, A. SALOMSKAS, M. STANKEVICIENE, (formerly Chlamydia psitaci) infection among R. MOCKELIUNAS and I. ALEKSEJUNIENE humans (psitacosis) and pet birds. J. Am. Vet. Med. (2009): Chlamydia prevalence in sick dogs with Assoc. 226, 532-539. uro-genital and/or conjunctival lesions. Lithuanian 21. SPRAGUE, L. D., E. SCHUBERT, H. HOTZEL, S. Veterinary Academy. SCHARF and K. SACHSE (2009): The detection of 15. MOULDER, J. W. (1984): Chlamydiales, In: D. Chlamydophila psitaci genotype C infection in H. Bergey, J. G. Holt, N. R. Krieg (Eds): Bergey’s dogs. Vet. J. 181, 274-279. manual of systematic bacteriology, Williams & 22. TAKAČOVA, D., M. HALANOVA, L. CISLAKOVA, Wilkins, Baltimore, pp. 729-739. D. KOVACOVA, M. HALAN and P. JARČUŠKA 16. Ofcial Gazete of the Bosnia and Herzegovina no. (2010): Long-term monitoring for ornithosis - 316/09. htp://vfs.unsa.ba/web/images/dokumenti/ psitacosis and mammals chlamydiosis in animals Zako_o_zastiti_i_dobrobiti_zivotinja.pdf in Slovakia. Acta Vet. (Beograd) 60, 241-248. 17. OWEN, W. M., C. P. STURGESS, D. A. HARBOUR, 23. WU, S. M., S. Y. HUANG, M. J. XU, D. H. ZHOU, K. EGAN and T. J. GRUFFYDD-JONES (2003): H. Q. SONG and X. Q. ZHU (2013): Chlamydia felis Efcacy of azithromycin for the treatment of feline exposure in companion dogs and cats in Lanzhou, chlamydophilosis. J. Feline Med. Surg. 5, 305-311. China: a public health concern. BMC Vet. Res. 9: 104.

Istraživanje klamidioze u populaciji pasa lutalica u Bosni i Hercegovini Dr. sc. Benjamin ČENGIĆ, dr. med. vet., dr. sc. Amel ĆUTUK, dr. med. vet., dr. sc. Edin ŠATROVIĆ, dr. med. vet., dr. sc. Nazif VARATANOVIĆ, dr. med. vet., dr. sc. Lejla VELIĆ, dr. med. vet., Veterinarski fakultet Univerziteta u Sarajevu, Bosna i Hercegovina; Renata LINDTNER KNIFIC, dr. med. vet., dr. sc. Brigita SLAVEC, dr. med. vet., dr. sc. Alenka DOVČ, dr. med. vet., Veterinarski fakultet Univerziteta u Ljubljani, Slovenija Naše istraživanje opisuje epidemiološko cegovine u suradnji odjela za zarazne bolesti prisuće klamidioze u različitim kategorijama Veterinarskog fakulteta u Sarajevu i Veteri- pasa u Bosni i Hercegovini. Problem pasa lu- narskog fakulteta u Ljubljani, Slovenija. Cilj talica koji nisu redovito pregledavani i necije- je istraživanja bio uporabom metoda serolo- pljenih jedan je od najkompleksnijih proble- gije i molekularne dijagnostike odrediti pri- ma među građanima, problema nevladinih suće klamidijalnih infekcija među različitim organizacija i institucija u Bosni i Hercegovi- kategorija pasa. Krvni serumi su uzorkovani ni. Klamidioza je zoonotska bolest koju izazi- tijekom 2012./2013. godine. Ukupno je pre- va Gram-negativna, intracelularna bakterija, gledavano 294 uzoraka na prisuće specifčnih a koja uključuje sojeve: Chlamydophilia felis, klamidijalnih protutijela metodom indirektne Chlamydophilia abortus, Chlamydophilia psitaci fuoroscence, dok je RT-PCR metoda korište- i Chlamydophilia caviae. Bolest je endemičnog na za određivanje antigena. Nakon pregleda karaktera i malo je dostupnih informacija o 294 uzoraka seruma, 2,04 % (6 uzorka) su bila prirodnim infekcijama u pasa, koje su obič- pozitivna na Cp. psitaci. Većina pozitivnih no bile povezivane s konjuktivitisom, ence- uzoraka je imala podrijetlo od uličnih pasa. falitisom i simptomima karakterističnim za Od serološki pozitivnih životinja, uzimani pneumoniju. Mali je broj europskih zemalja su brisevi nosne sluznice i pregledani upo- istraživao klamidiozu u pasa. Istraživali su rabom RT-PCR metode. Prisuće nukleinske ju: Njemačka, Slovačka, Švedska i Litva. Ovo kiseline od Cp. psitaci nije potvrđena niti u je prvo istraživanje ovog tipa, provedeno na jednoj od životinja. psima u Bosni i Hercegovini. Istraživanje je Ključne riječi: klamidioza, antigen, provedeno u dvanaest gradova Bosne i Her- protutijelo, psi, Sarajevo

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