Research of Chlamydiosis Presence in Dogs Population in Bosnia and Herzegovina
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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