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PREVALENCE AND MOLECULAR CHARACTERIZATION OF CHLAMYDIA SP. IN FREE-RANGE POULTRY FLOCKS IN CROATIA AND IN THE PHILIPPINES

The study aimed to assess the prevalence and perform molecular characterization of bacteria genus Chlamydia especially C. psittaci, C. gallinacea and C. avium in free-range poultry raised in Croatia and in the Philippines, and assess environmental contamination specifically in water and litter/soil. Altogether 70 samples from Croatia (54 flocks) and from Bohol, Philippines (16 flocks) were included in this study. Collected samples were triple swabs (conjunctival, pharyngeal and cloacal), litter- feces and water. Detection at Chlamydiaceae and species-specific level was done through quantitative polymerase chain reaction (qPCR) assay targeting 23S rRNA, incA and enoA genes revealed that flock prevalence rate in Croatia for C. psittaci and C. gallinacea and non- classified avian Chlamydia (NCC) was 7.41% (4/54), 46.30% (25/54) and 9.26% (5/54), respectively. For Philippines, the prevalence rate was 75 % (12/16) for C. gallinacea, 18.75% (3/16) for NCC and zero for C. psittaci and C. avium. Isolate HR 425/13 clustered with C. psittaci CP3, genotype B based on ompA gene sequence. For C. gallinacea, isolates HR 623/13, HR 7/16 and HR 11/16 clustered with C. gallinacea 08-1274/3 isolated from a chicken in France. The isolates-PH 11/17 and PH 21/17, grouped with Chlamydia isolate 11-2521 X001 from China and with Chlamydia isolate from France, respectively. Isolate PH 2/17 grouped with Chlamydiae bacterium clone d98 while PH 9/17 clustered with Criblamydia sequanensis strain CRIB-18, based on 16S rRNA gene sequence. The presence of Chlamydiaceae in water samples was significantly associated with the presence of the organism in chickens (p=.049). Handwashing before and after handling chickens and coughing experienced by farmworkers for the past six months at the time of survey were significantly associated with the presence of C. gallinacea in the flock. Certainly, C. gallinacea is the most prevalent avian Chlamydia species found in free-range chicken flocks in Croatia and in Bohol, Philippines. Chlamydia-like organisms were also identified, thus supporting the concept of high diversity among chlamydia group. It is highly important to implement effective biosecurity and hygienic measures in the farm and increase awareness on avian chlamydiosis especially among farmworkers and medical practitioners considering the zoonotic potential of avian chlamydia.

Chlamydia sp., avian chlamydiosis, free-range chicken, qPCR, phylogenetic

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