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Impact of Way of Life and Environment on the Prevalence of Chlamydia Felis in Cats As Potentional Sources of Infection for Humans

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Impact of Way of Life and Environment on the Prevalence of Chlamydia Felis in Cats As Potentional Sources of Infection for Humans Annals of Agricultural and Environmental Medicine 2019, Vol 26, No 2, 222–226 www.aaem.pl ORIGINAL ARTICLE Impact of way of life and environment on the prevalence of Chlamydia felis in cats as potentional sources of infection for humans Monika Halánová1,A,C-F , Lenka Petrová1,A-D,F , Miloš Halán2,B-C,E , Alexandra Trbolová2,A,C,E , Ingrid Babinská1,C-D,F , Tatiana Weissová2,B-C 1 Faculty of Medicine, Pavol Jozef Šafárik University, Košice, Slovakia 2 University of Veterinary Medicine and Pharmacy, Košice, Slovakia A – Research concept and design, B – Collection and/or assembly of data, C – Data analysis and interpretation, D – Writing the article, E – Critical revision of the article, F – Final approval of article Halánová M, Petrová L, Halán M, Trbolová A, Babinská I, Weissová T. Impact of way of life and environment on the prevalence of Chlamydia felis in cats as potentional sources of infection for humans. Ann Agric Environ Med. 2019; 26(2): 222–226. doi: 10.26444/aaem/100655 Abstract Introduction and objectives. Chlamydia (C.) felis can cause infection which may be associated with conjunctivitis and/or respiratory tract disease, particularly in kittens, but could also be the cause of the disease in adult cats. Infection is more common in multi-cat environments. The zoonotic potential of C. felis appears low, but exposure to this microorganism is possible by handling the affected cats, by contact with their aerosol, and also via fomites. Materials and method. In the study, 140 cats of various breeds from Košice region in Slovakia were studied. Conjunctival samples were obtained from 71 clinically healthy cats (50.7%) and 69 cats with clinical signs of conjunctivitis and upper respiratory tract impairment (49.3%). Cats were divided into 4 groups according to breed and type of environment in which they lived. In the 1st group were cats kept inside only (n=33), in the 2nd group, free-roaming cats (n=50), the 3rd group comprised stray cats, taken from the streets (n=28), and the 4th group included cats kept in shelters or deposit devices (n=29). Molecular method PCR and DNA sequencing was used as the diagnostic method. Results. Overall positivity was 17.1%. Of the 24 positive cats, the highest positivity was detected in the population of stray cats (35.7%) and shelter cats (31%). In the group of free-roaming cats, 10% had positivity. No positive animals were detected in the group of cats kept inside only. It was also found that the risk of C. felis in cats with clinical signs of disease was more than 7-fold higher than in cats without clinical signs of conjunctivitis and respiratory tract. Conclusion. The obtained results show that cats, especially stray and shelter cats, can be important sources of feline chlamydiosis, and due to their close contact with people they can present a risk for transmission. Key words Chlamydia felis, cat, prevalence, zoonosis, PCR, public health INTRODUCTION especially in the case of emerging, respectively, re-emerging diseases. The interaction between human and animal health is not a One of the most common microorganisms in nature new phenomenon. However, the scope, scale, and worldwide which have a zoonotic character are chlamydiae from the impact of zoonoses we are facing today have no historical family Chlamydiaceae. Chlamydiaceae has only a single precedent. Of the 1,415 identified infectious pathogens for genus, Chlamydia that comprises 12 species [3, 4], including humans 868 (61%) have a zoonotic character [1], and these Chlamydia felis. Chlamydia felis is an obligate intracellular diseases have an increasing trend, not only in developing bacterium that seems to have a predilection for conjunctival but also in developed countries. The ecological changes epithelial cells, and is the most common cause of feline caused due to the destruction of the natural environment chlamydiosis and different diseases in humans. increase the exposure of animals and humans to the vectors Feline chlamydiosis is the most common cause of and reservoirs of unknown pathogenic microorganisms. In pneumonia and acute or chronic conjunctivitis, particularly addition to this, weak public health systems, other natural in kittens [5, 6, 7], but could also be the cause of the disease disasters, absence of better surveillance and controlling in adult cats [8]. Clinical signs of infection with C. felis in systems against the disease and unhygienic living conditions cats are sneezing, transient fever, inappetence, weight lost, are the main factors for the emergence and re-emergence of nasal discharge, vaginal discharge and lethargy. Chlamydial the bacterial zoonotic diseases in many countries worldwide disease may be complicated by coinfection with other [2]. The rapid detection and response to diagnostic diseases microorganisms. Feline chlamydiosis is more common in is crucial, because on many occasions there is an unobserved multi-cat environments, particularly breeding catteries spreading and infiltration of diseases to new territorial units, where animals share a common space [9]. C. felis spreads through direct contact between cats, and cats and humans. Transmission between cats is due to direct Address for correspondence: Lenka Petrová, Pavol Jozef Šafárik University in Košice, Faculty of Medicine, Košice, Slovakia contact with infectious eye secretions. This pathogen is highly e-mail: [email protected] unstable outside the host and remains infectious for a short Received: 05.11.2018; accepted: 11.12.2018; first published: 09.01.2019 time in the environment. Annals of Agricultural and Environmental Medicine 2019, Vol 26, No 2 223 Monika Halánová, Lenka Petrová, Miloš Halán, Alexandra Trbolová, Ingrid Babinská, Tatiana Weissová. Impact of way of life and environment on the prevalence… The zoonotic potential of these bacteria seems to be 300μl saline solution for 60 seconds. Samples thus obtained low; however, C. felis may be exposed to the manipulation were stored at -80 °C until testing. of infected cats. In particular, cat owners and employees working with cats (veterinarians, breeders‘staff) are at Molecular analysis. For isolation of the DNA, samples increased risk, especially where poor hygiene conditions resulting from mixing 150μl samples from both eyes were are present. The risk of zoonotic transmission is likely to used. DNA isolation was carried out according to the be greater in immunocompromised individuals [10, 11, 12, manufacturer‘s instructions using a commercial diagnostic 13, 14]. In general, acute conjunctivitis is a frequent human kit DNA-Sorb/AM (Nucleic Acid Extraction Kit, AmpliSens, condition, with an estimated six million cases per year in the Federal State Institution of Science, Moscow, Russia). In USA, and Chlamydiae being the cause in 1.8–5.6% of all cases cases where the amplification was not been carried out [15]. However, only few human cases of C. felis conjunctivitis immediately, the samples were stored at -20 °C until their have been reported, despite its high incidence in cats and the next use. Positive specimens for Chlamydiaceae spp. were large number of pet cats in society. Actual prevalence may subsequently amplified using a set of primers U23Fa (5‘-GAT be much higher because human infection is predominantly GCC TTG GCA TTG ATA GGC GAT GAA GGA-3‘) and asymptomatic [16]. 23 SIGR (5‘-TGG CTC ATC ATG CAA AAG GCA-3‘) that amplify domain I of 23S rRNA gene. The reaction mixture for one reaction was prepared by mixing 0.2 μl 10 pmol of primer OBJECTIVE U23Fa, 0.2 μl 10 pmol of primer 23 SIGR, 9.6 μl of purified water, 4 μl of Taq polymerase (HOT FIREPol Blend Master Because the control of incidence of chlamydial infections Mix, Solis BioDyne, Estonia), and 6 μl of template (isolated between animals and humans is the key to a better DNA). Electrophoresis was carried out for 20 minutes at understanding of the relationship between chlamydiae 300V. PCR products were analysed by electrophoresis of 10 µl and their hosts, the aim of this study was to investigate the of each 20 µl reaction mixture on a 1.5% agarose gel stained incidence of C. felis in cats, according to breeding and the using GoldView stain (SBS Genetech, China) and visualized type of environment in which they live, and to compare the fragments were compared with the positive control and the incidence in cats with and without clinical signs of infection, 100 bp DNA ladder. and assess the zoonotic potential. Positive products Chlamydiaceae spp. were compared against the positive control for the length of 600bp under UV light at a wavelength of 254nm. The identity of the MATERIALS AND METHOD obtained sequences was examined by a BLAST search [17]. DNA sequence alignments and phylogenetic analysis Samples were collected from cats examined at the Department were conducted using software MEGA5. Phylogenetic of Small Animals at the University of Veterinary Medicine trees were created using alignments performed with the and Pharmacy in Kosice, Slovakia, and various deposit BioEditSequence Aligment Editor as a distance method and centres (shelters) in Kosice, Slovakia. 140 cats of various NJ (neighbour joining) as the tree construction method. breeds, aged from 1 month to 10 years were examined. Most All ambiguous positions were removed from each sequence of the examined cats were European shorthair cat breeds pair. The reliability of branches in trees was assessed using (ESH); lesser represented breeds were Maine Coon, British bootstrap analysis with 1,000 pseudoreplicates, with values blue and Sphinx. Based on the type of environment, the above 70% being noted. animals were divided into 4 groups for examination: Analysis of sequences for constructing the phylogenetic 1) cats kept inside human habitations, without possible tree also included 9 reference strains, the partial sequence of contact with other cats and other animals. A requirement the 23S rRNA gene of different chlamydial species from the for inclusion in this group was also internal breeding.
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