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Laboratory Diagnostics of Selected Feline Respiratory Pathogens and Their Prevalence in the Czech Republic

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Laboratory Diagnostics of Selected Feline Respiratory Pathogens and Their Prevalence in the Czech Republic Veterinarni Medicina, 64, 2019 (01): 25–32 Original Paper https://doi.org/10.17221/93/2017-VETMED Laboratory diagnostics of selected feline respiratory pathogens and their prevalence in the Czech Republic Dana Lobova1,2*, Vendula Kleinova1, Jarmila Konvalinova3, Petra Cerna1,4, Dobromila Molinkova1,2 1Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic 2CEITEC, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic 3Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Brno, Czech Republic 4Vets Now Referral and Emergency Hospital Glasgow, Glasgow, United Kingdom *Corresponding author: [email protected] Citation: Lobova D, Kleinova V, Konvalinova J, Cerna P, Molinkova D (2019): Laboratory diagnostics of selected feline respiratory pathogens and their prevalence in the Czech Republic. Veterinarni Medicina 64, 25–32. Abstract: Respiratory problems in cats have a multifactorial character. Therapy without the detection of pathogen is often ineffective. Our study was therefore focused on the detection of important feline respiratory bacterial pathogens such as Mycoplasma felis, Chlamydia felis and Bordetella bronchiseptica and viral pathogens such as Felid alphaherpesvirus-1 and feline calicivirus. The goal of this study was to map the occurrence of these patho- gens in cat populations in the Czech Republic with the aim of introducing rapid and highly sensitive methods into routine diagnostics and to provide consulting services to animal health professionals based on the acquired data. A total of 218 cats were investigated in the study: 69 were outdoor and 149 were indoor cats. Three groups of animals were compared: up to one year of age (60 cats), one to three years of age (68 cats) and more than three years of age (90 cats). Samples were taken from conjunctiva and/or the oropharynx. Samples originated from cats with various forms of respiratory disease or from healthy cats from different parts of the Czech Republic. Real-Time RT-PCR, multiplex Real-Time PCR, nested PCR and sequencing analyses were performed. Outdoor cats were infected more often (84 detected pathogens in 69 cats) than indoor cats (110 detected infections in 149 cats). More than one pathogen was detected in a total of 38 cats, and six cats were infected with more than two pathogens. The difference was statistically significant in the case of co-infections, but not for mono-infections (P < 0.05). Kittens and young adults up to the age of one year were the most common reservoirs of respiratory infections (only 19 cats out of 60 were negative and positive cats often harboured coinfections). The difference in age groups were not statistically significant (P > 0.05). Concerning the site of the sampling, feline calicivirus, M. felis and B. bronchiseptica were detected more often from oropharynx than from conjunctival swabs. M. felis was slightly more common in clinically diseased animals (39.6%) than in healthy ones (26.1%). The obtained results reveal the frequency of individual pathogens and their co-infections in cats kept on the territory of the Czech Republic, data which can be used to make the treatment of respiratory infections and breeding measures more effective. Therefore, the diagnostic methods are now available to veterinary surgeons with the possibility of con- sultation and discussion of the results. Keywords: respiratory disease complex; polymerase chain reaction; diagnostics; Mycoplasma felis; Chlamydia felis; Bordetella bronchiseptica; Felid alphaherpesvirus-1; FHV-1; feline calicivirus; FCV Supported by the Internal Grant Agency (IGA) VFU Brno, Czech Republic (Project No. IG161242). 25 Original Paper Veterinarni Medicina, 64, 2019 (01): 25–32 https://doi.org/10.17221/93/2017-VETMED Respiratory infections in cats are considered to in cats (Pudjiatmoko et al. 1997). Using other DNA be multifactorial diseases with nonspecific clinical analyses, a highly conserved gene for the ompA signs that very often present as conjunctivitis, in- protein (major outer membrane protein) has been flammation of the upper respiratory tract and less discovered among the different isolates of C. felis commonly also of the lower respiratory tract. They (Di Francesco and Baldelli 2002). are characterised by serous to mucopurulent dis- The disease is often complicated by co-infections charge from conjunctiva or the upper respiratory with other pathogens such as FCV, which causes tract (Konvalinova et al. 2016). The feline respiratory ulceration of the oral cavity together with conjunc- disease complex is thought to be mainly caused by tival infections. Feline immunodeficiency virus feline herpesvirus (Felid alphaherpesvirus 1, FHV- often complicates recovery and prolongs therapy. 1) and feline calicivirus (FCV). The main bacterial Moreover, Mycoplasma felis and Bordetella bron- pathogens involved in feline respiratory disease chiseptica are also involved in the worsening of complex are Mycoplasma felis, Chlamydia felis and clinical signs. The prevalence of C. felis in asymp- Bordetella bronchiseptica (Sykes 2014; Litster and tomatic cats is very low (< 5%) (Di Francesco et Leutenegger 2015). Some of these pathogens have al. 2004). C. felis is sensitive to tetracycline anti- zoonotic and epidemiological potential (B. bron- biotics, erythromycin, rifampicin, fluoroquinolo- chiseptica, M. felis, C. felis), mainly in immunosup- nes and azithromycin. Doxycycline is the drug of pressed individuals (Hartley et al. 2001). choice but is contraindicated in pregnant queens Mycoplasma felis (M. felis), class Mollicutes, and kittens. The immune response after C. felis family Mycoplasmataceae, genus Mycoplasma, infection is usually short and weak, and there are is one the most common pathogens of the upper different types of monovalent and polyvalent vac- respiratory tract in cats. Most feline-associated cines. Bordetella bronchiseptica (B. bronchiseptica), mycoplasma species are common microflora of class Betaproteobacteria, family Alcaligenaceae, ge- conjunctiva and the upper respiratory tract, and nus Bordetella is a strictly aerobic Gram-negative, they usually exercise pathogenicity as a secondary motile, rod-shaped bacteria. Clinical and experi- source of infection in co-infection with other path- mental studies have shown that B. bronchiseptica ogens of the upper respiratory tract. It is disputable is a primary pathogen of the respiratory tract of if Mycoplasma felis is only present in the upper cats (Hoskins et al. 1998). Bordetella bronchisep- respiratory tract as a secondary pathogen; how- tica is sensitive to doxycycline and other possible ever, in the lower respiratory tract, M. felis causes drugs of choice are co-trimoxazole, enrofloxacin, primary infection resulting in the inflammation pradofloxacin and cefovecin. It is also possible to of the lungs and pleural space. However, M. felis use macrolide antibiotics such as erythromycin has not been isolated from the lower respiratory or clarithromycin; however, these drugs are not tract in healthy individuals (Lee-Fowler 2014). A licensed for use in cats (Konvalinova et al. 2016). vaccination against M. felis infections has not yet Live intranasal vaccine (Nobivac BB) can be used been developed, and, therefore, therapy is based on as prophylaxis, mainly in animals in shelters. prolonged antibiotic treatment and on the improve- Felid alphaherpesvirus 1, feline herpesvirus also ment of cat welfare. The antibiotics of choice for M. known as feline rhinotracheitis virus (FHV-1) be- felis infections of the conjunctiva are tetracyclines longs to family Herpesviridae, (enveloped DNA such as eye ointments containing oxytetracycline, viruses), subfamily Alphaherpesvirinae, genus chloramphenicol or fluoroquinolones. Varicellovirus, and represents a typical alphaher- Chlamydia felis (C. felis) belongs to class Chla- pesvirus. Clinical signs are most obvious during mydiales (family Chlamydiaceae, genus Chlamydia) the primary infection of kittens. The disease is which number among the obligatory intracellular also described as “cat flu” and can reoccur and af- bacteria with a unique developmental cycle, and fect one or both eyes (Stiles 2014). Treatment of the pathogenesis of C. felis infections in cats is still disease caused by FHV-1 is mainly symptomatic, not fully clear. Serological studies have shown the even though there are studies showing promising existence of a few different strains of C. felis with results with the use of immunomodulators (Fiorito differing virulence (Kuroda-Kitagawa et al. 1993). et al. 2016) and antivirals (Thomasy et al. 2016). DNA fingerprinting has been used to reveal the Felid alphaherpesvirus 1 is commonly present in existence of two important strains of this species most polyvalent vaccines together with feline par- 26 Veterinarni Medicina, 64, 2019 (01): 25–32 Original Paper https://doi.org/10.17221/93/2017-VETMED vovirus and FCV, but no vaccine can protect the cats. Three groups of animals were compared: animal from infection and they only help to reduce up to one year of age (60 cats), one to three years its consequences. of age (68 cats) and more than three years of age Feline calicivirus (FCV) is small non-enveloped (90 cats). A total of 177 oropharyngeal swabs and RNA virus belonging to family Caliciviridae, genus 70 conjunctival swabs were collected. In 189 cats, Vesivirus. A simple genome consisting of single- we swabbed only one site, while in 29 cats both sites stranded
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