Serological evidence of canine respiratory virus infections in shelter dogs in Yozgat in TürkiyeABSTRACT
This study aimed to determine the seroprevalence of Canine Adenovirus (CAV), Canine Parainfluenza Virus (CPIV), Canine Influenza Virus (CIV), Canine Herpesvirus 1 (CHV-1), and Canine Distemper Virus (CDV) infections among shelter dogs in Yozgat Province, Türkiye. A total of 182 blood samples were collected from dogs housed in shelters located in Yozgat city center and the districts of Sorgun and Sefaatli. The sera were tested using commercial ELISA kits to detect antibodies against CAV, CPIV, CIV, CHV-1, and CDV. Among the samples analyzed, 28 (15.38%) were seropositive for CAV, 17 (9.34%) for CPIV, 16 (8.79%) for CIV, 10 (5.4%) for CHV-1, and 13 (7.14%) for CDV. Breed-based evaluation was not performed due to the inclusion of mixed-breed dogs of various genders and ages. The findings indicate that these viral infections are present at varying levels among shelter dogs in Yozgat Province. The study highlights that CAV had the highest seroprevalence, followed by CPIV, CIV, CDV, and CHV-1. In conclusion, these results demonstrate that the examined canine viruses are circulating in the region, and further studies involving larger populations are needed to better understand their epidemiology and to raise awareness among local veterinarians.
Keywords:
canine infectious respiratory disease complex; dog; ELISA; respiratory viruses; seroprevalence
RESUMO
Este estudo teve como objetivo investigar a soroprevalência das infecções por adenovírus canino (CAV), vírus parainfluenza canino (CPIV), vírus da influenza canina (CIV), herpesvírus canino tipo 1 (CHV-1) e vírus da cinomose canina (CDV), em cães de abrigos na província de Yozgat. Coletaram-se 182 amostras de sangue de cães alojados no abrigo central de Yozgat, bem como nos abrigos localizados nos distritos de Sorgun e Sefaatli. As amostras foram analisadas utilizando-se kits comerciais de ELISA para a detecção de CAV, CPIV, CIV, CHV-1 e CDV. Os resultados mostraram que 28 das 182 amostras de soro (15,38%) foram positivas para CAV, 17 (9,34%) para CPIV, 16 (8,79%) para CIV, 10 (5,4%) para CHV-1 e 13 (7,14%) para anticorpos contra CDV. Não foi realizada avaliação baseada em raça, devido à participação de animais de diferentes raças e sexos no estudo. Concluiu-se que as prevalências das infecções por CAV, CPIV, CIV, CHV-1 e CDV em cães de abrigos na província de Yozgat foram de 15,38%, 9,34%, 8,79%, 5,4% e 7,14%, respectivamente, indicando que esses vírus estão amplamente circulando na região. Recomenda-se a realização de estudos adicionais com populações maiores, a fim de compreender melhor as infecções causadas por esses vírus, e que os médicos-veterinários da região sejam informados sobre essas doenças.
Palavras-chave:
complexo de doenças respiratórias infecciosas caninas; cão; ELISA; vírus respiratórios; soroprevalência
INTRODUCTION
Canine Infectious Respiratory Disease Complex (CIRDC) (previously referred to as infectious tracheobronchitis or Kennel Cough) arises from the involvement of various pathogens, either individually or in combination. CIRDC is an endemic syndrome worldwide, primarily caused by viruses such as Canine Adenovirus type 2 (CAV-2), Canine Parainfluenza Virus (CPIV), Canine Influenza Virus (CIV), Canine Herpes Virus 1 (CHV-1), Canine Distemper Virus (CDV), as well as bacteria such as Bordetella bronchiseptica, Streptococcus zooepidemicus, and Mycoplasma spp. (Mochizuki et al., 2008; Kawakami et al., 2010; Day et al., 2020; Reagan and Sykes, 2020; Berliner, 2021). This disease complex, reported globally, is more frequently observed in environments where large dogs are housed together, such as animal shelters, boarding facilities, and daycare centers (Barrell et al., 2010; Anderson et al., 2012; Mitchell et al., 2017). In this study, antibodies specific to the major viral pathogens causing CIRDC were investigated. Additionally, the zoonotic potential of most of the viruses examined in this study adds to its significance.
To effectively combat viral infections in a region, it is essential first to determine the presence and prevalence of viruses in that area. To enable advanced virological research and to facilitate eradication efforts, the present study investigated the seroprevalence of CAV, CPIV, CIV, CHV-1, and CDV infections. Numerous seroprevalence studies on these infections have been conducted worldwide and continue to be reported (Ellis et al., 2011; Wright et al., 2013; Krogenæs et al., 2012; Kwasnik et al., 2020; Ruffalo et al., 2020; Joonè et al., 2024; Allen et al., 2025; Ramos-Martínez et al., 2025; Wheelhouse et al., 2025). Several seroprevalence studies on these viruses have also been carried out in Türkiye (Gur and Acar, 2007, 2009; Yesilbag et al., 2012; Bulut et al., 2013; Sahna and Aslan, 2014, 2015a; Yapici et al., 2018; Sayin, 2019; Yildirim et al., 2023). However, to the best of the authors’ knowledge, no study has simultaneously investigated all five viruses. Moreover, no research has specifically focused on the Yozgat region. It is worth emphasizing that, in Türkiye, CIV and CPIV infections have each been the subject of only one seroprevalence study (Gur and Acar, 2008; Aydin et al., 2018). Previous studies have demonstrated that CIV can be transmitted to humans (Su et al., 2019; Jurado-Tarifa et al., 2020). Given its zoonotic potential, further research on this infection is urgently needed.
Considering the geographical location of Yozgat within Türkiye and its role as a transit hub, the city is thought to play an important role in the spread of viral pathogens. The findings of this study will contribute valuable data to the literature regarding the prevalence of five different viral agents.
ETHICS ASPECTS
The research was submitted to the Ethics Committee on Animal Use of the University Erciyes, and approved under the number No. 22/039.
MATERIALS AND METHODS
A total of 182 blood samples were collected from three different shelters in Yozgat province between August 2022 and August 2024. Due to the relatively lower number of animals in shelters in Yozgat compared to other provinces, the study focused on sample size rather than considering parameters such as breed, gender, and age to achieve the main objective of the study.
The collected blood samples were centrifuged at 3000 rpm for 10 minutes, and the separated serum was stored in stock tubes at -20°C until testing. The obtained serum samples were analyzed using specific ELISA antibody test kits (AFG Bioscience ELISA) for five different viruses (CPIV, CIV, CAV, CHV-1, CDV) following the instructions provided by the manufacturer (Pratelli et al., 2002; De Benedictis et al., 2010; Ellis et al., 2011; Yesilbag et al., 2012; DiGangi et al., 2019). The plates used for the analysis were read at a wavelength of 450nm using an ELISA reader, and the results were evaluated according to the kit's assessment procedure.
Statistical analyses were performed using SPSS-25. The seroprevalence of each virus from the Yozgat Central Shelter was calculated as frequency and percentage, supported by 95% confidence intervals (CI). The Chi-square test was used to compare the differences between the seropositivity rates of the five viruses. Statistical significance was set at p<0.05.
RESULTS
The seroprevalence of five different viruses (CPIV, CIV, CAV, CHV-1, CDV) was studied in the serum of 182 dog blood samples collected from three shelters in Yozgat province. According to the indirect ELISA results performed with the serum obtained from the blood samples, 28 of the 182 blood sera were positive for CAV (15.38%), 17 for CPIV (9.34%), 16 for CIV (8.79%), 10 for CHV-1 (5.4%), and 13 for CDV (7.14%) antibodies. Statistical analysis revealed that the seroprevalence of CAV (15.38%) was significantly higher than that of CHV-1 (5.49%) (p<0.05). No statistically significant differences were observed between the prevalence rates of the other viruses (CPIV, CIV, CDV) (p>0.05). The detailed seroprevalence data and antibody distribution for these viruses are presented in Table 1.
DISCUSSION
In this study, specific antibodies against the primary viral pathogens causing CIRDC were investigated. In recent years, research on viral agents has gained momentum. In this context, the seroprevalence of the five main viruses affecting dogs in Yozgat province, for which no data had previously been entered, was studied. Having foundational data, like those provided in this study, is crucial for conducting further research.
The first study on CAV in Turkey was conducted in 1975 (Okuyan, 1975). Until 2018, studies have focused on seroprevalence and prevalence (Gur and Acar, 2009; Yildirim et al., 2009; Bulut et al., 2013; Sahna and Aslan, 2015a, 2015b). Since 2018, molecular analyses have been used in studies, and partial sequences obtained from some studies have been registered in the GenBank database (Timurkan et al., 2018; Dincer et al., 2020; Sarac et al., 2021; Abayli et al., 2023). As observed, the number of studies conducted is quite limited. According to previous seroprevalence studies, CAV-specific antibodies have been detected in various provinces at rates ranging from 14% to 87.2%. In this study, 28 out of 182 dogs (15.38%) were positive for CAV-specific antibodies (Table 1). The seroprevalence rate of 15.38% obtained in this study falls within the range of 14% to 87.2% previously detected, supporting the results of other studies. Considering the current situation in our country, it is recommended that further studies be urgently initiated to control this virus, which has a high seroprevalence. Additionally, it is advised that vaccination programs be restructured and dogs be vaccinated against this virus.
Only two studies on CPIV have been conducted previously in Turkey. Only one of these is a seroprevalence study. A seropositivity rate of 46.8% was detected (Gur and Acar, 2008). The other study was antigen-based, but no positive results were obtained (Avci et al., 2013). Given that this is the third study on this topic in Turkey and the first conducted in Yozgat province, the data obtained from this study are of significant importance. The seropositivity rate found in this study was determined to be 9.34% (Table 1). Since there is only one study in our country that has obtained findings other than this study, it is necessary to continue studies in this field and to determine programs to combat the virus according to the situation in our country.
To date, only one study has been conducted on the CIV virus in our country. In this study, which first identified the presence of CIV, a seropositivity rate of 5.8% was found in Erzurum province (Aydin et al., 2018). In this study conducted in Yozgat province, a seropositivity rate of 8.79% was observed (Table 1). The fact that so few studies have been conducted on this topic, which contains zoonotic information, is highly insufficient. It is of great importance that necessary studies on the prevalence of this virus be carried out immediately and routinely. If possible, research should be conducted nationwide.
One of the other viruses targeted in this study is CHV-1. To date, a total of five studies have been conducted on this topic in Turkey, with seropositivity rates ranging from 25.2% to 68.79% (Gur and Acar, 2007; Yesilbag et al., 2012; Sahna and Aslan, 2014; Yapici et al., 2018; Yildirim et al., 2023). According to the results of this study, the lowest seropositivity rate of 5.4% was determined (Table 1). Considering that Yozgat province is relatively smaller and has less animal movement compared to the other provinces where the studies were conducted, this result is considered normal. Despite the low rate, eradication efforts should be made to prevent the continued presence of the virus. Since this virus is transmitted through mating, it is recommended that, in addition to vaccination, sterilization activities be carried out with care and precision.
Due to its high morbidity and mortality, there are many studies on CDV in our country (Esin, 2013; Sayin, 2019; Saltik and Atli, 2023; Saltik and Kale, 2023). However, no studies on this virus have been conducted in Yozgat province previously. Through this study, it has been determined that CDV has a seroprevalence of 7.14% in Yozgat (Table 1). To control this widely prevalent infection in our country, vaccination is recommended.
CONCLUSIONS
In this study, seroprevalence positivity rates of 15.38% for CAV, 9.34% for CPIV, 8.79% for CIV, 5.4% for CHV-1, and 7.14% for CDV were detected in shelter dogs in Yozgat province (Table 1). Based on the findings obtained from this study, it is suggested that continuous monitoring of the five viruses studied (CPIV, CIV, CAV, CHV-1, CDV) for control and eradication efforts, conducting periodic seroepidemiological studies, and particularly increasing vaccination efforts could be beneficial in the fight against these viruses. The persistence of these viruses in the environment always poses an infection risk. For this reason, it is important to focus on studies in virology to facilitate the fight against viruses.
ACKNOWLEDGEMENTS
Thanks to the Yozgat Bozok University Scientific Research Projects Coordination Unit. We are grateful to all the researchers who contributed to this study.
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The research data are available within the article itself.
