Rural dog populations have long been recognized to be inadequately managed in terms of disease control and prevention. In this study we consider dog management in rural Shanghai and its implications for rabies control in the entire metropolitan area of Shanghai. The prerequisite to improve rabies vaccination coverage in rural Shanghai depends on a proper enumeration of the total rural dog population. In this study we selected one of the nine administrative districts in Shanghai (Jiading), within which there are 7 towns and 2 industrial zones (township-level division) that contain agricultural areas. A total of 9 villages (rabies model villages) were chosen from each township-level division in Jiading, and an additional 3 non-model villages were also included in the study. A household questionnaire survey was implemented in all 12 villages recruited. In 3 of the model villages and the 3 non-model villages chosen as a comparison, two methods of enumeration—a sight-resight survey and a household census survey—were implemented. Results from the household survey in these 6 villages showed that among the total 1,560 owned dogs, 80.4% were Chinese Garden Dogs, 69.1% were aged 1 to 3 years, 49.2% were homebred, and 88.3% were kept for the purpose of guarding the house. However, only 3.7% of the owned dogs were desexed. There was a higher proportion of chained or confined dogs in model compared to non-model villages. The model villages had an absolute rabies vaccination coverage of 100% among its owned dog population and a smaller number of stray dogs. It was also identified that the two enumeration methods yielded similar counts (P = 0.12), particularly within smaller villages. From the questionnaire survey implemented within all 12 villages and based on the average human-to-dog ratio, the total rural dog population of Jiading district was estimated to be 24,058. This study generated information on the general demographics of the rural dog population in Jiading, and demonstrates an approach to the study of rural dog populations within the context of a megacity. In such a context, rural dog populations need to be considered as a critical component of animal and public health.

Urban stray cats are cats without owners that survive in the wild for extended periods of time. They are one of the most common stray animals in cities, and as such, monitoring the pathogens carried by urban stray cats is an important component of urban epidemiological surveillance. In order to understand the prevalence of respiratory diseases in urban stray cats in Shanghai and provide scientific evidence for the development of targeted prevention and control strategies for respiratory diseases in stray cats, we collected 374 ocular, nasal, and oropharyngeal swabs from urban stray cats in Shanghai from January 2022 to December 2022. After RNA extraction, we used real-time PCR to detect six respiratory pathogens, including influenza A virus, feline calicivirus, feline herpesvirus type 1, Mycoplasma, Chlamydia, and Bordetella bronchiseptica. The results showed that among the 374 samples, 146 tested positive, with a positivity rate of 39.04%. The highest positivity rate was observed for Mycoplasma felis at 18.72% (70/374), followed by Chlamydia felis at 11.76% (44/374), feline calicivirus at 3.74% (14/374), feline herpesvirus 1 at 3.48% (13/374), Bordetella bronchiseptica at 1.34% (5/374), and influenza A virus was not detected. The highest positivity rate for Mycoplasma felis was in Minhang District at 31.94% (23/72), while Chlamydia felis and Bordetella bronchiseptica had the highest positivity rates in Jiading District at 23.53% (8/34) and 5.88% (2/34), respectively. The highest positivity rates for feline calicivirus and feline herpesvirus 1 were both observed in Qingpu District, at 14.46% (12/83) and 9.64% (8/83), respectively. A total of 36 samples showed mixed infections with two or more pathogens, with Mycoplasma felis being involved in 32 of these mixed infections, with the highest number of mixed infections being with Chlamydia felis at 25 samples. Respiratory pathogen positivity was detected throughout the year, with peak detection rates in summer and winter. The positivity rates of cat respiratory pathogens in different seasons showed statistical differences (χ² = 27.73, p < 0.01). There was no statistical difference in the positivity rates of respiratory pathogens between cats of different genders (χ² = 0.92, p > 0.05). The positivity rates of respiratory pathogens in cats of different age groups showed statistical differences (χ² = 44.41, p < 0.01). Mycoplasma felis and Chlamydia felis were the main pathogens causing respiratory infections in stray cats, with Mycoplasma felis showing a much higher positivity rate than other respiratory pathogens and often co-infecting with Chlamydia felis and feline calicivirus. The positivity rate of Mycoplasma felis was high in summer, autumn, and winter, with no statistical difference between seasons. These results indicate a serious overall prevalence of respiratory pathogens in urban stray cats in the Shanghai area, showing seasonal trends and mixed infections with other pathogens. These findings suggest the need for comprehensive prevention and control measures to address respiratory pathogen infections in urban stray cats in the Shanghai area.

Chlamydia felis is an important zoonotic agent for humans and various animals. A recombinase-aided amplification (RAA) assay was developed for detecting C. felis. RAA can be performed in a closed tube at 39°C within 30 min. The detection limit was 10.6 copies of the C. felis plasmid DNA per reaction. No positive signals for other pathogens were detected. The coincidence rate of RAA and conventional PCR was 95.24% (20/21) and 100% (96/96) for positive and negative samples, respectively. The established RAA assay is a simple, rapid, highly sensitive, and specific method for detecting C. felis.