Household Pets as Reservoirs of Zoonotic Pathogens: Prevalence, Risk Factors, and Prevention Strategies within a One Health Framework

63 Nadar Khan1, Attiq Ur Rehman2*, Hina Ali Ahmed3, Usman Khalid4, Muhammad Faizan Elahi Bhatti5, Shahzada Khurram Adrian Shah6, Hafiz Atif Khurshid7, Hina Faiqa8, Sharoz Munawar9, Saba Ashraf10 https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) ISSN Online: 3007-1941 ISSN Print: 3007-1933 Household Pets as Reservoirs of Zoonotic Pathogens: Prevalence, Risk Factors, and Prevention Strategies within a One Health Framework Article Details A B S T R A C T Keywords: Zoonotic Diseases, Companion Animals, Campylobacter Spp., Toxocara Canis, Salmonella Spp, Hygiene Practices, Vaccination, One Health, Public Health, Risk Factors Nadar Khan Department of Livestock and Dairy Development, Khyber Pakhtunkhwa, Pakistan Attiq Ur Rehman* Department of Broiler Production, Season Foods Supreme Farms, Pakistan Email: d[email protected]m Hina Ali Ahmed Department of Zoology, Faculty of Life Sciences, Sardar Bahadur Khan Women's University, Quetta, Baluchistan, Pakistan Usman Khalid Oklahoma State University, College of Veterinary Medicine, Stillwater Oklahoma, USA Muhammad Faizan Elahi Bhatti Department of Epidemiology and Public Health, University of Veterinary and Animal Sciences, Lahore, Punjab, Pakistan Shahzada Khurram Adrian Shah Department of Clinical Studies, Faculty of Veterinary Sciences, The University of Veterinary and Animal Sciences (UVAS), Swat, Khyber Pakhtunkhwa, Pakistan Hafiz Atif Khurshid Department of Veterinary Pathology, University of Veterinary and Animal Sciences, Lahore, Punjab, Pakistan Hina Faiqa Department of Epidemiology and Public Health, University of Veterinary and Animal Sciences, Lahore, Punjab, Pakistan Sharoz Munawar Department of Veterinary Medicine, University of Veterinary and Animal Sciences, Lahore, Punjab, Pakistan Saba Ashraf1 Livestock and Dairy Development Department, University of Veterinary and Animal Sciences, Lahore, Punjab, Pakistan Zoonotic diseases spread by domestic animals pose an increasing threat to the global One Health framework. This cross-sectional study examined the frequency of significant zoonotic diseases in domestic animals and identified the related behavioral and environmental risk factors in pet-owning households. Three hundred families possessing dogs, cats, or small mammals were surveyed using structured questionnaires, supplemented by laboratory examinations of fecal, oral, and hair samples. The findings indicated that 27.3% of pets harbored at least one zoonotic pathogen, with Campylobacter spp. (10.7%), Toxocara canis (8.3%), and Salmonella (5.7%) were the most identified pathogens. Statistical analysis using SPSS (v26) revealed significant correlations between infection identification and owner habits, namely inadequate handwashing after pet interaction (p = 0.002), inconsistent utensil sanitation (p = 0.001), and lack of regular immunization (p = 0.004). The logistic model revealed that inadequate utensil cleaning (AOR = 2.46; 95% CI: 1.38–4.38) and absence of immunization (AOR = 2.15; 95% CI: 1.18–3.91) were independent predictors of infection, whereas regular handwashing diminished the risk (AOR = 0.54; 95% CI: 0.31–0.92). The results highlight that insufficient hygiene, restricted veterinary services, and peri-urban living situations significantly elevate pet-related zoonotic hazards. Enhancing awareness, fostering responsible pet ownership, and improving veterinary surveillance are crucial for mitigating these hazards using a comprehensive One Health strategy. https://msra.online/index.php/Journal/about https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 64 INTRODUCTION: Zoonotic diseases are one of the most relevant global public health issues, with animal origins accounting for >60% of all human infectious diseases. With the growing involvement of companion animals in human households, the risk of pet-associated zoonotic transmission has become a growing concern, given the shared living, sleeping, and recreational areas of pets and their owners (Stull et al., 2015). Dogs, cats, and small mammals harbor a variety of bacterial, viral, and parasitic pathogens that cause substantial morbidity in humans (Damborg et al., 2016; Varela et al., 2022). Increased urbanization and the linked growth of periurban settlements represent a third key step in contributing to what is described as a global trend towards closer interactions between humans and animals. This trend has, in turn, created more opportunities for zoonotic agents to be maintained and spread within domestic settings (Gamble et al., 2023). With increasing human–animal interaction, knowledge of the epidemiology of zoonotic infections associated with pets is crucial for shaping public health and veterinary interventions (Overgaauw et al., 2020). Asymptomatic carriage in apparently healthy pets represents a potential reservoir for household exposure to common pathogens such as Salmonella spp., Campylobacter spp., and Toxocara canis (Bhat, 2021; O’Neil, 2018), a fact recognized in multiple studies. Direct contact, contaminated environments, and shared household surfaces are methods of transmission with particular risk in children, the elderly, and immunocompromised individuals (Whitfield & Smith, 2014). In addition, differing levels of hygiene among pet owners and sporadic veterinary monitoring (especially in lowand middle-income countries) only exacerbate the situation (Damborg et al., 2016). Companion animals have also been recently identified as possible bridging hosts connecting human populations with wildlife and environmental reservoirs in peridomestic and backyard settings (Langlois et al., 2025). These findings confirm the One Health paradigm, which acknowledges the integrated nature of human, animal, and environmental health systems in controlling zoonotic transmission (Desvars-Larrive et al., 2024). The potential for zoonoses (diseases transmissible between humans and pets) transmitted via household animals currently exceeds awareness of these risks. Therefore, understanding the effects of owner behavior, sanitation, and preventive care on the prevalence of pathogens in pet animals is essential. Previous surveys have reported inadequate awareness and poor hygiene practices among pet owners, even in high-income areas, indicating the need for more targeted education and risk communication (Stull et al., 2012). Research on animal-assisted intervention programs has supported our understanding of how easily zoonotic pathogens are transmitted through close human-animal contact (Boyle et al., 2019). In addition, studies conducted on non-traditional pet species have provided evidence of new zoonotic risks arising from altered ecological and socioeconomic drivers. Together, these trends indicate an urgent need for integrated epidemiological studies linking owner behavior, pet health care, and environmental determinants of health. Therefore, we conducted the present study to provide information about the prevalence of important zoonotic pathogens in household pets, their associated behavioral and environmental risk factors, and recommendations for health messages on risk reduction in a One Health context. 2. Research Methodology Study Design This cross-sectional study aimed to assess the role of domestic pets in zoonotic transmission routes and to correlate socio-behavioral and environmental factors with preventive measures related to zoonotic risk among pet owners. The cross-sectional design was chosen because it enables the simultaneous measurement of exposure variables and outcomes in each population, allowing for the detection of significant associations between pet-related practices and zoonotic diseases. A mixed methods approach was used to collect both quantitative and qualitative data, integrating epidemiological data with context-specific owner awareness and prevention practices. https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 65 Study Area and Population It was conducted in three urban centers in Punjab, Pakistan that represented a mix of socioeconomic status and pet-keeping styles. These locations were selected to ensure a variety of living environments, from intensely dense residential districts to suburban developments. The study population consisted of households with at least one owned domestic pet, such as a dog, cat, or small mammal like a rabbit or guinea pig. Households that owned pets for less than six months were excluded to reduce potential zoonotic interactions during the necessary exposure period. The participating dog owners were adults who provided informed consent. Sampling Technique and Sample Size We adopted a multistage random sampling procedure for this study. Stage 1: Select veterinary clinics using stratified random sampling to represent the proportions of the population in the three urban study areas. The second stage involved the random selection of households with pets from clinic registries. The final sample size of 300 households was derived using the formula for prevalence studies, with an expected zoonotic exposure of 30%, a 95% Confidence Level, and a 5% margin of error. The sample size was deemed sufficient to attain statistical power for both descriptive and inferential analyses. Data Collection Procedures Data collection consisted of a standardized questionnaire survey and laboratory examination of biological materials from domestic animals. Responses were collected through face-to-face interactions with pet owners using a structured questionnaire. It collected data on demographic characteristics, pet type, feeding and hygiene practices, lifestyle and vaccination details, knowledge of zoonotic diseases, and public visits to veterinarians. To check the clarity of the items and internal consistency (test-retest) reliability, the instrument was pretested on 10% of the sample in a similar environment. The initial analysis of the survey tool was conducted using a pretest to reveal its reliability, resulting in Cronbach’s alpha coefficient of 0.83. Trained veterinarians collected biological samples from pets under stringent aseptic conditions. Depending on the species, fecal swabs, oral samples, and fur were collected in sterile containers for laboratory analysis. Samples were individually coded to preserve the confidentiality of personal details while allowing unique identification. Following collection, all samples were transported to the microbiology laboratory under cold chain conditions for processing. Laboratory Analysis Our laboratory analysis focused on common zoonotic pathogens related to household pets, such as Salmonella spp., Campylobacter spp., Toxocara canis, and Microsporum canis. Detection was performed using standard microbiological and molecular techniques. Bacterial pathogens were isolated using selective culture media, biochemical confirmation, and polymerase chain reaction (PCR) testing for species identification. Parasitological investigations were performed using flotation and sedimentation methods. Fungal species were identified by culturing on Sabouraud dextrose agar and characterized morphologically and molecularly. Positive controls, negative controls, and 10% retested samples were included to ensure strict quality control and analytical reproducibility of the results. Data Management and Statistical Analysis All quantitative data collected from questionnaires and laboratory analyses were entered into a database and analyzed using the Statistical Package for Social Sciences (SPSS) version 26.0. We first performed descriptive statistical analyses on the data to describe a few main features of pet owners and animals (i.e., frequencies, means, and standard deviations). Finally, inferential statistics were used to examine the associations between pet practices and the detection of zoonotic pathogens, focusing on the complete Proportions and Prevalence of Zoonotic Infection. Risk Chi-square tests and Fisher's exact tests were used https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 66 for categorical variables. Logistic regression models were employed to identify significant predictors of zoonotic infection risk, with other potential confounding factors (i.e., age, education, type of pet) included as covariates in the model. Statistical significance was set at P < 0.05. Open-ended responses were transcribed, and thematic analysis was conducted on the qualitative data to identify key themes in knowledge, attitudes, and preventative practices related to zoonotic disease transmission. Ethical Considerations The Institutional Ethical Review Committee approved the study protocol for Animal and Human Research. Written informed consent was obtained from all participants prior to enrollment in the study, and the study was explained to all test individuals. All procedures for animal handling and sample collection were performed under veterinary supervision and in accordance with national and international codes and guidelines for the care and use of animals. We ensured participant confidentiality by removing all personal identifiers from the records and keeping them secure in a location with limited access. This study was conducted in accordance with the ethical standards of research involving humans and animals. Results Socio-Demographic Characteristics of Respondents A total of 300 pet-owning households participated in the study. Of these, 58.7% were male and 41.3% were female respondents, with a mean age of 36.5 ± 10.8 years. Educational attainment varied, with 44.0% having completed tertiary education, 37.3% secondary education, and 18.7% primary education or below. Urban residents represented 62.0% of the sample, while 38.0% resided in peri-urban areas. These findings are summarized in Table 1. The gender composition of the study population is visually illustrated in Figure 1, which demonstrates the predominance of male respondents in the survey. Table 1: Socio-Demographic Characteristics of Respondents (n = 300) Variable Category Frequency (n) Percentage (%) Gender Male 176 58.7 Female 124 41.3 Age group (years) 18–30 84 28.0 31–45 134 44.7 >45 82 27.3 Education level Primary or below 56 18.7 https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 67 Secondary 112 37.3 Tertiary 132 44.0 Residence Urban 186 62.0 Peri-urban 114 38.0 Figure 1: Gender distribution of respondents showing higher male participation (58.7%) compared to female respondents (41.3%). Pet Ownership Patterns and Hygiene Practices Dogs were the most commonly owned pets (49.3%), followed by cats (38.7%) and small mammals such as rabbits and guinea pigs (12.0%). A majority of owners (67.0%) housed their pets indoors, whereas 33.0% maintained outdoor confinement. Handwashing after pet contact was reported by 71.7% of owners, but only 54.3% practiced regular utensil disinfection. Furthermore, 21.0% of pet owners had not maintained a routine vaccination schedule for their animals. These descriptive statistics are detailed in Table 2. A stacked bar chart (Figure 2) compares compliance and non-compliance across major hygiene indicators, demonstrating relatively lower adherence to utensil cleaning compared with hand hygiene and vaccination. Table 2: Pet Ownership and Hygiene Practices Among Respondents (n = 300) Variable Category Frequency (n) Percentage (%) https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 68 Type of pet Dog 148 49.3 Cat 116 38.7 Small mammals 36 12.0 Pet housing Indoors 201 67.0 Outdoors 99 33.0 Handwashing after contact Yes 215 71.7 No 85 28.3 Regular utensil cleaning Yes 163 54.3 No 137 45.7 Routine vaccination Yes 237 79.0 No 63 21.0 https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 69 Figure 2: Comparison of compliance (blue bars) and non-compliance (gray bars) rates for handwashing, utensil cleaning, and vaccination among pet owners. Utensil disinfection shows the lowest adherence rate (54.3%). Laboratory Detection of Zoonotic Pathogens Out of 300 pet specimens examined, 82 (27.3%) tested positive for at least one zoonotic pathogen. The most frequently identified species was Campylobacter spp. (10.7%), followed by Toxocara canis (8.3%), Salmonella spp. (5.7%), and Microsporum canis (2.6%). The detailed pathogen distribution by pet species is shown in Table 3. The prevalence of each detected pathogen is also depicted in Figure 3, where Campylobacter spp. demonstrates the highest overall prevalence across sampled pets. Table 3: Distribution Of Zoonotic Pathogens Isolated from Pet Samples (n = 300) Pathogen detected Dogs (n = 148) Cats (n = 116) Small mammals (n = 36) Total positive (n) Prevalence (%) Campylob acter spp. 21 8 3 32 10.7 Salmonell a spp. 11 5 1 17 5.7 https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 70 Toxocara canis 14 8 3 25 8.3 Microspo rum canis 2 4 2 8 2.6 Total positive 48 27 9 82 27.3 Figure 3: Prevalence of zoonotic pathogens detected in pet samples, showing Campylobacter spp. as the most frequently isolated organism. Association between Owner Practices and Pathogen Detection Chi-square tests revealed statistically significant associations between hygiene behaviors and pathogen positivity. Lack of handwashing (χ² = 9.86; p = 0.002), irregular utensil cleaning (χ² = 11.14; p = 0.001), and absence of vaccination (χ² = 8.47; p = 0.004) were strongly correlated with infection detection (Table 4). Residence in peri-urban areas was also significantly associated with higher infection prevalence (χ² = 6.29; p = 0.012). No significant relationships were found for gender or education level (p > 0.05). Table 4: Association Between Hygiene Practices and Zoonotic Pathogen Detection Variable Category Pathogen Pathogen χ² p-value https://msra.online/index.php/Journal/about Volume 3, Issue 4 (2025) 71 Positive (%) Negative (%) Handwashing after contact Yes 21.9 78.1 9.86 0.002 No 41.2 58.8 Utensil cleaning Reg ular 20.2 79.8 11.14 0.001 Irre gular 37.9 62.1 Vaccination status Yes 22.8 77.2 8.47 0.004 No 44.4 55.6 Residence Urb an 22.0 78.0 6.29 0.012 Peri -urban 35.9 64.1 Logistic Regression Analysis of Risk Factors Multivariate logistic regression identified key predictors of zoonotic infection in household pets. After adjusting for confounders, irregular utensil cleaning (AOR = 2.46; 95% CI = 1.38–4.38; p = 0.002) and absence of vaccination (AOR = 2.15; 95% CI = 1.18–3.91; p = 0.011) emerged as independent risk factors. Conversely, consistent handwashing reduced infection odds by nearly half (AOR = 0.54; 95% CI = 0.31– 0.92; p = 0.024). Residence in peri-urban areas also remained a significant predictor (AOR = 1.67; 95% CI = 1.01–2.76; p = 0.045). These findings are presented in Table 5 and visually summarized through a foresttype plot in Figure 4, highlighting the magnitude and confidence intervals of each variable’s adjusted odds ratio. Table 5: Logistic Regression Analysis of Factors Associated with Zoonotic Pathogen Positivity Variable Adjusted Odds Ratio (AOR) Confidence Interval p-value No handwashing after 1.88 1.08 – 3.27 0.026