Assessment of the Implementation of Pakistan's National Action Plan on Antimicrobial Resistance in the Agriculture and Food Sectors.

The agriculture and food (agrifood) sectors play key roles in the emergence, spread, and containment of antimicrobial resistance (AMR). Pakistan's first National Action Plan (NAP) on AMR was developed to guide One Health interventions to combat AMR through 2017-2022. To improve subsequent iterations, we assessed the implementation of Pakistan's NAP in the agrifood sectors (NAPag) in October 2022, using the Progressive Management Pathway on AMR tool developed by the Food and Agriculture Organization of the United Nations (FAO). The assessment tool addressed four crucial focus areas of the NAPag: governance, awareness, evidence, and practices. Each focus area contains multiple topics, which involve four sequential stages of activities to progressively achieve systematic management of AMR risk in the agrifood sectors. High-level representatives of the NAPag stakeholders provided information for the assessment through pre-event documentary review and workshop discussions. The assessment results showed that Pakistan's NAPag had an overall moderate coverage (59%) of the anticipated activities. Gaps were particularly notable in strengthening governance, good practices, and interventions in non-livestock sectors. Furthermore, only 12% of the evaluated activities were fully executed and documented, consistently remaining at the planning and piloting stages in the livestock sector across all the examined topics. Insufficient attention to non-livestock sectors, inadequate regulation and enforcement capacity, and resource constraints have hindered scalable and sustainable interventions under the current plan. This assessment provides valuable insights to strengthen the inclusiveness and contribution of the agrifood sectors in the next NAP iteration. In the short-to-medium term, strategic prioritization is necessary to optimize the use of limited resources and target the most critical gaps, such as improving awareness among key stakeholders and fortifying regulations for prudent antimicrobial use. In the long term, integration of AMR into the country's broader health, development, and agricultural transformation agendas will be needed to generate sustainable benefits.

Future directions for One Health research: Regional and sectoral gaps.

Implementation of a One Health approach varies considerably between different geographical regions and remains challenging to implement without greater inclusivity of different disciplinary capacity and expertise. We performed comparative analyses of abstracts presented at the 1st World One Health Congress (WOHC 2011) and 6th WOHC (2020) to explore and describe the evolving demographics and disciplinary scope of One Health research. We classified abstracts into six One Health research categories and twenty-three subcategories. We also recorded corresponding authors' country and regional affiliation as well as study country (i.e., the country in which the research was conducted) to explore potential asymmetries between funding recipients and study subjects. The WOHC has seen a significant expansion in participation over the last 10 years. The numbers of abstracts accepted to the Congress increased threefold over the last decade (i.e., 302 abstracts in 2010, and 932 abstracts in 2020). At both Congresses, "Disease Surveillance" accounted for the largest proportion (105/302 (35%) and 335/932 (36%) in 2010 and 2020, respectively) of all abstracts accepted. However, "Environmental and Ecological Issues" (33/302 (10%) and 94/932 (11%)), and "Sustainable Food Systems" (19/302 (6%) and 44/932 (4%)) were less well-represented categories of One Health research in both 1st and 6th WOHC respectively. In contrast, "Antimicrobial Resistance" related research increased substantially over time (4/302 (1%) in 2011) and (119/932 (13%) in 2020). There were also differences in the type of research by authors based in "Very High Human Development" index countries compared to "Medium and Low Human Development. "Public Policy" dominated the former, whereas "Disease Surveillance" dominated the latter, suggesting potential regional differences regarding One Health research priorities. The results of the study highlight potential regional gaps and differences in One Health research priorities, with respect to emphasis on operational (surveillance) versus strategic (policy) One Health activities.

A cross-sectional survey on fruit bat-human interaction in Pakistan; one health perspective.

OBJECTIVE: Several factors, such as residential area topography, population density, and lack of infrastructure, were hypothesized to contribute toward respondents' knowledge, attitude, and practice regarding disease transmission. The present study was designed to investigate the knowledge, attitudes, and perception of human-fruit bat interaction by student respondents located in ten districts within the Punjab and Khyber Pakhtunkhwa provinces in Pakistan. METHOD: A cross-sectional survey was conducted by trained enumerators in academic institutions using a structured questionnaire among student respondents (n = 1466), living in two topographically distinct (Mountainous and Plain) residential regions of the Punjab and Khyber Pakhtunkhwa (KPK) provinces in Pakistan regarding their history of bat encounters. RESULTS: Our study revealed that 71.4% of the 1466 respondents had observed bats in their geographic region. 21% of our survey respondents reported bat bites incidents over their lifetime, but only 40% actively sought medical care for wound management despite reporting they had a close family member that had contracted rabies (27-35%). Our generalized linear models (GLMs) highlighted that a respondent residing in a residential region had a greater association with reporting a suspected bat bite over their lifetime and reported rabies victims in both near and extended family members (OR = -0,85, p-value = 0.03, 95% CI). This appeared to be due to delaying consulting a doctor or medical facility for treatment following a suspected bat bite in the topographic residential group as compared to the respondents in the provincial residential group (OR 1.12, p-value = 0.04, 95% CI). CONCLUSION: Our findings indicate the necessity of a One Health comprehensive surveillance system in Pakistan for emerging and re-emerging zoonotic pathogens in Pteropodidae.

Infection Dynamics of Clostridium perfringens Fingerprinting in Buffalo and Cattle of Punjab Province, Pakistan.

Clostridium perfringens produces core virulence factors that are responsible for causing hemorrhagic abomasitis and enterotoxemia making food, animals, and humans susceptible to its infection. In this study, C. perfringens was isolated from necropsied intestinal content of buffalo and cattle belonging to four major bovine-producing regions in the Punjab Province of Pakistan for the purpose offind out the genetic variation. Out of total 160 bovine samples (n: 160), thirty-three (n: 33) isolates of C. perfringens were obtained from buffalo (Bubales bubalis) and cattle (Bos indicus) that were further subjected to biochemical tests; 16S rRNA based identification and toxinotyping was done using PCR (Polymerase Chain Reaction) and PFGE (Pulse Field Gel Electrophoresis) pulsotypesfor genetic diversity. Occurrence of C. perfringens was found to be maximum in zone-IV (Bhakkar and Dera Ghazi Khan) according to the heatmap. Correlation was found to be significant and positive among the toxinotypes (α-toxin, and ε-toxin). Response surface methodology (RSM) via central composite design (CCD) and Box-Behnken design (BBD) demonstrated substantial frequency of C. perfringens based toxinotypes in all sampling zones. PFGE distinguished all isolates into 26 different pulsotypes using SmaI subtyping. Co-clustering analysis based on PFGE further decoded a diversegenetic relationship among the collected isolates. This study could help us to advance toward disease array of C. perfringens and its probable transmission and control. This study demonstrates PFGE patterns from Pakistan, and typing of C. perfringens by PFGE helps illustrate and mitigate the incidence of running pulsotypes.

Quantification and Trends of Antimicrobial Use in Commercial Broiler Chicken Production in Pakistan.

Antimicrobial resistance (AMR) is a global health challenge and antimicrobial use (AMU) in the livestock sector has been considered as one of the contributing factors towards the development of AMR in bacteria. This study summarizes the results of a point prevalence survey conducted to monitor farm-level AMU in commercial broiler chicken farms in Punjab and Khyber Pakhtunkhwa (KPK) provinces of Pakistan. A cross-sectional study was conducted to quantify AMU and to check seasonal variations of AMU in 12 commercial broiler chicken farms (six from each province) during the summer and winter seasons of the year 2020-2021. AMU was recorded using three AMU metrics: kg, mg per population correction unit (mg/PCU), and mg/kg of final flock weight. A total of 22 antimicrobial drugs (348.59 kg) were used for therapeutic or prophylactic purposes in surveyed broiler chicken farms. The total combined AMU for all the broiler chicken farms was 462.57 mg/PCU. The use of most of the antimicrobials increased during winter flocks compared to summer. The top three antimicrobial drugs used during the summer were neomycin (111.39 mg/PCU), doxycycline (91.91 mg/PCU), and tilmicosin (77.22 mg/PCU), whereas doxycycline (196.81 mg/PCU), neomycin (136.74 mg/PCU), and amoxicillin (115.04 mg/PCU) during the winter. Overall, 60% of the antibiotics used in broiler chicken were critically important antimicrobial classes (CIA) for human medicine as characterized by the World Health Organization. Our findings showed high AMU in broiler chicken production and a call for urgent actions to regulate CIA use in food animals in Pakistan. This baseline survey is critical for the design and implementation of a subsequent national level AMU surveys that can include additional farming types, animals' species, and geographical locations over a longer period of time.

Knowledge, attitude and practice (KAP) survey of canine rabies in Khyber Pakhtunkhwa and Punjab Province of Pakistan.

BACKGROUND: This study aimed to assess the extent of knowledge and understanding of rabies disease in rural and urban communities of Pakistan. It also identified malpractices after suspected dog bite that might pose a risk for humans contracting rabies. METHODS: A cross-sectional study was conducted (n = 1466) on people having different age groups and educational levels in four different geographic regions of Punjab and Khyber Pakhtunkhwa provinces in Pakistan. Knowledge, attitude, and practices of people were assessed using a structured questionnaire. We used a bivariate and multivariate analysis to study the association between rabies related mortalities in near or extended family members and different risk behaviors. RESULTS: Our results demonstrate that the majority of the juvenile population (less than 18 years of age) were not aware of the clinical signs of rabies in animals. 75% of the total respondents were not vaccinated against rabies, 60% did not seek a doctor's advice after a suspected animal bite, and 55% had inadequate health care facilities for rabies patients in local hospitals. Respondents that had pets at home had not vaccinated (38%; p < 0.05; odds ratio 1.58) themselves against rabies due to lack of knowledge and awareness of pre-exposure prophylaxis for rabies (51%; p < 0.05; odds ratio 1.25). They also tend to not visit doctor after suspected bite (52%; p < 0.05; odds ratio 1.97), which may had resulted in more deaths (65%; p < 0.05; odds ratio 1.73) of someone in their near or extended family due to rabies. CONCLUSIONS: Lack of knowledge about the nature of rabies disease and prophylaxis has contributed to increase of rabies related deaths. Inadequate health care facilities and poor attitude of not seeking medical attention after suspected dog bite are the major reasons of rabies related deaths. These findings could help in devising a targeted management strategy and awareness program to control and reduce the incidence of human rabies related deaths in Pakistan.

Seroprevalence and risk factors of avian influenza H9 virus among poultry professionals in Rawalpindi, Pakistan.

BACKGROUND: Avian influenza H9 is endemic in commercial and backyard poultry in Pakistan and is a serious occupational health hazard to industry workers. This study aimed to determine the seroprevalence of avian influenza H9 infection in people working with poultry in Rawalpindi, Pakistan and assess the measures they took to protect themselves from infection. METHODS: A cross-sectional study was conducted from December 2016 to May 2017 of 419 people working with poultry in Rawalpindi Division, including farm workers, vaccinators, field veterinarians, butchers and staff working in diagnostic laboratories. Potential participants were randomly approached and gave written consent to participate. Data were collected using a standardized questionnaire and serum samples were processed to detect H9 antibodies using the haemagglutination inhibition test. RESULTS: Of the 419 participants, 406 (96.9%) were male. The mean age of the participants was 36.4 (SD 10.86) years. A total of 332 participants agreed to a blood test, 167 of whom were positive for A(H9) antibodies, giving an overall seroprevalence of 50.3%. Laboratory staff had the highest seroprevalence (100%) and veterinarians the lowest (38.5%). Vaccinators, butchers and farm workers had a seroprevalence of 83.3%, 52.4% and 45.5% respectively. Personals who used facemasks had significantly lower (P<0.002) seroprevalence (29.6%) than those who never used them (90.6%). Similarly, those who always used gloves and washed their hands with soap had a seroprevalence of 32.8% compared with 89.0% in those who never took these precautions. Of the participants who handled antigens, 92.3% were seropositive. CONCLUSION: Laboratory staff and vaccinators are exposed to viral cultures and influenza vaccines respectively which may explain their high seroprevalence.