Prevalence of Salmonella spp. and Staphylococcus aureus in Chicken Meat and Pork from Cambodian Markets.

Salmonella spp. and Staphylococcus aureus are two of the most common foodborne bacteria in animal-source foods (ASF) that cause illness worldwide. This study aimed to determine the prevalence of Salmonella spp. and S. aureus in chicken meat and pork in markets in Cambodia. Sampling was done in 52 traditional markets and 6 supermarkets in 25 provinces of Cambodia between October 2018 and August 2019. In total, 532 samples were obtained: chicken meat and pork (n = 408, 204 of each), chicken and pork cutting board swabs (n = 124, 62 of each). All samples were analyzed for the presence of Salmonella spp. and S. aureus; colony-forming units per gram (CFU/g) of coagulase-positive Staphylococci (CPS) were counted, and a subset of samples was also analyzed for the most probable number (MPN, n = 136) of Salmonella. The overall prevalence of Salmonella spp. and S. aureus were 42.1% (224/532) and 29.1% (155/532), respectively, with 14.7% (78/532) of samples containing both bacteria. The prevalence of Salmonella spp. in chicken meat was 42.6%, on chicken cutting board it was 41.9%, on pork it was 45.1%, and the pork cutting board 30.6%. Chicken meat had a significantly (p-value < 0.05) higher prevalence of S. aureus, 38.2%, compared to the chicken cutting board, 17.7%, pork 28.9%, and pork cutting board 11.3%. Mean MPN-Salmonella was 10.6 MPN/g in chicken and 11.1 MPN/g in pork samples. Average Log CFU/g of CPS in chicken and pork samples were 2.6 and 2.5, respectively. The results indicate that chicken meat and pork in Cambodia were highly contaminated with Salmonella spp. and S. aureus, posing risks to consumers' health. Urgent interventions are necessary to improve hygiene for safer meat in Cambodian markets.

Detection of African swine fever virus in free-ranging wild boar in Southeast Asia.

African Swine Fever (ASF) is a highly contagious and fatal viral disease affecting both domestic and wild suids. The virus was introduced to Southeast Asia in early 2019 and has since spread rapidly throughout the region. Although significant efforts have been made to track and diagnose the disease in domestic pigs, very little is known about ASF in free-ranging wild boar and their potential role in maintaining the disease within Southeast Asia. Through a collaboration between government and non-government actors in Laos, Viet Nam, and Cambodia, investigations were conducted to (a) characterize the interface between domestic pigs and wild boar, (b) document risk factors for likely ASF spillover into wild boar populations by way of this interface, and (c) determine whether ASF in wild boar could be detected in each country. An extensive overlap between wild boar habitat and domestic pig ranging areas was found around villages bordering forests in all three countries, creating a high-risk interface for viral spillover between domestic pig and wild boar populations. Fifteen and three wild boar carcasses were detected through passive reporting in Laos and Viet Nam, respectively, in 2019 and early 2020. Four of five carcasses screened in Laos and two of three in Viet Nam were confirmed positive for African swine fever virus using real-time PCR. There were no confirmed reports of wild boar carcasses in Cambodia. This is the first confirmation of ASF in wild boar in Southeast Asia, the result of a probable viral spillover from domestic pigs, which highlights the importance of early reporting and monitoring of ASF in wild boar to enable the implementation of appropriate biosecurity measures.

Antibiotic use in pig farms at different levels of intensification-Farmers' practices in northeastern Thailand.

Understanding the patterns and drivers of antibiotic use in livestock is crucial for tailoring efficient incentives for responsible use of antibiotics. Here we compared routines for antibiotic use between pig farms of two different levels of intensification in Khon Kaen province in Thailand. Among the 113 family-owned small-scale farms (up to 50 sows) interviewed did 76% get advice from the pharmacy about how to use the antibiotics and 84% used it primarily for treating disease. Among the 51 medium-scale-farms (100-500 sows) belonging to two companies did 100% get advice about antibiotic use from the company's veterinarian (P<0.0001) and 94% used antibiotics mostly as disease preventive measure (P<0.0001). In 2 small scale farms 3rd generation cephalosporins, tylosin or colistin were used; antibiotics belonging to the group of highest priority critically important antimicrobials for human medicine. Enrofloxacin, belonging to the same group of antimicrobials, was used in 33% of the small-scale and 41% of the medium-scale farms. In the latter farms, the companies supplied 3-4 antibiotics belonging to different classes and those were the only antibiotics used in the farms. The median and mean estimated expenditure on antibiotics per sow was 4.8 USD (IQR = 5.8) for small-scale farms and 7 USD and 3.4 USD for the medium-scale farms belonging to the two respective companies. Our observations suggest to target the following areas when pig farming transitions from small-scale to medium-scale: (i) strengthening access to professional animal health services for all farmers, (ii) review of the competence and role of veterinary pharmacies in selling antibiotics and (iii) adjustment of farming company animal health protocols towards more medically rational use of antibiotics.

Carriage of carbapenemase- and extended-spectrum cephalosporinase-producing Escherichia coli and Klebsiella pneumoniae in humans and livestock in rural Cambodia; gender and age differences and detection of blaOXA-48 in humans.

OBJECTIVES: This study investigates the frequency and characteristics of carbapenemase-producing Escherichia coli/Klebsiella pneumoniae (CPE/K) and extended-spectrum cephalosporinase-producing E. coli/K. pneumoniae (ESCE/K) in healthy humans and livestock in rural Cambodia. Additionally, household practices as risk factors for faecal carriage of ESCE/K are identified. METHODS: Faecal samples were obtained from 307 humans and 285 livestock including large ruminants, pigs and poultry living in 100 households in rural Cambodia in 2011. Each household was interviewed, and multilevel logistic model determined associations between household practices/meat consumption and faecal carriage of ESCE/K. CPE and ESCE/K were detected and further screened for colistin resistance genes. RESULTS: CPE/K isolates harbouring blaOXA-48 were identified in two humans. The community carriage of ESCE/K was 20% in humans and 23% in livestock. The same ESBL genes: blaCTX-M-15 , blaCTX-M-14 , blaCTX-M-27 , blaCTX-M-55 , blaSHV-2 , blaSHV-12 , blaSHV-28 ; AmpC genes: blaCMY-2 , blaCMY-42, blaDHA-1 ; and colistin resistance genes: mcr-1-like and mcr-3-like were detected in humans and livestock. ESCE/K was frequently detected in women, young children, pigs and poultry, which are groups in close contact. The practice of burning or burying meat waste and not collecting animal manure indoors and outdoors daily were identified as risk factors for faecal carriage of ESCE/K. CONCLUSIONS: Faecal carriage of E. coli and K. pneumoniae harbouring extended-spectrum cephalosporinase genes are common in the Cambodian community, especially in women and young children. Exposure to animal manure and slaughter products are risk factors for intestinal colonization of ESCE/K in humans.

Emergence of Influenza A(H7N4) Virus, Cambodia.

Active surveillance in high-risk sites in Cambodia has identified multiple low-pathogenicity influenza A(H7) viruses, mainly in ducks. None fall within the A/Anhui/1/2013(H7N9) lineage; however, some A(H7) viruses from 2018 show temporal and phylogenetic similarity to the H7N4 virus that caused a nonfatal infection in Jiangsu Province, China, in December 2017.

Antimicrobial resistance in Cambodia: a review.

OBJECTIVES: Following the launch of the Global Antimicrobial Resistance Surveillance System (GLASS), antimicrobial resistance (AMR) rates in many countries remain poorly described. This review provides an overview of published AMR data from Cambodia in the context of recently initiated national human and food-animal surveillance. METHODS: PubMed and the Cochrane Database of Systematic Reviews were searched for articles published from 2000 to 2018, which reported antimicrobial susceptibility testing (AST) data for GLASS specific organisms isolated from Cambodia. Articles were screened using strict inclusion/exclusion criteria. AST data was extracted, with medians and ranges of resistance rates calculated for specific bug-drug combinations. RESULTS: Twenty-four papers were included for final analysis, with 20 describing isolates from human populations. Escherichia coli was the most commonly described organism, with median resistance rates from human isolates of 92.8% (n=6 articles), 46.4% (n=4), 55.4% (n=8), and 46.4% (n=5) to ampicillin, 3rd generation cephalosporins, fluoroquinolones, and gentamicin respectively. CONCLUSIONS: Whilst resistance rates are high for several GLASS organisms, there were insufficient data to draw robust conclusions about the AMR situation in Cambodia. The recently implemented national AMR surveillance systems will begin to address this data gap.

Detection of Low Pathogenicity Influenza A(H7N3) Virus during Duck Mortality Event, Cambodia, 2017.

In January 2017, an estimated 3,700 (93%) of 4,000 Khaki Campbell ducks (Anas platyrhynchos domesticus) died in Kampong Thom Province, Cambodia. We detected low pathogenicity avian influenza A(H7N3) virus and anatid herpesvirus 1 (duck plague) in the affected flock; however, the exact cause of the mortality event remains unclear.

Manure management and public health: Sanitary and socio-economic aspects among urban livestock-keepers in Cambodia.

Livestock manure is a valuable source of nutrients for crop production, but can also pose a public health hazard and have negative environmental impacts. This study investigated manure management practices among urban and peri-urban livestock keepers in Cambodia, to identify risk behaviours and socio-economic aspects associated with the handling of manure. A survey including 204 households was conducted, using a structured questionnaire with questions on demographics, socio-economic characteristics and household practices related to manure management. Faecal samples were obtained from pig pens and pig manure storage units for analysis of the potential zoonotic pathogens Salmonella enterica (Polymerase Chain Reaction (PCR)), Ascaris suum and Trichuris suis (McMaster flotation technique). The survey revealed a difference in management between cattle and pig manure. Cattle manure was most commonly used as fertiliser for crop production (66%) (p<0.001), whereas pig manure was most commonly dumped in the environment (46%) (p<0.001). Logistic regression models showed that households with a lower socio-economic position were more likely to dump pig manure (p<0.001), with scarcity of agricultural land (p<0.001) and lack of carts for transportation of manure (p<0.01) being identified as contributing factors. Salmonella enterica was detected in 9.7% of manure samples, while Ascaris suum and Trichuris suis were detected in 1.6% and 2.4% of the samples, respectively. The results presented in this study indicate that manure management by urban and peri-urban households may pose a public health threat and an environmental hazard. There is evidently a need for further knowledge support to the livestock keepers to promote good management practices.

Detection of Campylobacter in human and animal field samples in Cambodia.

Campylobacter are zoonotic bacteria and a leading cause of human gastroenteritis worldwide with Campylobacter jejuni and C. coli being the most commonly detected species. The aim of this study was to detect Campylobacter in humans and livestock (chickens, ducks, pigs, cattle, water buffalo, quail, pigeons and geese) in rural households by routine culturing and multiplex PCR in faecal samples frozen before analysis. Of 681 human samples, 82 (12%) tested positive by PCR (C. jejuni in 66 samples and C. coli in 16), but none by routine culture. Children were more commonly Campylobacter positive (19%) than adult males (8%) and females (7%). Of 853 livestock samples, 106 (12%) tested positive by routine culture and 352 (41%) by PCR. Campylobacter jejuni was more frequent in chickens and ducks and C. coli in pigs. In conclusion, Campylobacter proved to be highly prevalent by PCR in children (19%), ducks (24%), chickens (56%) and pigs (72%). Routine culturing was insufficiently sensitive in detecting Campylobacter in field samples frozen before analysis. These findings suggest that PCR should be the preferred diagnostic method for detection of Campylobacter in humans and livestock where timely culture is not feasible.

Household practices related to disease transmission between animals and humans in rural Cambodia.

BACKGROUND: Zoonotic diseases are disproportionately affecting poor societies in low-income countries and pose a growing threat to public health and global food security. Rural Cambodian households may face an increased likelihood of exposure to zoonotic diseases as people there live in close association with livestock. The objectives of the study was to identify practices known to influence zoonosis transmission in rural Cambodian households and relate the practices to agro-ecological region, socio-economic position, demographics, livestock management and zoonosis awareness. METHODS: The study was conducted in three different agro-ecological regions of Cambodia; 10 villages each in the central lowlands, north-west wetlands and on the south coast, where information was obtained in questionnaires administered to 300 households, and 30 village heads and animal health workers. RESULTS: Descriptive analysis revealed a gender difference in responsibility for livestock and that the main purpose of raising livestock was for sale. Few respondents (6%) perceived a likelihood of disease transmission in their village between livestock, humans and wildlife, despite household practices related to zoonosis transmission being common. More than one-forth of households practised behaviours such as culling sick animals for consumption, eating animals found dead and allowing animals to enter sleeping and food preparation areas. Associations between household practices and possible explanatory factors were analysed with multivariable models using generalised estimation equations to account for clustering of practices within villages. Factors found to influence household practices were agro-ecological region, socio-economic position, number of people in the household, livestock species reared and awareness of zoonoses. CONCLUSIONS: Cambodia has experienced numerous fatal human cases of zoonotic influenza and extensive influenza information campaigns have been run, yet only a few of the households surveyed here reported the threat of zoonosis to be a concern in their village. Zoonosis awareness was positively related to hand washing behaviour, but other practices associated with an increased or decreased likelihood of exposure to zoonotic pathogens were unaffected by awareness. The findings indicate a knowledge-to-action gap among rural farmers and highlight the necessity for reconstructed interventions in zoonotic disease control.