Evaluation of the efficacy of commercial live vaccines against the local Thai QX field strain for the protection of specific pathogen-free chicks.

Background and Aim: The high prevalence of QX-like variant among Thai isolates poses a significant threat to poultry production. In this study, we evaluated the protective efficacy of commercially available heterologous infectious bronchitis virus (IBV) vaccines against the local Thai QX-like strain in specific-pathogen-free (SPF) chicks from Thailand. Materials and Methods: The experiment involved 100 SPF chicks divided into 4 arms. Arms I and II received the TAbic IB VAR (233A) and Ibird (1/96) vaccines, respectively, on day 1. After 10 days, both arms received the H120 vaccine. Arms III and IV were non-vaccinated positive and negative controls. Challenge infection was local Thai QX-like virus on birds of Arms I, II, and III, and negative control of Arm IV. Clinical signs of infectious bronchitis (IB) and IBV detection using reverse transcription polymerase chain reaction were assessed at 2, 4, and 6 days post-challenge (dpc). At 6 dpc, the birds were humanely euthanized for post-mortem examination with the ciliostasis test and histopathological analysis of the tracheas, lungs, and kidneys. Results: Virus shedding started at 4 dpc (33.3% positive) and reached 100% positivity at 6 dpc with obvious clinical respiratory symptoms in non-vaccinated-challenged birds. No detection of IBV in vaccinated-challenged arms. Ciliary activity scores were significantly lower in non-vaccinated-challenged birds at 23.64 (standard deviation [SD] ± 1.74) and 96.50 (SD ± 1.91) and 95.64 (SD ± 1.77), respectively (p = 0.05) than in vaccinated-challenged birds. The most remarkable histopathological changes were observed in non-vaccinated-challenged birds, with moderately severe changes in the trachea, lungs, and kidneys. On the other hand, birds in vaccinated-challenged arms showed no significant changes. Conclusion: This study demonstrated the efficacy of TAbic IB VAR (233A) or Ibird (1/96) vaccine combined with a Massachusetts serotype vaccine (H120) against the local Thai QX-like strain in SPF chicks, contributing valuable insights to the selection of suitable commercially available vaccines to combat the prevalent local QX-like strains in Thailand.

A review and meta-analysis of non-typhoidal Salmonella in Vietnam: Challenges to the control and antimicrobial resistance traits of a neglected zoonotic pathogen.

Non-typhoidal Salmonella (NTS) is a food-borne zoonotic pathogen with important implications for human health. Despite its importance, the burden of NTS infections in Vietnam is unknown. We conducted a systematic review and a meta-analysis to describe the prevalence and phenotypic antimicrobial resistance (AMR) for NTS over time in Vietnam. Following PRISMA guidelines, we identified 72 studies from PubMed and Google Scholar containing data relevant to 'Salmonella', 'Salmonellosis', and 'Vietnam', or 'Viet Nam'. Of those, forty-two papers reporting prevalence of NTS, and twenty-six including data on phenotypic resistance were selected for meta-analysis. The prevalence of NTS ranged from 2% to 5% in humans and from 30% to 41% in samples from animals and the farm/slaughterhouse environment. Between 2000 and 2020 we observed a 27.3% (p = 0.044) increase in absolute terms in prevalence among individuals with enteric diseases and a 12.0% (p = 0.047) increase in aquaculture/seafood samples. The most commonly serovars identified across studies included S. Weltevreden (12.0%), followed by S. Typhimurium (10.1%), S. Derby (6.4%), S. London (5.8%), S. Anatum (4.4%), S. Rissen (3.9%), S. Enteritidis (3.7%), S. Albany (3.3%) and S. 4,[5],12:i:- (3.0%). Over the same period, there was an increasing trend in the pooled AMR prevalence for quinolones (15.6% relative increase), 3rd-, 4th-, and 5th-generation cephalosporins (23.7%), penicillins (16.1%), tetracyclines (12.9%), sulfonamides (8.8%), amphenicol (17.8%), and multidrug resistance (11.4%) (all p ≤ 0.014). A broad range of AMR genes (ARGs) were detected in both human and animal populations. The observed prevalence and AMR trends in Vietnam underscore the need of adopting a One Health strategy encompassing surveillance systems, research initiatives, and awareness campaigns to effectively address the challenges posed by NTS.

Economic assessment of an intervention strategy to reduce antimicrobial usage in small-scale chicken farms in Vietnam.

Antimicrobials are a core aspect of most livestock production systems, especially in low-and middle-income countries. They underpin the efficient use of scarce feed resources and stabilize returns on capital and labor inputs. Antimicrobial use (AMU) contributes to the production of healthy animals, yet AMU in livestock is linked to antimicrobial resistance (AMR) in animals, humans and the environment. The Vietnamese Platform for Antimicrobial Reduction in Chicken Production was implemented during 2016-2019 and was one of Southeast Asia's first interventions focused on AMU reductions in livestock production. The project targeted small-scale commercial poultry farms in the Mekong Delta region of Vietnam using a "randomized before-and-after controlled" study design. It provided farmers with a locally adapted support service (farmer training plan, advisory visits, biosecurity, and antimicrobial replacement products) to help them reduce their reliance on antimicrobials. A partial budget analysis was performed comparing the control group (status-quo) and intervention group (alternative). The median net farm-level benefit of the intervention strategies with the project's support was VND 6.78 million (interquartile range (IR) VND -71.9-89 million) per farm. Without project support the benefit was reduced to VND 5.1 million (IR VND -69.1-87.2 million) to VND 5.3 million (IR -VND 68.9-87.5 million) depending on the antimicrobial alternative product used. At the project level with a focus on AMU and its reduction, subsequently influence on the resistance reduction, our results showed that achieving resistance reduction benefits with the current knowledge and technologies required investment of at least VND 9.1 million (US$ 395.10) per farm during the project's lifetime. The results highlight the positive net profit for the majority of enrolled farms and a reasonable investments from the project. The recommendation focuses on the implementation of policies on financial support, legislation, and information as potential solutions to facilitate the application of intervention strategies to reduce AMU in poultry production.

Molecular characterization of Campylobacter spp. isolates obtained from commercial broilers and native chickens in Southern Thailand using whole genome sequencing.

Chickens are the primary reservoirs of Campylobacter spp., mainly C. jejuni and C. coli, that cause human bacterial gastrointestinal infections. However, genomic characteristics and antimicrobial resistance of Campylobacter spp. in low- to middle-income countries need more comprehensive exploration. This study aimed to characterize 21 C. jejuni and 5 C. coli isolates from commercial broilers and native chickens using whole genome sequencing and compare them to 28 reference Campylobacter sequences. Among the 26 isolates, 13 sequence types (ST) were identified in C. jejuni and 5 ST in C. coli. The prominent ST was ST 2274 (5 isolates, 19.2%), followed by ST 51, 460, 2409, and 6455 (2 isolates in each ST, 7.7%), while all remaining ST (464, 536, 595, 2083, 6736, 6964, 8096, 10437, 828, 872, 900, 8237, and 13540) had 1 isolate per ST (3.8%). Six types of antimicrobial resistance genes (ant(6)-Ia, aph(3')-III, blaOXA, cat, erm(B), and tet(O)) and one point mutations in the gyrA gene (Threonine-86-Isoleucine) and another in the rpsL gene (Lysine-43-Arginine) were detected. The blaOXA resistance gene was present in all isolates, the gyrA mutations was in 95.2% of C. jejuni and 80.0% of C. coli, and the tet(O) resistance gene in 76.2% of C. jejuni and 80.0% of C. coli. Additionally, 203 virulence-associated genes linked to 16 virulence factors were identified. In terms of phenotypic resistance, the C. jejuni isolates were all resistant to ciprofloxacin, enrofloxacin, and nalidixic acid, with lower levels of resistance to tetracycline (76.2%), tylosin (52.3%), erythromycin (23.8%), azithromycin (22.2%), and gentamicin (11.1%). Most C. coli isolates were resistant to all tested antimicrobials, while 1 C. coli was pan-susceptible except for tylosin. Single-nucleotide polymorphisms concordance varied widely, with differences of up to 13,375 single-nucleotide polymorphisms compared to the reference Campylobacter isolates, highlighting genetic divergence among comparative genomes. This study contributes to a deeper understanding of the molecular epidemiology of Campylobacter spp. in Thai chicken production systems.

A systematic review and meta-analysis of the global prevalence and relationships among Burkholderia pseudomallei sequence types isolated from humans, animals, and the environment.

Background and Aim: Burkholderia pseudomallei, a highly pathogenic bacterium responsible for melioidosis, exhibits ecological ubiquity and thrives within soil and water reservoirs, posing significant infection risks to humans and animals through direct contact. The aim of this study was to elucidate the genetic diversity and prevalence patterns of B. pseudomallei sequence types (STs) across a global spectrum and to understand the relationships between strains isolated from different sources. Materials and Methods: We performed a systematic review and meta-analysis in this study. Extensive research was carried out across three comprehensive databases, including PubMed, Scopus, and ScienceDirect with data collected from 1924 to 2023. Results: A total of 40 carefully selected articles contributed 2737 B. pseudomallei isolates attributed to 729 distinct STs and were incorporated into the systematic review. Among these, ST46 emerged as the most prominent, featuring in 35% of the articles and demonstrating a dominant prevalence, particularly within Southeast Asia. Moreover, ST51 consistently appeared across human, animal, and environmental studies. Subsequently, we performed a meta-analysis, focusing on nine specific STs: ST46, ST51, ST54, ST70, ST84, ST109, ST289, ST325, and ST376. Surprisingly, no statistically significant differences in their pooled prevalence proportions were observed across these compartments for ST46, ST70, ST289, ST325, and ST376 (all p > 0.69). Conversely, the remaining STs, including ST51, ST54, ST84, and ST109, displayed notable variations in their prevalence among the three domains (all p < 0.04). Notably, the pooled prevalence of ST51 in animals and environmental samples surpassed that found in human isolates (p < 0.01). Conclusion: To the best of our knowledge, this study is the first systematic review and meta-analysis to investigate the intricate relationships between STs and their sources and contributes significantly to our understanding of B. pseudomallei diversity within the One Health framework.

The characterization and correlation between the phenotypic and genotypic resistance of Campylobacter spp. isolates from commercial broilers and native chickens in the south of Thailand.

RESEARCH HIGHLIGHTS: High Campylobacter prevalence in chickens; C. jejuni more prevalent than C. coli.Susceptibility to macrolides but resistance to quinolones/tetracyclines in isolates.Homogeneous resistance patterns within farms; higher in broilers than in native birds.Partial association between phenotypic and genotypic resistance among isolates.

Prevalence and factors associated with long COVID and mental health status among recovered COVID-19 patients in southern Thailand.

Mental health disorders have become a growing public health concern among individuals recovering from COVID-19. Long COVID, a condition where symptoms persist for an extended period, can predict psychological problems among COVID-19 patients. This study aimed to investigate the prevalence of long COVID and mental health status among Thai adults who had recovered from COVID-19, identify the association between the mental health status and long COVID symptoms, and investigate the risk factors associated with the correlation between long COVID and mental health outcomes. A cross-sectional study was conducted among 939 randomly selected participants in Nakhon Si Thammarat province, southern Thailand. The Depression, Anxiety, and Stress Scale-21 was used to investigate mental health symptoms, and a checklist comprised of thirteen common symptoms was used to identify the long COVID among participants. Logistic regression models were used to investigate the risk factors associated with mental health status and long COVID symptoms among participants. Among the 939 participants, 104 (11.1%) had depression, 179 (19.1%) had anxiety, and 42 (4.8%) were stressed. A total of 745 participants (79.3%) reported experiencing at least one symptom of long COVID, with fatigue (72.9%, SE±0.02), cough (66.0%, SE±0.02), and muscle pain (54.1%, SE±0.02) being the most frequently reported symptoms. All long COVID symptoms were significantly associated with mental health status. Shortness of breath, fatigue, and chest tightness were the highest risk factors for mental health status among COVID-19 patients. The final multivariable model indicated that female patients (OR = 1.89), medical history (OR = 1.92), and monthly income lower than 5,000 Thai baht (OR = 2.09) were associated with developing long COVID symptoms and mental health status (all p<0.01). This study provides valuable insights into the potential long-term effects of COVID-19 on mental health and enhances understanding of the mechanisms underlying the condition for predicting the occurrence of mental health issues in Thai COVID-19 patients.

Impact of antimicrobial use on abundance of antimicrobial resistance genes in chicken flocks in Vietnam.

Objectives: We investigated longitudinally Vietnamese small-scale chicken flocks in order to characterize changes in antimicrobial resistance gene (ARG) content over their life cycle, and the impact of antimicrobial use (AMU) on an intervention consisting of veterinary advice provision. Methods: AMU data and faecal samples were collected from 83 flocks (25 farms) at day-old, mid- and late-production (∼4 month cycle). Using high-throughput real-time PCR, samples were investigated for 94 ARGs. ARG copies were related to 16S rRNA and ng of DNA (ngDNA). Impact of AMU and ARGs in day-olds was investigated by mixed-effects models. Results: Flocks received a mean (standard error, SE) animal daily dose (ADD) of 736.7 (83.0) and 52.1 (9.9) kg in early and late production, respectively. Overall, ARGs/16S rRNA increased from day-old (mean 1.47; SE 0.10) to mid-production (1.61; SE 0.16), further decreasing in end-production (1.60; SE 0.1) (all P > 0.05). In mid-production, ARGs/16S rRNA increased for aminoglycosides, phenicols, sulphonamides and tetracyclines, decreasing for polymyxins ß-lactams and genes that confer resistance to mutiple classes (multi-drug resistance) (MDR). At end-production, aminoglycoside resistance decreased and polymyxin and quinolone resistance increased (all P < 0.05). Results in relation to ngDNA gave contradictory results. Neither AMU nor ARGs in day-olds had an impact on subsequent ARG abundance. The intervention resulted in 74.2% AMU reduction; its impact on ARGs depended on whether ARGs/ngDNA (+14.8%) or ARGs/16S rRNA metrics (-10.7%) (P > 0.05) were computed. Conclusions: The flocks' environment (contaminated water, feed and residual contamination) is likely to play a more important role in transmission of ARGs to flocks than previously thought. Results highlight intriguing differences in the quantification of ARGs depending on the metric chosen.

A systematic review and meta-analysis of integrated studies on antimicrobial resistance in Vietnam, with a focus on Enterobacteriaceae, from a One Health perspective.

Vietnam is a low- and middle-income country (LMIC), a primary food producer, and an antimicrobial resistance (AMR) hotspot. AMR is recognized as a One Health challenge since it may transfer between humans, animals and the environment. This study aimed to apply systematic review and meta-analysis to investigate the phenotypic profiles and correlations of antimicrobial-resistant Enterobacteriaceae across three compartments: humans, animals and the environment in Vietnam. A total of 89 articles found in PubMed, Science Direct, and Google Scholar databases were retrieved for qualitative synthesis. E. coli and non-typhoidal Salmonella (NTS) were the most common bacterial species in studies of all compartments (60/89 studies). Among antimicrobials classified as critically important, the resistance levels were observed to be highest to quinolones, 3rd generation of cephalosporins, penicillins, and aminoglycosides. Of 89 studies, 55 articles reported the resistance prevalence of E. coli and NTS in healthy humans, animals and the environment against ciprofloxacin, ceftazidime, ampicillin, gentamicin, sulfamethoxazole-trimethoprim, chloramphenicol was used for meta-analysis. The pooled prevalence was found highest in E. coli against ampicillin 84.0% (95% CI 73.0-91.0%) and sulfamethoxazole-trimethoprim 66.0% (95% CI 56.0-75.0%) while in NTS they were 34.0% (95% CI 24.0-46.0%), 33.0% (95% CI 25.0-42.0%), respectively. There were no significant differences in the pooled prevalence of E. coli and NTS to these antimicrobials across healthy humans, animals and the environment, except for ceftazidime-resistant E. coli (χ2 = 8.29, p = 0.02), chloramphenicol-resistant E.coli (χ2 = 9.65, p < 0.01) and chloramphenicol-resistant NTS (χ2 = 7.51, p = 0.02). Findings from the multiple meta-regression models indicated that the AMR levels in E. coli (ß = 1.887, p < 0.001) and the North (ß = 0.798, p = 0.047) had a higher fraction of AMR than NTS and other regions of Vietnam. The outcomes of this study play an important role as the baseline information for further investigation and follow-up intervention strategies to tackle AMR in Vietnam, and more generally, can be adapted to other LMICs.

Impact of Freeze Storage on the Estimation of Phenotypic Antimicrobial Resistance Prevalence in Escherichia coli Collected from Faecal Samples from Healthy Humans and Chickens.

Many studies on phenotypic antimicrobial resistance (AMR) of bacteria from healthy populations are conducted on freeze-stored samples. However, the impact of this practice on phenotypic AMR is not known. We investigated the prevalence of phenotypic AMR in Escherichia coli from chicken (n = 10) and human (n = 11) faecal samples collected from healthy subjects, subject to freeze storage (-20 °C and -80 °C) for 1, 2, 3, and 6 months. We compared counts of E. coli and prevalence of phenotypic resistance against five antimicrobials commonly used in chicken farming (ciprofloxacin, enrofloxacin, doxycycline, gentamicin, and florfenicol) with samples processed within 24 h of collection. Prevalence of phenotypic AMR was estimated by performing differential counts on agar media with and without antimicrobials. At -20 °C, there was a considerable reduction in E. coli counts over time, and this reduction was greater for human samples (-0.630 log10 units per 100 days) compared with chicken samples (-0.178 log10 units per 100 days). For most antimicrobials, AMR prevalence estimates decreased in freeze-stored samples both in humans and chickens over time. Based on these results, we conclude that results on the prevalence of phenotypic AMR on samples from freeze-stored samples are unreliable, and only fresh samples should be used in such studies.

Antimicrobial resistance in commensal Escherichia coli from humans and chickens in the Mekong Delta of Vietnam is driven by antimicrobial usage and potential cross-species transmission.

Objectives: To investigate phenotypic antimicrobial resistance (AMR) in relation to antimicrobial use (AMU) and potential inter-species transmission among Escherichia coli from humans and chickens located in the same households in the Mekong Delta of Vietnam. Methods: We collected data on AMU and faecal swabs from humans (N = 426) and chickens (N = 237) from 237 small-scale farms. From each sample, one E. coli strain was isolated and tested for its susceptibility against 11 antimicrobials by Sensititre AST. The association between AMR and AMU was investigated by logistic regression modelling. Using randomization, we compared the degree of similarity in AMR patterns between human and chicken E. coli from the same farms compared with isolates from different farms. Results: The AMU rate was ∼19 times higher in chickens (291.1 per 1000 chicken-days) than in humans (15.1 per 1000 person-days). Isolates from chickens also displayed a higher prevalence of multidrug resistance (63.3%) than those of human origin (55.1%). AMU increased the probability of resistance in isolates from human (ORs between 2.1 and 5.3) and chicken (ORs between 1.9 and 4.8). E. coli from humans and chickens living on same farms had a higher degree of similarity in their AMR patterns than isolates from humans and chicken living on different farms. Conclusions: We demonstrated the co-influence of AMU and potential transmission on observed phenotypic AMR patterns among E. coli isolates from food-producing animals and in-contact humans. Restricting unnecessary AMU alongside limiting interspecies contact (i.e. increasing hygiene and biocontainment) are essential for reducing the burden of AMR.

Prevalence of carbapenem resistance and its potential association with antimicrobial use in humans and animals in rural communities in Vietnam.

Background: Vietnam and Southeast Asia are hotspots for antimicrobial resistance; however, little is known on the prevalence of carriage of carbapenem resistance in non-hospitalized humans and in animals. Carbapenem-resistant Enterobacteriaceae (CRE), particularly Escherichia coli (CREC) and Klebsiella pneumoniae (CRKP) and also Acinetobacter baumannii (CRAB) are emerging threats worldwide. Methods: We investigated healthy humans (n = 652), chickens (n = 237), ducks (n = 150) and pigs (n = 143) in 400 small-scale farms in the Mekong Delta of Vietnam. Samples (rectal swabs, faecal swabs) were investigated for carriage of CRE/CRAB and were further characterized phenotypically and genotypically. Results: In the Mekong Delta of Vietnam, the prevalence of CRE isolates in human rectal swabs was 0.6%, including 4 CREC and 1 CRKP. One pig was infected with CREC (prevalence 0.7%). CRAB was isolated from chickens (n = 4) (prevalence 2.1%) and one duck (prevalence 0.7%). CRKP was isolated from a human who was also colonized with CREC. The CRKP strain (ST16), from an 80 year-old person with pneumonia under antimicrobial treatment, genetically clustered with clinical strains isolated in a hospital outbreak in southern Vietnam. The prevalence of CRE was higher among humans that had used antimicrobials within 90 days of the sampling date than those had not (4.2% versus 0.2%) (P = 0.005). All CRE/CRAB strains were MDR, although they were susceptible to colistin and neomycin. The carbapenemase genes identified in study strains were bla NDM and bla OXA. Conclusions: The finding of a CRKP strain clustering with previous hospital outbreak raises concerns about potential transmission of carbapenem-resistant organisms from hospital to community settings or vice-versa.

Feasibility study of a field survey to measure antimicrobial usage in humans and animals in the Mekong Delta region of Vietnam.

BACKGROUND: Development of antimicrobial use (AMU) surveillance systems in humans and animals is a priority for many low- and middle-income countries; however accurate estimations are hampered by a diversity of animal production systems and metrics. The Mekong Delta region of Vietnam is a 'hotspot' of antimicrobial resistance and is home to a high density of humans and animal populations. OBJECTIVES: To measure and compare AMU using different metrics (standing population, biomass and population correction unit) in the Mekong Delta, and to explore the potential of field-based data collection methods in the design of AMU surveillance systems. METHODS: We collected AMU data from humans and animals (chickens, ducks, Muscovy ducks, pigs) from 101 small-scale farms in the Mekong Delta over a fixed period (90 days in humans, 7 days in animals). RESULTS: Humans used 7.1 DDDkg, or 175.9 mg of antimicrobial active ingredients (AAIs) per kg of standing body mass annually; animals consumed 60.9 ADDkg or 1324 mg. In the Mekong Delta humans represented 79.3% of the total body mass but consumed 29.6% of AAIs by weight. AAIs regarded of critical importance by WHO represented 56.9% and 50.2% of doses consumed by animals and humans, respectively. CONCLUSIONS: Using a One Health approach, we show that AMU can potentially be estimated from cross-sectional surveys, although results are hypothetical due to small sample size and are sensitive to the chosen population denominator. The methodology proposed here can potentially be scaled up be applied to design AMU surveillance in low-resource settings, allowing AMU reduction efforts to be focused on particular animal species.

Effects of prophylactic and therapeutic antimicrobial uses in small-scale chicken flocks.

Antimicrobials are extensively used both prophylactically and therapeutically in poultry production. Despite this, there are little data on the effect of antimicrobial use (AMU) on disease incidence rate and per cent mortality. We investigated the relationships between AMU and disease and between AMU and mortality using data from a large (n = 322 flocks) cohort of small-scale chicken flocks in the Mekong Delta, Vietnam, that were followed longitudinally from day old to slaughter (5,566 observation weeks). We developed a parameterized algorithm to emulate a randomized control trial from observational data by categorizing the observation weeks into 'non-AMU', 'prophylactic AMU' and 'therapeutic AMU'. To evaluate the prophylactic AMU effect, we compared the frequencies of clinical signs in 'non-AMU' and 'prophylactic AMU' periods. To analyse therapeutic AMU, we compared weekly per cent mortality between the weeks of disease episodes before and after AMU. Analyses were stratified by clinical signs (4) and antimicrobial classes (13). Prophylactic AMU never reduced the probability of disease, and some antimicrobial classes such as lincosamides, amphenicols and penicillins increased the risk. The risk of diarrhoea consistently increased with prophylactic AMU. Therapeutic AMU often had an effect on mortality, but the pattern was inconsistent across the combinations of antimicrobial classes and clinical signs with 14/29 decreasing and 11/29 increasing the per cent weekly mortality. Lincosamides, methenamines and cephalosporins were the only three antimicrobial classes that always decreased the mortality when used therapeutically. Results were robust respective to the parameters values of the weeks categorization algorithm. This information should help support policy efforts and interventions aiming at reducing AMU in animal production.

Antimicrobial use through consumption of medicated feeds in chicken flocks in the Mekong Delta of Vietnam: A three-year study before a ban on antimicrobial growth promoters.

Antimicrobials are included in commercial animal feed rations in many low- and middle-income countries (LMICs). We measured antimicrobial use (AMU) in commercial feed products consumed by 338 small-scale chicken flocks in the Mekong Delta of Vietnam, before a gradual nationwide ban on prophylactic use of antimicrobials (including in commercial feeds) to be introduced in the country over the coming five years. We inspected the labels of commercial feeds and calculated amounts of antimicrobial active ingredients (AAIs) given to flocks. We framed these results in the context of overall AMU in chicken production, and highlighted those products that did not comply with Government regulations. Thirty-five of 99 (35.3%) different antimicrobial-containing feed products included at least one AAI. Eight different AAIs (avilamycin, bacitracin, chlortetracycline, colistin, enramycin, flavomycin, oxytetracycline, virginamycin) belonging to five classes were identified. Brooding feeds contained antimicrobials the most (60.0%), followed by grower (40.9%) and finisher feeds (20.0%). Quantitatively, chlortetracycline was consumed most (42.2 mg/kg SEM ±0.34; 50.0% of total use), followed by enramycin (18.4 mg/kg SEM ±0.03, 21.8%), bacitracin (16.4 mg/kg SEM ±0.20, 19.4%) and colistin (6.40 mg/kg SEM ± 4.21;7.6%). Other antimicrobials consumed were virgianamycin, avilamycin, flavomycin and oxytetracycline (each ≤0.50 mg/kg). Antimicrobials in commercial feeds were more commonly given to flocks in the earlier part of the production cycle. A total of 10 (9.3%) products were not compliant with existing Vietnamese regulation (06/2016/TT-BNNPTNT) either because they included a non-authorised AAI (4), had AAIs over the permitted limits (4), or both (2). A number of commercial feed formulations examined included colistin (polymyxin E), a critically important antimicrobial of highest priority for human medicine. These results illustrate the challenges for effective implementation and enforcement of restrictions of antimicrobials in commercial feeds in LMICs. Results from this study should help encourage discussion about policies on medicated feeds in LMICs.

Characterizing Antimicrobial Resistance in Chicken Pathogens: A Step towards Improved Antimicrobial Stewardship in Poultry Production in Vietnam.

In the Mekong Delta of Vietnam, farmers use large quantities of antimicrobials to raise small-scale chicken flocks, often including active ingredients regarded of "critical importance'" by the World Health Organization. Due to limitations in laboratory capacity, the choice of antimicrobials normally does not follow any empirical criteria of effectiveness. The aim of this study was to highlight non-critically important antimicrobials against which chicken pathogens are likely to be susceptible as a basis for treatment guidelines. Microtiter broth dilution method was performed to determine the minimal inhibitory concentration (MIC) of 12 commonly used antimicrobials for 58 isolates, including Ornithobacterium rhinotracheale (ORT) (n = 22), Gallibacterium anatis (n = 19), and Avibacterium endocarditidis (n = 17). Unfortunately, internationally accepted breakpoints for resistance in these organisms do not exist. We drew tentative epidemiological cut-offs (TECOFFs) for those antimicrobial-pathogen combinations where MIC distributions suggested the presence of a distinct non-wild-type population. Based on the observed results, doxycycline would be the drug of choice for A.endocarditidis (11.8% presumptive non-wild type) and G. anatis infections (5.3% presumptive non-wild type). A total of 13.6% ORT isolates were non-wild type with regards to oxytetracycline, making it the drug of choice against this pathogen. This study illustrates the challenges in interpreting susceptibility testing results and the need to establish internationally accepted breakpoints for veterinary pathogens.

Reducing Antimicrobial Usage in Small-Scale Chicken Farms in Vietnam: A 3-Year Intervention Study.

Indiscriminate antimicrobial use (AMU) in animal production is a driver of antimicrobial resistance globally. There is a need to define sustainable interventions to reduce AMU in small-scale production systems, which currently represent the most widespread farming systems in South East Asia and many low- and middle-income countries. We conducted a before-and-after intervention study on a random sample of small-scale chicken farms in the Mekong Delta of Vietnam from 2016 to 2019. The study included a baseline followed by an intervention phase where farmers were provided with regular veterinary advice on flock health and husbandry, as well as antimicrobial replacement products. Of 102 recruited farms (raising >100 chickens per flock cycle), thirty-five (34.2%) entered the intervention phase, whilst the rest stopped raising chickens, mainly due to suboptimal flock performance. Through the implementation of our intervention, chicken flocks reduced levels of AMU by 66% [adjusted hazard ratio (HR) = 0.34; p = 0.002) from a baseline of 343.4 Animal Daily Doses per 1,000 chicken-days and decreased weekly mortality by 40% (adjusted HR = 0.60; p = 0.005) from a baseline mortality of 1.60 per 100 birds. Chicken bodyweight increased by 100 g (p = 0.002) in intervention flocks. Our findings demonstrate that the provision of veterinary advice can achieve substantial reductions in AMU in small-scale production systems without compromising flock health and productivity.

Veterinary Drug Shops as Main Sources of Supply and Advice on Antimicrobials for Animal Use in the Mekong Delta of Vietnam.

In the Mekong Delta of Vietnam, small-scale poultry farmers use large amounts of antimicrobials to raise their flocks, and veterinary drug shops owners and their staff are a key source of advice to farmers on antimicrobial use (AMU). We described the network of veterinary drug shops (n = 93) in two districts within Dong Thap province (Mekong Delta). We also interviewed a randomly selected sample of chicken farmers (n = 96) and described their linkages with veterinary drug shops. Antimicrobials represented 15.0% [inter quartile range (IQR) 6.0-25.0] of the shops' income. Fifty-seven percent shop owners had been/were affiliated to the veterinary authority, 57% provided diagnostic services. The median number of drug shops supplying antimicrobials to each farm during one production cycle was 2 [IQR 1-2]. Visited shops were located within a median distance of 3.96 km [IQR 1.98-5.85] to farms. Drug shops owned by persons affiliated to the veterinary authority that did not provide diagnostic services had a higher fraction of their income consisting of antimicrobial sales (ß = 1.913; p < 0.001). These results suggest that interventions targeting veterinary drug shop owners and their staff aiming at improving their knowledge base on livestock/poultry diseases and their diagnosis may contribute to reducing overall levels of AMU in the area.

Labelling and quality of antimicrobial products used in chicken flocks in the Mekong Delta of Vietnam.

BACKGROUND: The Mekong Delta of Vietnam is a hotspot of antimicrobial use (AMU), but there is no information on the quality of the labelling and strength of antimicrobial products used in poultry production. METHODS: Based on a large random sample of farms, we identified the 20 most used antimicrobial products in the area, and investigated their antimicrobial active ingredient (AAI) content by UPLC-MS/MS (91 analytical tests). RESULTS: Only 17/59 (28.8%) batches contained all AAIs within 10% of the declared strength. Worryingly, 65.0% products provided in their label preparation guidelines for both therapeutic and prophylactic use. Withdrawal times for both meat and eggs were stated in 8/20 (40%) products. CONCLUSION: Results highlight deficiencies in quality and labelling contents that undermine authorities' efforts to discourage inappropriate use of antimicrobials.

High-Resolution Monitoring of Antimicrobial Consumption in Vietnamese Small-Scale Chicken Farms Highlights Discrepancies Between Study Metrics.

Chicken is, among farmed species, the target of the highest levels of antimicrobial use (AMU). There are considerable knowledge gaps on how and when antimicrobials are used in commercial small-scale chicken farms. These shortcomings arise from cross-sectional study designs and poor record keeping practiced by many such farmers. Furthermore, there is a large diversity of AMU metrics, and it is not clear how these metrics relate to each other. We performed a longitudinal study on a cohort of small-scale chicken farms (n = 102) in the Mekong Delta (Vietnam), an area regarded as a hotspot of AMU, from October 2016 to May 2018. We collected data on all medicine products administered to 203 flocks with the following aims: (1) to describe types and quantities of antimicrobial active ingredients (AAIs) used; (2) to describe critical time points of AMU; and (3) to compare AMU using three quantitative metrics: (a) weight of AAIs related to bird weight at the time of treatment (mg/kg at treatment); (b) weight of AAIs related to weight of birds sold (mg/kg sold); and (c) "treatment incidence" (TI), or the number of daily doses per kilogram of live chicken [Vietnamese animal daily dose (ADDvetVN)] per 1,000 days. Antimicrobials contained in commercial feed, administered by injection (n = N = 6), or antimicrobials for human medicine (n = N = 16) were excluded. A total of 236 products were identified, containing 42 different AAIs. A total of 76.2% products contained AAIs of "critical importance" according to the World Health Organization (WHO). On average, chickens consumed 791.8 (SEM ±16.7) mg/kg at treatment, 323.4 (SEM ±11.3) mg/kg sold, and the TI was 382.6 (SEM ±5.5) per 1,000 days. AMU was more common early in the production cycle and was highly skewed, with the upper 25% quantile of flocks accounting for 60.7% of total AMU. The observed discrepancies between weight- and dose-based metrics were explained by differences in the strength of AAIs, mortality levels, and the timing of administration. Results suggest that in small-scale chicken production, AMU reduction efforts should preferentially target the early (brooding) period, which is when birds are most likely to be exposed to antimicrobials, whilst restricting access to antimicrobials of critical importance for human medicine.