Evidence of SARS-CoV-2 Related Coronaviruses Circulating in Sunda pangolins (Manis javanica) Confiscated From the Illegal Wildlife Trade in Viet Nam.

Despite the discovery of several closely related viruses in bats, the direct evolutionary progenitor of SARS-CoV-2 has not yet been identified. In this study, we investigated potential animal sources of SARS-related coronaviruses using archived specimens from Sunda pangolins (Manis javanica) and Chinese pangolins (Manis pentadactyla) confiscated from the illegal wildlife trade, and from common palm civets (Paradoxurus hermaphroditus) raised on wildlife farms in Viet Nam. A total of 696 pangolin and civet specimens were screened for the presence of viral RNA from five zoonotic viral families and from Sarbecoviruses using primers specifically designed for pangolin coronaviruses. We also performed a curated data collection of media reports of wildlife confiscation events involving pangolins in Viet Nam between January 2016 and December 2020, to illustrate the global pangolin supply chain in the context of Viet Nam where the trade confiscated pangolins were sampled for this study. All specimens from pangolins and civets sampled along the wildlife supply chains between February 2017 and July 2018, in Viet Nam and tested with conventional PCR assays designed to detect flavivirus, paramyxovirus, filovirus, coronavirus, and orthomyxovirus RNA were negative. Civet samples were also negative for Sarbecoviruses, but 12 specimens from seven live pangolins confiscated in Hung Yen province, northern Viet Nam, in 2018 were positive for Sarbecoviruses. Our phylogenetic trees based on two fragments of the RdRp gene revealed that the Sarbecoviruses identified in these pangolins were closely related to pangolin coronaviruses detected in pangolins confiscated from the illegal wildlife trade in Yunnan and Guangxi provinces, China. Our curated data collection of media reports of wildlife confiscation events involving pangolins in Viet Nam between January 2016 and December 2020, reflected what is known about pangolin trafficking globally. Pangolins confiscated in Viet Nam were largely in transit, moving toward downstream consumers in China. Confiscations included pangolin scales sourced originally from Africa (and African species of pangolins), or pangolin carcasses and live pangolins native to Southeast Asia (predominately the Sunda pangolin) sourced from neighboring range countries and moving through Viet Nam toward provinces bordering China.

The prevalence of pre-treatment and acquired HIV drug resistance in Vietnam: a nationally representative survey, 2017-2018.

INTRODUCTION: Monitoring the population-level emergence and transmission of HIV drug resistance (HIVDR) is necessary for supporting public health programmes. This study provides a nationally representative prevalence estimate of HIVDR in people initiating antiretroviral therapy (ART) and estimates of acquired HIVDR and viral load (VL) suppression in people who have received it for 12 or ≥48 months in Vietnam. METHODS: The study was conducted between September 2017 and March 2018 following World Health Organization guidance. Thirty ART clinics were randomly sampled using probability proportional to size sampling from a total of 367 ART clinics in the country. RESULTS AND DISCUSSION: In total, 409 patients initiating ART were enrolled into the survey of pre-treatment HIVDR. The prevalence of any pre-treatment HIVDR was 5.8% (95% CI 3.4-9.5%), and the prevalence of non-nucleoside reverse transcriptase inhibitor resistance was 3.4% (95% CI 1.8-6.2%). Four hundred twenty-nine patients on ART for 12±3 months and 723 patients on ART for ≥48 months were enrolled into the surveys of acquired HIVDR. The prevalence of VL suppression (defined as <1000 copies/ml) in patients on ART for 12±3 and ≥48 months was 95.5% (95% CI 91.3-97.8%) and 96.1% (95% CI 93.2-97.8%), respectively. Among individuals with viral non-suppression, any HIVDR was detected in 11/14 (weighted prevalence 74.3%) of those on ART for 12±3 months and in 24/27 (weighted prevalence 88.5%) of those receiving ART for ≥48 months. CONCLUSIONS: This nationally representative study of HIVDR found high levels of VL suppression among those on ART for 12 and ≥48 months. Overall, high levels of VL suppression at both time points suggested good adherence among patients receiving ART and quality of treatment services in Vietnam. CLINICAL TRIAL NUMBER: Not applicable.

Viral load suppression and acquired HIV drug resistance in adults receiving antiretroviral therapy in Viet Nam: results from a nationally representative survey.

OBJECTIVE: The purpose of this survey was to estimate the prevalence of viral load (VL) suppression and emergence of HIV drug resistance (HIVDR) among individuals receiving antiretroviral therapy (ART) for 36 months or longer in Viet Nam using a nationally representative sampling method. METHODS: The survey was conducted between May and August 2014 using a two-stage cluster design. Sixteen ART clinics were selected using probability proportional to proxy size sampling, and patients receiving ART for at least 36 months were consecutively enrolled. Epidemiological information and blood specimens were collected for HIV-1 VL and HIVDR testing; HIVDR was defined by the Stanford University HIVDR algorithm. RESULTS: Overall, 365 eligible individuals were recruited with a mean age of 38.2 years; 68.4% were men. The mean time on ART was 75.5 months (95% confidence interval [CI]: 69.0-81.9 months), and 93.7% of the patients were receiving non-nucleoside reverse transcriptase inhibitor-based regimens. Of the 365 individuals, 345 (94.7%, 95% CI: 64.1-99.4%) had VL below 1000 copies/mL and 19 (4.6%, 95% CI: 2.8--7.5) had HIVDR mutations. DISCUSSION: Our nationally representative survey found a high level of VL suppression and a low prevalence of HIVDR among individuals who received ART for at least 36 months in Viet Nam. Continued surveillance for HIVDR is important for evaluating and improving HIV programs.

Toxocara malaysiensis infection in domestic cats in Vietnam--An emerging zoonotic issue?

Toxocara canis of canids is a parasitic nematode (ascaridoid) that infects humans and other hosts, causing different forms of toxocariasis. This species of Toxocara appears to be the most important cause of human disease, likely followed by Toxocara cati from felids. Although some studies from Malaysia and China have shown that cats can harbor another congener, T. malaysiensis, no information is available about this parasite for other countries. Moreover, the zoonotic potential of this parasite is unknown at this point. In the present study, we conducted the first investigation of domestic dogs and cats for Toxocara in Vietnam using molecular tools. Toxocara malaysiensis was identified as a common ascaridoid of domestic cats (in the absence of T. cati), and T. canis was commonly found in dogs. Together with findings from previous studies, the present results emphasize the need to explore the significance and zoonotic potential of T. malaysiensis in Vietnam and other countries where this parasite is endemic and prevalent in cats.

Association of Chicken Growth Hormones and Insulin-like Growth Factor Gene Polymorphisms with Growth Performance and Carcass Traits in Thai Broilers.

Molecular marker selection has been an acceptable tool in the acceleration of the genetic response of desired traits to improve production performance in chickens. The crossbreds from commercial parent stock (PS) broilers with four Thai synthetic breeds; Kaen Thong (KT), Khai Mook Esarn (KM), Soi Nin (SN), and Soi Pet (SP) were used to study the association among chicken growth hormones (cGH) and the insulin-like growth factor (IGF-I) genes for growth and carcass traits; for the purpose of developing a suitable terminal breeding program for Thai broilers. A total of 408 chickens of four Thai broiler lines were genotyped, using polymerase chain reaction-restriction fragment length polymorphism methods. The cGH gene was significantly associated with body weight at hatching; at 4, 6, 8, 10 weeks of age and with average daily gain (ADG); during 2 to 4, 4 to 6, 0 to 6, 0 to 8, and 0 to 10 weeks of age in PS×KM chickens. For PS×KT populations, cGH gene showed significant association with body weight at hatching, and ADG; during 8 to 10 weeks of age. The single nucleotide polymorphism variant confirmed that allele G has positive effects for body weight and ADG. Within carcass traits, cGH revealed a tentative association within the dressing percentage. For the IGF-I gene polymorphism, there were significant associations with body weight at hatching; at 2, 4, and 6 weeks of age and ADG; during 0 to 2, 4 to 6, and 0 to 6 weeks of age; in all of four Thai broiler populations. There were tentative associations of the IGF-I gene within the percentages of breast muscles and wings. Thus, cGH gene may be used as a candidate gene, to improve growth traits of Thai broilers.

Evaluation of four coproscopic techniques for detection of small trematode eggs in dog faeces.

The study was conducted to evaluate copro-diagnostic techniques for detection of small trematode eggs in dogs. FLOTAC, a novel flotation technique, and DBL, a sieving and sedimentation technique developed at the former Danish Bilharziasis Laboratory (DBL), were compared using 53 subsamples from four copro-positive dogs. Moreover, a modified version of the DBL technique and the Kato-Katz (KK) thick smear were later compared using faecal samples from 21 dogs. The four techniques were pair-wise compared regarding sensitivity, infection intensity and practical applicability. For the former two techniques, egg recovery subsequent to storage and reproducibility were also compared. The DBL technique detected all 53 subsamples positive for small trematode eggs. Based on 17 subsamples, mean infection intensity of 47±49 eggs per gram of faeces (EPG) was detected by the technique. Due to large amount of sediment, examination of a single subsample required an average of 3 hours. The FLOTAC technique was found less sensitive (82%) than the DBL technique and recovered significantly less eggs (4±6 EPG). Both sensitivity and intensity were further reduced following storage. As the FLOTAC technique requires specialised equipment, safety disposal and personal protective equipment, it was found less suited than the DBL technique for a basic laboratory. Additionally, poor reproducibilities were found for both the DBL and FLOTAC techniques (30±15% and 38±33%, respectively). Based on the 21 faecal samples, the modified version of the DBL technique was found more sensitive (85%) than the KK technique (68%), whereas egg counts were significantly higher for the latter (23±26 EPG vs. 482±909 EPG). By modifying the DBL technique, it was possible to diminish the retained sediment and examination time to a maximum of an hour, which was also the time required by the KK technique, although the latter was faster and more easily processed. Based on the results obtained in this study, none of the techniques evaluated were found applicable in their current form for detection of small trematode eggs in faeces from dogs in Vietnam.

Field trial of praziquantel for control of fishborne zoonotic trematodes in reservoir hosts in Vietnam.

This field trial was conducted to determine whether 40 or 75 mg/kg of praziquantel is suitable for treatment of fishborne zoonotic trematodes (FZT) in naturally infected dogs (n=10) and cats (n=11). Three days after treatment all animals at either dose were negative for small trematode eggs. In two cats and one dog treated with 75 mg/kg, however, a few damaged eggs were found 3 days post-treatment; no small trematode eggs were seen in these animals at day 14 post-treatment. In addition, at the 75 mg dose, two cats and two dogs experienced vomiting or diminished appetite. Therefore a praziquantel dose of 40 mg/kg is suggested for treatment of FZT in dogs and cats.

Poultry as reservoir hosts for fishborne zoonotic trematodes in Vietnamese fish farms.

Fishborne zoonotic trematodes (FZT) are widespread in Vietnam and Southeast Asia. It is now recognized that the risk of being infected from eating raw fish dishes applies not only to humans, but also to domestic animals (e.g., cats, dogs, and pigs) and fish-eating birds. The role of ducks and chicken, commonly raised on fish farms, as reservoir hosts, however, has not been adequately investigated. To study this question, chickens and ducks from integrated poultry-fish farms in Nghia Lac and Nghia Phu communes, Nam Dinh province, Vietnam were surveyed for FZT infections. A total of 50 ducks and 50 chickens from each commune were examined. Results revealed that 12% of chickens and 30% of ducks were infected with various species of trematodes, including two zoonotic species, Centrocestus formosanus and Echinostoma cinetorchis. Both occurred in chickens whereas only E. cinetorchis was found in ducks. Prevalence of these zoonotic species was 12% and 7% in ducks and chickens, respectively. Among other trematodes, Hypoderaeum conoideum, also a zoonotic fluke, was the most prevalent (20-30%). The feeding of snails and fish remains to poultry, either intentionally or by discharge of waste from the slaughter of ducks and chickens into the ponds, was identified as risk factors for trematode infection. The FZT species and low prevalence found in poultry in these communes indicate their role as reservoir hosts is minor.

Evaluation of techniques for detection of small trematode eggs in faeces of domestic animals.

This study was conducted to evaluate techniques for detection of small trematode eggs in faeces of dogs, cats and pigs. Faecal samples from dogs (n=80), cats (n=35) and pigs (n=114) were examined by Kato-Katz technique (KK), formalin-ether sedimentation technique (FE) and a method of combining: filtration, sedimentation and centrifugation, developed at DBL - Centre for Health Research and Development (former Danish Bilharziasis Laboratory) (DBL). Necropsy was performed on 38 dogs, 25 cats and 16 pigs and was considered as a gold standard method for evaluation of the techniques. The results showed 100% specificity for the three techniques. Lower sensitivity was seen for the KK-technique in dog samples in comparison to that for DBL- and FE-technique. The sensitivity of the three techniques was similar in cats and pigs. Based on these findings and practical issues, DBL-technique was chosen as most suitable because the eggs were easily detected and quantified. No toxic chemicals or special equipment were required in comparison with FE-technique that needs ether solution and thus fume cupboards which are often unavailable in local veterinary centres in Vietnam and other developing countries.