Unexpected cases in field diagnosis of African swine fever virus in Vietnam: The needs consideration when performing molecular diagnostic tests

Background: The first confirmed case of African swine fever (ASF) in Vietnam was reported officially in February 2019. To date, ASF virus (ASFV) have been detected in 63/63 provinces in Vietnam. Currently, real-time polymerase chain reaction (PCR) is considered to be a powerful tool for viral detection in field samples, including ASFV. However, some recent reports have suggested that mismatches in primer and probe binding regions may directly affect real-time PCR qualification, leading a false-negative result.Aim: This study aims to further examine a conflicting result obtained from two OIE recommended methods, conventional PCR and real-time PCR, for ASFV detection.Methods: Two ASF suspected pigs from different provinces in the north of Vietnam were selected for this study based on clinical signs and postmortem lesions. The different results obtained by OIE-recommended conventional PCR and real-time PCR were further analyzed by the Sanger sequencing method and virus isolation in combination with hemadsorption (HAD) test using porcine alveolar macrophages cells.Results: The results showed that when the primer sequence matched perfectly with the sequences of field isolates, a mutation in probe binding region was found, indicating that a single mismatch in the probe binding site may cause a false-negative result by real-time PCR in detecting ASFV in clinical samples in Vietnam. An agreement between conventional PCR, using PPA1/PPA2 primers and two golden standard methods, virus isolation in combination with HAD assay, and sequencing method was observed in this study.Conclusion: A single mismatch in the probe binding site caused a failse-negative result by realtime PCR method in field diagnosis of ASFV. The needs consideration when selecting the appropriate molecular diagnostic methods is based on the current databases of ASFV sequences,  particularly for epidemiological surveillance of ASF. Keywords: African swine fever, PCR, Pigs, Real-time PCR, Vietna

Brza serotipizacija virusa afričke kuge svinja zasnovana na analizi kratkih fragmenata EP402R gena koji kodira CD2-like protein

The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.Prvi potvrđen slučaj afričke kuge svinja (ASF) u Vijetnamu bio je zvanično prijavljen u februaru 2019. godine. Do danas, ASF virus (ASFV) je detektovan u 63 provincije Vijetnama. Radi boljeg razumevanja potencijalne uloge EP402R gena u grupisanju ASFV serotipova, analizirano je trideset ASFV sekvenci EP402R gena (pristupni brojevi: MN711757-86) poreklom iz Centralne severne obale Vijetnama i 68 referentnih uzoraka serotip grupa koji su dobro poznati iz prethodnih studija. Interesantan je bio nalaz da je kratak fragment od 90 nukleotida bio tipičan za 8 seroloških grupa ASF virusa. Dizajniran je set prajmera sa ciljem amplifi kacije kratkih fragmenata (90 nukleotida), korišćenjem Primer3 programa, a za uspostavljanje jednostavne metode serotipiziranja ASFV. Rezultati ukazuju da je fi logenetska analiza kratkih fragmenata (90 nukleotida) EP402R gena je veoma specifična i predstavlja korisnu metodu serotipizacije ASFV u poređenju sa prethodno opisanim metodama koje su koristile duge fragmente (816 nukleotida) ovog gena kao i dobro poznate referentne serotipove dobijene metodom inhibicije hemadsorbcije

Prospects for Food Fermentation in South-East Asia, Topics From the Tropical Fermentation and Biotechnology Network at the End of the AsiFood Erasmus+Project

Fermentation has been used for centuries to produce food in South-East Asia and some foods of this region are famous in the whole world. However, in the twenty first century, issues like food safety and quality must be addressed in a world changing from local business to globalization. In Western countries, the answer to these questions has been made through hygienisation, generalization of the use of starters, specialization of agriculture and use of long-distance transportation. This may have resulted in a loss in the taste and typicity of the products, in an extensive use of antibiotics and other chemicals and eventually, in a loss in the confidence of consumers to the products. The challenges awaiting fermentation in South-East Asia are thus to improve safety and quality in a sustainable system producing tasty and typical fermented products and valorising by-products. At the end of the “AsiFood Erasmus+ project” (www.asifood.org), the goal of this paper is to present and discuss these challenges as addressed by the Tropical Fermentation Network, a group of researchers from universities, research centers and companies in Asia and Europe. This paper presents current actions and prospects on hygienic, environmental, sensorial and nutritional qualities of traditional fermented food including screening of functional bacteria and starters, food safety strategies, research for new antimicrobial compounds, development of more sustainable fermentations and valorisation of by-products. A specificity of this network is also the multidisciplinary approach dealing with microbiology, food, chemical, sensorial, and genetic analyses, biotechnology, food supply chain, consumers and ethnology

Safety and efficacy of fluoxetine on functional outcome after acute stroke (AFFINITY): a randomised, double-blind, placebo-controlled trial

Background Trials of fluoxetine for recovery after stroke report conflicting results. The Assessment oF FluoxetINe In sTroke recoverY (AFFINITY) trial aimed to show if daily oral fluoxetine for 6 months after stroke improves functional outcome in an ethnically diverse population. Methods AFFINITY was a randomised, parallel-group, double-blind, placebo-controlled trial done in 43 hospital stroke units in Australia (n=29), New Zealand (four), and Vietnam (ten). Eligible patients were adults (aged ≥18 years) with a clinical diagnosis of acute stroke in the previous 2–15 days, brain imaging consistent with ischaemic or haemorrhagic stroke, and a persisting neurological deficit that produced a modified Rankin Scale (mRS) score of 1 or more. Patients were randomly assigned 1:1 via a web-based system using a minimisation algorithm to once daily, oral fluoxetine 20 mg capsules or matching placebo for 6 months. Patients, carers, investigators, and outcome assessors were masked to the treatment allocation. The primary outcome was functional status, measured by the mRS, at 6 months. The primary analysis was an ordinal logistic regression of the mRS at 6 months, adjusted for minimisation variables. Primary and safety analyses were done according to the patient's treatment allocation. The trial is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12611000774921. Findings Between Jan 11, 2013, and June 30, 2019, 1280 patients were recruited in Australia (n=532), New Zealand (n=42), and Vietnam (n=706), of whom 642 were randomly assigned to fluoxetine and 638 were randomly assigned to placebo. Mean duration of trial treatment was 167 days (SD 48·1). At 6 months, mRS data were available in 624 (97%) patients in the fluoxetine group and 632 (99%) in the placebo group. The distribution of mRS categories was similar in the fluoxetine and placebo groups (adjusted common odds ratio 0·94, 95% CI 0·76–1·15; p=0·53). Compared with patients in the placebo group, patients in the fluoxetine group had more falls (20 [3%] vs seven [1%]; p=0·018), bone fractures (19 [3%] vs six [1%]; p=0·014), and epileptic seizures (ten [2%] vs two [<1%]; p=0·038) at 6 months. Interpretation Oral fluoxetine 20 mg daily for 6 months after acute stroke did not improve functional outcome and increased the risk of falls, bone fractures, and epileptic seizures. These results do not support the use of fluoxetine to improve functional outcome after stroke

Rapid Identification for Serotyping of African Swine Fever Virus Based on the Short Fragment of the EP402R Gene Encoding for CD2-Like Protein

The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.Prvi potvrđen slučaj afričke kuge svinja (ASF) u Vijetnamu bio je zvanično prijavljen u februaru 2019. godine. Do danas, ASF virus (ASFV) je detektovan u 63 provincije Vijetnama. Radi boljeg razumevanja potencijalne uloge EP402R gena u grupisanju ASFV serotipova, analizirano je trideset ASFV sekvenci EP402R gena (pristupni brojevi: MN711757-86) poreklom iz Centralne severne obale Vijetnama i 68 referentnih uzoraka serotip grupa koji su dobro poznati iz prethodnih studija. Interesantan je bio nalaz da je kratak fragment od 90 nukleotida bio tipičan za 8 seroloških grupa ASF virusa. Dizajniran je set prajmera sa ciljem amplifi kacije kratkih fragmenata (90 nukleotida), korišćenjem Primer3 programa, a za uspostavljanje jednostavne metode serotipiziranja ASFV. Rezultati ukazuju da je fi logenetska analiza kratkih fragmenata (90 nukleotida) EP402R gena je veoma specifična i predstavlja korisnu metodu serotipizacije ASFV u poređenju sa prethodno opisanim metodama koje su koristile duge fragmente (816 nukleotida) ovog gena kao i dobro poznate referentne serotipove dobijene metodom inhibicije hemadsorbcije

Depth Profiles of Microplastics in Sediment Cores from Two Mangrove Forests in Northern Vietnam

Plastics are essential materials that play critical roles in people’s lives in the modern world. However, plastic pollution in the ocean has become a critical environmental problem due to the fact of its durability and long life span. In the present study, we analyze the contamination of microplastics in sediment cores from mangrove forests of the Red River Delta and Tien Yen Bay in Northern Vietnam. The results showed that the concentration of microplastics in sediment cores ranged from 0 to 4941 particles/kg, with four types of microplastics: microfiber, microfragment, microfoam, and microfilm. Microplastics were not observed in sediment samples lower than 65 and 70 cm core depth in sediment cores from the Tien Yen Bay and Red River Delta. The microplastic concentration in sediment cores from the Red River Delta was significantly higher than those of Tien Yen Bay, reflecting the influence of anthropogenic activities on microplastics pollution. The present results highlight that the accumulation of microplastics may have occurred a long time ago and tends to increase in the future. Further investigation on microplastic accumulation rates, sources of microplastics, and reducing plastic pollution is necessary to minimize the impacts of microplastic pollution on aquatic life and the environment

Depth Profiles of Microplastics in Sediment Cores from Two Mangrove Forests in Northern Vietnam

Plastics are essential materials that play critical roles in people&rsquo;s lives in the modern world. However, plastic pollution in the ocean has become a critical environmental problem due to the fact of its durability and long life span. In the present study, we analyze the contamination of microplastics in sediment cores from mangrove forests of the Red River Delta and Tien Yen Bay in Northern Vietnam. The results showed that the concentration of microplastics in sediment cores ranged from 0 to 4941 particles/kg, with four types of microplastics: microfiber, microfragment, microfoam, and microfilm. Microplastics were not observed in sediment samples lower than 65 and 70 cm core depth in sediment cores from the Tien Yen Bay and Red River Delta. The microplastic concentration in sediment cores from the Red River Delta was significantly higher than those of Tien Yen Bay, reflecting the influence of anthropogenic activities on microplastics pollution. The present results highlight that the accumulation of microplastics may have occurred a long time ago and tends to increase in the future. Further investigation on microplastic accumulation rates, sources of microplastics, and reducing plastic pollution is necessary to minimize the impacts of microplastic pollution on aquatic life and the environment

Simultaneously Utilizing Excited Holes and Electrons for Piezoelectric-Enhanced Photoproduction of H2O2 from S-Scheme 2D S-doped VOx/g-C3N4 Nanostructures

The simultaneous use of excited holes and electrons in photocatalytic research has become a strong driving force for rationalizing materials in design and development. As a promising strategy, step-scheme (S-scheme) photocatalysts have enormously ignited the photocatalytic community in recent decades. Introducing piezoelectric fields under light irradiations to generate valuable chemical products, such as hydrogen peroxide (H2O2), has also gained current interest in enhancing the large-scale production of these chemicals. In the research, we reported the fabrication of 2D S-doped VOx deposited on 2D g-C3N4 to produce H2O2 via the piezo-photocatalytic process. The finding pointed out that adding sulfur to VOx can help improve the catalytic outcomes by modifying the electronic and morphological properties of pristine VOx. In addition, when coupled with g-C3N4, the presence of sulfur limits the formation of graphene in the VOx/g-C3N4 composites, causing shielding effects in the materials. Besides, the research also sheds light on the charge transportation between g-C3N4 and S-VOx under irradiation and how the composites work to trigger the formation of H2O2. It turned out that the formation of H2O2 significantly relies on the reduction of oxygen to generate oxygenic radical species at g-C3N4 sites. Meanwhile, S-VOx provides oxidative sites in the composites to oxidize water molecules to generate H2O2. This study confirms the validation of S-scheme piezo-photocatalysts, thus encouraging further research on developing heterojunction materials with high catalytic efficiency, which can be used in practical conditions