Strongyloidiasis in northern Vietnam: epidemiology, clinical characteristics and molecular diagnosis of the causal agent.

BACKGROUND: Strongyloidiasis is a health problem in Vietnam, but appropriate information is still limited. The aim of this study was to determine the prevalence, geographical distribution, epidemiological aspects, symptoms and other health indicators of Strongyloides stercoralis infections in patients from 27 provinces of northern Vietnam attending the Hanoi Medical University Hospital during 2016 and 2017. METHODS: Blood samples of 2000 patients were analyzed for S. stercoralis infection with an IgG ELISA test. Seroprevalence was analyzed by gender, age group, locality of origin (rural or urban areas) and symptoms. Stools from the seropositive patients were examined for the detection of worms which were subsequently used for species identification by morphology and rDNA ITS1 sequencing. RESULTS: A seroprevalence of 20% was detected, showing an increasing prevalence from young to older age groups but without significant gender difference. Seroprevalence was higher in rural areas than in urban areas, both in general and individually in all provinces without exception, and lower in the mountainous areas than in the large valley lowlands. The follow-up of the 400 patients showed eosinophilia in 100% of cases, diarrhoea in 64.5%, digestion difficulties in 58.0%, stomachache in 45.5%, stomach and duodenal ulcers in 44.5%, itching in 28.0% and fever in 9.5%. The prevalence of symptoms and signs were also higher in older age groups than in younger age groups. Worms were detected in stools of 10.5% of the patients. Sequencing of a 501-bp nuclear ribosomal DNA ITS1 fragment allowed for the verification of infection by Strongyloides stercoralis. CONCLUSIONS: To our knowledge, this study is the largest survey of human strongyloidiasis in Vietnam so far and the first molecular identification of this nematode species in this country. Long-term chronicity may probably be usual in infected subjects, mainly in the older age groups.

The first report of two cases of cystic echinococcosis in the lung by Echinococcus ortleppi infection, in Vietnam.

In 2013, two cases of infection by Echinococcus that caused cystic echinococcosis in the lungs were reported. In the first case, there was a cyst of 6 × 7 cm in diameter, and in the second case, there were four cysts of 5 × 6 cm, 4 × 4 cm, 3.5 × 3 cm and 2.5 × 2 cm in diameter, respectively. In both cases, Echinococcus larvae were collected from the cysts. The larvae were identified as Echinococcus ortleppi by morphology and a molecular method (using reduced nicotinamide adenine dinucleotide hydrogenase [NADH] with 99%-100% homology compared with E. ortleppi in GenBank). This is the first time that this species has been found in humans in Vietnam.

Glioma-Derived Platelet-Derived Growth Factor-BB Recruits Oligodendrocyte Progenitor Cells via Platelet-Derived Growth Factor Receptor-α and Remodels Cancer Stroma.

Glioma is an aggressive and incurable disease, and is frequently accompanied by augmented platelet-derived growth factor (PDGF) signaling. Overexpression of PDGF-B ligand characterizes a specific subclass of glioblastoma multiforme, but the significance of the ligand remains to be elucidated. For this end, we implanted a glioma-cell line transfected with PDGF-BB-overexpressing vector (GL261-PDGF-BB) or control vector (GL261-vector) into wild-type mouse brain, and examined the effect of glioma-derived PDGF on the tumor microenvironment. The volume of GL261-PDGF-BB rapidly increased compared with GL261-vector. Recruitment of many PDGF receptor (PDGFR)-α and Olig2-positive oligodendrocyte precursor cells and frequent hemorrhages were observed in GL261-PDGF-BB but not in GL261-vector. We then implanted GL261-PDGF-BB into the mouse brain with and without Pdgfra gene inactivation, corresponding to PDGFRα-knockout (KO) and Flox mice, respectively. The recruitment of oligodendrocyte precursor cells was largely suppressed in PDGFRα-KO than in Flox, whereas the volume of GL261-PDGF-BB was comparable between the two genotypes. Frequent hemorrhage and increased IgG-leakage were associated with aberrant vascular structures within the area where many recruited oligodendrocyte precursor cells accumulated in Flox. In contrast, these vascular phenotypes were largely normalized in PDGFRα-KO. Increased matrix metalloproteinase-9 in recruited oligodendrocyte precursor cells and decreased claudin-5 in vasculature may underlie the vascular abnormality. Glioma-derived PDGF-B signal induces cancer stroma characteristically seen in high-grade glioma, and should be therapeutically targeted to improve cancer microenvironment.

Trichinellosis in Vietnam.

Trichinellosis is a zoonotic parasitic disease with a worldwide distribution. The aim of this work was to describe the epidemiological and clinical data of five outbreaks of trichinellosis, which affected ethnic minorities living in remote mountainous areas of northwestern Vietnam from 1970 to 2012. Trichinellosis was diagnosed in 126 patients, of which 11 (8.7%) were hospitalized and 8 (6.3%) died. All infected people had consumed raw pork from backyard and roaming pigs or wild boar at wedding, funeral, or New Year parties. The short incubation period (average of 9.5 days), the severity of the symptoms, which were characterized by diarrhea, abdominal pain, fever, myalgia, edema, weight loss, itch, and lisping, and the high mortality, suggest that patients had ingested a high number of larvae. The larval burden in pigs examined in one of the outbreaks ranged from 70 to 879 larvae/g. These larvae and those collected from a muscle biopsy taken from a patient from the 2012 outbreak were identified as Trichinella spiralis. Data presented in this work show that the northern regions of Vietnam are endemic areas for Trichinella infections in domestic pigs and humans.

Anti-Trichinella IgG in ethnic minorities living in Trichinella-endemic areas in northwest Vietnam: study of the predictive value of selected clinical signs and symptoms for the diagnosis of trichinellosis.

The objective of this study was to assess the presence of anti-Trichinella IgG in the serum of persons from ethnic minorities from northwest Vietnam with clinical signs and symptoms that are compatible with trichinellosis. A total of 645 persons were enrolled, of which 200 people lived in two villages where outbreaks of human trichinellosis had been documented in 2004 and 2008, and 445 people who were hospitalized in the Dien Bien and Son La provincial hospitals without a definitive diagnosis. Presence of anti-Trichinella IgG was demonstrated in serum samples by a standardized Enzyme-linked Immunosorbant Assay (ELISA); positive serum samples were subjected to Western blot (WB) for confirmation. Seven (3.5%; 95% CI: 1.4-7.1) persons from the villages and seven (1.6%; 95% CI: 0.6-3.2) hospitalized patients, tested positive by both ELISA and WB. Fever (N=13), eosinophilia (N=12), myalgia (N=9), facial edema (N=9) and leukocytosis (N=8) were the most common clinical signs and symptoms in the serologically positive persons. The concomitant occurrence of facial edema and myalgia among the enrolled persons from the villages, accounted for 75% of the positive predictive value (PPV) and 99.5% of the negative predictive value (NPV), suggesting that they could be used for suspecting trichinellosis when serology is not available. The high prevalence (1.6-3.5%) of anti-Trichinella IgG in persons from Vietnamese provinces where Trichinella spiralis is circulating in pigs strongly supports the need to develop control programs to eliminate the infection from pigs and for consumers' education and protection.

Current status of taeniasis and cysticercosis in Vietnam.

Several reports on taeniasis and cysticercosis in Vietnam show that they are distributed in over 50 of 63 provinces. In some endemic areas, the prevalence of taeniasis was 0.2-12.0% and that of cysticercosis was 1.0-7.2%. The major symptoms of taeniasis included fidgeted anus, proglottids moving out of the anus, and proglottids in the feces. Clinical manifestations of cysticercosis in humans included subcutaneous nodules, epileptic seizures, severe headach, impaired vision, and memory loss. The species identification of Taenia in Vietnam included Taenia asiatica, Taenia saginata, and Taenia solium based on combined morphology and molecular methods. Only T. solium caused cysticercosis in humans. Praziquantel was chosen for treatment of taeniasis and albendazole for treatment of cysticercosis. The infection rate of cysticercus cellulosae in pigs was 0.04% at Hanoi slaughterhouses, 0.03-0.31% at provincial slaughterhouses in the north, and 0.9% in provincial slaughterhouses in the southern region of Vietnam. The infection rate of cysticercus bovis in cattle was 0.03-2.17% at Hanoi slaughterhouses. Risk factors investigated with regard to transmission of Taenia suggested that consumption of raw meat (eating raw meat 4.5-74.3%), inadequate or absent meat inspection and control, poor sanitation in some endemic areas, and use of untreated human waste as a fertilizer for crops may play important roles in Vietnam, although this remains to be validated.

Molecular diagnosis of an ocular toxocariasis patient in Vietnam.

An ocular Toxocara canis infection is reported for the first time in Vietnam. A 34-year-old man residing in a village of Son La Province, North Vietnam, visited the National Eye Hospital (NEH) in August 2011. He felt a bulge-sticking pain in his left eye and loss of vision occurred over 3 months before visiting the hospital. The eye examination in the hospital showed damage of the left eye, red eye, retinal fibrosis, retinal detachment, inflammation of the eye tissues, retinal granulomas, and a parasitic cyst inside. A larva of Toxocara was collected with the cyst by a medical doctor by surgery. Comparison of 264 nucleotides of internal transcribed spacer 2 (ITS2) of ribosomal DNA was done between our Vietnamese Toxocara canis and other Toxocara geographical isolates, including Chinese T. canis, Japanese T. canis, Sri Lankan T. canis, and Iranian T. canis. The nucleotide homology was 97-99%, when our T. canis was compared with geographical isolates. Identification of a T. canis infection in the eye by a molecular method was performed for the first time in Vietnam.

An outbreak of trichinosis with molecular identification of Trichinella sp. in Vietnam.

The 5th outbreak of trichinosis occurred in a mountainous area of North Vietnam in 2012, involving 24 patients among 27 people who consumed raw pork together. Six of these patients visited several hospitals in Hanoi for treatment. Similar clinical symptoms appeared in these patients within 5-8 days after eating infected raw pork, which consisted of fever, muscle pain, difficult moving, edema, difficult swallowing, and difficult breathing. ELISA revealed all (6/6) positive reactions against Trichinella spiralis antigen and all cases showed positive biopsy results for Trichinella sp. larvae in the muscle. The larvae detected in the patients were identified as T. spiralis (Vietnamese strain) by the molecular analysis of the mitochondrial cytochrome c oxidase subunit III (cox3) gene.

Prevalence and intensity of fish-borne zoonotic trematodes in cultured freshwater fish from rural and urban areas of northern Vietnam.

Consumption of raw freshwater fish produced in both rural farm and urban wastewater ponds is a common practice in Vietnam. The present study assessed the risk of fish-borne zoonotic trematode (FZT) infection from fish raised in both these aquaculture systems in northern Vietnam. The diversity, prevalence, and infection intensity of FZT metacercariae in 1,500 freshwater fish collected from 6 sites located in rural and urban areas in northern Vietnam were investigated. The specific diagnosis of species was made by morphologic methods. The overall FZT prevalence in fish from both urban wastewater ponds and rural farm ponds was 11.2%. In wastewater ponds, the overall prevalence was 5.1%, ranging from 2.0% in tilapia to 7.3% in common and grass carp. In fish from farm ponds, the prevalence was 17.3%, and ranged from 6.7% in mud carp to 26.7% in common carp. The mean intensity of FZT infection was also higher in fish from farm ponds than that in fish from wastewater ponds (6.0% and 8.4%, respectively). The FZT species recovered from infected fish included both liver (Clonorchis sinensis) and intestinal flukes (Haplorchis taichui, Haplorchis pumilio, and Centrocestus formosanus). The prevalence of FZT in fish raised in these common farm systems represents a significant public health risk for a population with a strong cultural preference for consuming raw or inadequately prepared fish. These research results should encourage the public health and agriculture sectors to conduct the risk factor research required to develop control programs for FZT.

Genotypic characterization and species identification of Fasciola spp. with implications regarding the isolates infecting goats in Vietnam.

Ribosomal RNA sequences (361 or 362bp) of the second internal transcribed spacer 2 (ITS-2) and a portion of mitochondrial cox1 (423bp) for Fasciola spp. obtained from specimens collected in indigenous and hybrid goats and sheep in Vietnam were characterized for genotypic status and hybridization/introgression. Alignment of 48 ITS-2 sequences (also those from goats and sheep in this study) indicates that F. gigantica and F. hepatica differ typically from each other at seven sites whereas one of these is a distinguishing deletion (T) at the 327th position in F. gigantica relative to F. hepatica. The isolates from the mountainous goats in the North of Vietnam (Yen Bai province) showed the ITS-2 composition relatively identical to that of F. hepatica. The ITS-2 sequences from populations of Fasciola isolates in goats had probably experienced introgression/hybridization as reported previously in other ruminants and humans. All Vietnamese goat-of-origin specimens had high pairwise percentage of mitochondrial cox1 sequences to F. gigantica (97-100%), and very low identity to F. hepatica (91-93%), suggesting their maternal linkage to be traced to F. gigantica. The presence of hybrid and/or introgressed populations of liver flukes bearing genetic material from both F. hepatica and F. gigantica in the goats/sheep in Vietnam, regardless of indigenous or imported hosts, appears to be the first demonstration from a tropical country.

Fishborne zoonotic intestinal trematodes, Vietnam.

Although fishborne zoonotic trematodes that infect the liver are well documented in Vietnam, intestinal fishborne zoonotic trematodes are unreported. Recent discoveries of the metacercarial stage of these flukes in wild and farmed fish prompted an assessment of their risk to a community that eats raw fish. A fecal survey of 615 persons showed a trematode egg prevalence of 64.9%. Infected persons were treated to expel liver and intestinal parasites for specific identification. The liver trematode Clonorchis sinensis was recovered from 51.5%, but > or =1 of 4 intestinal species of the family Heterophyidae was recovered from 100%. The most numerous were Haplorchis spp. (90.4% of all worms recovered). These results demonstrate that fishborne intestinal parasites are an unrecognized food safety risk in a country whose people have a strong tradition of eating raw fish.

Paragonimus heterotremus Chen and Hsia (1964), in Vietnam: a molecular identification and relationships of isolates from different hosts and geographical origins.

Paragonimus heterotremus Chen and Hsia (1964), and paragonimiasis caused by this species is a newly detected disease in Vietnam. Twelve samples of Paragonimus (Platyhelminthes: Trematoda: Digenea: Paragonimidae) from different life-stages (eggs, miracidia, metacercariae, adults from natural and experimental hosts) and host species (crab, dog, cat and human) were collected in different geographical locations in Vietnam. DNA sequences were obtained from each for partial mitochondrial cytochrome c oxidase subunit 1 (cox1) (387 bp) and the entire second ribosomal internal transcribed spacer (ITS-2) (361 bp). The ITS-2 sequences were identical among all specimens, including those previously reported in GenBank. For cox1, there were sequence differences between specimens from Vietnam (four provinces, different locations) and those from Guangxi (China) and Saraburi (Thailand). Phylogenetic trees inferred from cox1 and ITS-2 sequences using sequence data for 15 P. heterotremus and for other Paragonimus spp. revealed that all P. heterotremus originating from Vietnam, Thailand and China form a distinct group. This information also confirms the identity of the Vietnamese specimens as P. heterotremus.

Clonorchis sinensis and Opisthorchis viverrini: development of a mitochondrial-based multiplex PCR for their identification and discrimination.

We report a single, one-step PCR approach for detection and discrimination of Clonorchis sinensis and Opisthorchis viverrini in different life-stage forms (adults, metacercariae, and eggs) from fish intermediate hosts and from infected patients. Primers designed for species-specific PCR, amplifying portions of the mitochondrial (mt) genome, were also suitable for a multiplex PCR. The latter was a single, one-step reaction under high stringency conditions, using simultaneously 2 pairs of primers (1 pair for C. sinensis--product size 612 bp, and 1 pair for O. viverrini--product size 1357 bp). Assays using serially diluted templates demonstrated that as little as 0.78 ng of genomic DNA of either species could yield amplicons. Genomic DNA extracted from different life-stage forms including adult worms (of both species), eggs (of O. viverrini), eggs possibly of several trematode species (collected from patients infected with C. sinensis in Vietnam) and mixed metacercariae of common trematodes (collected from fishes in the C. sinensis endemic areas), yielded specific bands of the correct size and their identity was confirmed by sequence analysis. The multiplex PCR approach described here proved to be a species-specific, sensitive and fast tool for accurate diagnosis of clonorchiasis and/or opisthorchiasis, permitting the detection of their metacercariae in infected fishes or adult/eggs from patients in endemic areas.

Taenia solium taeniosis/cysticercosis in Asia: epidemiology, impact and issues.

Several reports of patients with cysticercosis from many countries in Asia such as India, China, Indonesia, Thailand, Korea, Taiwan and Nepal are a clear indicator of the wide prevalence of Taenia solium cysticercosis and taeniosis in these and other Asian countries. However, epidemiological data from community based studies are sparse and available only for a few countries in Asia. Cysticercosis is the cause of epilepsy in up to 50% of Indian patients presenting with partial seizures. It is also a major cause of epilepsy in Bali (Indonesia), Vietnam and possibly China and Nepal. Seroprevalence studies indicate high rates of exposure to the parasite in several countries (Vietnam, China, Korea and Bali (Indonesia)) with rates ranging from 0.02 to 12.6%. Rates of taeniosis, as determined by stool examination for ova, have also been reported to range between 0.1 and 6% in the community in India, Vietnam, China, and Bali (Indonesia). An astonishingly high rate of taeniosis of 50% was reported from an area in Nepal populated by pig rearing farmers. In addition to poor sanitation, unhealthy pig rearing practices, low hygienic standards, unusual customs such as consumption of raw pork is an additional factor contributing to the spread of the disease in some communities of Asia. Undoubtedly, cysticercosis is a major public health problem in several Asian countries effecting several million people by not only causing neurological morbidity but also imposing economic hardship on impoverished populations. However, there are wide variations in the prevalence rates in different regions and different socio-economic groups in the same country. It is important to press for the recognition of cysticercosis as one of the major public health problems in Asia that needs to be tackled vigorously by the governments and public health authorities of the region.

Taenia solium cysticercosis in a village in northern Viet Nam: seroprevalence study using an ELISA for detecting circulating antigen.

An enzyme-linked immunosorbent assay (ELISA) for detecting circulating Taenia solium antigen was evaluated in Viet Nam; 12 of 210 people gave a positive result, including 5 persons with epilepsy. Cysticercosis was confirmed in 9 persons. Agreement between the ELISA, computerized tomography scanning and biopsy examination was high.