Molecular characterization of Cryptosporidium and Giardia in environmental samples and faecal samples from biogas users in Bac Giang, Vietnam.

Little is known about Cryptosporidium and Giardia in biogas waste and humans in Vietnam. There is a potential risk of infections during or after using the biogas system. The detected protozoan genotypes are zoonotic pathogens, and contamination of vegetables may relay through runoff to the surface waters and soil. The objective of this study was to understand the role of the environment in the epidemiology of human infections in Bac Giang province, Vietnam, with a focus on investigating the presence of Cryptosporidium spp. genotypes and Giardia assemblages among 239 environmental samples and 94 faecal samples of biogas users. PCR and sequencing analysis were used to identify the occurrence and genotypes of Cryptosporidium and Giardia in these samples. Results showed that 13/333 (3.9 %) and 9/333 (2.7 %) samples were positive for Cryptosporidium oocyst and Giardia cysts, respectively. Characterization revealed the presence of Cryptosporidium scrofarum, C. suis, C. meleagridis, C. bailey and Giardia intestinalis assemblage A and E. C. scrofarum and Giardia assemblage E were identified for the first time in humans in Bac Giang. The current information from the above investigations will be valuable for protozoan source tracking and control interventions against Cryptosporidium and Giardia infection associated with biogas wastes in Vietnam.

Comparison of rapid methods for detection of Giardia spp. and Cryptosporidium spp. (oo)cysts using transportable instrumentation in a field deployment.

Reliable, sensitive, quantitative, and mobile rapid screening methods for pathogenic organisms are not yet readily available, but would provide a great benefit to humanitarian intervention units in disaster situations. We compared three different methods (immunofluorescent microscopy, IFM; flow cytometry, FCM; polymerase chain reaction, PCR) for the rapid and quantitative detection of Giardia lamblia and Cryptosporidium parvum (oo)cysts in a field campaign. For this we deployed our mobile instrumentation and sampled canal water and vegetables during a 2 week field study in Thailand. For purification and concentrations of (oo)cysts, we used filtration and immunomagnetic separation. We were able to detect considerably high oo(cysts) concentrations (ranges: 15-855 and 0-240 oo(cysts)/liter for Giardia and Cryptosporidium, respectively) in 85 to 300 min, with FCM being fastest, followed by PCR, and IFM being slowest due to the long analysis time per sample. FCM and IFM performed consistently well, whereas PCR reactions often failed. The recovery, established by FCM, was around 30% for Giardia and 13% for Cryptosporidium (oo)cysts. It was possible to track (oo)cysts from the wastewater further downstream to irrigation waters and confirm contamination of salads and water vegetables. We believe that rapid detection, in particular FCM-based methods, can substantially help in disaster management and outbreak prevention.

Epidemiological and comparative genomic analysis of Bacillus anthracis isolated from northern Vietnam.

To understand the epidemiological and genetic background of anthrax cases occurring in Vietnam from 2011 to 2015, we surveilled and genetically analyzed Bacillus anthracis isolated in the north of the country. Epidemiological surveillance showed that most human cutaneous anthrax cases occurred in association with animal dissection. Whole-genome sequences were obtained from six B. anthracis strains from human patients with cutaneous anthrax in the endemic area. Comparative genomic analysis showed that the genetic homogeneity among Vietnamese B. anthracis strains was very high. All Vietnamese B. anthracis strains belonged to the canSNP lineage of A.Br.011/009, which mostly consists of strains of the trans-Eurasian (TEA) group, including the most closely related strain, Carbosap. To clarify the genetic diversity of Vietnamese strains and strains belonging to A.Br.011/009 and A.Br.008/011 canSNP lineages, we applied a reference genome-based single-nucleotide polymorphism (SNP) and gene-by-gene genomic analysis (whole-genome MLST) strategy. The phylogeny from core genome SNPs revealed that the Vietnamese strains were positioned close to each other; moreover, several SNPs specific to Vietnamese B. anthracis were identified. Whole-genome MLST analysis revealed the differences in the number of SNPs between Vietnamese strains, which could enable discrimination at the strain level.

Faecal and protozoan parasite contamination of water spinach (Ipomoea aquatica) cultivated in urban wastewater in Phnom Penh, Cambodia.

OBJECTIVE: To identify the level of contamination with thermotolerant coliforms (ThC), intestinal helminth eggs and protozoan parasites in water spinach (Ipomoea aquatica) cultivated in a wastewater-fed lake in Phnom Penh, Cambodia. METHODS: The investigation was carried out from July 2004 to May 2005 in Boeng Cheung Ek lake, a main recipient of wastewater from the city of Phnom Penh. Water spinach samples were collected near two wastewater entry points in the northern part of the lake and near a wastewater exit point in the southern part of the lake, where water is discharged from the lake into a stream. Water spinach samples were harvested at each of the three locations with and without wastewater contact according to the normal practices of farmers, and analysed for ThC, protozoan parasites (Giardia, Cryptosporidium and Cyclospora), and helminth eggs (Ascaris lumbricoides, Trichuris trichura and hookworm) using standard methods. Wastewater samples were also collected at each of the three locations and analysed for ThC and helminth eggs. RESULTS: High concentrations of ThC (approximately 10(5)-10(7)/g) were found in water spinach samples. ThC mean counts did not differ significantly, neither between sample types at each location, nor between locations. ThC mean concentration of water spinach samples collected in the rainy season of 2005 was higher than that in the dry season of 2004. Protozoan parasites were detected in the samples from all three locations and in water spinach harvested with and without wastewater contact. Giardia spp. were present in 56% (20) of samples with an average cyst concentration of 6.6/g. Mean counts of Giardia were not significantly associated with wastewater contact status of water spinach. Cryptosporidium oocysts were found in 17% (six) of samples with an average concentration of 0.5 oocyst per gram of water spinach. Cyclospora oocysts were detected in 8% (three) of samples. Four (11%) water spinach samples contained helminth eggs (average concentration: 0.1 egg per gram). Water samples contained approximately 10(4)-10(7) ThC per 100 ml and a mean of 27.4 helminth eggs per litre. There was a 2-3 log reduction of ThC between the water samples collected at the wastewater entry and exit point(s) of the lake. CONCLUSIONS: Water spinach cultivated in the lake was highly contaminated with feces, as indicated by high ThC concentrations and the presence of protozoan parasites. A reduction in ThC numbers almost to WHO guideline levels for irrigation water occurred through natural biological and physical processes in the lake, as shown by differences in ThC counts at wastewater entry and exit points.