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Additional file 2: Cryptosporidium spp. low-volume microscopy panel and bulk water PCR results.

Additional file 4: Filtered sediments of 1 L bulk water samples from sampling sites. Filtration of low-volume (1 L) bulk water with 1.2 µm pore size glass microfiber filter is efficient in recovering dispersed sediments from surface water samples and can be used for high turbidity aquatic matrices by simply changing the filter once full. Different sampling sites show different density and physical characteristics of suspended solids.

Additional file 1: Consolidated details of Study sites, sampling sites, and sampling points in the Philippines.

Additional file 3: Raw data on Cryptosporidium spp. oocyst counts in 200 microscopic fields per 1 cm diameter (25µL) smear.

Introduction: This paper reviews the current knowledge and understanding of Cryptosporidium spp. an d Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results: Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices and detection methodologies used. Conclusion: Upgrading and making the diagnosis/detection procedures more uniform is recommended throughout the region. Public health authorities should actively work towards increasing reporting and standardising reporting practices as these prerequisites for the reported data to be valid and therefore necessary for appropriate control plans.

Figure S1. Sequence alignment of QH1 to QH11 and AB477011.1 of E. multilocularis cox 1 gene. QH1 to QH11: Sequences isolated from Qinghai voles; AB477011.1: Reference sequence downloaded from the GenBank database. (DOCX 44Â kb)

Multilingual abstracts in the five official working languages of the United Nations. (PDF 698Â kb)

Three methods, microscopy, nested polymerase chain reaction (nPCR), and loop-mediated isothermal amplification (LAMP) have been applied for malaria diagnosis in 105 human blood samples collected in Northern Thailand. Only Plasmodium falciparum and Plasmodium vivax infections were detected. A total number of 57 positives (54%) could be detected for P. falciparum and 25 (24%) for P. vivax when all samples that were positive in any of the three methods are counted together. The nPCR was used as a reference standard for comparison with the other methods, microscopy and LAMP. The sensitivity of LAMP for P. falciparum was 100%. All nPCR-negative samples for P. falciparum were also negative by both microscopy and LAMP (specificity, 100%). For diagnosis of P. vivax, microscopy detected 15 of 23 nPCR-positive samples (sensitivity, 65%). LAMP detected 22 of 23 nPCR-positives (sensitivity, 96%). Among the 82 nPCR-negative samples microscopy detected two samples (specificity, 98%). All 82 nPCR-negative were also negative by the LAMP method (specificity, 100%). Both Plasmodium genus- and species-specific LAMP primer sets yielded the same results in all samples. There were no significant differences in the prevalence detected by each method. We assume that LAMP was as reliable as nPCR and more reliable than microscopy in the detection of Plasmodium DNA tested in the examined Thai field blood samples. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of malaria cases and together with nPCR can also be used as supplementary methods for clinical and epidemiological use.

Here, we present the first time findings regarding the occurrence of Cryptosporidium and Giardia in sewage waters and the first molecular characterization of Cryptosporidium species in Greece. Biological treatment plants from three regions in Greece have been investigated. The detection of Cryptosporidium oocysts was by modified Ziehl-Neelsen acid fast (MZN-AF) and by immunofluorescence microscopy (IFT) for Cryptosporidium and Giardia (oo)cysts, whereas nested PCR based on the SSU rDNA assay was used for molecular detection of Cryptosporidium followed by sequencing for the genetic characterization of the species. In total, 73 samples (37 raw sewage samples and 38 of treated water samples) were collected and analyzed. Of the 73 water samples, 4 samples were Cryptosporidium-positive by IFT and staining, 12 samples were Cryptosporidium-positive by nested PCR; 9 samples were Giardia-positive by IFT. We showed that Cryptosporidium cysts are found both in the input and the discharge of the biological treatment plants. Molecular characterization of Cryptosporidium based on the small subunit ribosomal DNA gene resulted in the determination of Cryptosporidium parvum and Cryptosporidium muris Greek isolates. This is the first report of Cryptosporidium and Giardia occurrence in wastewaters and the first molecular identification of Cryptosporidium species in Greek environments. As the treated water is used for irrigation, or it is discharged into the sea, our findings indicate that biological treatment facilities constitute a possible risk for public health because the related species are prevalent in humans; the results invite for further epidemiological investigations to evaluate the real public health risk in Greece.

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