Phylogenetic analysis of Cryptosporidium isolates from captive reptiles using 18S rDNA sequence data and random amplified polymorphic DNA analys

Sequence alignment of a polymerase chain reaction-amplified 713-base pair region of the Cryptosporidium 18S rDNA gene was carried out on 15 captive reptile isolates from different geographic locations and compared to both Cryptosporidium parvum and Cryptosporidium muris isolates. Random amplified polymorphic DNA (RAPD) analysis was also performed on a smaller number of these samples. The data generated by both techniques were significantly correlated (P < 0.002), providing additional evidence to support the clonal population structure hypothesis for Cryptosporidium. Phylogenetic analysis of both 18S sequence information and RAPD analysis grouped the majority of reptile isolates together into 1 main group attributed to Cryptosporidium serpentis, which was genetically distinct but closely related to C. muris. A second genotype exhibited by 1 reptile isolate (S6) appeared to be intermediate between C. serpentis and C. muris but grouped most closely with C. muris, as it exhibited 99.15% similarity with C. muris and only 97.13% similarity with C. serpentis. The third genotype identified in 2 reptile isolates was a previously characterized 'mouse' genotype that grouped closely with bovine and human C. parvum isolates

Giardia duodenalis cysts of genotype a recovered from clams in the Chesapeake Bay subestuary, Rhode River

Filter-feeding molluscan shellfish can concentrate zoonotic and anthroponotic waterborne pathogens. Cysts of Giardia sp. were detected by immunofluorescent antibodies in tissues of the clams Macoma balthica and M. mitchelli from Rhode River, a Chesapeake Bay (Maryland) subestuary. Molecular tests identified the cysts as Giardia duodenalis Genotype A, the most common genotype recovered from humans. Macoma clams are burrowers in mud or sandy- mud substrata and preferentially feed on the surface sediment layer. Waterborne Giardia cysts settle rapidly to the bottom in slow-moving waters and contaminate the sediment. Macoma clams do not have economic value, but can serve as biologic indicators of sediment contamination with Giardia sp. cysts of public health importance. These clams can be used for sanitary assessment of water quality

Molecular markers and sentinel organisms for environmental monitoring

Molecular methods are useful for both to monitor anthropogenic viral, bacterial, and protozoan enteropathogens, and to track pathogen specific markers in a complex environment in order to reveal sources of these pathogens. Molecular genetic markers for fecal viruses, bacteria, and protozoans hold promise for monitoring environmental pollution and water quality. The demand for microbiologically safe waters grows exponentially due to the global demographic rise of the human population. Economically important shellfish, such as oysters, which are harvested commercially and preferentially consumed raw can be of public health importance if contaminated with human waterborne pathogens. However, feral molluscan shellfish which do not have an apparent economic value serve as indicators in monitoring aquatic environments for pollution with human waterborne pathogens and for sanitary assessment of water quality. Current technology allows for multiplexed species-specific identification, genotyping, enumeration, viability assessment, and source-tracking of human enteropathogens which considerably enhances the pathogen source-tracking efforts

First detection of microsporidia in raised pigeons in Poland

Abstract Microsporidia are single-celled intracellular parasites which occur in a number of animals, both vertebrates and invertebrates. Several species of microsporidia can cause disease in humans in both immunocompromised and immunocompetent individuals. However, the sources of human infection and the routes of transmission of microsporidia have not yet been fully determined, although more and more researchers are of the opinion that microsporidia in humans is zoonotic. The aim of the presented study was to identify the most common microsporidial species in the droppings of raised and wild pigeons in Poland. A total of 139 collective samples of droppings (33 samples of droppings from feral pigeons and 106 samples from raised pigeons) were examined using conventional staining and molecular techniques. Using chromotrope staining and multiplex FISH techniques, the microsporidial spores were found in 12 (8.6%) of the 139 samples of droppings. The spores of Encephalitozoon hellem were detected in five samples of pigeon droppings (3.6%), spores of E. intestinalis in four samples of droppings (2.9%), while spores of E. cuniculi and E. bieneusi were only detected in two samples (1.4%). Furthermore, a mixed infection of E. bieneusi and E. cuniculi was found in a single sample of droppings (0.7%). The presence of microsporidial spores in droppings collected from raised pigeons indicates a risk of infection to humans, mainly pigeon fanciers

Human zoonotic enteropathogens in a constructed free-surface flow wetland

Effluents from a small-scale free-surface flow constructed wetland, used for polishing of secondary treated wastewater, contained significantly higher concentrations of potentially viable Giardia duodenalis cysts and Enterocytozoon bieneusi spores than did wetland influents consisting of secondary treated wastewater. Zoonotic Assemblage A of G. duodenalis cysts was identified in wetland inflows, while Assemblage A and two nonhuman infective Assemblages (i.e., C, and E) were present in wetland effluents. E. bieneusi spores represented genotype K based on DNA sequencing analysis of internal transcribed spacer. The study demonstrated that: (1) free-surface flow small-scale constructed wetlands may not provide sufficient remediation for human zoonotic protozoa and fungi present in secondary treated wastewater; (2) dogs and livestock can substantially contribute human-pathogenic protozoan and fungal microorganisms to engineered vegetated wetland systems; and (3) large volumes of wetland effluents can contribute to contamination of surface waters used for recreation and drinking water abstraction and therefore represent a serious public health threat

Immunological survey of babesiosis (

Babesia peircei was extracted from nucleated erythrocytes of naturally infected Jackass penguin (Spheniscus demersus) from South Africa (SA). Babesia peircei glycoprotein-enriched fractions were obtained by concanavalin A-Sepharose affinity column chromatography and separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). At least 14 protein bands (9, 11, 13, 20, 22, 23, 24, 43, 62, 90, 120, 204, and 205 kDa) were observed, with the major protein at 25 kDa. Blood samples of 191 adult S. demersus were tested by enzyme-linked immunosorbent assay (ELISA) utilizing B. peircei glycoprotein-enriched fractions to detect anti-B. peircei IgG. The samples originated from three groups of free-ranging penguins (n = 110), 1 group of penguins (n = 66) which were rescued after offsh ore oil-spill contaminations and rehabilitated at the South African National Foundation for the Conservation of Coastal Birds (SANCCOB), and the final group from SANCCOB-resident penguins (n = 15). The overall B. peircei seroprevalence was 65 %, and the mean seropositive ranged from 60 to 71 % among the five penguin groups. The ELISA appeared to be specific for B. peircei IgG as tested against Haemoproteus columbae IgG and avian malaria (Plasmodium relictum, and P. elongatum) IgG. Toxoplasma gondii antibody (Ab) were detected by the direct agglutination test using killed T. gondii tachyzoites. All birds were seronegative for T. gondii Ab. The lack of T. gondii-positive penguins was due to the appropriate sanitary conditions and anti- Toxoplasma prevention procedures utilized by the SANCCOB