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

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

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

The first case of Enterocytozoon bieneusi infection in Poland

Microsporidia are intracellular parasites that cause opportunistic infections in humans of various immunological status. Only a few case reports exist on microsporidial infection in solid organ transplant recipients worldwide. The presented study demonstrates the first case in Poland of Enterocytozoon bieneusi infection in a liver transplant patient. Parasites were diagnosed in stool samples using both modified trichrome staining and PCR

Prevalência de Cryptosporidium serpentis em serpentes de cativeiro Prevalence of Cryptosporidium serpentis in captive snakes

Cryptosporidium é um protozoário encontrado em uma grande variedade de espécies animais como responsável por casos de gastrite e enterite, porém com epidemiologia pouco conhecida em animais silvestres. A presente investigação teve como objetivo avaliar a prevalência de Cryptosporidium serpentis em lavado gástrico de serpentes mantidas em cativeiro no serpentário do Instituto Butantan (São Paulo, Brasil). A coleta foi realizada uma semana após alimentação, evitando, assim, a regurgitação devido à manipulação. Foram realizados esfregaços do sedimento do lavado gástrico, obtido por centrifugação, corados pela técnica de coloração de Kinyoun. Parte do sedimento foi submetido à técnica de RFLP-PCR para identificação da espécie de Cryptosporidium. O serpentário é dividido em três seções, por espécie - a primeira com oito jibóias (Boa constrictor amarali), a segunda com dez jararacas (Bothropoides jararaca) e a última com sete cascavéis (Caudisona durissa). A prevalência de C. serpentis encontrada neste estudo para as serpentes C. durissa, B. jararaca e Boa c. amarali, foi de 57,14% (04/07), 40% (04/10) e 37,5% (03/08), respectivamente, revelando importante ocorrência desse protozoário em serpentes de cativeiro. Apesar da alta prevalência encontrada, apenas as jiboias apresentaram sintomas como perda de peso e regurgitação, refletindo uma sensibilidade diferente da espécie para C. serpentis.<br>Cryptosporidium is a protozoan found in a wide variety of animal species which is responsible for gastritis and enteritis, but its epidemiology is poorly known in wild animals. The present investigation aimed to evaluate the prevalence of Cryptosporidium serpentis in gastric aspirate of captive snakes from the public serpentarium of the Butantan Institute (São Paulo, Brazil). Sampling was performed preferably one week after feeding, thereby preventing regurgitation due to manipulation. Smears were done from the gastric aspirate sediment obtained by centrifugation and stained by Kinyoun technique. Part of the pellet was submitted to RFLP-PCR technique for amplification of Cryptosporidium segment (833bp, CSP01) of SSU rRNA gene. The serpentarium was divided in three sections by species - the first housing eight Amaral&acute;s Boa (Boa constrictor amarali), the second with ten jararacas (Bothropoides jararaca) and the last one with seven south american rattlesnakes (Caudisona durissa). The prevalence of C. serpentis found in this study for the snakes C. durissa, B. jararaca and B. constrictor was 57.14% (04/07), 40% (04/10) and 37.5% (03/08), respectively, thus revealing the high occurrence of this protozoan among captive snakes. Despite the high prevalence found, only B. constrictor amarali presented symptoms as regurgitation and weight loss, probably due to a different sensibility of this species to C. serpentis