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.

The marsupial trypanosome Trypanosoma copemani is not an obligate intracellular parasite, although it adversely affects cell health.

BACKGROUND: Trypanosoma cruzi invades and replicates inside mammalian cells, which can lead to chronic Chagas disease in humans. Trypanosoma copemani infects Australian marsupials and recent investigations indicate it may be able to invade mammalian cells in vitro, similar to T. cruzi. Here, T. cruzi 10R26 strain (TcIIa) and two strains of T. copemani [genotype 1 (G1) and genotype 2 (G2)] were incubated with marsupial cells in vitro. Live-cell time-lapse and fluorescent microscopy, combined with high-resolution microscopy (transmission and scanning electron microscopy) were used to investigate surface interactions between parasites and mammalian cells. RESULTS: The number of parasites invading cells was significantly higher in T. cruzi compared to either genotype of T. copemani, between which there was no significant difference. While capable of cellular invasion, T. copemani did not multiply in host cells in vitro as there was no increase in intracellular amastigotes over time and no release of new trypomastigotes from host cells, as observed in T. cruzi. Exposure of host cells to G2 trypomastigotes resulted in increased host cell membrane permeability within 24 h of infection, and host cell death/blebbing was also observed. G2 parasites also became embedded in the host cell membrane. CONCLUSIONS: Trypanosoma copemani is unlikely to have an obligate intracellular life-cycle like T. cruzi. However, T. copemani adversely affects cell health in vitro and should be investigated in vivo in infected host tissues to better understand this host-parasite relationship. Future research should focus on increasing understanding of the T. copemani life history and the genetic, physiological and ecological differences between different genotypes.

Urban environments alter parasite fauna, weight and reproductive activity in the quenda (Isoodon obesulus).

Some wildlife species are capable of surviving in urbanised environments. However, the implications of urbanisation on wildlife health, and public health regarding zoonoses, are often unknown. Quenda (syn. southern brown bandicoots, Isoodon obesulus) survive in many areas of Perth, Australia, despite urbanisation. This study investigated differences in gastrointestinal and macroscopic ecto-parasitic infections, morphometrics and reproductive status between bushland and urban dwelling quenda. 287 quenda in the greater Perth region were captured and sampled for faeces (to detect gastrointestinal parasites), blood (to detect Toxoplasma gondii antibodies), ectoparasites, and morphometrics. Data were analysed using multivariable logistic and linear regression. Most parasitic infections identified in quenda were of native parasite taxa that are either not known to, or considered highly unlikely to, infect humans or domestic animals. However, stickfast fleas (Echidnophaga spp.) were present at low prevalences and intensities, and Giardia spp., Cryptosporidium spp. and Amblyomma spp. infections require further investigation to clarify their anthropozoonotic significance. Quenda captured in urbanised environments had differing odds of or intensity of certain parasitic infections, compared to those in bushland - likely attributable to quenda population density, and in some cases the availability of other host species or anthropogenic sources of infection. Urbanised environments were associated with an increase in net weight of adult male quenda by 189.0g (95% CI 68.6-309.5g; p=0.002; adjusted R2=0.06) and adult female quenda by 140.1g (95% CI 3.9-276.3g; p=0.044; adjusted R2=0.07), with study findings suggesting a tendency towards obesity in urbanised environments. Adult female quenda in bushland had increased odds of an active pouch (adjusted OR=4.89, 95% CI 1.7-14.5), suggesting decreased reproductive activity in quenda from urbanised environments. These results highlight the subtle, yet extensive impacts that urbanised environments may have on wildlife ecology, even for those species which apparently adjust well to urbanisation.

Emerging technologies for the detection and genetic characterization of protozoan parasites.

The development and adaptation of new technologies for the genetic characterization and identification of parasites continue to accelerate, providing an increasing number of research and analytical tools. We review emerging technologies that have applications in this area, including real-time PCR and microarrays, and discuss the fundamental principles of some of these technologies and how they are applied to characterize parasites. We give special consideration to the application of genetic data to biological questions, where selection of the most appropriate technique depends on the biological question posed by the investigator.

Parasites and biosecurity--the example of Australia.

Biosecurity issues as they impact on agriculture, health and the environment are becoming increasingly important. Surveillance is crucial in maintaining a country's biosecurity and this article looks at some of the issues in the context of Australia and its neighbors in the Asia-Pacific region, and emerging threats from parasitic diseases. The important threats to agriculture, public health and wildlife are highlighted, and attention is given to how future surveillance activities must be undertaken on a regional basis and involve neighboring countries if they are to be effective and lead to the protection of both the trade and health of the population.

Echinococcus granulosus cattle strain identification in an autochthonous case of cystic echinococcosis in central Mexico.

Echinococcosis is a frequent hepatic parasitic disease in several countries but it is practically absent in Mexico. A cattle strain of Echinococcus granulosus was identified by RAPD, PCR-RFLP and mitochondrial CO1 gene analysis in an autochthonous case. The parasite was obtained after a laparoscopic excision of a liver cyst from a patient that was symptomatic for 6 years but mis-diagnosed before hospitalization.

Reevaluating the evidence for Toxoplasma gondii-induced behavioural changes in rodents.

The ubiquitous protozoan parasite Toxoplasma gondii has been associated with behavioural changes in various hosts, including humans. In rodents, these behavioural changes are thought to represent adaptive manipulation by T. gondii to enhance transmission from intermediate hosts to the feline definitive host. In this review, we have tabulated evidence of changes in motor coordination, learning, memory, locomotion, anxiety, response to novelty and aversion to feline odour in rodents experimentally infected with T. gondii. In general, there was no consistent indication of the direction or magnitude of behavioural changes in response to infection. This may be due to the use, in these experimental studies, of different T. gondii strains, different host species and sexes and/or different methodologies to measure behaviour. A particular problem with studies of behavioural manipulation is likely to be the validity of behavioural tests, that is, whether they are actually measuring the traits that they were designed to measure. We suggest that future studies can be improved in three major ways. First, they should use multiple tests of behaviour, followed by multivariate data analysis to identify behavioural constructs such as aversion, anxiety and response to novelty. Second, they should incorporate longitudinal measurements on the behaviour of individual hosts before and after infection, so that within-individual and between-individual variances and covariances in behavioural traits can be estimated. Finally, they should investigate how variables such as parasite strain, host species and host sex interact with parasite infection to alter host behaviour, in order to provide a sound foundation for research concerning the proximate and ultimate mechanism(s) responsible for behavioural changes.

Multi-locus analysis of Giardia duodenalis intra-Assemblage B substitution patterns in cloned culture isolates suggests sub-Assemblage B analyses will require multi-locus genotyping with conserved and variable genes.

Recent research concerning Giardia duodenalis has focused on resolving possible sub-assemblages within Assemblages A and B to better understand host-specific and zoonotic relationships. In the present study nine cloned, cultured, Assemblage B isolates were used to investigate the intra-Assemblage B substitution patterns of conserved (ssrDNA, ef, h2b, h4) and variable (tpi, gdh, bg) genes to assess their suitability for further application to sub-assemblage analyses. The resolution of each gene was found to be proportional to its substitution rate and for the genetically narrow sample set examined, the variable genes best represented the consensus phylogeny while the conserved genes only established fractions. However it was demonstrated that the spectra of conserved and variable genes were required to ensure accuracy of inferred phylogeny and it was therefore concluded that further research into sub-Assemblage B groups would require a mixture of conserved and variable genes for the multi-locus analyses of this genetically broad assemblage.

Emergence of sylvatic Echinococcus granulosus as a parasitic zoonosis of public health concern in an indigenous community in Canada.

Within a remote Canadian Indigenous community, at least 11* of people had antibodies against Echinococcus granulosus and E. granulosus eggs were detected in 6* of environmentally collected canine fecal samples. Dog ownership, hunting, and trapping were not risk factors for seropositivity, suggesting that people are most likely exposed to E. granulosus through indirect contact with dog feces in the environment. In this situation, human exposure could be most effectively curtailed by preventing consumption of cervid viscera by free-roaming dogs.

The prevalence, intensities and risk factors associated with geohelminth infection in tea-growing communities of Assam, India.

OBJECTIVE: To determine the prevalence, intensity and associated risk factors for infection with Ascaris, hookworms and Trichuris in three tea-growing communities in Assam, India. METHODS: Single faecal samples were collected from 328 individuals and subjected to centrifugal flotation and the Kato Katz quantitation technique and prevalence and intensities of infection with each parasite calculated. Associations between parasite prevalence, intensity and host and environmental factors were then made using both univariate and multivariate analysis. RESULTS: The overall prevalence of Ascaris was 38% [95% confidence interval (CI): 33, 43], and the individual prevalence of hookworm and Trichuris was 43% (95% CI: 38, 49). The strongest predictors for the intensity of one or more geohelminths using multiple regression (P < or = 0.10) were socioeconomic status, age, household crowding, level of education, religion, use of footwear when outdoors, defecation practices, pig ownership and water source. CONCLUSION: A universal blanket treatment with broad-spectrum anthelmintics together with promotion of scholastic and health education and improvements in sanitation is recommended for helminth control in the communities under study.

COVID Moonshot: Open Science Discovery of SARS-CoV-2 Main Protease Inhibitors by Combining Crowdsourcing, High-Throughput Experiments, Computational Simulations, and Machine Learning

The COVID-19 pandemic is a stark reminder that a barren global antiviral pipeline has grave humanitarian consequences. Future pandemics could be prevented by accessible, easily deployable broad-spectrum oral antivirals and open knowledge bases that derisk and accelerate novel antiviral discovery and development. Here, we report the results of the COVID Moonshot, a fully open-science structure-enabled drug discovery campaign targeting the SARS-CoV-2 main protease. We discovered a novel chemical scaffold that is differentiated from current clinical candidates in terms of toxicity, resistance, and pharmacokinetics liabilities, and developed it into noncovalent orally-bioavailable nanomolar inhibitors with clinical potential. Our approach leveraged crowdsourcing, high-throughput structural biology, machine learning, and exascale molecular simulations. In the process, we generated a detailed map of the structural plasticity of the main protease, extensive structure-activity relationships for multiple chemotypes, and a wealth of biochemical activity data. In a first for a structure-based drug discovery campaign, all compound designs (>18,000 designs), crystallographic data (>500 ligand-bound X-ray structures), assay data (>10,000 measurements), and synthesized molecules (>2,400 compounds) for this campaign were shared rapidly and openly, creating a rich open and IP-free knowledgebase for future anti-coronavirus drug discovery.