Using Molecular Epidemiology to Track Toxoplasma gondii from Terrestrial Carnivores to Marine Hosts: Implications for Public Health and Conservation

Background

Environmental transmission of the zoonotic parasite Toxoplasma gondii, which is shed only by felids, poses risks to human and animal health in temperate and tropical ecosystems. Atypical T. gondii genotypes have been linked to severe disease in people and the threatened population of California sea otters. To investigate land-to-sea parasite transmission, we screened 373 carnivores (feral domestic cats, mountain lions, bobcats, foxes, and coyotes) for T. gondii infection and examined the distribution of genotypes in 85 infected animals sampled near the sea otter range.

Methodology/Principal Findings

Nested PCR-RFLP analyses and direct DNA sequencing at six independent polymorphic genetic loci (B1, SAG1, SAG3, GRA6, L358, and Apico) were used to characterize T. gondii strains in infected animals. Strains consistent with Type X, a novel genotype previously identified in over 70% of infected sea otters and four terrestrial wild carnivores along the California coast, were detected in all sampled species, including domestic cats. However, odds of Type X infection were 14 times higher (95% CI: 1.3–148.6) for wild felids than feral domestic cats. Type X infection was also linked to undeveloped lands (OR = 22, 95% CI: 2.3–250.7). A spatial cluster of terrestrial Type II infection (P = 0.04) was identified in developed lands bordering an area of increased risk for sea otter Type II infection. Two spatial clusters of animals infected with strains consistent with Type X (P≤0.01) were detected in less developed landscapes.

Conclusions

Differences in T. gondii genotype prevalence among domestic and wild felids, as well as the spatial distribution of genotypes, suggest co-existing domestic and wild T. gondii transmission cycles that likely overlap at the interface of developed and undeveloped lands. Anthropogenic development driving contact between these cycles may increase atypical T. gondii genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans, domestic animals, and wildlife.     

Aquatic invertebrates in final void water bodies at an open-cut coal mine in central Queensland

Abstract  We describe the diversity of aquatic invertebrates colonising water-filled final voids produced by an open-cut coal mine near Moura, central Queensland. Ten disused pits that had been filled with water from <1 year to 22 years prior to the survey and three nearby 'natural' water bodies were sampled in December 1998 and again in March 1999. All invertebrates collected were identified to family with the exception of oligochaetes, cladocerans, ostracods and copepods, which were identified to these coarser taxonomic levels. Sixty-two taxa were recorded from >20 000 individuals. The greatest familial richness was displayed by the Insecta (33 families) followed by the mites (Acari) with 12 families. While natural water bodies held the greatest diversity, several mine pits were almost as rich in families. Classification analyses showed that natural sites tended to cluster together, but the groupings did not clearly exclude pit sites. Mining pits that supported higher diversity tended to be older and had lower salinity (<2000 µS/cm); however, salinity in all water bodies varied with rainfall conditions. We conclude that ponds formed in final voids at this mine have the potential to provide habitat for many invertebrate taxa typical of lentic inland water bodies in central Queensland.     

Reemergence of Guadalupe fur seals in the U.S. Pacific Northwest: The epidemiology of stranding events during 2005–2016

Guadalupe fur seals (GFS), Arctocephalus philippii townsendi, an U.S. Endangered Species Act threatened pinniped, have recently reappeared in their historic range along the western seaboard of the continental United States. Starting 2005 through 2016, 169 GFSs stranded in Washington and Oregon, involving two designated unusual mortality events. The circumstances surrounding GFS strandings, mortality, and their increased presence in Oregon and Washington were analyzed during this study. Detailed necropsies, histopathology (n = 93), and epidemiological analysis found three main causes of death (COD): emaciation (44%), trauma (29%), and infectious disease (19%) and the factors associated with overall strandings and emaciation. Trauma included many cases found associated with fisheries interactions and clustered near the mouth of the Columbia River, where high levels of commercial fishing occur. The most common pathogens found associated with disease were Toxoplasma gondii, Sarcocystis neurona, and gastrointestinal helminths. Seasonality and upwelling were associated with higher stranding numbers regardless of COD. Seasonal migration into the region, coinciding with postweaning, suggests young GFSs are in search of prey and habitat resources. Reemergence of GFSs is likely due to conservation efforts, which have been critical for species recovery in the region. Continued monitoring is needed as this vulnerable species continues to rebound.     

Compound loci and coincident mutation inNeurospora

Summary To explain the high frequency of partial and intermediate back-mutants and the back-mutation of deletions, a model of mutation production based on unequal sister strand exchange in multi-repeat loci is proposed.The recovery of complementary products of mutation which the model predicts should be produced from single nuclei is cited as evidence for its plausability.Back-mutation of the mutant used (ahis-3 mutant,Gg59) is associated with a high frequency of simultaneous switch mutations of other characters in the same genome.It is suggested that unequal sister strand exchange may also contribute to the production of spontaneous mutations.The validity of citing ability to back-mutate as critical evidence for classifying a mutant as a point (single base pair substitution?) mutation is queried.With 2 Figures in the Text     

Interpretation of Changes in Circulating Tumor Cell Counts

The presence of circulating tumor cells (CTCs) in the blood of cancer patients may guide the use of therapy. We investigated how to evaluate a reduction in the number of CTCs after administration of therapy. CTCs were enumerated with the CellSearch system in 111 metastatic breast and 185 metastatic prostate cancer patients before start of a new line of chemotherapy and after initiation of therapy. Different means to express changes in CTC counts were evaluated with respect to overall survival (OS). A static CTC cutoff is the best method to determine whether a therapy is effective. This is exemplified by the highest Cox hazard ratio of 2.1 for OS; three methods to express relative differences performed worse. A lookup table is provided from which the significance of a change in CTCs can be derived. The aim of therapy should be the elimination of all CTCs. A period of 10 to 12 weeks of therapy is needed to reach the treatment effect on CTCs.     

Body temperature in captive long-beaked echidnas (Zaglossus bartoni)

The routine occurrence of both short-term (daily) and long-term torpor (hibernation) in short-beaked echidnas, but not platypus, raises questions about the third monotreme genus, New Guinea's Zaglossus. We measured body temperatures (T(b)) with implanted data loggers over three and a half years in two captive Zaglossus bartoni at Taronga Zoo, Sydney. The modal T(b) of both long-beaks was 31 degrees C, similar to non-hibernating short-beaked echidnas, Tachyglossus aculeatus, in the wild (30-32 degrees C) and to platypus (32 degrees C), suggesting that this is characteristic of normothermic monotremes. T(b) cycled daily, usually over 2-4 degrees C. There were some departures from this pattern to suggest periods of inactivity but nothing to indicate the occurrence of long-term torpor. In contrast, two short-beaked echidnas monitored concurrently in the same pen showed extended periods of low T(b) in the cooler months (hibernation) and short periods of torpor at any time of the year, as they do in the wild. Whether torpor or hibernation occurs in Zaglossus in the wild or in juveniles remains unknown. However, given that the environment in this study was conducive to hibernation in short-beaks, which do not easily enter torpor in captivity, and their large size, we think that torpor in wild adult Zaglossus is unlikely.     

Sexual recombination and clonal evolution of virulence in Toxoplasma

The protozoan parasite Toxoplasma gondii is endemic worldwide. For such a widespread pathogen that has few geographic or host boundaries, it possess an unexpected population structure comprised principally of three clonally propagated lineages. The origin and the evolutionary dynamics of these three lines are unclear. Recent population genetic analyses suggest that a meiotic recombination between two discrete gene pools produces a pandemic outbreak of three super-successful lines, which have recently come to dominate most other strains worldwide.     

Surface antigens of Toxoplasma gondii: variations on a theme

Toxoplasma gondii is an obligate intracellular protozoan parasite with an exceptionally broad host range. Recently, it has become apparent that the number of surface antigens (SAGs) it expresses may rival the number of genera it can infect. Most of these antigens belong to the developmentally regulated and distantly related SAG1 or SAG2 families. The genes encoding the surface antigens are distributed throughout the T. gondii genome, with remarkably little polymorphism observed at each locus. Results from a number of studies have suggested that the surface antigens play an important role in the biology of the parasite. For example, SAG3 null mutants generated by targeted disruption provide convincing evidence that this surface antigen, at least, functions during parasite attachment. Analyses of a SAG1 knockout in rodents, however, indicate that this surface antigen may play a crucial role in immune modulation or virulence attenuation. The current understanding of the SAG1 and SAG2 families will be discussed here.     

Measurement of the spatial redistribution of water in rabbit Achilles tendon in response to static tensile loading

The redistribution of water in response to static tensile loading was investigated in rabbit Achilles tendons in vitro. The distribution of water was measured along a radially oriented line, using a one-dimensional proton-density map created from fits to diffusion-weighted magnetic resonance (MR) data. Water movements were measured during application of tensile loads of 5N (N=7) and 10N (N=6). Water distribution along the line was measured before loading and up to 42 min after load application. Static loading with either 5 or 10N loads caused a steady increase in proton density in the outside edge (rim) of the tendon. The 10N load lowered the proton density in the core of the tendon, but did so in a single step that was observed when the load was applied. The 5N load caused no change in proton density in the core region. The immediate redistribution from the core was statistically significant for the 10N load, but not the 5N load application. Statistically significant within-group proton-density increases were observed in the rim after 42 min postload for all tendons irrespective of load condition. The rate of proton-density postload increase at the rim region did not depend upon load. The rate for the 5N load case was 0.010 +/- 0.002 min(-1) and 0.007 +/- 0.002 min(-1) in the 10N case. Thus, while generally consistent with an extrusion model, the data show other features that argue for a more complex model.