In this study, we evaluate the MM3-COPRO method for detection of Fasciola coproantigens in human fecal samples, and the usefulness of a new preservative/diluent, CoproGuard, developed for preservation of Fasciola coproantigens. The MM3-COPRO assay was evaluated with 213 samples from healthy patients, 30 Fasciola positive fecal samples (according to the Kato-Katz method), and 83 samples from patients with other parasitic infections. All Fasciola positive specimens were detected with the MM3-COPRO assay (100% sensitivity) and there was no cross-reactivity with other common parasites present in the clinical specimens analyzed (100% specificity). The use of CoproGuard enhanced coproantigen extraction without affecting the detection limit of the assay, and the antigenicity of Fasciola coproantigens in fecal samples stored at 37 degrees C was retained throughout the entire observation period (120 days). We concluded that the MM3-COPRO ELISA combined with the use of CoproGuard may be a very useful tool for the diagnosis of human fascioliasis.

Cryptosporidium is an apicomplexan protozoan that lives in most vertebrates, including humans. Its gp60 gene is functionally involved in its attachment to host cells, and its high level of genetic variation has made it the reference marker for sample typing in epidemiological studies. To understand the origin of such high diversity and to determine the extent to which this classification applies to the rest of the genome, we analysed the patterns of variation at gp60 and nine other nuclear loci in isolates of three Cryptosporidium species. Most loci showed low genetic polymorphism (πS <1%) and similar levels of between-species divergence. Contrastingly, gp60 exhibited very different characteristics: (i) it was nearly ten times more variable than the other loci; (ii) it displayed a significant excess of polymorphisms relative to between-species differences in a maximum-likelihood Hudson-Kreitman-Aguadé test; (iii) gp60 subtypes turned out to be much older than the species they were found in; and (iv) showed a significant excess of polymorphic variants shared across species from random expectations. These observations suggest that this locus evolves under balancing selection and specifically under negative frequency-dependent selection (FDS). Interestingly, genetic variation at the other loci clusters very well within the groups of isolates defined by gp60 subtypes, which may provide new tools to understand the genome-wide patterns of genetic variation of the parasite in the wild. These results suggest that gp60 plays an active and essential role in the life cycle of the parasite and that genetic variation at this locus might be essential for the parasite's long-term success.

Red algae contain agglutinins that show activity against animal and human erythrocytes. Five red seaweeds are described whose hemagglutinic activity are not previously reported: Schyzimenia dubyi, Gelidium canilagineum, Callithamnion tetragonum, Chondria tenuissima and Polysiphonia brodiaei. The presence of agglutinins was also demonstrated in another twelve algal species, at least against one of the different erythrocyte types assayed. The highest titre was obtained with Callithamnion tetragonum against rabbit erythrocytes (2 22 ). Agglutinic activity is shown to be useful in chemotaxonomy of seaweeds.

We present a case of a severe orbital myiasis focusing on its management and life-threatening nature.

Production of chlamydospores on casein agar at 24°C for 48 h provides a simple means for differentiating Candida dubliniensis from Candida albicans based on chlamydospore production. Of 109 C. dubliniensis isolates tested on this medium, 106 (97.2%) produced abundant chlamydospores and three produced few chlamydospores. In contrast, of the 120 C. albicans isolates tested, 111 (92.5%) failed to produce any chlamydospores, whereas the remaining nine isolates produced few chlamydospores. These findings indicate that abundant chlamydospore production on casein agar is a useful test for discriminating between C. dubliniensis and C. albicans.Since its first description in 1995, Candida dubliniensis has been isolated from a variety of specimens from humans in countries all over the world (6,13,15,(18)(19)(20). As a consequence of the increasing number of reports on the isolation of C. dubliniensis, it is important to be able to rapidly and accurately identify this species in most clinical mycology laboratories. However, identification of C. dubliniensis is hampered by its close relationship with Candida albicans, a situation that has sometimes led to the misidentification of isolates of C. dubliniensis as C. albicans (19). At present, the most accurate differentiation between isolates of the two species is performed in reference laboratories with the use of molecule-based techniques such as PCR or DNA fingerprinting with repetitive sequence-containing DNA probes (5,17,19). However, these sophisticated techniques are not suitable and often not readily applicable for use in small clinical mycology laboratories, where simple and rapid methods are needed. Reliable phenotypic methods for the identification of C. dubliniensis isolates include carbohydrate assimilation profile analysis by using commercially available yeast identification systems and detection of differential antigen expression by immunofluorescence microscopy (2,3,11,12,20). Furthermore, a variety of other ancillary tests have been developed for discriminating between C. dubliniensis and C. albicans isolates, including the inability of C. dubliniensis to grow at 45°C (12). However, whereas these tests are useful for the presumptive identification of C. dubliniensis, they are not definitive. One of the key features employed in the initial description of C. dubliniensis was its ability to produce abundant chlamydospores on cornmeal agar and rice-agar-Tween-agar (20). Chlamydospore production by C. dubliniensis on Staib agar and caffeic acid-ferric citrate agar has also been used recently for the differentiation of C. dubliniensis from C. albicans (1,17). In the present study, the production of chlamydospores by C. dubliniensis and C. albicans on casein agar was investigated as an additional means for differentiating the two species.The reference and clinical isolates used in this study are shown in Table 1. Conventional morphological and physiologic methods, as well as molecular techniques, were employed to confirm the identity of all isolates (1,2,4,5,11,19). ...