Animal rhinosporidiosis in Brazil. Report of three additional cases

Fifteen cases of animal rhinosporidiosis seen in Brazil are briefly reviewed. Additional cases of the disease in two horses and one mule are reported.     

In vitro studies on the mechanisms of endospore release by Rhinosporidium seeberi

Studies of Rhinosporidium seeberi have demonstrated that this organism has a complex life cycle in infected tissues. Its in vivo life cycle is initiated with the release of endospores into a host's tissues from its spherical sporangia. However, little is known about the mechanisms of sporangium formation and endospore release since this pathogen is intractable to culture. We have studied the in vitro mechanisms of endospore release from viable R. seeberi's sporangia. It was found that watery substances visibly stimulates the mature sporangia of R. seeberi to the point of endospore discharge. The internal rearrangement of the endospores within the mature sporangia, the opening of an apical pore in R. seeberi's cell wall, and the active release of the endospores were the main features of this process. Only one pore per sporangium was observed. The finding of early stages of pore development in juvenile and intermediate sporangia suggested that its formation is genetically programed and that it is not a random process. The stimulation of R. seeberi's sporangia by water supports the epidemiological studies that had linked this pathogen with wet environments. It also explains, in part, its affinities for mucous membranes in infected hosts. The microscopic features of endospore discharge suggest a connection with organisms classified in the Kingdom Protoctista. This study strongly supports a recent finding that placed R. seeberi with organisms in the protoctistan Mesomycetozoa clade. This revised version was published online in June 2006 with corrections to the Cover Date.     

Rhinosporidium seeberi: Light,phase contrast,fluorescent and scanning electron microscopic study

Phase contrast microscopic study indicated the multilayered structure of the sporangial wall of R. seeberi while the scanning electronmicroscopic study revealed a trilaminated wall compared to a thick double walled light microscopic structure. The scanning electronmicroscopy revealed the spores of varying sizes which were found either discretely or in groups interconnected and seen attached to the inner aspect of the sporangial wall. Autofluorescence of sporangia and spores was observed under microscope. Acridine orange staining revealed the presence of DNA materials in the spore and sporangia.     

Cell-mediated immune responses (CMIR) to Rhinosporidium seeberi in mice

There is no published data on Cell Mediated Immune Responses in experimental animals to Rhinosporidium seeberi the causative agent of human and animal rhinosporidiosis. The quantitative mouse foot-pad model was used to assay the Delayed-Type Hypersensitivity (DTH) cell-mediated immune response to extracts of purified endospores and sporangia of R. seeberi. Histological examination was used to confirm that the foot-pad reactions were compatible with DTH reactions in the mouse. We report that sonically disintegrated rhinosporidial endospores/sporangia induced DTH responses in the foot-pads of sensitized mice which were comparable in intensity and histological profile to that induced by sheep red blood cells in SRBC sensitized mice. Anti-rhinosporidial antibody was also induced. Filtrates of the soluble antigens in sonicated suspensions failed to evoke a DTH-foot-pad (DTH-FP) response in sensitized mice although an anti-rhinosporidial antibody response to this preparation was detected. Prolonged pre-treatment with sonicated suspensions of endospores and sporangia resulted in a decrease of DTH reactivity as compared with reactions following pre-treatment of a shorter duration. This revised version was published online in June 2006 with corrections to the Cover Date.     

A simple filtration device for separating nonadherent microorganisms from mammalian cells in in vitro studies on microbial adherence

In adherence studies, the removal of nonadherent microorganisms is essential for the valid enumeration of microorganisms that adhere to host cells. Although filtration devices are available commercially for the removal of nonadherent microorganisms, these are expensive and not reusable. In this article, we describe a simple, inexpensive, and reusable filtration device composed of two chambers of nylon, a nylon membrane of desired pore size, a rubber washer, and supporting stainless steel mesh. The device was effective in in vitro adherence assays for removing nonadherent endospores of Rhinosporidium seeberi from human buccal epithelial cells, providing valid counts of adherent microorganisms.     

Crassula helmsii: attempts at elimination using herbicides

The high resistance to control by herbicides of stands of the aggressively-invasive water plant Crassula helmsii (Australian swamp stone crop or New Zealand Pygmy Weed, also sold as Tillaea recurva) was shown during a series of tank and field trials aimed at:firstly, selecting the most appropriate UK-approved herbicide showed that diquat, either directly or in alginate form was effective on submerged plants particularly at low biomass, whereas for emergent stands, although glyphosate was initially selected as effective, diquat was subsequently recommended;secondly, the efficacy of the herbicides selected under a range of conditions of biomass, season of application and, particularly, field conditions showed that whilst low biomasses could be controlled and the plant could probably be eliminated, elevated or multiple applications would be necessary at the very high biomasses (up to 45 kg fresh weight per m²) achieved by this plant, unless the bulk of the biomass could be physically reduced prior to herbicide application; further trials were considered necessary to meet legal current constraints.     

Chromosome elimination in Hordeum vulgare x H. bulbosum hybrids

Summary The hybrid progeny from a stable amphidiploid of H. vulgare x H. bulbosum involving the cultivar Vada and an unstable amphidiploid involving the cultivar Emir were studied. The genotypes examined contained two genomes from Vada or one from Vada and one from Emir, with one or two genomes from H. bulbosum. Comparisons between the chromosome numbers in root-tips and anthers revealed that there was no chromosome elimination in most plants, whether there was one or two Vada genomes present. The one plant in which chromosome elimination was positively identified had Emir as opposed to Vada cytoplasm. It also had a high incidence of degraded or fragmented chromosomes in the PMCs. Differences in stability between a 27 chromosome plant and other hypotetraploids suggest that Vada contains both elimination genes and elimination suppressor genes. Upon selfing, again irrespective of the number of Vada genomes present, circa triploid hybrids gave rise to diploid H. vulgare offspring while hypotetraploids produced hybrid-like plants. These included diploids, triploids and tetraploids. There was evidence that suggested that H. vulgare as well as H. bulbosum chromosomes had been eliminated.     

Rhinosporidiosis in man: Case reports

Three cases of rhinosporidiosis with complications, hitherto not reported, are presented. Frog-faced deformity and seizures were observed in one case, excessive bleeding, loss of olfactory sensation and septal perforation were noticed in the second case, while the third one exhibited a disseminated type of the disease. Clinical findings of these cases are discussed.     

High crossability of wild barley (Hordeum spontaneum C. Koch) with bread wheat and the differential elimination of barley chromosomes in the hybrids

Four bread wheat (Triticum aestivum L.) cultivars, Aobakomugi, Chinese Spring, Norin 61 and Shinchunaga, were pollinated with five barley lines/cultivars consisting of three cultivated barley (Hordeum vulgare L.) lines, Betzes, Kinai 5 and OHL089, and two wild barley (Hordeum spontaneum C. Koch) lines, OUH602 and OUH324. Crossability, expressed as the percentage of embryo formation, varied from 0 to 55.4% among the cross combinations. The two wild barley lines generally had a higher crossability than the previously reported best pollinator, Betzes, and some Japanese wheat cultivars were better as the female parent than Chinese Spring. Ninety four hybrid plants were obtained from 250 embryos cultured, and their somatic chromosome numbers ranged from 21 to 36. Eighteen plants were mosaic in chromosome number. Twenty one-chromosome plants appeared most frequently (45.7%) followed by 28-chromosome plants (14.9%). C-banding analysis revealed that elimination of barley chromosomes was mainly responsible for the occurrence of aneuploid plants. In hypoploids derived from Betzes-crosses, chromosome 5 was preferentially eliminated as previously reported, while in hypoploids derived from OUH602-crosses, chromosome 4 was preferentially eliminated. The wild barley line OUH602 may be a useful parent for producing a new wheat-barley addition set because of its high crossability with wheat and a different pattern of chromosome elimination.     

Premeiotic chromosome doubling after genome elimination during spermatogenesis of the species hybrid Rana esculenta

Summary Gamete production in the hybridogenetic species hybrid Rana esculenta (Rana ridibunda X Rana lessonae) is preceded by a premeiotic elimination of the R. lessonae genome and subsequent duplication of the remaining R. ridibunda genome, so that only ridibunda chromosomes enter a quasi normal meiosis, and only ridibunda gametes are formed. This is demonstrated by differences in genome specific centromere fluorescence and electrophoretic patterns between somatic and gonadal tissue.     

Asynchronous meiotic rhythm as the cause of selective chromosome elimination in an interspecific <Emphasis Type="Italic">Brachiaria</Emphasis> hybrid

This paper reports the occurrence of chromosome elimination during microsporogenesis in an interspecific hybrid between a sexual diploid accession (SEX) of Brachiaria ruziziensis (2n=2x=18) and an apomictic tetraploid accession (APO) of B. brizantha (2n=4x=36). Meiosis was very abnormal in the triploid hybrid (2n=3x=27); we observed a distinct asynchrony from metaphase I to the end of meiosis. The APO and the SEX genomes did not show the same meiotic rhythm. When the former, with nine bivalents, was in metaphase I, the nine SEX univalents were not yet aligned; when the latter reached the plate, the APO genome was already in anaphase. In subsequent stages, the APO genome had reached the poles while the SEX was undergoing sister-chromatid segregation. As the SEX genome always remained temporally behind, it gave rise to one extra-nucleus in each pole. In the second division, the behavior was the same but anaphase II did not occur for the SEX genome, and only one extra-nucleus was observed in each cell in telophase II. Chromosome elimination for the SEX genome ranged from partial to total. The importance of these findings with respect to Brachiaria breeding programmes is discussed.     

Glycerol-, inositol-, and reducing end hexose-containing oligosaccharides in human urine

Ten previously unreported oligosaccharides have been purified from the urines of human subjects using a combination of gel filtration, ion exchange, and thin-layer chromatographies. Their structures were determined by direct probe mass spectrometry, methylation analysis, and proton NMR spectroscopy of the permethylated oligosaccharide alditols.On the basis of composition, the oligosaccharides could be divided into three groups. Five oligosaccharides containing glycerol were characterized as glucosyl1-1glycerol; glucosyl1-1glycerol; galactosyl1-1glycerol; glucosyl-1-1(fucosyl-1-2)glycerol and/or fucosyl-1-1(glucosyl-1-2)glycerol; and glucosyl-1-1(galactosyl-1-2)glycerol or galactosyl-1-1(glucosyl-1-2)glycerol. Four inositol-containing oligosaccharides were characterized as galactosyl1 (fucosyl1)inositol,N-acetylgalactosaminyl1 (fucosyl1)inositol, fucosyl1-2galactosyl1 (N-acetylgalactosaminyl1)inositol and fucosyl1-2galactosyl1-4-N-acetylglucosaminyl1(N-acetylgalactosaminyl1)inositol. Finally, galactosyl1-3(fucosyl1-2)galactosyl1-6galactosyl1-4(fucosyl1-3)glucose, an oligosaccharide with glucose at its reducing end, was tentatively identified. The significance and possible origins of the carbohydrate structures are discussed.     

Histopathology of an aphid,Schoutedenia lutea,parasitized by a gall midge,Pseudendaphis sp

Pseudendaphis sp. (Barnes) [Diptera: Cecidomyiidae] is a larval endoparasitoid of the aphidSchoutedenia lutea (van der Goot) [Aphididae: Greenideinae]. A brief investigation of the biology ofPseudendaphis sp. revealed that parasitization by this species occurs only in viviparous instar III to adult life stages of the host aphid. Comparative histology of parasitized, unparasitized and starved aphids was used to evaluate proposed mechanisms of pathogenesis. Parasitization byPseudendaphis sp. was found to cause the destruction of host embryos and pathological changes in the host fat body, yet resulted in an increase in the number of host mycetocytes. There was no evidence of injury to host tissues through direct physical attack, a cytolytic enzyme or starvation-like factors. Remaining hypotheses for altered host histologies in parasitized aphids are selective nutrient depletion and production of a non-cytolytic bioactive substance by thePseudendaphis larvae.      

Metabolism and elimination of ingested allelochemicals in a holometabolous and a hemimetabolous insect

Experiments were conducted to test the hypothesis that hemimetabolous insects such as Orthoptera utilize physical barriers in the gut to provide protection from ingested toxins, whereas holometabolous Lepidoptera metabolize such toxins more efficiently. Migratory grasshoppers, Melanoplus sanguinipes Fab. (Acrididae), eliminated the thiophene -terthienyl in their feces more quickly and metabolized it to a lesser degree than did variegated cutworms, Peridroma saucia Hübner (Noctuidae). Xanthotoxin, a phototoxic linear furanocoumarin, and digitoxin, a cardiac glycoside, underwent similar metabolic modification in both species. Rates of fecal elimination of xanthotoxin were similar, whereas digitoxin was voided more rapidly by P. saucia. Overall, our results fail to provide convincing evidence in support of the test hypothesis because the two species investigated do not differ markedly in the manner in which they metabolize and eliminate the allelochemicals utilized in these studies.     

Differential organ infection studies, potyvirus elimination, and field performance of virus-free garlic plants produced by tissue culture

Presence of potyvirus in single garlic (Allium sativum L.) cloves from the same bulb, and in five single leaves excised from commercial field-grown individual plants was studied using ELISA. It was found that the viruses were not present in all organs of the same plant, since some cloves of the same bulb were infected with potyvirus but some others were potyvirus-free. Analyzed leaves from a given plant also exhibited irregular distribution of potyvirus. This study also aimed to obtain potyvirus-free plants from two commercial garlic cultivars (Taiwan and Chileno) using cloves subjected to thermotherapy, chemotherapy or meristematic dissection followed by in vitro culture. Thermotherapy (sequential treatment at 32°C for a week, 36°C for 2 weeks, and 38°C for 3 weeks) was found to affect survival of explants and 36.5% cloves from Taiwan and 26.8% from Chileno cultivars were recovered after the treatment. ELISA tests showed that 63% of the cloves of Taiwan that survived the treatment and 70.9% of Chileno explants were potyvirus-negative. Regarding chemotherapy (205 μM Ribavirin solution), the explants (cloves) survived, but only an average of 27.0–34.8% were negative for the presence of potyvirus. When meristematic dissection was applied, an average of 41.7% explants of Taiwan and 34.2% of Chileno survived the treatment, and approximately 64% of these explants from both cultivars were potyvirus-negative. Potyvirus-free garlic plants grown in field conditions showed longer stems with a major fresh and dry weight per bulb, and also exhibited a higher yield than non-treated plants.     

Aphanomyces infection in juvenile soft-shelled turtle,Pelodiscus sinensis,imported from Singapore

Aphanomyces sp. was isolated from the carapaces of two juvenile soft-shelled turtles with fungal infections imported from Singapore. Their sizes were 2.9–3.5 cm in carapace length. Lesions with integumental necrosis and ulceration looked like white cotton. The fungus exhibited slow growth, hyphae were 7.5–15 μm in diam, coarse, and abundantly branched. Zoosporangia observed in the isolate were complex, its entire thallus being converted into zoosporangial units, with short or long lateral evacuation tubes, and isodiametric, 100–500 μm in length. Clusters of zoospores were also produced at the terminals of hyphae. The production of the primary zoospores was achlyoid. The primary encysted zoospores were spherical, 10–15 μm in diam. No sexual stages were observed on a hemp seed incubated in sterile tap water. The optimal temperature for the fungus was 30° C.     

A novel transposon-based method for elimination of large bacterial plasmids

Elimination or modification of large plasmids of bacteria is often an essential step in functional analysis of these replicons. However, the conventional plasmid-curing procedures such as ethidium bromide and heat treatment are insufficient in many cases. For instance, curing of the large virulence plasmid of Salmonella Enteritidis 2,102 has failed when these treatments were applied. To overcome the difficulties, a two-step transposon-based curing method has been developed. First, a Tn10-based transposable unit carrying a Km(R) marker gene and the joined IS30 ends transposes from a replication deficient conjugative plasmid into the target replicon. Then, the inducible IS30 transposase, using the highly reactive joined IS30 ends, mediates deletions or gives rise to the loss of the target plasmid. The efficiency of the method has been monitored by the frequency of Km(S) colonies after induction of IS30 transposase, and it was shown that the Km(S) phenotype often accompanied the complete loss of the virulence plasmid or the formation of deletion derivatives. The procedure has been successfully applied also in removing the large virulence plasmid from enterotoxigenic Escherichia coli (ETEC O147), suggesting that the transposon-based method can be a useful tool for eliminating native plasmids in several bacteria.     

The relations between morphology, asymmetry of mutual articular facets, and range of movement, in the human hand joints

Surface area measurements were performed on the articular facets in the human hand joints in order to calculate the area-asymmetry (incongruency) between mutual articular facets in every one of the hand joints, and to learn about the relations between the asymmetry in the joint, the joint's morphology, and the range of movement exhibited by it. Several different trials have led to the conclusion that the asymmetry in a joint is strongly influenced by its morphological type (plane; sellar; condyloid; bicondylar). No such correlation was found between the asymmetry in a joint and its characteristic range of movement. The results contradict the intuitive assumption that the greater the incongruency in a joint, the wider the movement it can perform.     

Geographical distribution of human Schistosoma japonicum infection in The Philippines: tools to support disease control and further elimination

Schistosoma japonicum infection is believed to be endemic in 28 of the 80 provinces of The Philippines and the most recent data on schistosomiasis prevalence have shown considerable variability between provinces. In order to increase the efficient allocation of parasitic disease control resources in the country, we aimed to describe the small-scale spatial variation in S. japonicum prevalence across The Philippines, quantify the role of the physical environment in driving the spatial variation of S. japonicum, and develop a predictive risk map of S. japonicum infection. Data on S. japonicum infection from 35,754 individuals across the country were geo-located at the barangay level and included in the analysis. The analysis was then stratified geographically for the regions of Luzon, the Visayas and Mindanao. Zero-inflated binomial Bayesian geostatistical models of S. japonicum prevalence were developed and diagnostic uncertainty was incorporated. Results of the analysis show that in the three regions, males and individuals aged ?20 years had significantly higher prevalence of S. japonicum compared with females and children <5 years. The role of the environmental variables differed between regions of The Philippines. Schistosoma japonicum infection was widespread in the Visayas whereas it was much more focal in Luzon and Mindanao. This analysis revealed significant spatial variation in the prevalence of S. japonicum infection in The Philippines. This suggests that a spatially targeted approach to schistosomiasis interventions, including mass drug administration, is warranted. When financially possible, additional schistosomiasis surveys should be prioritised for areas identified to be at high risk but which were under-represented in our dataset.