Identification of zoonotic Cryptosporidium and Giardia genotypes infecting animals in Sydney's water catchments

To identify the animal sources for Cryptosporidium and Giardia contamination, we genotyped Cryptosporidium and Giardia spp. in wildlife from Sydney’s water catchments using sequence analysis at the 18S rRNA locus for Cryptosporidium and 18S rRNA and glutamate dehydrogenase (gdh) for Giardia. A total of 564 faecal samples from 16 different host species were analysed. Cryptosporidium was identified in 8.5% (48/564) samples from eight host species and Giardia was identified in 13.8% (78/564) from seven host species. Eight species/genotypes of Cryptosporidium were identified. Five G. duodenalis assemblages were detected including the zoonotic assemblages A and B.     

Flow cytometric analysis of glyoxalase-1 expression in human leukocytes

Altered glyoxalase‐1 (GLO‐1) activity and expression is associated with the development of late diabetic complications, malignancy and oxidative stress‐ and aging‐related diseases. In the present study, we developed a flow cytometry method for GLO‐1 detection in human leukocytes isolated from peripheral blood samples to investigate GLO‐1 expression in leukocyte subsets from type 1 and 2 diabetes mellitus patients (n = 11) and healthy subjects (n = 8). The flow cytometry analysis of GLO‐1 in leukocytes showed that expression index of GLO‐1‐positive cells was slightly increased in mononuclear leukocytes from diabetic patients. This result correlated with the increase in GLO‐1 activity in the whole blood samples of type 2 diabetes patients. In conclusion, the present study demonstrates that flow cytometry is suitable for the detection of the GLO‐1 enzyme in human leukocytes and that this method could be used to investigate the fast adaptation of the glyoxalase system related to the pathogenesis of late complications of diabetes mellitus and other glycation stress‐related disorders. Copyright © 2011 John Wiley & Sons, Ltd.     

Foodborne cryptosporidiosis

Foodborne illness, the majority of which is caused by enteric infectious agents, costs global economies billions of dollars each year. The protozoan parasite Cryptosporidium is particularly suited to foodborne transmission and is responsible for >8?million cases of foodborne illness annually. Procedures have been developed for sensitive detection of Cryptosporidium oocysts on fresh produce and molecular diagnostic assays have been widely used in case linkages and infection source tracking, especially during outbreak investigations. The integrated use of advanced diagnostic techniques with conventional epidemiological studies is essential to improve our understanding of the occurrence, source and epidemiology of foodborne cryptosporidiosis. The implementation of food safety management tools such as Good Hygienic Practices (GHP), Hazard Analysis and Critical Control Points (HACCP), and Quantitative Microbial Risk Assessment (QMRA) in industrialised nations and Water, Sanitation, and Hygiene (WASH) in developing countries is central for prevention and control and foodborne cryptosporidiosis in the future.     

Fenestrated endothelium of the adrenal gland: Freeze-fracture studies

Little is known of how adrenal hormones pass from the interstitial to the vascular space. We have begun to examine the adrenal endothelium as a barrier to hormone passage, by the freeze-fracturing technique. The endothelium of both cortex and medulla is fenestrated. Fractures from both regions show endothelial cells to be extremely thin in regions where fenestrations are abundant. En face fractures show fenestrae disposed in tracts; the fenestrae reaching a distribution of 35/μ². In both cortex and medulla there are areas of continuous endothelium which contain caveolae. Structures believed to represent fenestra diaphragms contain randomly disposed particles and occasional pits. We have not identified in replicas the central ring and pore described in thin-sectioned material (Elfvin, 1965). The main differences between freeze-fractured aspects of cortical and medullary endothelium are the greater abundance of caveolae in the medulla and the size of the fenestrae (fenestra rims in the medulla are 525–780 Å in diameter; in the cortex 570–1660 Å). These differences may reflect the different embryological origins of the medulla and cortex. While caveolae may participate in hormone transport, there is no evidence for this. In the medulla the caveolae are more numerous and may have a function not necessarily related to transport. Possibly, caveolae play a role in processing hormones and related substances. For example, ATP and specific proteins are released as well as epinephrine during exocytosis from chromaffin cells. Epinephrine enters the vascular space but ATP does not. ATPase enzymes are a common feature of caveolae of other endothelia and may occur as well in adrenal endothelium.     

FREEZE-ETCH STUDIES OF THE PLASMA MEMBRANE OF PULMONARY ENDOTHELIAL CELLS

Pulmonary endothelial cells are capable of metabolizing a variety of circulating hormonal substances. Indirect evidence indicates that some of the relevant enzymes are located on the plasma membrane. The associated caveolae are of special interest as globular subunits, possibly enzyme clusters, are evident in their membranes. In the present study, freeze-etch techniques were used to improve understanding of the fine structure of endothelial cells and to extend our investigations of possible sites of enzymes capable of metabolizing circulating vasoactive agents. As in other cells studied by freeze-etching, intramembranous particles are found on both inner aspects of the plasma membrane. In undifferentiated areas of plasma membrane, the particles appear to have a random distribution. These areas fracture such that approximately equal proportions of the particles adhere to the cytoplasmic aspect of the outer leaflet and the extracellular aspect of the inner leaflet. However, the particles organize into rosettes and plaques at the base of caveolae, and, after fracture, the rosettes and plaques adhere predominantly to the cytoplasmic aspect of the outer leaflet. The peculiar organization of particles in association with caveolae supports the concept that caveolae have a stomal skeletal structure and raises the possibility that the organization may be in some way related to pinocytosis.     

Identification of novel Babesia and Theileria genotypes in the endangered marsupials, the woylie (Bettongia penicillata ogilbyi) and boodie (Bettongia lesueur)

Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood samples were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood samples (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these individuals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus).