Molecular evidence for Toxoplasma gondii in the brain of striped dolphins (Stenella coeruleoalba) stranded along the Ligurian Sea coast of Italy

Toxoplasma gondii is a protozoan agent of increasing concern for both free-ranging and captive cetaceans. Nonetheless, information on epidemiology, biology, genetics and pathogenic potential of T. gondii for aquatic mammals is incomplete. This work provides molecular evidence for T. gondii in the brain tissue from five striped dolphins (Stenella coeruleoalba) found stranded in 2007-2008 along the Ligurian Sea coast of Italy. These animals showed a more or less severe, subacute to chronic, non purulent, multifocal meningo-encephalitis. Additionally, the cerebral parenchyma of 3 animals harboured protozoan cysts and zoites which were immunohistochemically linked to T. gondii. Consequently, a preliminary molecular investigation was undertaken to confirm the presence of the protozoan in these dolphins. Genomic DNA was extracted from the brain tissue of the animals and the extracts were examined with a nested PCR specific for the B1 gene of T. gondii. Three out of the 5 animals scored positive upon B1-PCR. Sequences were subsequently searched against the GenBankTM Database using the Basic Local Alignment Search Tool. The three sequences were identical to each other and showed 100% homology with the corresponding B1 sequence of T. gondii, with no insertions/deletions nor substitutions. The other 2 dolphins were negative upon B1-specific PCRs and also upon PCRs targeting other T. gondii genes (i.e. GRA and UPRT). To explore more in depth the genetic make-up of this protozoan, complementary studies are presently ongoing on GRA and UPRT genes of the 3 animals which were molecularly confirmed (i.e. B1-PCR) as infected by T. gondii. In conclusion, further studies are needed to elucidate a number of epidemiological and biological features of T. gondii infection in free-ranging cetaceans, with special emphasis on “pelagic” species such as striped dolphins. 

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Application of ESPI-Method for Strain Analysis in Thin Wall Cylinders

The Centre for Industrial and Engineering Optics, DIT, Ireland, have recently developed and applied new optical techniques for the measurement of mechanical strain, one of which was based on electronic speckle pattern interferometry (ESPI). The accuracy of this optical technique when used on flat surfaces is well established. In this research the technique is tested on curved surfaces, and results are compared with the theoretical hoop strain as predicted by the bi-axial strain equation modified for the thin cylinder, and by those obtained by electrical resistance strain gauge (ERSG) undoubtedly the principal method of measuring mechanical strain. For testing procedures, a special unit was designed and produced for holding and loading the thin cylinder. Also the basic equation for hoop strain by ESPI for flat surfaces was modified for applications on curved surfaces. Thin cylinder hoop strain obtained by ESPI and calculated by a modified equation show remarkably good correlation to the predicted theoretical value as well as to the results obtained by ERSG. Since the ESPI theory was originally developed for use on flat surfaces, there are reasonable grounds for further investigating this relationship especially where curved surfaces are involved

Fate of novel Quasi reverse steroidal substrates by Aspergillus tamarii KITA: Bypass of lactonisation and an exclusive role for the minor hydroxylation pathway

The fungus Aspergillus tamarii transforms progesterone to testololactone in high yield through a flexible four-step enzymatic pathway. To date no studies have investigated the effect of transposition of steroidal functionality between ring-A and ring-D in order to determine the effect on steroidal metabolism. A series of novel quasi reverse steroids (7–9) were synthesised through Linz and Schäfer oxidation where 14-en-16-one functionality is generated on ring-D of the steroid. To retain parity with the normal series ring-D functionality was substituted onto ring-A of the analogues. All of the analogues (7–9) were handled through a minor 11β-hydroxylation pathway with no lactones being formed. In previous studies testololactone is produced within the first 12 h of metabolism. A time course experiment demonstrated that the transformation of these steroids initiated with the formation of a 3β-hydroxy group after which (48–96 h) hydroxylation through a minor pathway occurred, indicating that this hydroxylase was only then being induced. This is in contrast to the normal fungal metabolism of xenobiotic steroidal substrates where they are primarily hydroxylated. Furthermore, ring-D hydrogenation is reported for the first time as is reverse metabolism on this pathway. All metabolites were isolated by column chromatography and were identified by 1H and 13C NMR spectroscopy, DEPT analysis and other spectroscopic and crystallographic data