Successful Treatment of Capture Myopathy and Satellite Transmitter Injury in an Atlantic Yellow-nosed Albatross (Thalassarche chlororhynchos).

An Atlantic yellow-nosed albatross (Thalassarche chlororhynchos) was found on shore 3 days after having been captured at sea by researchers. It presented very lethargic, moderately dehydrated, and in poor body condition. It was mildly hypothermic, with moderate pediculosis, and dark malodorous feces with yellow urates. The bird had a 48-g satellite transmitter attached with a backpack-style chest harness, which caused an ulcerative lesion on the interscapular area. The bird was severely anemic (packed cell volume, 18%), and plasma chemistry results were suggestive of a severe rhabdomyolysis (aspartate transaminase, 3620 U/L; creatine kinase, 100 400 U/L). We hypothesized that capture myopathy occurred because of a combination of capture stress and prolonged physical restraint by researchers, stress associated with repeated attempts by the bird to remove the satellite-transmitter harness, and a lengthy road transport to the rehabilitation center. A treatment protocol, which relied on a combination of aggressive fluid therapy, selenium, vitamins E and B12, and multivitamin supplementation, was administered after the initial physical assessment of the albatross. Isoflurane inhalation anesthesia was used to minimize stress associated with the performed medical procedures (eg, physical examination, removal of the satellite transmitter harness, blood collection, and wound management). Measures were adopted while the bird was hospitalized to reduce stress (eg, quiet and comfortable environment with visual barriers and restricting handling of the patient to experienced staff). Clinical and hematological monitoring was used to assess the patient's condition as the bird gradually recovered while hospitalized. The albatross was successfully released 28 days after the initial presentation, suggesting that the medical protocol employed in this case may be useful for the treatment of capture myopathy in albatrosses and other birds.

New insights in Trichoderma harzianum antagonism of fungal plant pathogens by secreted protein analysis.

Trichoderma harzianum ALL42 were capable of overgrowing and degrading Rhizoctonia solani and Macrophomina phaseolina mycelia, coiling around the hyphae with formation of apressoria and hook-like structures. Hyphae of T. harzianum ALL42 did not show any coiling around Fusarium sp. hyphae suggesting that mycoparasitism may be different among the plant pathogens. In this study, a secretome analysis was used to identify some extracellular proteins secreted by T. harzianum ALL42 after growth on cell wall of M. phaseolina, Fusarium sp., and R. solani. The secreted proteins were analyzed by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. A total of 60 T. harzianum ALL42 secreted proteins excised from the gel were analyzed from the three growth conditions. While seven cell wall-induced proteins were identified, more than 53 proteins spots remain unidentified, indicating that these proteins are either novel proteins or proteins that have not yet been sequenced. Endochitinase, ß-glucosidase, α-mannosidase, acid phosphatase, α-1,3-glucanase, and proteases were identified in the gel and also detected in the supernatant of culture.

Light-dependent roles of the G-protein alpha subunit GNA1 of Hypocrea jecorina (anamorph Trichoderma reesei).

BACKGROUND: The filamentous ascomycete Hypocrea jecorina (anamorph Trichoderma reesei) is primarily known for its efficient enzymatic machinery that it utilizes to decompose cellulosic substrates. Nevertheless, the nature and transmission of the signals initiating and modulating this machinery are largely unknown. Heterotrimeric G-protein signaling represents one of the best studied signal transduction pathways in fungi. RESULTS: Analysis of the regulatory targets of the G-protein alpha subunit GNA1 in H. jecorina revealed a carbon source and light-dependent role in signal transduction. Deletion of gna1 led to significantly decreased biomass formation in darkness in submersed culture but had only minor effects on morphology and hyphal apical extension rates on solid medium. Cellulase gene transcription was abolished in Deltagna1 on cellulose in light and enhanced in darkness. However, analysis of strains expressing a constitutively activated GNA1 revealed that GNA1 does not transmit the essential inducing signal. Instead, it relates a modulating signal with light-dependent significance, since induction still required the presence of an inducer. We show that regulation of transcription and activity of GNA1 involves a carbon source-dependent feedback cycle. Additionally we found a function of GNA1 in hydrophobin regulation as well as effects on conidiation and tolerance of osmotic and oxidative stress. CONCLUSION: We conclude that GNA1 transmits a signal the physiological relevance of which is dependent on both the carbon source as well as the light status. The widespread consequences of mutations in GNA1 indicate a broad function of this Galpha subunit in appropriation of intracellular resources to environmental (especially nutritional) conditions.

The influence of N-glycosylation on biochemical properties of Amy1, an alpha-amylase from the yeast Cryptococcus flavus.

The yeast Cryptococcus flavus secretes a glycosylated alpha-amylase (Amy1) when grown in a starch-containing medium. The effects of N-glycosylation on secretion, enzyme activity, and stability of this glycoprotein were studied. Addition of tunicamycin (TM) to the medium at a concentration higher than 0.5 microg mL(-1) affected C. flavus growth. Amy1 activity increased by 55% in the intracellular fraction after C. flavus growth in the presence of 0.5 microg mL(-1) TM. SDS-PAGE and gel activity detection showed that native enzyme and deglycosylated enzyme had apparent molecular mass of 68 and 64.5 kDa, respectively. The N-glycosylation process did not affect either optimum pH or optimum temperature. The K(M) values of native and non-glycosylated alpha-amylases were 0.052 and 0.098 mg mL(-1), and V(max) values were 0.038 and 0.047 mg min(-1), respectively. However, the non-glycosylated form was more sensitive to inactivation by both the proteolytic enzyme trypsin and high temperature. Furthermore, the activity of the non-glycosylated enzyme was affected by Hg(2+) and Cu(2+) suggesting that N-glycosylation is involved in the folding of Amy1.

Involvement of G-alpha protein GNA3 in production of cell wall-degrading enzymes by Trichoderma reesei (Hypocrea jecorina) during mycoparasitism against Pythium ultimum.

The involvement of the G-alpha protein GNA3 in the production of cell wall-degrading enzymes (CWDEs) by Trichoderma reesei during antagonism against Pythium ultimum was investigated. cAMP content was 2.8-fold higher in the T. reesei mutant gna3QL than in the parental TU-6. The gna3QL, like TU-6, inhibited the growth of P. ultimum in dual culture assays. Scanning electron microscopy showed that the gna3QL promoted more morphological alterations of P. ultimum cell wall than TU-6. In general, gna3QL produced higher activities of CWDEs than TU-6. We therefore suggest that CWDEs production during mycoparasitism by T. reesei against P. ultimum may be associated with the level of GNA3 activity.