Biodegradation of wheat straw lignocarbohydrate complexes (LCC)

Summary In laboratory and semi-industrial scale experiments the influence of the substrate water content, temperature, and incubation time on the progress of solid state fermentation of straw colonized by white rot fungi was investigated. The parameters used to evaluate the fermentation process were degradation of total organic matter and lignin, in vitro digestibility, the content of water soluble substances in the substrate and the pH.The degradation of total organic matter was species specific. Only Trametes hirsuta enhanced the degradation at elevated temperature (30 °C). With Abortiporus biennis, Ganoderma applanatum, and Pleurotus serotinus, elevated temperature had and adverse effect. Prolonged incubation only improved degradation of straw by the relatively slowgrowing fungi Ganoderma applanatum, Lenzites betulina, and Pleurotus sajor caju.Elevated temperature and prolonged incubation shifted the relative degradation rates in favour of total organic matter degradation. With Ganoderma applanatum, Pleurotus ostreatus, and Pleurotus serotinus lignin degradation, even on an absolute scale, was less at 30 °C than at 22 °C.In general, the in vitro digestibility also decreased, when the incubation time and temperature were raised. With Ganoderma applanatum the in vitro digestibility dropped below the value of the sterile straw control.Solid state fermentation of straw was at an optimum at a medium water content of 75 ml/25 g of substrate. However, most of the fungi tested could digest straw over a wide range of water content. At higher water contents (125–150 ml/25 g of substrate) an increased production of aerial mycelium was observed.In semi-industrial batch experiments (40 kg) with Abortiporus biennis the in vitro digestibility dropped below the reference value for sterile straw during the first 19 days of incubation. Later, the in vitro digestibility again rose and reached its optimum after about 60 days. The in vitro digestibility in the semi-industrial experiments was always lower than in the laboratory experiments (+9% and +25%, respectively).In long term experiments (2.5 kg batches, 8 months of incubation) very different values for the in vitro digestibility were found, and these depended on the fungus used (Abortiporus biennis, +16%; Pleurotus ostreatus, +4%; and Ganoderma applanatum, –27%).     

Mast cell histamine release induced by Portuguese man-of-war (Physalia) venom

Nematocyst venom from Portuguese Man-of War ($\text{Physalia}$ sp.) tentacles causes isolated rat peritoneal mast cells to release histamine. Extent of histamine release is dose-dependent (K_0.5 = 6.1 μg venom/ml) and attains 100% at high doses of venom. Release is independent of intra- and extracellular calcium levels and does not depend upon a cellular supply of ATP. The rate of histamine release is temperature-dependent and the extent of release is maximized broadly over the range of 10–30°C. The cytoplasmic marker lactate dehydrogenase, is released concomitantly with histamine but is more sensitive to the venom (K_0.5 = 2.1 μg/ml). Antimycin A, while it does not significantly affect venom-induced histamine release, increases the sensitivity of lactate dehydrogenase release (K_0.5 = 0.2 μg/ml). We conclude that $\text{Physalia}$ nematocyst venom induces the release of histamine from mast cells by a cytolytic mechanism and that this action is antagonized by an intracellular, energy-requiring process.     

Synthesis of some long-chain acylamidoalkyl glucosides

4-O-β-D-Galactopyranosyl-α,β-D-glucopyranosylamine (lactosylamine), β-D-gluco-, α- and β-D-manno-pyranosylamines were bound to the carbodiimide-activated groups of lysozyme. Of the 11 free carboxyl groups of the protein, ≈3 were substituted by α,β-6-lactosylamine, and ≈2 by the monohexo-sylamines. One of the 4 glycopeptides isolated from the tryptic digest of the lysozyme-lactosylamine conjugate was identical to synthetic l-N-L-leucinoyl-4-O-β-D-galactopyranosyl-β-D-glucopyranosylamine, indicating the substitution of the carboxyl group of the C-terminal leucine residue. The isolation of a glycopeptide containing the aspartic acid residue in position 117 indicates that the second α,β-lactosylamine residue is linked to the carboxyl group of this amino acid. Both of the 2 other glycopeptides contain the same free carboxyl groups (one glutamic and two aspartic acid residues in positions 35, 48, and 52, respectively). The third α,β-lactosylamine residue seems to be linked to one of these carboxyl groups.     

Global protein expression analysis in apicomplexan parasites: current status

Members of the phylum Apicomplexa are important protozoan parasites that cause some of the most serious, and in some cases, deadly diseases in humans and animals. They include species from the genus Plasmodium, Toxoplasma, Eimeria, Neospora, Cryptosporidium, Babesia and Theileria. The medical, veterinary and economic impact of these pathogens on a global scale is enormous. Although chemo- and immuno-prophylactic strategies are available to control some of these parasites, they are inadequate. Currently, there is an urgent need to design new vaccines or chemotherapeutics for apicomplexan diseases. High-throughput global protein expression analyses using gel or non-gel based protein separation technologies coupled with mass spectrometry and bioinformatics provide a means to identify new drug and vaccine targets in these pathogens. Protein identification based proteomic projects in apicomplexan parasites is currently underway, with the most significant progress made in the malaria parasite, Plasmodium falciparum. More recently, preliminary two-dimensional gel electrophoresis maps of Toxoplasma gondii and Neospora caninum tachyzoites and Eimeria tenella sporozoites, have been produced, as well as for micronemes in E. tenella. In this review, the status of proteomics in the analysis of global protein expression in apicomplexan parasites will be compared and the challenges associated with these investigations discussed.     

Metabolic fate of [14C]-labeled meal protein amino acids in Aedes aegypti mosquitoes

We developed a method to follow the metabolic fate of [(14)C]-labeled Euglena gracilis protein amino acids in Aedes aegypti mosquitoes under three different adult nutritional regimes. Quantitative analysis of blood meal protein amino acid metabolism showed that most of the carbon of the amino acids was either oxidized to CO(2) or excreted as waste. Under the three different adult nutritional regimes, no significant differences in the metabolism of amino acids were found, which indicated that the female A. aegypti mosquitoes possess a substantial capacity of maintaining metabolic homeostasis during a gonotrophic cycle. The amount of maternal glycogen and lipid after egg laying were significantly lower in the mosquitoes that underwent a partial starvation before a blood meal and/or starvation after the blood meal. The content of egg lipid or protein or the number of eggs laid did not show a significant difference among the three different regimes, which indicates that stable fecundity of A. aegypti under the partial starvation before a blood meal and/or starvation after the blood meal seemed to result from a trade-off between current fecundity and future survival after the eggs laid. The methods described in this paper can be applied to a wide range of questions about the effects of environmental conditions on the utilization of blood meal amino acids.     

Effects of triclosan on growth, viability and fatty acid synthesis of the oyster protozoan parasite Perkinsus marinus

Perkinsus marinus, a protozoan parasite of the Eastern oyster Crassostrea virginica, has severely impacted oyster populations from the Mid-Atlantic region to the Gulf of Mexico coast of North America for more than 30 yr. Although a chemotherapeutic treatment to reduce or eliminate P. marinus from infected oysters would be useful for research and hatchery operations, an effective and practical drug treatment does not currently exist. In this study, the antimicrobial drug triclosan 5-chloro-2-(2,4 dichlorophenoxy) phenol, a specific inhibitor of Fab1 (enoyl-acyl-carrier-protein reductase), an enzyme in the Type II class of fatty acid synthetases, was tested for its effects on viability, proliferation and fatty acid synthesis of in vitro-cultured P. marinus meronts. Treatment of P. marinus meront cell cultures with concentrations of > or = 2 microM triclosan at 28 degrees C (a temperature favorable for parasite proliferation) for up to 6 d stopped proliferation of the parasite. Treatment at > or = 5 microM at 28 degrees C greatly reduced the viability and fatty acid synthesis of meront cells. Oyster hemocytes treated with > or = 20 microM triclosan exhibited no significant (p < 0.05) reduction in viability relative to controls for up to 24 h at 13 degrees C. P. marinus meronts exposed to > or = 2 microM triclosan for 24 h at 13 degrees C exhibited significantly (p < 0.05) lower viability relative to controls. Exposure of P. marinus meronts to triclosan concentrations of > or = 20 microM resulted in > 50% mortality of P. marinus cells after 24 h. These results suggest that triclosan may be effective in treating P. marinus-infected oysters.     

Urinary excretion of collagen degradation markers by sows during postpartum uterine involution

Incomplete uterine involution is the putative cause of the increased embryo mortality and reproductive failure often exhibited by sows that lactate for less than 21 days. Since such short lactation lengths are common in American swine production, an effective technique to monitor the postpartum involution process and test this hypothesis might be valuable. Rapid and extensive catabolism of uterine collagen is essential for normal postpartum involution. The objective of this study was to characterize postpartum excretion of two biochemical markers of collagen degradation. In experiment I, urine samples were collected from five sows every other day from the day before parturition (day -1), through a 21-day lactation, to day 8 postweaning. The collagen crosslinks hydroxylysyl pyridinoline (HP), which is present in many tissues, and lysyl pyridinoline (LP), which is primarily concentrated in bone, were assayed by both ELISA and HPLC. Urinary levels of both free (ELISA) and total (HPLC) HP and LP increased (P < 0.001) approximately two-fold during lactation. The mean molar ratio of total HP:LP increased (P < 0.001) from 6.6 +/- 1.6 at day 1 to a maximum of 10.2 +/- 1.5 at day 7 postpartum and averaged 9.1 +/- 0.3 for the entire sampling period. These data are consistent with a postpartum increase of soft tissue collagen catabolism since bone has a low HP:LP ratio of 4 and soft tissues like the uterus have a high HP:LP ratio of >/=20 because they contain only trace amounts of LP. Since HPLC (total) and ELISA (free) crosslinks estimates were highly correlated (r = 0.85-0.91, P < 0.001) in experiment I, only the less technical ELISA technique was used in experiment II. Urine samples were collected from 21 sows every third day from day 1 to 19 of lactation. Sows from this second group exhibited one of four distinct crosslinks excretion patterns: peak on day 1 (n = 3), peak on day 7 (n = 4), peak on day 10, 13 or 16 (n = 7), or no peak (n = 7). This variation of postpartum crosslinks excretion among sows was not related to parity, body weight, lactation body weight change, litter size, or litter birth weight. Overall, data from experiments I and II indicate that urinary HP does increase postpartum in a pattern temporally consistent with uterine involution. However, significant variation among sows in the magnitude and timing of peak HP excretion was evident.     

Grafting raises the salt tolerance of tomato through limiting the transport of sodium and chloride to the shoot

With the aim of determining whether grafting could improve salinity tolerance of tomato (Lycopersicon esculentum Mill.), and what characteristics of the rootstock were required to increase the salt tolerance of the shoot, a commercial tomato hybrid (cv. Jaguar) was grafted onto the roots of several tomato genotypes with different potentials to exclude saline ions. The rootstock effect was assessed by growing plants at different NaCl concentrations (0, 25, 50, and 75 mM NaCl) under greenhouse conditions, and by determining the fruit yield and the leaf physiological changes induced by the rootstock after 60 d and 90 d of salt treatment. The grafting process itself did not affect the fruit yield, as non-grafted plants of cv. Jaguar and those grafted onto their own root showed the same yield over time under non-saline conditions. However, grafting raised fruit yield in Jaguar on most rootstocks, although the positive effect induced by the rootstock was lower at 25 mM NaCl than at 50 and 75 mM NaCl. At these higher levels, the plants grafted onto Radja, Pera and the hybrid VolgogradskijxPera increased their yields by approximately 80%, with respect to the Jaguar plants. The tolerance induced by the rootstock in the shoot was related to ionic rather than osmotic stress caused by salinity, as the differential fruit yield responses among graft combinations were mainly related to the different abilities of rootstocks to regulate the transport of saline ions. This was corroborated by the high negative correlation found between fruit yield and the leaf Na(+) or Cl(-) concentrations in salt-treated plants after 90 d of salt treatment. In conclusion, grafting provides an alternative way to enhance salt tolerance, determined as fruit yield, in the tomato, and evidence is reported that the rootstock is able to reduce ionic stress.