The production of polypeptides containing a high percentage of 2-methylalanine residues by a number of isolates of Trichoderma spp. has been examined. It has been shown that good yields (0.5-1.0 g L-1) can be achieved on synthetic media provided an insoluble carbohydrate is included and provided single-spore isolates that have this production ability are selected from time to time. Such yields could not be obtained on any single nitrogen source investigated, but a mixture of potassium nitrate, glutamine, and 2-methylalanine was effective. It was shown that at least eight polypeptides were produced in shake-flask or tank fermentation and that the proportions of these metabolites depended on the fermentation temperature, its pH, age, and aeration. Fermentation conditions for enhancing the production (independently) of two of the metabolites at the expense of the others are given. These two metabolites have been obtained in crystalline form and details of some of their physical and chemical properties are given.

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Simple SummaryThe utilization of animal donors of rumen fluid for laboratory experiments can raise ethical concerns due to invasive methods of collection (rumen cannulated or intubated animals). Societies are strongly oriented to support cruelty free experiments and alternatives to the collection of rumen fluids from live animals are urgently requested from the scientific community. Thus, in order to attenuate the dependence of laboratories on animal donors, this study compared the rumen inoculum collected at slaughter with the fermentation liquid from a rumen continuous fermenter and both rumen inoculum were used fresh or preserved (by refrigeration, chilling and freeze-drying). The results support the possibility of using continuous fermenters to generate inoculum for in vitro purposes, and short-term refrigeration is confirmed to be a valuable storage system to facilitate transfer inoculum from the collection sites. These findings should attenuate the need for laboratories’ frequent collections from animals while continuing research in ruminant nutrition.AbstractThe utilization of animal donors of rumen fluid for laboratory experiments can raise ethical concerns, and alternatives to the collection of rumen fluids from live animals are urgently requested. The aim of this study was to compare the fresh rumen fluid (collected at slaughter, W) with that obtained from a continuous fermenter (RCF) and three methods of rumen fluid preservation (refrigeration, R, chilling, C, and freeze-drying, FD). The fermentability of different inoculum was evaluated by three in vitro tests (neutral detergent fiber (NDF) and crude protein (CP) degradability and gas production, NDFd, RDP and GP, respectively) using six feeds as substrates. Despite the two types of inoculum differed in terms of metabolites and microbiota concentration, the differences in vitro fermentability between the two liquids were less pronounced than expected (−15 and 20% for NDFd and GP when the liquid of fermenter was used and no differences for RDP). Within each in vitro test, the data obtained from rumen and from fermenter liquids were highly correlated for the six feeds, as well as between W and R (r: 0.837–0.985; p < 0.01). The low fermentative capacity was found for C and, particularly, FD for liquids. RCF could be used to generate inoculum for in vitro purposes and short-term refrigeration is a valuable practice to manage inoculum.

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