Grazing alters network architecture during interspecific mycelial interactions

The changes that occur in mycelial architecture of Phanerochaete velutina interacting with Hypholoma fasciculare mycelium in soil microcosms in the presence and absence of the collembola Folsomia candida are investigated employing tools developed in graph theory and statistical mechanics. There was substantially greater overgrowth of H. fasciculare by P. velutina mycelium when grazed than when un-grazed. There was a marked disappearance of hyphal links in all un-grazed systems between 8 d and 34 d, predominantly in areas distant from the interaction, but this was much less evident in grazed systems. Further, new tangential cross-links connecting radial cords distant from the inoculum formed in grazed systems. The thickness of cords increased with time, and more so in grazed systems. There was no significant difference in transport efficiency between the grazed and un-grazed systems. The ability of the mycelial network to modify dynamically link strengths is crucial to achieving a balance between transport capacity/robustness to damage and overall cost of production.     

Incorporating sheep into dryland grain production systems: I. Impact on over-wintering larva populations of wheat stem sawfly,Cephus cinctus Norton (Hymenoptera: Cephidae)

Wheat stem sawfly (WSS), Cephus cinctus Norton (Hymenoptera: Cephidae) is the most damaging insect pest to Montana's $ 1 billion dollar per year grain industry. Current WSS control methods are either expensive, reduce wheat yields, or are not effective. Our objective was to compare burning, grazing, tilling, trampling and clipping wheat stubble fields on over-wintering WSS larval populations. Treatments were evaluated in three experiments using a randomized complete block design and four replications at each site. Eight, six, and two sites were used for Experiments 1, 2, and 3, respectively. Contrast statements were used to make pre-planned comparisons among treatments. For Experiment 1, treatments were fall tilled, fall grazed, spring grazed, fall and spring combined (Fall/Spr) grazed, and an untreated control. Five mature ewes were confined with electric fence to 111 m² plot for 24 h in the fall and spring grazed treatments resulting in a stocking rate of 452 sheep d/ha. For Fall/Spr, the stocking rate was 904 sheep d/ha. For Experiment 2, treatments were fall grazed, fall burned, fall tilled, and an untreated control. In Experiment 3, treatments were fall trampled, spring trampled, Fall/Spr trampled, hand clipped to a stubble height of 4.5 cm, and an untreated control. Trampled treatments were done at the same stocking rates as grazing treatments but sheep were muzzled to prevent intake. Wheat stem sawfly larval numbers were collected in the fall and spring, pre- and post-treatment, respectively, by collecting all plant material from three, 0.46 m lengths of row and counting the number of live larvae present. In Experiment 1, WSS mortality was greater (P < 0.01) for the mean of all grazed treatments (68.4%) than either control (43%) or tilled (47%) plots. Mortality did not differ (P = 0.75) between fall (67%) and spring (64%) grazed plots but was greater (P = 0.02) for Fall/Spr (74%). In Experiment 2, larva mortality was greater (P < 0.01) for fall grazed (63%) than burned plots (52%). In Experiment 3, WSS mortality was greater (P < 0.01) for the mean of all trampling treatments (57%) than either control (33%) or clipped (32%) plots. Mortality did not differ (P > 0.25) between fall (54%) and spring trampling (47%) but was greater (P = 0.01) for Fall/Spr (70.6%). No differences (P > 0.25) were detected for WSS mortality when grazing was compared to trampling. These results indicate the potential for using grazing sheep to control wheat stem sawfly infestations in cereal grain production systems.     

Effects of preservation procedures of rumen inoculum on in vitro microbial diversity and fermentation

Sheep rumen contents were used as inoculum for an in vitro semi-continuous incubation system to study whether preservation method affects microbial fermentation pattern. Rumen fluid was filtered and either used immediately as inoculum (CTL) or dispensed into 110 mm × 16 mm tubes, that were stored refrigerated at 6 °C for 4 h (REF) or frozen at ?20 °C (FRZ), frozen in liquid N (FLN) or added with 0.04 glycerol and frozen in liquid N (FGL) for 48 h. Frozen inocula were thawed at 39 °C for 2 min before use (16 ml per bottle). Two 24 h incubations with four bottles per treatment were completed. The microbial utilisation of added glycerol after thawing in FGL increased total gas production (P<0.05) and 24 h volatile fatty acid (VFA) production (P<0.05), and also increased propionate and butyrate proportions at the expense of acetate. The other freezing inocula (i.e., FLN and FRZ) reduced the rate of gas production (as ml/g dry matter per hour), compared with CTL in the first 2 and 4 h of incubation (P<0.05), but this was compensated by increased fermentation at 8 and 12 h, respectively. Differences in gas production did not manifest a different VFA pattern at either 6 or 24 h incubation. Bacterial diversity was slightly affected by the preservation process, and the similarity index between untreated inocula and the 24 h incubated CTL samples was 0.690–0.724. Similarity between bacterial communities in FRZ and FLN with that in CTL after incubation was 0.678. The freezing preservation method of rumen inocula for subsequent in vitro gas production studies does not affect microbial fermentation pattern or bacterial biodiversity, provided that processing is rapid enough by using a high surface to volume ratio. Freezing in liquid N is more appropriate than at ?20 °C.     

The collembola Lobella sokamensis juvenile as a new soil quality indicator of heavy metal pollution

We propose Lobella sokamensis Deharveng and Weiner, 1984 juvenile as a new soil quality indicator to assess heavy metal polluted soils. L. sokamensis is a collembola commonly found with earthworms, and it plays a key role in decomposing dead earthworms. The soil quality assessment of cadmium (Cd), copper (Cu), antimony (Sb), and zinc (Zn) on the survival of L. sokamensis adults were performed in artificial soil. The LC50 values for L. sokamensis adults exposed to Cd, Cu, Sb and Zn for 5 days were calculated to be 4729, 4472, 4702, and 2521 mg kg⁻¹, respectively. L. sokamensis juveniles (10–12 days) were also exposed to Cd, Cu, Sb, and Zn, and 5 days-LC50 values for Cd, Cu, Sb, and Zn were 282, 229, 447, and 163 mg kg⁻¹, respectively. Both adult and juvenile survivals of L. sokamensis in metal spiked soil were inhibited. We found that the soil quality assessment using L. sokamensis juveniles were sensitive enough to evaluate soil pollution, compared to conventional Folsomia candida assay which is widely used. In addition, L. sokamensis has advantage for easy detection in soil due to the big and orange-colored body. The 5 days-soil quality assessment using L. sokamensis juveniles appears to be rapid protocols for the collembola assay. This is the first report on the collembola assay of heavy metal contaminated soils using the L. sokamensis as a soil indicator.     

Processing characteristics of submerged fermentation of Antrodia cinnamomea in airlift bioreactor

Three 5-L airlift bioreactors including airlift reactor with solid draft tube (ALs), airlift reactor with net draft tube (ALn) and bubble column reactor (BC) were investigated for their suitability for cultivating Antrodia cinnamomea, and a stirred tank reactor (ST) was used for comparison. Results indicated that after 7 days fermentation, ALs yielded the highest mycelium content (313 mg/100 mL) and had the lowest dissolved oxygen in the broth. Among different aeration rates (0.025, 0.05, 0.1, 0.5, 1 vvm) used during cultivation of A. cinnamomea in ALs, the aeration rate 0.1 vvm resulted in a volumetric oxygen transfer coefficient of 10.8 h⁻¹ and produced the highest mycelium content. When the optimal conditions were used for the fermentation of A. cinnamomea in an industrial 500-L ALs, the mycelium content in the broth reached 542 mg/100 mL in 28 days. The IC₅₀ values of the ethanol extracts of A. cinnamomea mycelium cultivated in 5-L and 500-L ALs for 28 days were 23 and 17 μg/mL, respectively, for hepatocellular carcinoma cells HepG2. And after 42 days cultivation in 500-L ALs, the IC₅₀ value of the mycelium ethanol extract was reduced to 10 μg/mL.     

High-level production of lycopene in metabolically engineered E. coli

Recombinant lycopene was generated by utilizing metabolically engineered Escherichia coli with yields being dependent upon inocula state. Yields were especially low in the case of cultures harboring high-copy plasmids that were established with inocula at the stationary growth phase. On the other hand, cultures derived using low-copy plasmid, however, yielded high amounts of lycopene irrespective of inocula state. Nevertheless, it showed still an inocula dependence pattern in lycopene productivity (mg/l/h). To further increase lycopene productivity, we applied a temperature-shift culture technique (37 → 25 °C). Using this method, we effectively enhanced lycopene productivity without any problematic phenomena. As a result, we were able to increase lycopene yield by approximately 20% compared to previous culture methods. In the present study, we were able to reach a final lycopene yield up to 260 mg/l for 60 h, which corresponds to the highest titer to date for the production of lycopene in E. coli.     

Fungal network responses to grazing

Mycelial networks operate on scales from microscopic to many m² and naturally persist for extended periods. As fungi exhibit highly adaptive development, it is important to test behavioural responses on natural substrata with realistic nutrient levels across a range of spatial scales and extended time periods. Here we quantified network responses over 7.5 months in large (57 × 57 cm) microcosms to test whether grazing shifts the network to a more resilient architecture. Resource limitation constrained any ability to respond at all, with both grazed and ungrazed networks gradually thinning out over time. Added resources sustained further exploratory growth, but only transiently increased cross-connectivity and network resilience, when tested by simulated damage in silico. Grazed networks were initially weaker and emergence of new exploratory growth was curtailed. However, increased interstitial proliferation led to new cross-links, consolidating the existing mycelial network and increasing the resilience of the network to further attack.     

Incorporating sheep into dryland grain production systems: II. Impact on changes in biomass and weed density

Weed control in fallow management to conserve soil moisture and nutrients is the largest variable cost to dryland grain production. Our objective was to compare burning, grazing, tilling, trampling and clipping wheat stubble fields on changes in total aboveground biomass and weed density. Treatments were evaluated in three experiments using a randomized complete block design for each experiment with four replications at each site. Contrasts statements were used to make pre-planned comparisons. For experiment 1, treatments were fall tilled, fall grazed, spring grazed, fall and spring combined (Fall/Spr) grazed, and an untreated control. For grazing treatments, five mature ewes were confined with electric fence to 111 m² plot for 24 h for fall and spring resulting in a stocking rate of 452 sheep day/ha. For Fall/Spr the stocking rate was 904 sheep day/ha. For experiment 2, treatments were fall grazed, fall burned, fall tilled, and an untreated control. In experiment 3, treatments were fall trampling by sheep, spring trampling by sheep, fall and spring combined (Fall/Spr) trampling by sheep, stubble hand clipped to a height of 4.5 cm, and an untreated control. Trampling treatments were applied at the same stocking rates as grazing treatments but sheep were muzzled to prevent intake. Data were collected in the fall, prior to treatment imposition, and spring, after treatments had been removed. Post treatment biomass and weed density were greater (P < 0.05) in either control or tilled plots when compared to grazed plots. Post treatment biomass and weed density were greater (P < 0.01) for control than burned plots. Post treatment biomass, weed density, and percent change in these variables, did not differ (P > 0.08) between burned and tilled, and burned and grazed treatments. These results indicate the potential for using grazing sheep as a component in fallow management to reduce biomass and control weeds.     

Grazing of cover crops in integrated crop-livestock systems

Conventional agriculture is specializing rapidly into the management of few monoculture crops, threatening crop diversity and questioning the sustainability of extensive cropping systems. The grazing of cover crops in integrated crop-livestock systems could be a feasible biologically based technology to restore crop diversity and mitigate ecological issues in cropping systems. However, there is limited evidence on plausible synergies or trade-offs for the practice, and about how grazing plans could affect the herbage production and services from cover crops. This work assessed the effects of cattle grazing on the primary and secondary production of annual ryegrass (Lolium multiflorum) in an integrated ryegrass–soybean rotation system. Specifically, the prediction for synergistic effects of cattle grazing on the ryegrass herbage production, residual crop cover and animal performance were tested in a 2-year (2014 and 2015) study comprising a randomized complete block design of four grazing intensity treatments, replicated three times. A no-cattle grazing treatment (NG), used as control, or continuous grazing with Holstein heifers (~ 220 kg live weight) at targeted sward heights of 5, 10, 15 and 20 cm (hereafter referred as G₅, G₁₀, G₁₅ and G₂₀, respectively) was applied to ryegrass plots. The herbage production and residual herbage cover of ryegrass, and the average daily gain (ADG, kg/day) and live weight gain per hectare (LWG, kg/ha) of heifers were analyzed by ANOVA (P < 0.05) and compared by the Tukey_HSD test (P < 0.05). Regression models were used to estimate relationships between herbage production, animal performance and sward height. The herbage production was 60% higher (P < 0.01) for the grazing treatments compared to NG. The residual herbage for G₁₅ and G₂₀ was not different than that for NG and increased linearly as sward heights increased, reaching highest values for G₁₅ and G_20. Maximum ADG was 1.10 kg/day for ryegrass grazed at a 20.6 cm height, whereas maximum LWG was 427 kg/ha for ryegrass grazed to a 16.1 cm height. The results support the hypothesis for synergistic effects of using annual ryegrass as a dual forage and service cover crop. Moderate grazing intensity to sward height of 12–18 cm with continuous stocking led to optimized forage production and utilization by dairy heifers.     

Nutritional supplementation improves the sexual response of bucks exposed to long days in semi-extensive management and their ability to stimulate reproduction in goats

In confined management systems, well-nourished bucks rendered sexually active by exposure to long days are efficient in fertilizing out-of-season goats. However, underfeeding is common in semi-extensive management systems and may reduce the reproductive efficiency of bucks. The objective of the present study was to determine whether nutritional supplementation improved the sexual activity of bucks submitted to long days in semi-extensive management systems and their ability to stimulate the reproduction of goats in semi-extensive or confined conditions. In experiment 1, three groups of bucks were placed in different flocks and grazed daily with females for 7 h. Each day after grazing, males were separated from females and moved into open pens. One group did not receive any treatment (control group; n = 6). Two other groups were submitted to artificially long days from 15 November to 15 January. From 16 January, one group did not receive nutritional supplementation (long-day group; n = 5), whereas bucks from the other group each received 600 g of a commercial concentrate (long-day + supplementation group; n = 5). The fourth group was kept in confined conditions, exposed to long days and fed alfalfa hay (long-day confined group; n = 6). On 26 March, anovulatory goats from other flocks were assigned to four groups (n = 27 each) and confined separately in open pens. Three bucks of each group were housed with the females. Pregnancy rates were greater in the goats housed with the long-day group than those housed with the control group (P < 0.01). However, pregnancy rates did not differ between the long-day confined group (89%) and long-day + supplementation group (70%; P = 0.09), but these rates were greater than those from the long-day (37%) and control groups (0%; P < 0.05). In experiment 2, two groups of males (n = 3 each) were incorporated into two flocks under semi-extensive management and grazed daily with females for 7 h. One group of males did not receive any treatment (control group). The other group was submitted to long days and nutritional supplementation as in experiment 1 (long-day + supplementation group). Males remained with females during the whole study. The pregnancy rate was greater in the goats joined by males of the long-day + supplementation group (78%) than in those from the control group (0%; P < 0.001). We conclude that long days and nutritional supplementation improve the ability of bucks kept in semi-extensive management to stimulate reproduction of out-of-season goats in confined or semi-extensive management systems.     

Inulinase biosynthesis using immobilized mycelium of Aspergillus niger

Conidia of Aspergillus niger 20 Osm producing extracellular inulinase were immobilized on pumice stones or polyurethane sponge and used in repeated-batch processes. Some factors affecting inulinase biosynthesis by the mycelium A. niger immobilized on pumice stones were investigated. Maximal inulinase production occurred in 50 ml of medium containing 0.5 g of carrier at 30 °C, pH 6.0 and at an agitation speed of 200 rpm. This procedure enabled repeated-batch enzyme production and as many as six subsequent 24 h batches could be fermented by using the same carrier. This is the first report on inulinase biosynthesis by mycelium of A. niger immobilized on polyurethane sponge using unconventional oxygenation of culture which ensures that the dissolved oxygen concentration remains constant.     

Correlation of pigment production with mycelium morphology in extractive fermentation of Monascus anka GIM 3.592

Extractive fermentation with nonionic surfactants is a potential method for producing Monascus pigments. In this study, the correlation between mycelium morphology and pigment production was investigated in extractive fermentation of Monascus anka GIM 3.592. The results demonstrated that pigment biosynthesis was associated with mycelial morphology and the accumulation of granular inclusions in cells. The physiological status in terms of hyphal and pellet diameters exhibited an excellent correlation with pigment accumulation, especially the yield of extracellular pigment in extractive fermentation (r > 0.85, p < 0.05). Nonionic surfactants could reduce pigment yield by influencing the morphology of hyphae and mycelium pellets. High yields of both intracellular and extracellular pigments could be achieved by controlling variations in hyphal diameters in two-stage extractive fermentation. Moreover, continuous extractive fermentation led to stable pigment production, with a relatively high productivity of total pigments reaching 72.3 AU/day. This study proposed a simple method for monitoring pigment biosynthesis in extractive fermentation using mycelium morphology as an indicating factor.     

Incorporating sheep into dryland grain production systems: III. Impact on changes in soil bulk density and soil nutrient profiles

Changes in soil bulk density and soil nutrient profiles are a major concern of dryland grain producers considering grazing sheep on cereal stubble fields. Our objective was to compare burned, grazed, tilled, trampled and clipped wheat stubble fields on changes in soil bulk density and soil nutrient profiles. Treatments were evaluated in a series of three experiments using a randomized complete block design and four replications at each site. Contrast statements were used to make pre-planned comparisons among treatments. For Experiment 1, treatments were fall tilled, fall grazed, spring grazed, fall and spring combined (Fall/Spr) grazed, and an untreated control. Five mature ewes were confined with electric fence to a 111 m² plot for 24 h for fall and spring grazed plots resulting in a stocking rate of 452 sheeps d/ha. For Fall/Spr, the stocking rate was 904 sheeps d/ha. For Experiment 2, treatments were fall grazed, fall burned, fall tilled, and an untreated control. In Experiment 3, treatments were fall trampling by sheep, spring trampling by sheep, fall and spring combined (Fall/Spr) trampling by sheep, hand clipping to a stubble height of 4.5 cm, and an untreated control. Trampling treatments were done at the same stocking rates as grazing treatments but sheep were muzzled to prevent intake. In Experiment 1, post-treatment organic matter tended to be greater (P = 0.09) in the mean of the grazed treatments than control plots. In all of the experiments, change in soil bulk density, and soil nutrient profiles did not consistently differ (P > 0.07) among treatments in any manner that would suggest a detrimental impact of grazing sheep on small grain residue. These results indicate a strong potential for grazing sheep on grain stubble without adversely impacting soil bulk density or nutrient profiles.     

Effect of inoculum and pepsin–pancreatin hydrolysis on fibre fermentation measured by the gas production technique in pigs

Two experiments were undertaken to adapt the in vitro gas production technique in syringes, used for ruminants, to fibre fermentation studies in the large intestine of pigs.In a first experiment, two inocula (faeces and large intestine content) were compared at four dilution levels in a buffer solution (0.025, 0.05, 0.1 and 0.2 g ml⁻¹) with two substrates: wheat bran and sugar–beet pulp. The accumulated gas produced over 72 h was modelled and the kinetics parameters compared. The time to half asymptote was lower for the intestinal inoculum (5.5 versus 8.0 h, P<0.02), but the 2 inocula yielded similar fractional rates of degradation (0.16 h⁻¹) and gave equal final gas production (252 ml g⁻¹ substrate). No interaction (P>0.05) was observed between inocula and substrates. The dilution of the samples in the buffer solution increased (P<0.001) the lag time (from 0.9 to 2.1 h for dilution rates ranging from 0.2 to 0.025 g ml⁻¹, respectively) and decreased (P<0.001) the rates of substrate degradation (from 0.18 to 0.13 h⁻¹).A second experiment aimed to study the effect of an in vitro pepsin–pancreatin hydrolysis of the sample prior to the gas test. Six substrates were tested: maize, wheat bran, sugar–beet pulp, lupins, peas and soybean meal. The enzymatic hydrolysis affected (P<0.001) the kinetics parameters and the ranking order of the fermented substrates. The lag times also increased for all ingredients. The rate of degradation decreased when peas, lupins, maize and wheat bran were hydrolysed (P<0.001) but it increased with soybean meal (P=0.014) and sugar–beet pulp (P<0.001). Final gas production increased with peas and soybean meal (P<0.001), remained unchanged for lupins and decreased for the other substrates (P<0.001).In conclusion, the method using faeces as a source of microbial inoculum is reliable to characterise the fermentation kinetics of ingredients in the large intestine of pigs. However, it is important to hydrolyse the substrates with pepsin and pancreatin before the gas tests.     

Beef cattle pasture to wetland reconversion: Impact on soil organic carbon and phosphorus dynamics

There is a major need to understand the historical condition and chemical/biological functions of the ecosystems following a conversion of wetlands to agricultural functions. To better understand the dynamics of soil total organic carbon (TOC) and phosphorus (P) during beef cattle pastures to wetland reconversion, soil core samples were collected from the beef cattle pasture and from the natural wetland at Plant City, FL, during five summer seasons (2002–2007). The levels of TOC and soil P were significantly affected by changing land use and hydrology. Draining natural wetlands to grazed pastures resulted in very pronounced reduction of TOC from 180.1 to 5.4 g g^?1. Cumulative concentrations of total phosphorus (TP) in soils (1134 mg kg^?1) under drained condition are two to three times lower than those in soils (2752 mg kg^?1) under flooded condition over the periods of land use reconversion. There was a declining trend (r = 0.82**; p ≤ 0.01) in total soil P from natural wetland (763 mg kg^?1) to altered pastures (340 mg kg^?1), largely as organic-bound P (natural wetland, 48%; grazed pastures, 44%; altered pastures, 29%). These results are important in establishing baseline information on soil properties in pasture and wetland prior to restoring and reconverting pasture back to wetland conditions. The results further suggest that changes in soil properties due to changing land use and hydrologic conditions (drying and re-wetting) could be long lasting.     

Decay and termite resistance,water absorption and swelling of thermally compressed wood panels

This study evaluated decay and termite resistance of thermally compressed pine wood panels under pressure at either 5 or 7 MPa and either 120 or 150 °C for 1 h. Wood specimens from the panels were exposed to laboratory decay resistance tests by using the wood degrading fungi, Gloeophyllum trabeum and Trametes versicolor. The thermal compression process caused increases in density and decreases in thickness of the panels; however, laboratory decay resistance tests revealed that thermally compressed wood was not resistant against the wood degrading fungi tested. More interesting results were found in laboratory termite resistance tests by using the Eastern subterranean termites, Reticulitermes flavipes. As pressure and temperature applied to the specimens increased to 7 MPa and 120 °C, mass losses in the specimens gradually decreased in comparison with control specimens. However, the specimens compressed at 7 MPa and 150 °C showed higher mass losses when compared to the specimens compressed at 7 MPa and 120 °C. The lowest water absorption and swelling rates were seen in the specimens exposed to a pressure of 7 MPa at 120 °C. The thermal compression process at 7 MPa and 150 °C resulted in the highest water absorption and swelling in the specimens.     

Effect of pig faecal donor and of pig diet composition on in vitro fermentation of sugar beet pulp

Two experiments were undertaken to investigate the influence of (1) pig bodyweight and (2) dietary fibre content of the diet on the in vitro gas production of sugar beet pulp fibre using faecal inoculum.In the first experiment, inocula prepared from young pigs (Y; 16–50 kg), growing pigs (G; 62–93 kg) and sows (S; 216–240 kg) were compared. Sugar beet pulp, hydrolysed in vitro with pepsin and then pancreatin, was used as the fermentation substrate. The cumulated gas productions over 144 h were modelled and the kinetics parameters compared. Lag times (Y: 4.6 h; G: 6.4 h; S: 9.2 h) and half-times to asymptote (Y: 14.7 h; G: 15.9 h; S: 20.8 h) increased with pig bodyweight (P<0.001) and the fractional degradation rates of the substrate differed between the pig categories (Y: 0.110 h⁻¹; G: 0.115 h⁻¹; S: 0.100 h⁻¹; P<0.001). The final gas production was not affected (P=0.10) by the inoculum source.In the second experiment hydrolysed sugar beet pulp was fermented with four inocula prepared from pigs fed diets differing in their total and soluble dietary fibre contents, i.e. low fibre diet rich in soluble fibre (LOW-S) or in insoluble fibre (LOW-I) or high fibre diet rich in soluble fibre (HIGH-S) or in insoluble fibre (HIGH-I). The total and the soluble dietary fibres influenced the kinetics of gas production. The presence of soluble fibres decreased the lag times, whatever the total dietary fibre content (2.7 h for LOW-S versus 3.5 h for LOW-I, 4.0 h for HIGH-S versus 4.4 h for HIGH-I; P<0.001). The half-times to asymptote were higher with the low fibre diets (P<0.001) and, for similar total dietary fibre contents, they were lower when the proportion of soluble fibres increased (LOW-S: 9.9 h; LOW-I: 11.4 h; HIGH-S: 8.9 h; HIGH-I: 10.1 h; P<0.001). The fractional degradation rates of the substrate were the highest with the fibre-rich diet containing a high proportion of soluble fibres (0.158 h⁻¹; P<0.001).In conclusion, the bodyweight of the faeces donors and the dietary fibre composition of the pig diet influence the in vitro fermentation kinetics of hydrolysed sugar beet pulp, but not the final gas production.     

Preservation of Agaricus subrufescens strains at low temperature by using cultures on sorghum grains

BackgroundOne of the main problems for the preservation of genetics resources of Agaricus subrufescens is to maintain the viability of the strains because the mycelium is very sensitive to cooling and therefore it ages rapidly.AimsEvaluate the viability of A. subrufescens strains stored as cultures on sorghum grain (spawn) at different temperatures.MethodsEighteen strains of A. subrufescens and three strains of Agaricus bisporus were studied. Spawn's viability was evaluated under the following conditions: (1) control at 25 °C (C), (2) cooling to 4 °C (R) and (3) freezing in liquid nitrogen at ?196 °C (LN). Samples were recovered from week 4 every 2 weeks until week 12 and week 24 in C and R, whereas in LN samples were recovered at 4, 12 and 24 weeks. Viability was evaluated in 50 seeds, by strain and condition, recovering the mycelium in Petri dishes with potato dextrose agar medium (PDA). Mycelium growth was also evaluated on PDA after 14 days of recovery.ResultsMost strains showed 100% viability and they were recovered usually in 1 day. In LN the viability ranged between 84 and 100% depending on the strain, but in some cases recovery took more than 10 days. Mycelial growth decreased gradually over time and although the results show significant differences between treatments C and R, the decline is associated with ageing of the mycelium rather than the treatment itself.ConclusionsCulture on sorghum grain and storage at low temperature is an interesting way to preserve genetic resources of A. subrufescens.     

Heterologous expression of a newly screened β-agarase from Alteromonas sp. GNUM1 in Escherichia coli and its application for agarose degradation

The gene of agaG1 from Alteromonas sp. GNUM1 encoding a β-agarase (AgaG1) was heterologously expressed in E. coli BL21 (DE3). The recombinant strain was cultured at 37 °C and then AgaG1 was expressed at 25 °C and 0.5 mM IPTG. The optimum conditions for AgaG1 to hydrolyze agarose were pH 7.0 and 40 °C. The main products of agarose hydrolysis by AgaG1 were confirmed to be neoagarobiose and neoagarotetraose. A new agarose hydrolysis process using AgaG1 was developed, in which the reaction temperature was adjusted stepwise to avoid gelation problem with no chemical pretreatment step. The enzyme AgaG1 was found to be very effective and highly selective. When 10.0 g/L agarose was hydrolyzed, 98% of the agarose added was converted to 3.8 and 6.4 g/L of neoagarobiose and neoagarotetraose, respectively.     

Persistent efficacy of long-acting moxidectin for control of trichostrongylid infections of sheep

The objective was to evaluate long-acting moxidectin (Cydectin^® 2% LA for sheep) against trichostrongylids in sheep. We performed a blocked, parallel, controlled clinical trial. We included 45 ewe-lambs, allocated into treated (n = 30, 1.0 mg moxidectin per kg bw, subcutaneously at base of ear on D 0) or controls (n = 15). Animals had been naturally infected (pre-treatment geometric mean epg counts: 233 and 249, respectively) and throughout the study grazed at the same pasture. Fortnightly, we collected faecal samples and performed established parasitological techniques (epg counts, coprocultures). We used analysis of covariance for post-treatment results, using pre-treatment counts as covariate, treatment as fixed effect, block as random effect. Geometric mean epg counts in treated animals were 0 up to D 112, 1 on D 126, 4 on D 140 and 13 on D 161. Respective figures for controls were 369–820 up to D 140 and 168 on D 161 (p < 0.05). Persistent efficacy (>90%) was 140 d for Teladorsagia, 119 d for Haemonchus and 115 d for Trichostrongylus. This is the first reported clinical trial of efficacy of long-acting moxidectin in sheep in Europe. Cydectin 2% LA was found to be effective against trichostrongylids of sheep. The particularly long persistent efficacy can provide new possibilities in formulating anthelmintic programs for sheep.