Category Archives: BIOMASS — DETECTION, PRODUCTION AND USAGE

Future perspectives

Due to the predicted impacts of climate change, many farmers are increasingly concerned about severe soil compaction and water stagnation on their fields. To prevent the deterioration of arable soils, appropriate soil management stratigies have to be developed. Earthworms are an important component of the soil biodiversity and their positive effects on soil structure are well-known. A variety of functional groups of earthworms can be restored through decrease in a soil disturbance and occurence of crop residues in the upper

soil. Investigating earthworm biomass and population is a very complex process and therefore consists of various methods of sampling. However, it is difficult to conduct efficient investigations due to horizontally aggregated earthworm populations and their complex phenologies. Of the vaiorus methods, hand sorting which involves sorting through soil samples by hand is one of the most earliest popular sampling methods. The soil washing method is more effective in sorting out cocoons and smaller earthworms. This method consists of a combination of washing and sieving soil samples, with a possible flotation stage. Another method that is used for soil sampling is the electrical method which consists of inserting an electrode into the groud causing earthworms to surface due to the electrical pulse in the soil. These methods, however, usually result in disrupting earthworm biomass and population, killing and injuring them and affecting their habitats. Considering the relationships between heavy metals and earthworms inhabiting in contaminated soils, it needs to adjust study focus on the long-term effects of multiple elements, not one heavy metal, to earthworms. However, these methods make it difficult to consistantly study and investigate a selective biomass during long periods.

With future developements in terms of remote sensing used for detecting the small — or large-scale acquisition of information of an object or phenomenon, these issues will no longer serve as a problem because biomass can be studied without any need of disruption. Although underground remote sensing technologies are in use, they have not yet been applied to the investigation of living organisms, such as earthworms. For that reason, we believe that scientists and remote sensing developers should put their heads together to optimize remote sensing equipment to the investigation of underground living organisms. These advancements will significantly help researchers to consistantly study a select biomass and calculate the amount of toxic materials that are being inserted into the soil more accurately.

Treatmenta

Rateb

% mortality (mean

± SE) at weeks after treatment

4

8

16

MSS

12.5

0

6 ± 3.3

53 ± 6.0

25

3 ± 3.3

7 ± 6.7

93 ± 6.0

50

10 ± 10.0

13 ± 3.3

70 ± 5.2

ISS

12.5

37 ± 18.6

60 ± 5.8

87 ± 6.0

25

7 ± 6.7

97 ± 3.3

100

50

0

97 ± 3.3

100

LPS

12.5

0

30 ± 5.8

97 ± 3.0

25

0

3 ± 3.3

97 ± 3.0

50

0

27 ± 8.8

100

AFPS

12.5

0

0

33 ± 6.0

25

0

17 ± 3.3

20 ± 5.2

50

0

0

90 ± 9.0

PMC

12.5

0

0

17 ± 7.9

25

0

0

20 ± 0.0

50

3 ± 3.3

3 ± 3.3

3 ± 3.0

ab Tons of dry matter ha-1 year-1

2. Conclusion

The long-term applications of organic waste materials containing heavy metals and HEMs affected the establishment of Megascolecid and Moniligastrid earthworms in field. The biomass of earthworms in lysimeter and microcosm soil tests would provide valuable tools for establishing the integrated hazard assessment system for organic wastes. Future research is needed to establish additional soil physico-chemical characteristics, particularly those that might influence heavy-metal bioaccumulation and bioavailability and physical habitat such as compaction and soil water holding capacity across treatment through time course.

3. Acknowledgement

This work was supported by the Rural Development Administration and WCU (World Class University) programme (R31-10056) through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology.

Traditional range management and knowledge

Information was collected in three districts. Data collection procedures include interviews, focused group discussion, and transect walk in the range areas. Fifty to sixty key informants from each site were involved on broad issues like traditional knowledge of range management. Dialogues with the communities were made to assess the existing range — livestock system, grazing patterns, and related information. Main focused areas were pastoralist knowledge on plants, grazing patterns, and migration patterns, collection of plants for winter season, communal grazing, and livestock management. Range productivity was also measured on the community lands.

Strategies for improvement of carotenoid-synthesizing strains

2.4.1 Media compostion and cultivation mode

The production biotechnological process proceeds essentially in two stages: fermentation and product recovery. An important aspect of the fermentation process is the development of a suitable culture medium to obtain the maximum amount of desired product. In recent years, cheap raw materials and by-products of agro-industrial origin have been proposed as low-cost alternative carbohydrate sources for microbial metabolite production, with the view also of minimizing environmental and energetic problems related to residues and effluent disposal. For fermentation, seed cultures are produced from the original strain cultures and subsequently used in an aerobic submerged batch fermentation to produce a biomass rich in carotene pigment and other additional metabolites, e. g. ergosterol, metal ions etc. In the whole-cell strategy product isolation is not necessary and, moreover, complex biotechnological product in the form of slightly modified biomass could be obtained.

The traditional batch production system has the disadvantage of inducing the Crabtree effect (characterized by the synthesis of ethanol and organic acids as fermentation products), due to high concentrations of initial sugars, diminishing pigment and biomass yield. The strategy for solving this problem is the fed-batch culture. Maximum astaxanthin production (23.81 mg/l) by P. rhodozyma was achieved in fed-batch fermentation with constant pH = 6.0, 4.8 times greater that the one obtained in a batch culture and the biomass concentration (39.0 g/l) was 5.3 times higher than that in the batch culture (Ramirez et al., 2006). The maximum astaxanthin concentration by X. dendrorhous at fed-batch fermentation with pH-shift control strategy reached 39.47 mg/l, and was higher by 20.2 and 9.0% than that of the batch and fed-batch fermentation, respectively, with constant pH = 5.0. However, the maximal cell density at fed-batch fermentation with pH-shift control was 17.42 g dry cells/l, and was lower by 2.0% than that of fed-batch fermentation with constant pH = 5.0. As a result of the two stage fed-batch culture P. rhodozyma, cell and astaxanthin concentrations reached 33.6 g/l and 16.0 mg/l, respectively, which were higher when compared with batch culture. The final specific astaxanthin concentration (mg/ g dry wt of cells) in the second stage was ca. threefold higher than that in the first stage and 1.5-fold higher than that in the dissolved oxygen controlled batch culture, indicating that the astaxanthin production was enhanced mush more in the second stage than in the first stage (Hu et al., 2007).

The astaxanthin production was enhanced by a high initial C/N ratio in the medium (second stage), whereas a lower C/N ratio was suitable for cell growth (first stage). A significant increase (54.9%) in astaxanthin production by X. dendrorhous was achieved in pulse fed-batch process when compared with batch process. The astaxanthin concentration was 33.91 mg/l in pulse fed-batch when compared with 30.21 mg/l in constant glucose fed — batch and 21.89 mg/l in batch fermentation. In contrast with this strain producing high yields of biomass and astaxanthin in pulse fed-batch process, another strain of P. rhodozyma demonstrated high astaxanthin-synthesizing activity during continuous fed-batch process (Hu et al., 2005). The utilization of continuous feeding showed to be the most efficient feeding method in fed-batch processes, as it did not lead to a reduction in the cellular astaxanthin concentration, as observed in the pulsed feeding. In the pulsed and continuous fed-batch processes, a cellular astaxanthin concentration of 0.303 mg/ g biomass and 0.387 mg/ g biomass, an astaxanthin concentration of 5.69 and 7.44 mg/l, a biomass concentration of 18.7 and 19.3 g/l were obtained, respectively.

Temperature was reported to control changes in enzyme activities that regulate metabolic activity in microorganisms. For example, Rhodotorula glutinis biosynthesized |3-carotene more efficiently at lower temperature, whereas increased torulene formation was accompanied by higher temperature (Bhosale & Gadre, 2002). The reason might be found in y-carotene that acts as the branch point of carotenoid synthesis. Subsequent dehydrogenation and decarboxylation leading to torulene synthesis is known to be temperature dependent since the respective enzymes are less active at lower temperature compared to the activity of P-carotene synthase. This is probable reason for an increase in the proportion of P-carotene at lower temperature in Rhodotorula glutinis. The moderately psychrophilic yeast Xanthophyllomyces dendrorhous also displayed a 50% increase in total carotenoids at low temperatures with elevated levels of astaxanthin (Ducrey Sanpietro & Kula, 1998).

Fed-batch co-cultures R. glutinis-D. castellii gave a volumetric production of 8.2 mg total carotenoid/l, about 150% of that observed in batch co-cultures and biomass concentration of 9.8 g/l which was about two times higher when compared with batch fermentation (Buzzini, 2001). The fedbatch technique maximized the specific growth rate of R. glutinis, resulted in higher biomass and minimized substrate inhibition of pigment formation. Molasses in the fed-batch mode led to increased biomass by 4.4- and 7-fold in double — and triple-strength feed, respectively when compared with 12.2 g/l biomass in batch fermentation. R. glutinis also produced a very high carotenoid concentration for double — and triple-strength feed supplement (71.0 and 185.0 mg/l, respectively), and was higher 2- and 3.7-fold of that observed in batch fermentation (Frengova & Beshkova, 2009).

Scanning-electron microscopy (SEM)

SEM is a useful technique in the study of both the natural sorbent morphology and its modification derived from sorbate interactions. SEM is an electron microscope, which provides images of the sample surface by scanning it with a high-energy beam of electrons. The electron interactions with the atoms of the sample produce signals that contain information about topography, morphology, and composition of the sample surface. The samples must be electrically conductive, at least on their surface, for conventional SEM imaging. Nonconductive samples are coated with an ultra-thin layer of electrically — conducting material; this coating prevents the accumulation of static electric charges on the sample surface during electron irradiation. Magnification of the imaging can be controlled over a range of up to 6 orders of magnitude from about x25 to 250,000 times. When coupled with energy dispersive X-ray analysis (EDAX), the atom concentrations on the sorbent surface can be determined. This enables the confirmation of a mechanism of ion exchange, generally investigated by determining the concentration of alkaline and alkaline-earth metal ions released from the sorbent after metal sorption.

The total sugar content in potato tubers

The total sugar content in potato tubers was significantly modified by intercrop fertilization and straw fertilization (table 9). Intercrop fertilization significantly decreased the concentration of total sugars in potato tubers. The lowest its content was recorded in potato tubers fertilized with a mixture of white clover with Italian ryegrass and phacelia plowed down in the autumn and left till spring in the form of mulch. The content of reducing sugars in potato tubers fertilized with white clover and Italian ryegrass did not differ significantly from their concentrations observed in tubers fertilized with farmyard manure. However, on control object, the content of total sugars in potato tubers was significantly higher than in the potato fertilized with farmyard manure. Straw fertilization also significantly modified the content of total sugars in potato tubers. At the sub-block without straw content of total sugars in potato tubers was significantly lower than at the sub-block with straw.

3.3.6 Vitamin C content in potato tubers

The vitamin C content in potato tubers was significantly differentiated by the examined factors of experiment and their interaction (table 10). Intercrop fertilization in comparison with control object caused a significant increase of vitamin C content in potato tubers. Indeed, the highest concentration of vitamin C were characterized in potato tubers fertilized with phacelia in the form of mulch and white clover. The vitamin C content in potato tubers fertilized with a

mixture of white clover with Italian ryegrass and phacelia developed at a similar level as in the potato fertilized with farmyard manure. Straw fertilization also significantly differentiate the concentrations of vitamin C in potato tubers. On objects with straw the content of vitamin C in tubers was significantly higher than on the objects without straw. From the interaction between studied factors shows that the highest concentration of vitamin C were characterized by potato tubers fertilized with phacelia both plowed down in the autumn, and left till spring in the form of mulch, in combination without straw and with straw, and white clover and white clover with straw, and the lowest in potato tubers from control object.

Catch crop fertilization

Straw fertilization

Means

Subblock without straw

Subblock with straw

Control object

0.63

0.56

0.60

Farmyard manure

0.54

0.52

0.53

White clover

0.53

0.51

0.52

White clover + Italian ryegrass

0.48

0.42

0.45

Italian ryegrass

0.50

0.51

0.51

Phacelia

0.47

0.46

0.47

Phacelia-mulch

0.46

0.44

0.47

Means

0.52

0.49

LSD0.05

Catch crop ferilization

0.04

Straw fertilization

0.02

Interaction

n. s.

Table 9. The total sugar content in potato tubers, % (means from years 2005-2007)

Catch crop fertilization

Straw fertilization

Means

Subblock without straw

Subblock with straw

Control object

203.4

217.6

210.5

Farmyard manure

218.6

217.3

218.0

White clover

222.5

224.2

223.4

White clover + Italian ryegrass

219.4

222.5

221.0

Italian ryegrass

217.7

218.4

218.1

Phacelia

220.6

221.7

221.2

Phacelia-mulch

223.4

224.8

224.1

Means

217.9

220.9

SLD0.05

Catch crop ferilization

3.2

Straw fertilization

1.8

Interaction

4.3

Harvesting tops and branches after clear cutting

After clear cutting, tops and branches from felled trees are traditionally left on site together with small trees (Fig. 8). On nutrient-limited sites this slash should not be removed because that would reduce the nutrients present on site. The amount of biomass present in tops and branches is estimated to amount to 20-30 % of the total harvest. The supply of biomass from tops and branches is the main source of bioenergy production in Sweden.

image150

Fig. 8. Clear cut area with branches and tops (left) and stacks of branches and tops (right)

2. Fast-growing species

Besides conventional forestry management, there is increasing interest in management of so — called fast-growing species. Depending on geographical location, different species can be considered fast-growing. There are at least three types of tree suitable and frequently used for management in Europe, the USA and Canada: Salix clones, poplar and hybrid aspen. In areas with higher temperatures than northern Europe, species of Eucalyptus are also planted.

RGB imaging

Digital image analysis has been an important tool in biological research and also has been applied to satellite images, aerial photographs as well as macroscopic images (Nilsson,

1995) . The imaging method has been proposed to infer plant biomass accurately as a non­destructive and fast alternative to the conventional means of determining shoot dry weight. The approach predominantly cited in literature is the estimation of plant biomass as a linear function of the projected shoot area of plants using RGB images.

A relevant application of image analysis which has been used for decades is in the area of remote sensing forestry and precision agriculture in which the area of plant species cover and the biomass of the above-ground canopy are estimated from satellite and airborne images (Montes et al, 2000; Lamb and Brown, 2001).

These techniques have found a recent application in estimating the biomass of individual plants in a controlled environment and also in the field. There have been only a few reports on the application of image analysis techniques to estimate above-ground biomass of an individual plant. In these reports, the projected shoot area of the plants captured on two dimensional images was used as a parameter to predict the plant biomass (Tackenberg, 2007; Sher-Kaul et al, 1995; Paruelo et al, 2000).

Material and methods

1.1 Plant material and conditions of cultivation

In the three-year period 2005-2007 spring wheat (Triticum aestivum L.) "Vanek" variety (maintenance of variety: Lochow-Petkus, GmbH, Germany, producer: Selekta, Inc., Czech Republic) was cultivated at 10 m2 trial field plots outside environment (50°2’0"N, 14°36’54"E) on brown loamy soil. Every tested compound was applied in four replicates (10 m2 field plots). There were sowed 217 kg of seeds per hectare. As foregoing crop broad bean was cultivated on the trial field before wheat plants every year and before wheat sowing the field was fertilized with the dose 60 kg N ha-1 with nitrogen-phosphorus-potassium fertilizer. Average content of minerals in trial field soil is described in Table 1.

In another experiment spring wheat, Vanek variety (maintenance of variety: Lochow — Petkus, GmbH, Germany, producer: Selekta, Inc., Czech Rep.) was cultivated for two years (2006, 2007) in pots in the outside environment. Plants were cultivated in the soil anthropogenic contaminated with heavy metals from the location Pribram, Central Bohemia, historically polluted from metal ores mining and smelting activities. Average content of minerals in contaminated soil are given in Table 2. Sowing was performed into the pots of 5 L volume filled with 5 kg of homogenized soil. Each pot was fertilized with the same dose of NPK (1.43 g N in the NHNCb form, 0.16 g P and 0.40 g K in the K2HPO4 form). The final number of plants in a pot was twenty. Plants were irrigated with demineralised water.

Weather conditions in cultivation period (from April to July) were similar in both years. Mean air temperature in both years was higher compared with the long-term normal. Mean precipitation in 2006 was higher in April, May and June, lower in July compared with the long-term normal. In April and May 2007 mean precipitation was by 25 per cent lower than normal and in June and July was higher compared with the long-term normal.

Depth of mould

N

(NO3-)

N

(NH4+)

N

(total)

K

Mg

Ca

P

pHKCl

mg kg-1 DM

30 cm

21.1+2.1

0.4+0.04

21.5+2.2

264+13.2

132+6.6

3380+169

134+6.7

6.70+0.1

60 cm

4.8+0.5

0.4+0.04

5.2+0.5

185+9.3

141+7.1

2763+138

44+2.2

6.39+0.1

Table 1. Average content of minerals in the soil in field experiment

Soil

„Pribram"

Cation H+ exchange capacity

pHKCl

Cox

Zn

Cu

Cd

Pb

Unit

1 mmol kg-1

%

mg kg-1 DM

Value

123

4.52+0.02

1.91+0.006

187+8.0

42.7+2.0

3.60+0.17

1321+71

Table 2. Content of selected metals and characteristics of used soil contaminated with heavy metals from the district of Pribram, Czech Republic in outside environment pot experiment

Case study: august 2002

A prolonged SALLJ event that occurred in conjunction with biomass burning took place from 23 to 28 August. The low-level jet had an important latitudinal extent and strength with a pattern that varied according to a baroclinic synoptic environment.

3.1.1 Meteorological environment and SALLJ features

Figure 7 depicts the 1000 hPa geopotential height and the 500/1000 hPa thickness fields for selected days during the event. On 23 August, the western branch of the SASH was over an important extension of SA and the low-level flow was from the N as far as 40° S. In the southernmost edge of SA, a baroclinic region — oriented NW to SE — was present and deep low-pressure systems were moving south-eastward. During the following day, a geopotential trough developed over central Argentina. The thickness field showed the associated maximum depth. There was a persistent N-NW flow over south-eastern SA. On 25 August, a further deepening of the trough over central Argentina occurred. The baroclinic region related to the cold front was located between 30° S and 40° S and moved towards the northeast. The low-pressure system behind the cold front weakened. There was a strong channelling of the low-level flow between the trough and the western region of the SASH. Twenty-four hours later, the baroclinic zone approached the southern region of Buenos Aires. A deep thickness trough was present over the eastern Pacific Ocean. Central

Argentina was still with the minimum geopotential. The flow from the north at low levels persisted. On 27 August, the situation was almost similar, with the new system strengthening and moving eastward and starting to surpass the Andes barrier. The northern low-level flow was still present over south-eastern SA and the low pressure further deepened over central Argentina. On 28 August, the system was able to reach eastern Argentina. The associated cold front presented a nearly north-south orientation and moved eastward towards the Atlantic Ocean. The low-pressure system over Argentina deepened and the low-level north-western flow persisted. During 29 August the cold sector of the front moved past Buenos Aires and Uruguay and the related surface cyclone, centred near 40° S and 55° W, deepened. The near-surface airflow over south-eastern SA was from the N — NE sector and from the S in Buenos Aires. On 30 August, the baroclinic region in the 500/1000 hPa thickness field was located at 30° S, with zonal orientation. The surface low — pressure system had its maximum depth at 0600 UTC and then started to fill while travelling to the east over the Atlantic Ocean. Central Argentina had relatively higher surface pressure. The near surface flow was from the S over northern Argentina. During the final day of the study period (31 August) the baroclinic region was in southern Brazil, co­located with a surface col region. Argentina had near surface southerly winds. The surface cyclone was in the occlusion stage at 1800 UTC.

image235

Fig. 7. Daily fields of 1000 hPa geopotential height (red solid (positive), blue dot (negative) contours) and 500/1000 hPa thickness (green long dash contours) (both every 40 mgp), from 23 to 31 August. Terrain elevations higher than 1500 m are shaded.

Bolivia to north-eastern Argentina, part of Uruguay and southern Brazil. The jet core was located over northern Paraguay. The related flow was from the N-NW sector. During the following day, the SALLJ had increased strength and vertical shear, as well as more spatial extension. The southernmost edge was near 40° S. On 25 August, the NW-SE region associated with the SALLJ had a greater latitudinal extension and the low-level flow was from the northwest and stronger due to the westward displacement of the Atlantic anticyclone. During the next day, the SALLJ had a smaller southward penetration and reached only 35° S. This was due to the advance of the cold front that was located past 40° S at the 850-hPa level over the ocean. The flow was more northerly oriented. The jet core was over western Paraguay and northern Argentina.

image236

Fig. 8. Daily SALLJ fields from 23 to 31 August. Wind (vector); wind speed (shaded) at 850 hPa and wind shear between 850 hPa and 700 hPa (contours). Shaded: wind intensity stronger than 12 m s-1. Contours: wind shear greater than 6 m s-1. Terrain elevations higher than 1500 m are shown.

On 27 August, the SALLJ was present over northern and central Argentina. The flow was from the N-NE sector mostly governed by the western region of the anticyclone centred near 32.5° S and 40° W over the Atlantic Ocean. During 28 August, the jet strengthened and spread, reaching the latitudes near 45° S and extending from 65° W to 40° W. The SALLJ reinforced due to the new cold front that was located near 60° W at 1200 UTC with north — south orientation. On 29 August, the front reached Paraguay and south-eastern Brazil. The wind field at 850 hPa shows clearly the northwest wind ahead of the front whereas the winds behind were strong, from the southwest. The region spanned by the strongest winds

has the typical shape of the frontal zone but the wind field did not verify the Bonner’s criteria. During 30 August, the north-western edge of the frontal zone was over Sao Paulo, with the southerly winds blowing clear and dry air over South America up to 15° S. The convergence in the airflow is related to the surface cold front and the baroclinic region near Sao Paulo. The situation persisted on 31 August. SALLJ did not occur either. During this particular event, an important southward penetration of the low-level jet occurred and the associated moisture convergence at the exit region of the current favoured the development of convective systems south of 40° S, which strengthened mostly over the Atlantic Ocean. The interaction with the cold front further contributed to the convection.

Screening

Salinity and waterlogging co-exist in the lower reaches of several river basins throughout the world, affecting agricultural production and the livelihoods of the affected communities (Wichelns and Oster, 2006). Efforts being made to overcome salinity and waterlogging problems by consist of engineering solutions such as installation of a drainage system to manage the drainage effluent generated by irrigated agriculture. This is a long term strategy; however drainage installation is expensive. The areas under salt-affected and waterlogged soils are expanding because of inappropriate on-farm water and soil management. Selection and cultivation of high-yielding salt-tolerant varieties of different crops is a potential interim strategy to fulfill the needs of the communities relying on these soils for their livelihoods (Ayers and Westcot, 1989). Many crops show intraspecific variation in response to salinity. Sorghum is moderately salt-tolerant. Generally, substantial genotypic differences exist among sorghum cultivars in response to salinity stress (Sunseri et al., 2002; Netondo et al., 2004).

1.1.1 Screening methods based on growth or yield

Screening large numbers of genotypes for salinity tolerance in the field is difficult, due to spatial heterogeneity of soil chemical and physical properties, and to seasonal rainfall distribution. Frequently, short-term growth experiments have revealed little difference between genotypes that differ in long-term biomass production or yield. Many short-term growth experiments measuring whole shoot biomass revealed little difference between plant genotypes in their response to salinity, even between those known to differ in long-term biomass production or yield (Rivelli et al., 2002). Longer-term experiments are necessary to detect genotypic differences in the effects of salinity on growth: it is necessary to expose plants to salinity for at least two weeks, and sometimes several months (Munns et al., 1995). Even with rice, a fast growing and salt sensitive species, it is necessary to grow plants for several weeks to be confident of obtaining reproducible differences in salinity tolerance between genotypes (Zhu et al., 2001).