Preliminary Study on the Activity of Phycobiliproteins against Botrytis cinerea

Phycobiliproteins (PBPs) are proteins of cyanobacteria and some algae such as rhodophytes. They have antimicrobial, antiviral, antitumor, antioxidative, and anti-inflammatory activity at the human level, but there is a lack of knowledge on their antifungal activity against plant pathogens. We studied the activity of PBPs extracted from Arthrospira platensis and Hydropuntia cornea against Botrytis cinerea, one of the most important worldwide plant-pathogenic fungi. PBPs were characterized by using FT-IR and FT-Raman in order to investigate their structures. Their spectra differed in the relative composition in the amide bands, which were particularly strong in A. platensis. PBP activity was tested on tomato fruits against gray mold disease, fungal growth, and spore germination at different concentrations (0.3, 0.6, 1.2, 2.4, and 4.8 mg/mL). Both PBPs reduced fruit gray mold disease. A linear dose–response relationship was observed for both PBPs against disease incidence and H. cornea against disease severity. Pathogen mycelial growth and spore germination were reduced significantly by both PBPs. In conclusion, PBPs have the potential for being also considered as natural compounds for the control of fungal plant pathogens in sustainable agriculture.     

Effect of commercial lignosulfonate-humate on Zea mays L. metabolism

Lignosulfonate-humate a and lignosulfonate-humate b, derived by an industrial process from lignin, were studied chemically and biologically, and their effects on maize metabolism compared with the responses induced by humic substances obtained from leonardite. Lignosulfonate-humate a and lignosulfonate-humate b elicited hormonelike activity and leonardite displayed giberellin properties. To improve our understanding of their biological action, lignosulfonate-humate a, lignosulfonate-humate b and leonardite were supplied to maize plants and their effect was studied on growth, nitrogen metabolism and photosynthesis. All products increased root and leaf growth. Glutamine-synthetase, glutamate-synthase enzyme activities and protein content were all increased. The treatments also increased chlorophyll content, glucose, fructose and rubisco enzyme activity, suggesting a positive role of lignosulfonate-humate a, lignosulfonate-humate b and leonardite in the photosynthetic process. In addition, an increase in phenol content was observed. In light of these results, being environmentally friendly products, lignosulfonate-humate a and lignosulfonate-humate b could be used to increase crop yield.     

Cell therapy for tendinitis,experimental and clinical report

To compare cultured bone marrow mesenchymal cells (cBMSC), bone marrow mononucleated cells (BMMNCs), and placebo to repair collagenase-induced tissue damage in an equine model of experimental tendonitis, 6 Standardbred horses with no signs of previous SDF tendon injury have been recruited. Three weeks after collagenase treatment an average of either 5.5 × 10⁶ cBMSCs or 122.3 × 10⁶ BMMNCs, saline solution (placebo) or fibrin glue were injected intralesionally in random order. Horses were stall rested for 21 weeks, and tendon ultrasound scans performed before and during this period. Horses were euthanized and tendons harvested for histology and immunohistochemistry. Data observed in this study showed effectiveness of cBMSC and BMMNC in regenerating tendon tissue after collagenase -induced tendonitis. Both cBMSC and BMMNC transplantation resulted in qualitatively similar regeneration of tendon extracellular matrix in terms of type I/III collagen ratio, fiber orientation, and COMP expression. After this favourable results, 20 horses were recruited referred for spontaneous lesions of the flexor tendons or the suspensory ligament. Horses were treated with autologous graft of BMMNCs.After treatment the. the exercise program allowed was 8 weeks stall rest, 4 weeks hand walking, 4 weeks trotting, 4 weeks of gradually raising of exercise level then horses were gone back to race. US characteristics of lesions started to improve at T3. CSA-l, FPS and TLS were better in all patients, with an appreciable filling of lesions indicated by a decreasing of CSA-l and increasing of TLS. When horses started the exercise program T8 tendon architecture improved, demonstrated by their longitudinal alignment and length. At T6, and persistently in later follorw-up, no lameness was evident by clinical examination. At time of writing 12 patients (60%) were go back to races, while other 8 (40%) are under controlled exercise program. Re-injury rate was assessed at 25%. All the owners judged good to excellent the outcome in term of athletic success.

     

Effect of soil pH on the chemical composition of organic matter in physically separated soil fractions in two broadleaf woodland sites at Rothamsted,UK

Although acid soils are common in forest ecosystems, and there is documented evidence of pH influencing transformations of organic matter in soil, there are surprisingly few studies on the influence of soil pH on the chemical structure of physically fractionated soil organic matter (SOM). The aim of this study was to characterize the influence of pH on the chemical and physical processes involved in SOM stabilization. Forest soils of different pH (4.4 and 7.8) sampled from two long‐term experiments at Rothamsted Research (UK) were physically fractionated. The free light fraction (FLF), the intra‐aggregate light fraction and the fine silt and clay (S + C, <25 µm) were characterized using elemental, isotopic (δ¹³C), thermogravimetric, differential thermal, diffuse reflectance infrared Fourier transform spectroscopy and high‐resolution magic angle spinning ¹H nuclear magnetic resonance analyses. The quantitative distribution of carbon (C) between SOM fractions differed between the two soils. Carbon contents in the light fractions from the acid soil were significantly greater than in those of the alkaline soil. In contrast, in S + C fractions, C content was greater in the alkaline soil. FLF from the acid soil was characterized by a greater C:N ratio, smaller δ¹³C and greater content of thermo‐labile compounds compared with FLF from the alkaline soil. In contrast, there was only a weak effect of soil pH on the chemical composition of the organic matter in S + C fractions. Irrespective of soil pH, these latter fractions contained mainly aliphatic compounds such as carbohydrates, carboxylic acid, amide and peptide derivates. This suggested that physical mechanisms, involving the interactions between SOM and mineral surfaces, are of greater importance than the presence of chemically recalcitrant species in protecting SOM associated with the finest soil fractions.     

Investigating the effects of wettability and pore size distribution on aggregate stability: the role of soil organic matter and the humic fraction

Soil organic matter (SOM) is an important factor influencing aggregate stability. Interactions between SOM and soil structure are widely studied, although the subtle relationship between SOM content, pore size distribution and aggregate stability is not fully known. Here we investigate such a relationship by means of a long‐term experiment established in 1962 in northeastern Italy, which considers different fertilizer practices (organic, mineral and mixed) applied to a continuous maize crop rotation. We measured wet stability of 1–2 mm aggregates subjected to different pretreatments. Both soil physical properties (such as pore size distribution and hydrophobicity) and chemical properties (soil organic and humic carbon content) affecting aggregate stability were considered. The chemical structure of humic substances was characterized by thermal and spectroscopic analyses (TG‐DTA, DRIFT and ¹H HR MAS NMR). The Pore‐Cor network model was then applied to evaluate the contribution of hydrophobicity and porosity to aggregate wetting. Our study suggests that SOM and its humic fraction can affect aggregate wetting and consequently slaking by modifying the pore size distribution with a shift from micropores (5–30 µm) and mesopores (30–75 µm) to ultramicropores (0.1–5 µm); hydrophobicity was also increased as a result of different humic composition. Spectroscopic analysis showed that hydrophobic compounds were mostly associated with complex humic molecules. Models of fast wetting dynamics, however, suggest that the contribution that hydrophobicity makes to aggregate stability, especially to soils with large carbon inputs, may not be the most significant factor.     

Use of autologous bone marrow mononuclear cells and cultured bone marrow stromal cells in dogs with orthopaedic lesions

The aim of the study is to evaluate the clinical application in veterinary orthopedics of bone marrow mononuclear cells (BMMNCs) and cultured bone marrow stromal cells (cBMSCs) for the treatment of some orthopaedic lesions in the dog. The authors carried out a clinical study on 14 dogs of different breed, age and size with the following lesions: 1 bone cyst of the glenoid rime; 2 nonunion of the tibia; 3 nonunion of the femur; 2 lengthening of the radius; 1 large bone defect of the distal radius;1 nonunion with carpus valgus; 4 Legg-Calvé-Perthés disease. In 9 cases the BMMCNs were used in combination with a three dimensional resorbable osteogenic scaffold the chemical composition and size of which facilitates the ingrowth of bone. In these cases the BMMNCs were suspended in an adequate amount of fibrin glue and then distribuited uniformly on a Tricalcium-Phosphate (TCP) scaffold onto which were also added some drops of thrombin. In 1 case of nonunion of the tibia and in 3 cases of Legg-Calvè-Perthés (LCP) disease the cultured BMSCs were used instead because of the small size of the dogs and of the little amount of aspirated bone marrow. X-ray examinations were performed immediately after the surgery. Clinical, ultrasounds and X-ray examinations were performed after 20 days and then every month. Until now the treated dogs have shown very good clinical and X-ray results. One of the objectives of the study was to use the BMMNCs in clinical application in orthopaedic lesions in the dog. The advantages of using the cells immediately after the bone marrow is collected, are that the surgery can be performed the same day, the cells do not need to be expanded in vitro, they preserve their osteogenic potential to form bone and promote the proper integration of the implant with the bone and lastly, the technique is easier and the costs are lower.     

Cell therapy for tendinitis, experimental and clinical report

To compare cultured bone marrow mesenchymal cells (cBMSC), bone marrow mononucleated cells (BMMNCs), and placebo to repair collagenase-induced tissue damage in an equine model of experimental tendonitis, 6 Standardbred horses with no signs of previous SDF tendon injury have been recruited. Three weeks after collagenase treatment an average of either 5.5 x 10(6) cBMSCs or 122.3 x 10(6) BMMNCs, saline solution (placebo) or fibrin glue were injected intralesionally in random order. Horses were stall rested for 21 weeks, and tendon ultrasound scans performed before and during this period. Horses were euthanized and tendons harvested for histology and immunohistochemistry. Data observed in this study showed effectiveness of cBMSC and BMMNC in regenerating tendon tissue after collagenase -induced tendonitis. Both cBMSC and BMMNC transplantation resulted in qualitatively similar regeneration of tendon extracellular matrix in terms of type I/III collagen ratio, fiber orientation, and COMP expression. After this favourable results, 20 horses were recruited referred for spontaneous lesions of the flexor tendons or the suspensory ligament. Horses were treated with autologous graft of BMMNCs.After treatment the. the exercise program allowed was 8 weeks stall rest, 4 weeks hand walking, 4 weeks trotting, 4 weeks of gradually raising of exercise level then horses were gone back to race. US characteristics of lesions started to improve at T3. CSA-l, FPS and TLS were better in all patients, with an appreciable filling of lesions indicated by a decreasing of CSA-l and increasing of TLS. When horses started the exercise program T8 tendon architecture improved, demonstrated by their longitudinal alignment and length. At T6, and persistently in later follow-up, no lameness was evident by clinical examination. At time of writing 12 patients (60%) were go back to races, while other 8 (40%) are under controlled exercise program. Re-injury rate was assessed at 25%. All the owners judged good to excellent the outcome in term of athletic success.     

Soil pyrogenic organic matter characterisation by spectroscopic analysis: a study on combustion and pyrolysis residues

Purpose

Pyrogenic organic matter (PyOM) in the soil of a maritime pine forest in Central Italy, formed during a fire of high severity, was characterised by Fourier transform infrared (FT-IR) and ¹³C nuclear magnetic resonance (NMR). Furthermore, soil samples from burnt and unburnt sampling sites and natural charcoal collected from the ground were characterised after progressive heating under air and nitrogen atmosphere. The aim was to better understand the role fire plays on PyOM formation and oxidation.

Materials and methods

The top 10 cm of mineral soil and the above-lying charcoal particles were collected soon after the fire. Sampling was also performed on an adjacent unburnt portion of the forest. The bulk soil organic matter (SOM), its extractable fraction and charcoal particles were investigated by FT-IR and ¹³C NMR spectroscopies. They also underwent thermogravimetric analysis under air or N₂, stopping the thermal reactions at the end of the first exothermic reaction in the range 350–500 °C.

Results and discussion

The NMR investigation clearly revealed a significant enrichment in aromatic and alkyl C in the burnt soil compared to the unburnt one. Several clues led to hypothesise that SOM was not exposed to extreme heating during the fire, notwithstanding the high fire severity estimated by a vegetation-based visual scale. In the thermal treatment mimicking fire, charcoal lost much of its mass and carbon content. However, at 500 °C, it still maintained a significant recalcitrant fraction. Nitrogen concentration in the bulk soil increased after heating, particularly under air condition. This phenomenon could be due to the formation of heterocyclic nitrogen compounds in the charred material.

Conclusions

In the study area, PyOM is rich in aliphatic compounds presumably because of the understory sclerophyllous vegetation typically found in Mediterranean environments. A large fraction of the charcoal released to the soil during the fire is sensitive to oxidation by subsequent fires. On the other hand, charcoal preserves a significant fraction of C, the most recalcitrant one, with expected long residence time in soil. PyOM formed under high oxygen availability is richer in N than that formed in inert atmosphere, which might make PyOM more susceptible to biochemical degradation.

     

Mineral Content and Root Respiration of In Vitro Grown Kiwifruit Plantlets Treated with Two Humic Fractions

ABSTRACT

In vitro grown kiwifruit (Actinidia deliciosa, Liang and Ferguson) plantlets were treated with two humic fractions distinguished by two different relative molecular mass and characterized through their elemental composition and ¹³Carbon (C) nuclear magnetic resonance. The effects exerted on plant growth, root morphology, and nutrition were evidenced by means of leaves and roots mineral content, root respiration, and nitrate reductase activity. The two humic substances differentially influenced the studied parameters. The lower molecular fraction humic substance, endowed with a higher content of phenolic and carboxylic groups, caused an enhanced content of micro- and macro-elements, together with a higher root respiration at lower concentrations (0.5–1 mg C L^{? 1}). The high molecular fraction humic substance, needed higher concentrations (> 5 mg C L^{? 1}) in order to achieve similar effects. This fraction also caused major changes on root morphology. The ability to improve micronutrient assimilation, in particular iron, confirmed the agronomic importance of humic substances on soil fertility.