Entomopathogenic fungi as biological control agents for the vector of the laurel wilt disease,the redbay ambrosia beetle,Xyleborus glabratus (Coleoptera: Curculionidae)

The redbay ambrosia beetle (RAB), Xyleborus glabratus, is a wood-boring insect that vectors the fungal pathogen, Raffaelea lauricola, which causes laurel wilt, a lethal disease of avocado. The objective of this study was to determine the susceptibility of RAB to infection and subsequent death by exposure to three commercial strains of entomopathogenic fungi [two strains of Isaria fumosorosea (Ifr 3581 and PFR), and strain GHA of Beauveria bassiana]. RAB females were dipped in fungal spore solutions and their median survivorship times (MST) determined. Contact with any of the biopesticides resulted in death of all RAB females. MSTs of RAB females ranged from 3 days (B. bassiana) to 5 days (I. fumosorosea PFR). B. bassiana killed RAB females faster, followed by Ifr 3581 and PFR. RAB females dipped in B. bassiana suspensions had the highest number of viable spores attached to their bodies, followed by Ifr 3581. Beetles dipped in PFR suspension had significantly less viable spores attached to their bodies. No significant differences were observed in the mortality of beetles exposed to entomopathogenic fungi by dipping in a fungal suspension or walking on treated avocado bolts. Beetles bored into the logs and constructed galleries, but they were found dead inside the galleries a few days after exposure to the entomopathogens. Entomopathogenic fungal infection in dead beetles was confirmed through molecular techniques. This is the first study to demonstrate that entomopathogenic fungi are potential biological control agents against RAB.     

Partial purification and characterization of Helicoverpa armigera (Lepidoptera: Noctuidae) active aminopeptidase secreted in midgut

We report the partial purification to apparent homogeneity of a soluble aminopeptidase (EC 3.4.11.1) from midgut of Helicoverpa armigera larvae, which preferentially degraded Leucine p-nitroanilide (LpNA). After midgut isolation, extraction and precipitation of soluble proteins with acetone, proteins were purified in two consecutive steps including gel filtration and ion-exchange chromatographies. Aminopeptidase activity was increased 8.95 fold after gel filtration chromatography. The purified enzyme appeared as single band with a molecular mass of ～ 112 kDa in SDS-PAGE, with a pH optimum of 7.0. Zymogram analysis revealed two enzymatically active proteinases using LpNA as substrate. The optimal temperature of aminopeptidase activity was 50–60 °C. The enzyme was characterized as metalloprotease as it was strongly inhibited by 1,10 phenanthroline. Strong inhibition was also being observed using the specific aminopeptidase inhibitor bestatin. Heavy metal ions, EDTA and cysteine strongly inhibited the enzyme, while Ca^+ 2, Mn^+ 2 and Mg^+ 2 somewhat stimulated aminopeptidase activity. Besides LpNA, the purified aminopeptidase also cleaved with decreasing activity ApNA, VpNA and BApNA. Study could be helpful to understand the mechanism of action of N-terminal degrading enzymes and also important is to further study the differential interaction of Bacillus thuringiensis cry insecticidal toxin with midgut receptor of insects.     

Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition

Mucor circinelloides produces plant cell wall degrading enzymes that allow it to grow on complex polysaccharides. Although the genome of M. circinelloides has been sequenced, only few plant cell wall degrading enzymes are annotated in this species. We applied peptide pattern recognition, which is a non-alignment based method for sequence analysis to map conserved sequences in glycoside hydrolase families. The conserved sequences were used to identify similar genes in the M. circinelloides genome. We found 12 different novel genes encoding members of the GH3, GH5, GH9, GH16, GH38, GH47 and GH125 families in M. circinelloides. One of the two GH3-encoding genes was predicted to encode a β-glucosidase (EC 3.2.1.21). We expressed this gene in Pichia pastoris KM71H and found that the purified recombinant protein had relative high β-glucosidase activity (1.73 U/mg) at pH5 and 50 °C. The K_m and V_max with p-nitrophenyl-β-d-glucopyranoside as substrate was 0.20 mM and 2.41 U/mg, respectively. The enzyme was not inhibited by glucose and retained 84% activity at glucose concentrations up to 140 mM. Although zygomycetes are not considered to be important degraders of lignocellulosic biomass in nature, the present finding of an active β-glucosidase in M. circinelloides demonstrates that enzymes from this group of fungi have a potential for cellulose degradation.     

Soil organic carbon fractions are affected by different land uses in an agro-pastoral transitional zone in Northeastern China

Soil organic carbon (SOC) consists of various C fractions with different stabilities and chemical compositions that are differently affected by changes in land use. A better understanding of the responses of different C fractions to land uses is vital for maintaining soil quality and mitigating global warming. Using data from a short-term land use experiment in northeastern China, this paper investigated the effect of five land uses, corn cropland (Corn), alfalfa grassland (Alfalfa), artificial grassland of Lyemus chinensis (AG), Lyemus chinensis grassland for mowing (AG + Mow) and restored grassland (RG), on the dynamics of total SOC and four SOC fractions with increasing degrees of oxidizability at 0 to 50 cm depths. The results show that land use had a significant effect (P < 0.05) on the total SOC and SOC fractions of very labile C (F1), labile C (F2) and less labile C (F3), while the difference in recalcitrant C (F4) was less pronounced. SOC in the study area was characterized by a predominantly very labile C fraction, and the percentages of F1 to total SOC were more than 40% for all land uses. Compared with Corn, the treatments AG + Mow, AG and RG decreased the percentage of F1 to SOC (by 4.49%, 6.53% and 3.55%, respectively) and increased the percentages of F2 (by 3.32%, 2.77% and 6.60%, respectively) and F3 (by 4.47%, 3.46% and 0.3%, respectively) to SOC. These findings suggest that land-use type is a major factor that influences soil C fractions and that labile C fractions contribute a large part of the total SOC. In addition, grassland colonization of croplands improves soil C sequestration in northeastern China.     

Celery oil modulates DEHP-induced reproductive toxicity in male rats

The objective of the study was to investigate the protective effect of Apium graveolens (AP) against di-(2-ethylhexyl) phthalate (DEHP)-induced testes injury in rats. Adult rats were divided into nine groups: (1) control group (no treatment); (2) corn oil (60 μg/kg body weight – bwt); (3) AP (50 μg/kg bwt); (4) 300 mg DEHP/kg bwt; (5) 500 mg DEHP/kg bwt; (6) 1000 mg DEHP/kg bwt; (7) 300 mg DEHP/kg bwt + AP; (8) 500 mg DEHP/kg bwt + AP; and (9) 1000 mg DEHP/kg bwt + AP. Oral administration of treatments was performed daily for 6 weeks. DEHP decreased (p < 0.01) body weight, testis weight and serum concentrations of testosterone, cholesterol and total proteins. Moreover, DEHP increased (p < 0.001) total antioxidant capacity in the testis and plasma DEHP level. In addition, DEHP decreased mRNA expression of two testicular steroidogenic enzymes: 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase. DEHP also caused atrophy, vacuolar degeneration and aspermia of the seminiferous tubules. AP administered concurrently with DEHP effectively alleviated most of the DEHP-induced effects. In conclusion, in male rats, DEHP had adverse effects on the testis including inhibition of androgen production. A concurrent administration of A. graveolens (celery oil) protected the testis against DEHP-induced toxicity.     

The antifungal activity of fatty acids of all stages of Sarcophaga carnaria L. (Diptera: Sarcophagidae)

Fatty acids as components of cuticular lipids of insects play a significant role in antifungal in protection against fungal infection. The chemical composition of cuticular and internal extracts obtained from all developmental stages of flesh flies Sarcophaga carnaria was identified. The fatty acids were detected using gas chromatography coupled with mass spectrometry and the most abundant for all examined stages were: 18:1 > 16:0 > 16:1 > 18:0 > 18:2. Polyunsaturated fatty acids (PUFA) C20 were found in both, cuticular and internal extracts. GC–MS analysis showed higher relative content of PUFA in adults than in preimaginal stages.Fatty acids alone as well as their cuticular and internal extracts obtained from larvae, pupae male and female of S. carnaria were tested according to their potential antimicrobial activity against entomopathogenic fungi: Paecilomyces lilacinus, Paecilomyces fumosoroseus, Lecanicillium lecanii, Metarhizium anisopliae, Beauveria bassiana (Tve-N39) and B. bassiana (Dv-1/07). FA presented diverse antimicrobial activity depending on the length of the chain and the presence of unsaturated bonds. Short chain and unsaturated FA (6:0, 11:0, 13:0) have shown significantly stronger activity against fungi but they were detected in lower concentrations. PUFA inhibit fungal growth more effectively than unsaturated long chain fatty acids. Cuticular and internal extracts of all living forms of S. carnaria exhibited approximately equal activity against tested entomopathogenic fungi. We presumed that the most abundant saturated long chain FA and additionally PUFA founded in our analysis are involved in protecting the flies against fungal infection.     

Purification and characterization of an extracellular antifungal chitinase from Penicillium ochrochloron MTCC 517 and its application in protoplast formation

A highly chitinolytic strain Penicillium ochrochloron MTCC 517 was procured from MTCC, Chandigarh, India. Culture medium supplemented with 1% chitin was found to be suitable for maximum production of chitinase. Purification of extracellular chitinase was done from the culture medium by organic solvent precipitation and DEAE-cellulose column chromatography. The chitinase was purified 6.92-fold with 29.9% yield. Molecular mass of purified chitinase was found to be 64 kDa by SDS-PAGE. The chitinase showed optimum temperature 40 °C and pH 7.0. The enzyme activity was completely inhibited by Hg²⁺, Zn²⁺, K⁺ and NH₄⁺. The enzyme kinetic study of purified chitinase revealed the following characteristics, such as apparent K_m 1.3 mg ml^?1, V_max 5.523 × 10^?5 moles l^?1 min^?1 and K_cat 2.37 s^?1 and catalytic efficiency 1.82 s^?1 M^?1. The enzyme hydrolyzed colloidal chitin, glycol chitin, chitosan, glycol chitosan, N,N′-diacetylchitobiose, p-nitrophenyl N-acetyl-β-d-glucosaminide and 4-methylumbelliferyl N-acetyl-β-d-glucosaminide. The chitinase of P. ochrochloron MTCC 517 is an exoenzyme, which gives N-acetylglucosamine as the main hydrolyzate after hydrolysis of colloidal chitin. Protoplasts with high regeneration capacity were obtained from Aspergillus niger using chitinase from P. ochrochloron MTCC 517. Since it also showed antifungal activity, P. ochrochloron MTCC 517 seems to be a promising biocontrol agent.     

The interaction of two potential fungal bioherbicides and a sub-lethal rate of glyphosate for the control of shattercane

Greenhouse and laboratory experiments were conducted with the potential bioherbicides Colletotrichum graminicola (Cg) and Gloeocercospora sorghi (Gs) for control of shattercane weed. Single-spray tank mixture applications containing different ratios of the two fungi resulted in additive percent weed biomass losses. Intraspecific (Cg + Cg or Gs + Gs) and interspecific (Cg + Gs or Gs + Cg) sequential applications 1- or 7-days apart indicated antagonistic interactions in percent biomass loss. Application of either fungus with, or 1–3 days prior to, a sub-lethal concentration of glyphosate resulted in an antagonistic percent biomass loss; while application of glyphosate prior to either potential bioherbicide resulted in a synergistic weed disease response. Conidia germination studies conducted both in vitro on agar plates and with leaf impression peels suggest that antagonistic interactions observed in weed disease severity are probably due to the host–pathogen response following infection.     

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.     

Partial replacement of barley grain for corn grain: Associative effects on lambs’ growth performance

Our objective was to study the positive effects of partial replacement of barley grain for corn in high concentrate diets on growth performance of growing lambs, and to determine the minimum amount of corn needed to produce such effects. Thirty-three male Awassi lambs weaned at 60 days of age were divided into three groups of 11 according to their live weight and offered three isonitrogenous diets. The control diet (B) contained 81 and 14% barley grain and wheat straw, respectively (DM basis). Corn grain replaced barley grain at 10 and 20% of dietary DM for low (LC) and high (HC) corn diets, respectively. Barley had a higher (P < 0.05) rate of digestion (11.5%/h) compared with corn (8.3%/h). The in vitro 30-h digestion extent was also higher (P < 0.05) for barley. The digestion rate for the B diet was higher (P < 0.05) compared with the HC diet, whereas the value for the LC diet was intermediate. Lambs fed LC and HC diets consumed more (P > 0.05) DM (average = 855 g/day) compared with lambs fed B diet (757 g/day). DM and CP digestibilities were similar among diets and averaged 67.0 and 64.2%, respectively. Final BW, BW change and average daily gain (ADG) for lambs fed HC were higher (P < 0.05) compared with B and LC. Moreover, lambs that consumed LC tended (P = 0.12) to grow faster than lambs fed B. Feed to gain ratio was lower (P < 0.05) for lambs fed HC (4.6) compared with B and LC (5.2). In summary, positive associative effects of partial replacement of barley with corn in high concentrate diets for fattening sheep were detected. However, a minimum of 20% replacement of dietary DM from barley with corn was needed to positively improve both performance and feed efficiency.     

The endophytic fungal entomopathogens Beauveria bassiana and Purpureocillium lilacinum enhance the growth of cultivated cotton (Gossypium hirsutum) and negatively affect survival of the cotton bollworm (Helicoverpa zea)

The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum, were assessed on the growth of cultivated cotton (Gossypium hirsutum) and development of the cotton bollworm (Helicoverpa zea). In two replicate greenhouse trials, cotton plants were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia and evaluated for effects on both plant dry biomass, number of nodes and number of developing flowers (squares). We similarly treated cotton plants and evaluated H. zea performance using no-choice in planta assays starting at the 2nd larval instar. Treatment with both fungal endophytes resulted in a significant increases in plant dry biomass (ANOVA, P = 0.024). Plant developmental stage and number of squares were also significantly enhanced in the endophyte treated plants (ANOVA, P = 0.005 and P = 0.027, respectively). The survivorship of H. zea was significantly different among the endophyte treatment groups (Kaplan–Meier, P = 0.02), where insects feeding on control plants exhibited higher survival than insects on the endophyte treated plants. There were no significant endophyte treatment effects on larval or pupal weights of H. zea individuals. There was no endophyte effect on days to pupation among treatments, but there was a marginal effect on days to eclosion (Kaplan–Meier, P = 0.07). Overall, our results demonstrate (i) the positive plant growth enhancing effects of the target endophyes on cultivated cotton under greenhouse conditions and (ii) the negative effects of endophytic P. lilacinum and B. bassiana on H. zea survivorship and development using whole plant assays.     

Direct ethanol production from N-acetylglucosamine and chitin substrates by Mucor species

Chitin, which is a polymer of β-(1–4) linked N-acetyl-d-glucosamine (GlcNAc) residues, is one of the most abundant renewable resources in nature, after cellulose. In this study, we found some native Mucor strains, which can use GlcNAc and chitin substrates as carbon sources for growth and ethanol production. One of these strains, M. circinelloides NBRC 6746 produced 18.6 ± 0.6 g/l of ethanol from 50 g/l of GlcNAc after 72 h and the maximum ethanol production rate was 0.75 ± 0.1 g/l/h. Furthermore, M. circinelloides NBRC 4572 produced 6.00 ± 0.22 and 0.46 ± 0.04 g/l of ethanol from 50 g/l of colloidal chitin and chitin powder after 16 and 12 days, respectively. We also found an extracellular chitinolytic enzyme producing strain M. ambiguus NBRC 8092, and successfully improved ethanol productivity of NBRC 4572 from colloidal chitin using crude chitinolytic enzyme derived from NBRC 8092. The ethanol titer reached 9.44 ± 0.10 g/l after 16 days. These results were the first bioethanol production from GlcNAc and chitin substrates by native organisms, and also suggest that these Mucor strains have great potential for the simultaneous saccharification and fermentation (SSF) of chitin biomass.     

Anion inhibition profiles of α-, β- and γ-carbonic anhydrases from the pathogenic bacterium Vibrio cholerae

Among the numerous metalloenzymes known to date, carbonic anhydrase (CA, EC 4.2.1.1) was the first zinc containing one, being discovered decades ago. CA is a hydro-lyase, which catalyzes the following hydration–dehydration reaction: CO₂ + H₂O ⇋ HCO₃⁻ + H⁺. Several CA classes are presently known, including the α-, β-, γ-, δ-, ζ- and η-CAs. In prokaryotes, the existence of genes encoding CAs from at least three classes (α-, β- and γ-class) suggests that these enzymes play a key role in the physiology of these organisms. In many bacteria CAs are essential for the life cycle of microbes and their inhibition leads to growth impairment or growth defects of the pathogen. CAs thus started to be investigated in detail in bacteria, fungi and protozoa with the aim to identify antiinfectives with a novel mechanism of action. Here, we investigated the catalytic activity, biochemical properties and anion inhibition profiles of the three CAs from the bacterial pathogen Vibrio cholera, VchCA, VchCAβ and VchCAγ. The three enzymes are efficient catalysts for CO₂ hydration, with k_cat values ranging between (3.4 − 8.23) × 10⁵ s⁻¹ and k_cat/K_M of (4.1 − 7.0) × 10⁷ M⁻¹ s⁻¹. A set of inorganic anions and small molecules was investigated for inhibition of these enzymes. The most potent VchCAγ inhibitors were N,N-diethyldithiocarbamate, sulfamate, sulfamide, phenylboronic acid and phenylarsonic acid, with K_I values ranging between 44 and 91 μM.     

Effects of exposure to sublethal propiconazole on intestine-related biochemical responses in rainbow trout,Oncorhynchus mykiss

The effect of long-term (30 days) exposure to PCZ (0.2, 50, and 500 μg l^?1) on intestine-related biochemical markers in rainbow trout was investigated. Multiple biomarkers were measured, including digestive enzymes (proteolytic enzymes and amylase), antioxidant responses (TBARS, CP, SOD, CAT, GR and GPx) and energy metabolic parameters (RNA/DNA ratio, Na⁺-K⁺-ATPase). Exposure to 500 μg l^?1 PCZ led to significantly inhibited (p < 0.01) proteolytic enzyme and amylase activity. Activities of the antioxidant enzymes SOD, CAT, and GPx gradually increased at lower PCZ concentrations (0.2 and 50 μg l^?1). At the highest concentration (500 μg l^?1), oxidative stress was apparent as significant higher (p < 0.05) lipid peroxidation and protein carbonyls, associated with an inhibition of antioxidant enzymes activity. Moreover, energy metabolic parameters (RNA/DNA ratio, Na⁺-K⁺-ATPase) were significantly inhibited (p < 0.01) in the intestines of fish exposed to 500 μg l^?1 PCZ, compared with controls. We suggest that long-term exposure to PCZ could result in several responses in intestine-related biochemical markers, which potentially could be used as indicators for monitoring residual PCZ present in the aquatic environment.     

Management of Frankliniella occidentalis (Thysanoptera: Thripidae) with granular formulations of entomopathogenic fungi

Western flower thrips (WFT), Franklinella occidentalis, is a major pest of ornamentals. Mycotized millet grains with entomopathogenic fungi applied to soil of potted marigold plants were tested to target the soil-dwelling stages of thrips. Two experimental fungal isolates, (Beauveria bassiana [ARS7060] and Metarhizium anisopliae [ERL1171]), were compared with the registered B. bassiana strain GHA [commercialized as BotaniGard®] and untreated controls in greenhouse caged trials. Mycotized millet grains were mixed into the upper surface of the potting soil in pots of flowering ‘Hero Yellow’ marigolds (4 g/pot). One week after application five mated WFT females were released onto each plant (four plants per cage). At 8 weeks post-infestation, the mean total number of thrips per plant was 81% and 90% less in the ERL1171 and ARS 7060 treatments, respectively, than in the control. The mean numbers of thrips per plant for the control and GHA treatments were not significantly different. Plant damage was 60% less on plants treated with the experimental fungi than the control and GHA treatments. At 10 weeks post-application, 75–90% of WFT collected from the treatments of the experimental fungi were infected with the fungal species applied. These results demonstrate that soil applications of entomopathogenic fungi can reduce WFT populations significantly and prevent major damage.     

A new semi-selective medium for Fusarium graminearum,F. proliferatum,F. subglutinans and F. verticillioides in maize seed

Zea mays L., known also as corn and maize, is the most important crop according to the amount of tonnes produced each year. Fungi cause significant destruction of maize in the field as well as during storage rendering the grain unsuitable for human consumption by decreasing its nutritional value and by producing mycotoxins that are detrimental to both human and animal health. Fusarium species are widely distributed and are amongst the most frequently isolated fungal species by plant pathologists. Due to the fact that the Fusarium species involved in maize ear rot vary in fungicide sensitivity, pathogenicity as well as in their capability to produce mycotoxins, accurate quantification and identification is of paramount significance. Currently no method has been developed to test for Fusarium species in maize seed that has been validated and published by the International Seed Testing Association (ISTA). Malachite green agar 2.5 ppm (MGA 2.5) is a potent selective medium for isolation and enumeration of Fusarium spp. In this study, eight different media compositions, potato dextrose agar (PDA), PDA + malachite green oxalate, corn meal agar, 1/2 PDA + malachite green oxalate, 1% malt agar, carnation leaf agar supplemented with potassium chloride (KCLA), malachite green agar (MGA 2.5) and MGA 2.5 + sterile carnation leaf pieces were compared using four Fusarium species (F. graminearum, F. proliferatum, F. subglutinans and F. verticillioides) and five commonly encountered saprophytic fungi (Aspergillus niger, Penicillium crustosum, P. digitatum, Trichoderma harzianum and Rhizopus stolonifer). The maize kernels were surface disinfected using three concentrations of sodium hypochlorite (0.5%, 1% and 1.5% NaOCl) and for different time intervals (1 min, 3 min, 5 min and 10 min). The effect of black-blue light (365 nm) on sporulation of the fungi was also investigated. Surface disinfection of maize seeds with 1% NaOCl for 5 min provided consistent results. PDA, 1/2 PDA, 1% malt agar and KCLA allowed profuse growth of the Fusarium species as well as saprophytes. Media that contained malachite green oxalate was most inhibitory to the radial colony growth of the saprophytes and the Fusarium species. The Fusarium species growing on these media formed underdeveloped morphological structures, thereby obscuring accurate identification. MGA 2.5 showed better hindering of the saprophytes in some instances. MGA 2.5 amended with sterile carnation leaf pieces was the most satisfactory medium in hindering the growth of the saprophytes while allowing adequate sporulation by the four Fusarium species to permit accurate identification. The media also resulted in higher F. verticillioides and lower saprophytic fungal isolation frequency when compared to the other media tested.     

Involvement of polyamines,abscisic acid and anti-oxidative enzymes in adaptation of Blue Panicgrass (Panicum antidotale Retz.) to saline environments

A hydroponic experiment was conducted to assess the possible involvement of polyamines (PAs), abscisic acid (ABA) and anti-oxidative enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in adaptation of six populations of Panicum antidotale Retz. to selection pressure (soil salinity) of a wide range of habitats. Plants of six populations were collected from six different habitats with ECe ranging from 3.39 to 19.23 dS m⁻¹ and pH from 7.65 to 5.86. Young tillers from 6-month-old plants were transplanted in plastic containers each containing 10 l of half strength Hoagland's nutrient solution alone or with 150 mol m⁻³ NaCl. After 42 days growth, contents of polyamines (Put, Spd and Spm) and ABA, and the activities of anti-oxidative enzymes (SOD, POD and CAT) of all populations generally increased under salt stress. The populations collected from highly saline habitats showed a greater accumulation of polyamines and ABA and the activities of anti-oxidative enzymes as compared to those from mild or non-saline habitats. Moreover, Spm/Spd and Put/(Spd + Spm) ratios generally increased under salt stress. However, the populations from highly saline environments had significantly higher Spm/Spd and Put/(Spd + Spm) ratios as compared to those from mild or non-saline environments. Similarly, the populations adapted to high salinity accumulated less Na⁺ and Cl⁻ in culm and leaves, and showed less decrease in leaf K⁺ and Ca²⁺ under salinity stress. Higher activities of anti-oxidative enzymes and accumulation of polyamines and ABA, and increased Spm/Spd and Put/(Spm + Spd) ratios were found to be highly correlated with the degree of adaptability of Panicum to saline environment.     

Ruminal ochratoxin A degradation—Contribution of the different microbial populations and influence of diet

The mycotoxin ochratoxin A (OTA) is degraded extensively in the rumen. In this study, the relative contribution of different rumen microbial populations (MP) and the effect of diet on degradation of OTA were evaluated in a factorial design experiment. Degradation of OTA was quantified by using the Hohenheim gas test (HGT) in vitro fermentation system. Five different HGT diets were used (concentrate:forage proportions (C:F) – 10:90, 30:70, 50:50, 70:30, 90:10), and donor animals were fed diets with the respective ratio. Diets with the highest concentrate content were supplied with and without 10 g/kg sodium bicarbonate (70:30 BC and 90:10 BC). The MP included whole rumen fluid, fungi + protozoa, bacteria + protozoa, protozoa and bacteria + fungi. Protozoa numbers were counted after 24 h and OTA and ochratoxin alpha (OTα) analysed at 0, 4, 8, 12, 24 h. Area under the curve (AUC) and half-life were calculated for the latter two. The short average OTA half-life for whole rumen fluid of 2.6 h (1.3–4.5 h) demonstrates the high OTA degradation capacity of the rumen MP (i.e., standard HGT inoculum) and corresponds well with published in vivo results. Both MP and diet affected OTA degradation. Interactions among factors occurred (P<0.001), which made it necessary to do further comparisons within factor levels. Among MP, those with bacteria (bacteria + fungi and bacteria + protozoa) had lower AUC values (P<0.001) for OTA (196–673 ng/ml h, meaning higher degradation capacity, than those without bacteria (fungi + protozoa and protozoa; 701–1206 ng/ml h). Whole rumen fluid had the lowest AUC values (146–249 ng/ml h; P<0.05). Diet had a quadratic effect (P=0.001) on protozoal numbers with minimum values for the lowest and highest C:F ratios, for bacteria + protozoa, fungi + protozoa and protozoa, but no corresponding effect was found for OTA degradation parameters. While the generally high capacity to degrade OTA was confirmed, results for the contribution of different microbial groups shed new light on ruminal OTA degradation.     

Production of yeast chitin–glucan complex from biodiesel industry byproduct

Crude glycerol from the biodiesel industry was used as carbon source for high cell density fed-batch cultivation of Pichia pastoris aiming at producing a chitin–glucan complex (CGC). More than 100 g L^?1 biomass was obtained in less than 48 h. The yield of biomass on a glycerol basis was 0.55 g g^?1 during the batch phase and 0.63 g g^?1 during the fed-batch phase. The chitin–glucan complex was recovered from the yeast cell wall by hot alkaline extraction. CGC content in the cell wall was found to be relatively constant throughout the cultivation (18–26%) with a volumetric productivity of 1.28 g L^?1 h^?1 at the end of the fed-batch phase. The molar ratio of chitin:β-glucan in the extracted biopolymer was 16:84, close to other CGC extracted from Aspergillus biomass. The extracted polymer was characterized by Differential Scanning Calorimetry (DCS) and solid-state Nuclear Magnetic Resonance (NMR) spectroscopy and compared with commercial biopolymers, namely, crab shell chitin and/or chitosan, algal β-glucan (laminarin) and fungal chitin–glucan complex (kiOsmetine).