Fusarium proliferatum isolated from garlic in Spain: Identification, toxigenic potential and pathogenicity on related Allium species

Fusarium proliferatum has been reported on garlic in the Northwest USA, Spain and Serbia, causing water-soaked tan-colored lesions on cloves. In this work, Fusarium proliferatum was isolated from 300 symptomatic garlic bulbs. Morphological identification of Fusarium was confirmed using species-specific PCR assays and EF-1α sequencing. Confirmation of pathogenicity was conducted with eighteen isolates. Six randomly selected F. proliferatum isolates from garlic were tested for specific pathogenicity and screened for fusaric acid production. Additionally, pathogenicity of each F. proliferatum isolate was tested on healthy seedlings of onion (Allium cepa), leek (A. porrum), scallions (A. fistulosum), chives (A. schoenoprasum) and garlic (A. sativum). A disease severity index (DSI) was calculated as the mean severity on three plants of each species with four test replicates. Symptoms on onion and garlic plants were observed three weeks after inoculation. All isolates tested produced symptoms on all varieties inoculated. Inoculation of F. proliferatum isolates from diseased garlic onto other Allium species provided new information on host range and pathogenicity. The results demonstrated differences in susceptibility with respect to host species and cultivar. The F. proliferatum isolates tested all produced fusaric acid (FA); correlations between FA production and isolate pathogenicity are discussed. Additionally, all isolates showed the presence of the FUM1 gene suggesting the ability of Spanish isolates to produce fumonisins.

In vitro evaluation of commercial fibrolytic enzymes for improving the nutritive value of low-quality forages.

The aim of this work was to assess the effects of four doses of three commercial fibrolytic enzymes on ruminal fermentation of rice straw, maize stover and Pennisetum purpureum clon Cuba CT115 hay in batch cultures of ruminal micro-organisms from sheep. One enzyme was produced by Penicillium funiculosum (PEN) and two were from Trichoderma longibrachiatum (TL1 and TL2). Each liquid enzyme was diluted 200 (D1), 100 (D2), 50 (D3) and 10 (D4) - fold and applied to each substrate in quadruplicate over time and incubated for 120 h in rumen fluid. The D4 dose of each enzyme increased (P<0.05) the fractional rate of gas production and organic matter effective degradability for all substrates, and TL2 had similar effects when applied at D3. In 9 h incubations, PEN at D4, TL1 at all tested doses, and TL2 at D2, D3 and D4 increased (P<0.05) volatile fatty acid production and dry matter degradability for all substrates. The commercial enzymes tested were effective at increasing in vitro ruminal fermentation of low-quality forages, although effective doses varied with the enzyme.

New Schizosaccharomyces applications in red wines alcoholic fermentation

The Schizosaccharomyces strains consumed less primary amino nitrogen and produced less urea and more pyruvic acid than other Saccharomyces species. Further, three of the four Schizosaccharomyces strains completed the breakdown of malic acid by day 4 of fermentation. The main negative effect of the use of Schizosaccharomyces was strong acetic acid production. The Schizosaccharomyces strains that produced most pyruvic acid (938 and 936) were associated with better ?wine? colour than the remaining yeasts. The studied Schizosaccharomyces could therefore be of oenological interest.

Nutritive value for ruminants of winter oats-legume intercrops in organic cultivation

Winter oats were grown according to European organic farming regulations in monoculture (oats) and in intercropping with bard vetch (BAV), bitter vetch (BIV) or both legumes (MIX) to evaluate the effects of intercropping on forage yield and nutritive value for ruminants. The experiment was carried out as a randomised complete block design with four replications, and whole forage samples were obtained at two harvest dates (June and July). For both harvest times, all intercrops increased (P < 0.05) forage yield compared with oats, but forage crude protein content was only increased (P < 0.05) for BAV and MIX. Compared with oats, intercropping with BAV increased (P < 0.05) in vitro rate of gas production and total volatile fatty acid production, indicating a higher rate and extent of rumen degradation of BAV forage. In contrast, BIV forage harvested in June had lower (P < 0.05) rate of gas production and total volatile fatty acid production than June oats, but in general no differences in the in vitro rumen fermentation were detected between oats and BIV samples harvested in July. The results indicate that forage yield and quality can be enhanced by intercropping oats with BAV; however, intercropping with BIV increased yield but decreased nutritive value of the forage.

Biokerosene from Babassu and Camelina Oils: production and properties of their blends with fossil kerosene

Babassu and camelina oils have been transesterified with methanol by the classical homogeneous basic catalysis method with good yields. The babassu fatty acid methyl ester (FAME) has been subjected to fractional distillation at vacuum, and the low boiling point fraction has been blended with two types of fossil kerosene, a straight-run atmospheric distillation cut (hydrotreated) and a commercial Jet-A1. The camelina FAME has been blended with the fossil kerosene without previous distillation. The blends of babassu biokerosene and Jet-A1 have met some of the specifications selected for study of the ASTM D1655 standard: smoke point, density, flash point, cloud point, kinematic viscosity, oxidative stability and lower heating value. On the other hand, the blends of babassu biokerosene and atmospheric distillation cut only have met the density parameter and the oxidative stability. The blends of camelina FAME and atmospheric distillation cut have met the following specifications: density, kinematic viscosity at −20 °C, and lower heating value. With these preliminary results, it can be concluded that it would be feasible to blend babassu and camelina biokerosenes prepared in this way with commercial Jet-A1 up to 10 vol % of the former, if these blends prove to accomplish all the ASTM D1655-09 standards.

The influence of diet on the effectiveness of garlic oil and cinnamaldehyde to manipulate in vitro ruminal fermentation and methane production.

The objective of this study was to evaluate the effects of increasing doses [0 (control: CON), 20, 60, 180 and 540 mg/L incubation medium] of garlic oil (GO) and cinnamaldehyde (CIN) on in vitro ruminal fermentation of two diets. Batch cultures of mixed ruminal microorganisms were inoculated with ruminal fluid from four sheep fed a medium-concentrate diet (MC; 50 : 50 alfalfa hay : concentrate) or four sheep fed a high-concentrate diet (HC; 15 : 85 barley straw : concentrate). Diets MC and HC were representative of those fed to dairy and fattening ruminants, respectively. Samples of each diet were used as incubation substrates for the corresponding inoculum, and the incubation was repeated on 4 different days (four replicates per experimental treatment). There were GO × diet-type and CIN × diet-type interactions (P < 0.001–0.05) for many of the parameters determined, indicating different effects of both oils depending on the diet type. In general, effects of GO were more pronounced for MC compared with HC diet. Supplementation of GO did not affect (P > 0.05) total volatile fatty acid (VFA) production at any dose. For MC diet, GO at 60, 180 and 540 mg/L decreased (P < 0.05) molar proportion of acetate (608, 569 and 547 mmol/mol total VFA, respectively), and increased (P < 0.05) propionate proportion (233, 256 and 268 mmol/mol total VFA, respectively), compared with CON values (629 and 215 mmol/mol total VFA for acetate and propionate, respectively). A minimum dose of 180 mg of GO/L was required to produce similar modifications in acetate and propionate proportions with HC diet, but no effects (P > 0.05) on butyrate proportion were detected. Methane/VFA ratio was reduced (P < 0.05) by GO at 60, 180 and 540 mg/L for MC diet (0.23, 0.16 and 0.10 mol/mol, respectively), and by GO at 20, 60, 180 and 540 mg/L for HC diet (0.19, 0.19, 0.16 and 0.08 mol/mol, respectively), compared with CON (0.26 and 0.21 mol/mol for MC and HC diets, respectively). No effects (P = 0.16–0.85) of GO on final pH and concentrations of NH3-N and lactate were detected. For both diet types, the highest CIN dose decreased (P < 0.05) production of total VFA, gas and methane, which would indicate an inhibition of fermentation. Compared with CON, CIN at 180 mg/L increased (P < 0.05) acetate proportion for the MC (629 and 644 mmol/mol total VFA for CON and CIN, respectively) and HC (525 and 540 mmol/mol total VFA, respectively) diets, without affecting the proportions of any other VFA or total VFA production. Whereas for MC diet CIN at 60 and 180 mg/L decreased (P < 0.05) NH3-N concentrations compared with CON, only a trend (P < 0.10) was observed for CIN at 180 mg/L with the HC diet. Supplementation of CIN up to 180 mg/L did not affect (P = 0.18–0.99) lactate concentrations and production of gas and methane for any diet. The results show that effectiveness of GO and CIN to modify ruminal fermentation may depend on diet type, which would have practical implications if they are confirmed in vivo.

Combine use of Selected Schizosaccharomyces pombe and Lachancea thermotolerans Yeast Strains as an Alternative to the Traditional Malolactic Fermentation in Red Wine Production

Most red wines commercialized in the market use the malolactic fermentationprocess in order to ensure stability from a microbiological point of view. In this secondfermentation, malic acid is converted into L-lactic acid under controlled setups. Howeverthis process is not free from possible collateral effects that on some occasions produceoff-flavors, wine quality loss and human health problems. In warm viticulture regions suchas the south of Spain, the risk of suffering a deviation during the malolactic fermentationprocess increases due to the high must pH. This contributes to produce wines with highvolatile acidity and biogenic amine values. This manuscript develops a new red winemakingmethodology that consists of combining the use of two non-Saccharomyces yeast strains asan alternative to the traditional malolactic fermentation. In this method, malic acid is totallyconsumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilizationobjective, while Lachancea thermotolerans produces lactic acid in order not to reduce andeven increase the acidity of wines produced from low acidity musts. This technique reducesthe risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than thetraditional controls that have undergone the classical malolactic fermentation.

Amelioration of a degraded acid soil from SW Spain by no-tillage and Ca-amendment

En los suelos, el exceso de acidez lleva asociado deficiencias en ciertos nutrientes y una alta disponibilidad de aluminio, tóxico para los cultivos propios del ambiente mediterráneo. Su laboreo, provoca la pérdida de materia orgánica (MO), deteriora su estructura y reduce la actividad biológica, provocando en última instancia una menor calidad del suelo. Es de esperar pues que cuando se labran suelos ácidos, sus problemáticas particulares tiendan a agravarse. En nuestra zona de estudio, la “raña” de Cañamero (Extremadura, España), predominan los suelos muy ácidos y degradados por un laboreo inadecuado. Las rañas constituyen amplias plataformas casi horizontales, con unos suelos muy viejos (Palexerults), que se caracterizan por tener el complejo de cambio dominado por el aluminio, y un pH ácido que decrece en profundidad. Poseen un potente horizonte Bt rico en arcillas caoliníticas, que propicia que en periodos con exceso de lluvia, se generen capas colgadas de agua cercanas a la superficie. En torno a los años 1940’s estos suelos, que previamente sostenían un alcornocal, o su matorral de sustitución, se pusieron en cultivo. El laboreo aceleró la mineralización de la materia orgánica, agravó los problemas derivados del exceso de acidez y condujo al abandono de los campos cultivados por falta de productividad. Para recuperar la calidad de estos suelos degradados y obtener unos rendimientos compatibles con su uso agrícola es necesario, por un lado, aplicar enmiendas que eleven el pH y reduzcan la toxicidad del aluminio y, por otro, favorecer el incremento en el contenido en MO. En 2005 se implantó en esta raña un ensayo de campo para estudiar la influencia del no laboreo y de la utilización de una enmienda cálcica en parámetros relacionados con la calidad del suelo en un cultivo forrajero. El diseño experimental fue en parcelas divididas con cuatro repeticiones donde el factor principal fue el tipo de laboreo, no laboreo (NL) frente a laboreo convencional (LC), y el factor secundario el uso o no de una enmienda cálcica. La enmienda consistió básicamente en una mezcla de espuma de azucarería y yeso rojo y se incorporó al comienzo del ensayo hasta los 7 cm de profundidad. Desde el comienzo del ensayo el NL influyó positivamente en el contenido de carbono orgánico total (COT) y particulado (COP), mientras que la enmienda tuvo una ligera influencia al principio del ensayo en ambos pero su efecto positivo se desvaneció con el paso del tiempo. Los mayores contenidos en COT y POC se observaron cuando se combinó el NL con la enmienda. La enmienda incrementó con rapidez el pH, y el Ca, y disminuyó el contenido en aluminio hasta una profundidad de 50 cm, incluso en NL, y mejoró ligeramente la agregación del suelo. El NL por sí solo, gracias al aumento en POC, TOC y las proteínas del suelo relacionadas con la glomalina (PSRG), que son capaces de formar compuestos estables no tóxicos con el aluminio, también contribuyó a la reducción de la toxicidad de aluminio en la capa más superficial. Cuando en las campañas con exceso de precipitaciones se generaron capas colgadas de agua próximas a la superficie, el NL generó unas condiciones más favorables para la germinación y desarrollo del cultivo, resultando en una producción más alta que el LC. A ello contribuyó la mayor capacidad de almacenamiento de agua y la mayor transmisividad de esta hacia abajo, en la capa más superficial (0-5 cm) que propició una menor saturación por agua que el LC. Respecto a los parámetros relacionados con la agregación, el NL aumentó los macroagregados hasta los 10 cm de profundidad y favoreció la acumulación de CO y N en todas las fracciones de tamaño de agregados. Sin embargo, la recuperación del grado de macroagregación tras el cese del laboreo resulta lenta en comparación con otros suelos, posiblemente debido al bajo contenido en arcilla en el horizonte Ap. En comparación con el NL, la enmienda mostró también un efecto positivo, aunque muy ligero, en la agregación del suelo. En contradicción con otros estudios en suelos ácidos, nuestros resultados indican la existencia de una jerarquía de agregados, y destacan el papel importante de la MO en la mejora de la agregación. Tanto el NL como la enmienda favorecieron por separado varias propiedades químicas, físicas y biológicas del suelo, pero, en general, encontramos los mayores beneficios con su uso combinado. Además, a largo plazo el efecto positivo de NL en las propiedades del suelo fue en aumento, mientras que el efecto beneficioso de la enmienda se limitó básicamente a las propiedades químicas y se desvaneció en pocos años. Destacamos que las condiciones meteorológicas a lo largo del ensayo beneficiaron la producción de biomasa en NL, y en consecuencia las propiedades relacionadas con la materia orgánica, por lo que son un factor a tener en cuenta a la hora de evaluar los efectos de la enmienda y el laboreo sobre las propiedades del suelo, especialmente en zonas donde esas condiciones son muy variables entre una campaña y otra. Los resultados de este estudio han puesto de manifiesto que el NL no ha mermado la eficacia de la enmienda caliza, posiblemente gracias a la alta solubilidad de la enmienda aplicada, es más, el manejo con NL y enmienda es el que ha favorecido en mayor medida ciertos parámetros de calidad del suelo. Por el contrario el LC sí parece anular los beneficios de la enmienda en relación con las propiedades relacionadas con la MO. Por tanto, cabe concluir que la combinación de NL y la enmienda es una práctica adecuada para mejorar las propiedades químicas y físicas de suelos ácidos degradados por el laboreo. ABSTRACT Excessive acidity in soils is associated with deficiencies in certain nutrients and high concentrations of available aluminum, which is toxic for most Mediterranean crops. Tilling these soils results in the loss of soil organic matter (SOM), damages soil structure and reduces biological activity, ultimately degrading soil quality. It is expected, therefore, that when acid soils are tilled, their particular problems will tend to get worse. In our study area, the "Cañamero’s Raña” (Extremadura, Spain), acid soils degraded by an inappropriate tillage prevail. Rañas are large and flat platforms with very old soils (Palexerults), which are characterized by an exchange complex dominated by aluminum and an acid pH which decreases with depth. These soils have a strong Bt horizon rich in kaolinite clays, which encourages the formation of perched water-tables near the soil surface during periods of excessive rain. During the first third of the 20th century, these soils, that previously supported cork oak or its scrub replacement, were cultivated. Tillage accelerated the mineralization of the SOM, aggravating the problems of excessive acidity, which finally led to the abandonment of the land due to low productivity. To recover the quality of these degraded soils and to obtain consistent yields it is necessary, first, to apply amendments to raise the pH and reduce aluminum toxicity, and second to encourage the accumulation of SOM. In 2005 a field trial was established in the Raña to study the influence of no-tillage and the use of a Ca-amendment on soil quality related parameters in a forage crop agrosystem. The experimental design was a split-plot with four replicates where the main factor was tillage type, no-tillage (NT) versus traditional tillage (TT) and the secondary factor was the use or not of a Ca-amendment. The Ca-amendment was a mixture of sugar foam and red gypsum that was incorporated into the top 7 cm of the soil. Since the beginning of the experiment, NT had a positive influence on total and particulate organic carbon (TOC and POC, respectively), while the Ca-amendment had a small positive influence at the beginning of the study but its effect diminished with time. The highest TOC and POC contents were observed when NT and the Ca-amendment were combined. The Ca-amendment, even under NT, rapidly increased pH and Ca, and decreased the aluminum content to a depth of 50 cm, as well as improving soil aggregation slightly. NT, due to the increased POC, TOC and Glomalin-related soil proteins (GRSP), which can form stable non-toxic compounds with aluminum, also contributed to the reduction of aluminum toxicity in the upper layer. When perched water-tables near the soil surface were formed in campaigns with excessive rainfall, NT provided more favorable conditions for germination and crop development, resulting in higher yields compared with TT. This was directly related to the higher water storage capacity and the greater transmissivity of the water downwards from the upper layers, which led to lower water saturation under NT compared with TT. With regards to the aggregation-related parameters, NT increased macroaggregation to a depth of 10 cm and favored the accumulation of OC and N in all aggregate size fractions. However, the degree of recovery of macroaggregation after tillage ceased was slow compared with other soils, possibly due to the low clay content in the Ap horizon. Compared with NT, the Ca-amendment had a slight positive effect on soil aggregation. In contrast to other studies in acid soils, our results indicate the existence of an aggregate hierarchy, and highlight the important role of SOM in improving aggregation. Both NT and the Ca-amendment separately favored various chemical, physical and biological soil properties, but in general we found the greatest benefits when the two treatments were combined. In addition, the positive effect of NT on soil properties increased with time, while the beneficial effect of the Ca-amendment, which was limited to the chemical properties, vanished after a few years. It is important to note that the meteorological conditions throughout the experiment benefited biomass production under NT and, as a consequence, organic matter related properties. This suggests that meteorological conditions are a factor to consider when evaluating the effects of Ca-amendments and tillage on soil properties, especially in areas where such conditions vary significantly from one campaign to another. The results of this study show that NT did not diminish the effectiveness of the Ca-amendment, possibly due to the high solubility of the selected amendment. Moreover, the combination of NT and the Ca-amendment was actually the management that favored certain soil quality parameters the most. By contrast, TT seemed to nullify the benefits of the Ca-amendment with regards to the OM related properties. In conclusion, the combination of NT and the application of a Ca-amendment is an advisable practice for improving the chemical and physical properties of acid soils degraded by tillage.

Sunflower meal and spring pea ruminal degradation protection using malic acid or orthophosphoric acid-heat treatments

The effects of solutions of malic or orthophosphoric acids (0.752 Eqg/kg of feed) and heat to protect proteins of sunflower meal (SFM) and spring pea (SP) against ruminal degradation were studied using particle transit, 15N infusion, in situ and electrophoretic techniques. Three wethers fitted with rumen and duodenum cannulae were successively fed three isoproteic diets including SFM and SP, untreated or treated with malic or orthophosphoric acids. Incubations of tested meals were only performed while feeding the respective diet. Estimates of the ruminally undegraded fraction (RU) and its intestinal digestibility of dry matter, organic matter (only for RU), crude protein and starch (only in SP) were obtained considering ruminal microbial contamination and particle comminution and outflow rates. When corrected for microbial contamination, estimates of RU and intestinal digestibility decreased in all tested fractions for both feeds. All RU estimates increased with the protective treatments, whereas intestinal digestibility-dry matter also increased in SFM. Low intestinal digestibility-crude protein values suggested the presence of antitrypsin factors in SP. Protective treatments of both feeds led to consistent increases in the intestinal digested fraction of dry matter and crude protein, being only numerically different for SP-starch (60.5% as average). However, treatments also reduced the organic matter fermentation, which may decrease ruminal microbial protein synthesis. Electrophoretic studies showed albumin disappearance in both SFM and SP, whereas changes in other RU proteins were more pronounced in SP than SFM.

Effect of level of fiber of the rearing phase diets on egg production, digestive tract traits, and body measurements of brown egg-laying hens fed diets differing in energy concentration

This research studied the effects of additional fiber in the rearing phase diets on egg production, gastrointestinal tract (GIT) traits, and body measurements of brown egg-laying hens fed diets varying in energy concentration from 17 to 46 wk of age. The experiment was completely randomized with 10 treatments arranged as a 5 × 2 factorial with 5 rearing phase diets and 2 laying phase diets. During the rearing phase, treatments consisted of a control diet based on cereals and soybean meal and 4 additional diets with a combination of 2 fiber sources (cereal straw and sugar beet pulp, SBP) at 2 levels (2 and 4%). During the laying phase, diets differed in energy content (2,650 vs. 2,750 kcal AMEn/kg) but had the same amino acid content per unit of energy. The rearing diet did not affect any production trait except egg production that was lower in birds fed SBP than in birds fed straw (91.6 and 94.1%, respectively; P < 0.05). Laying hens fed the high energy diet had lower feed intake (P < 0.001), better feed conversion (P < 0.01), and greater BW gain (P < 0.05) than hens fed the low energy diet but egg production and egg weight were not affected. At 46 wk of age, none of the GIT traits was affected by previous dietary treatment. At this age, hen BW was positively related with body length (r = 0.500; P < 0.01), tarsus length (r = 0.758; P < 0.001), and body mass index (r = 0.762; P < 0.001) but no effects of type of diet on these traits were detected. In summary, the inclusion of up to 4% of a fiber source in the rearing diets did not affect GIT development of the hens but SBP reduced egg production. An increase in the energy content of the laying phase diet reduced ADFI and improved feed efficiency but did not affect any of the other traits studied.

Production of biodiesel from animal fat using supercritical ethanol

Biodiesel is currently produced from a catalytic transesterification reaction of various types of edible and non-edible oil with methanol. The use of waste animal tallow instead of edible oils opens a route to recycle this waste. This material has the advantage of lower costs but the problem of high content of free fatty acids, becoming necessary a pre-esterification reaction that increases the cost of the catalytic process. The production of biodiesel using supercritical alcohols is appropriate for materials with high acidity and water content, therefore the use of this process with animal fat is a promising alternative. Ethanol has been used because it can be produced from biomass via fermentation resulting in a complete renewable biodiesel, instead of methanol that derives from fossil feedstocks. Two different processes have been studied: first, the direct transesterification of animal fat using supercritical ethanol and second a two-step process where the first step is a hydrolysis of the animal fat and the second step is the esterification of the resulting fatty acids. The temperature, the molar ratio ethanol:fat and the time have been modified in the different reactions to study the effect in the final conversion and the degradation of the unsaturated fatty acid esters, main inconvenient of these high temperature and pressure processes.