Potential of Mortierellaceae for polyunsaturated fatty acids production: mini review.

The health benefits of polyunsaturated fatty acids (PUFAs) have encouraged the search for rich sources of these compounds. However, the supply chain of PUFAs from animals and plants presents environmental concerns, such as water pollution, deforestation, animal exploitation and interference in the trophic chain. In this way, a viable alternative has been found in microbial sources, mainly in single cell oil (SCO) production by yeast and filamentous fungi. Mortierellaceae is a filamentous fungal family world-renowned for PUFA-producing strains. For example, Mortierella alpina can be highlighted due to be industrially applied to produce arachidonic acid (20:4 n6), an important component of infant supplement formulas. Thus, the state of the art of strategies to increase PUFAs production by Mortierellaceae strains is presented in this review. Firstly, we have discussed main phylogenetic and biochemical characteristics of these strains for lipid production. Next, strategies based on physiological manipulation, using different carbon and nitrogen sources, temperature, pH and cultivation methods, which can increase PUFA production by optimizing process parameters are presented. Furthermore, it is possible to use metabolic engineering tools, controlling the supply of NADPH and co-factors, and directing the activity of desaturases and elongase to the target PUFA. Thus, this review aims to discuss the functionality and applicability of each of these strategies, in order to support future research for PUFA production by Mortierellaceae species.

Anti-Trypanosoma cruzi Properties of Sesquiterpene Lactones Isolated from Stevia spp.: In Vitro and In Silico Studies.

Stevia species (Asteraceae) have been a rich source of terpenoid compounds, mainly sesquiterpene lactones, several of which show antiprotozoal activity. In the search for new trypanocidal compounds, S. satureiifolia var. satureiifolia and S. alpina were studied. Two sesquiterpene lactones, santhemoidin C and 2-oxo-8-deoxyligustrin, respectively, were isolated. These compounds were assessed in vitro against Trypanosoma cruzi stages, showing IC50 values of 11.80 and 4.98 on epimastigotes, 56.08 and 26.19 on trypomastigotes and 4.88 and 20.20 µM on amastigotes, respectively. Cytotoxicity was evaluated on Vero cells by the MTT assay. The effect of the compounds on trypanothyone reductase (TcTR), Trans-sialidase (TcTS) and the prolyl oligopeptidase of 80 kDa (Tc80) as potential molecular targets of T. cruzi was investigated. Santhemoidin C inhibited oligopeptidase activity when tested against recombinant Tc80 using a fluorometric assay, reaching an IC50 of 34.9 µM. Molecular docking was performed to study the interaction between santhemoidin C and the Tc80 protein, reaching high docking energy levels. Plasma membrane shedding and cytoplasmic vacuoles, resembling autophagosomes, were detected by transmission microscopy in parasites treated with santhemoidin C. Based on these results, santhemoidin C represents a promising candidate for further studies in the search for new molecules for the development of trypanocidal drugs.

In vitro cytotoxic effect of a chitin-like polysaccharide produced by Mortierella alpina on adrenocortical carcinoma cells H295R, and its use as mitotane adjuvant.

This study presents an in vitro evaluation of the antitumor potential of a chitin-like exopolysaccharide (EPS, produced by Mortierella alpina) on Adrenocortical carcinoma cells (ACC) compared to mitotane, a commercial drug commonly used in ACC treatment, and known for its side effects. Techniques of cellular viability determination such as MTT and fluorescence were used to measure the cytotoxic effects of the EPS and mitotane in tumoral cells (H295R) and non-tumoral cells (VERO), observing high cytotoxicity of mitotane and a 10% superior pro-apoptotic effect of the EPS compared to mitotane (p < 0.05). The cytotoxic effect of the EPS was similar to the effect of 50 µM mitotane on tumoral cells (p < 0.05). A decrement of the lysosomal volume was also noted in tumoral cells treated with the EPS. To enhance the antitumor effect, a combination of mitotane at a lower dosage and the EPS (as adjuvant) was also tested, showing a slight improvement of the cytotoxicity effect on tumoral cells. Therefore, the results indicate a cytotoxic effect of the EPS produced by Mortierella alpina on adrenocortical carcinoma, and a possible application in biomedical formulations or additional treatments.

Production, characterization, and biological activity of a chitin-like EPS produced by Mortierella alpina under submerged fermentation.

The production of a chitin-like exopolysaccharide (EPS) was optimized through experimental design methods, evaluating the influence of urea, phosphate, and glucose. Under optimized conditions, up to 1.51 g/L was produced and its physicochemical characteristics were evaluated by chromatography, NMR, and FTIR spectroscopy, and rheological techniques. The results showed a homogeneous EPS (Mw 4.9 × 105 g mol-1) composed of chitin, linear polymer of ß-(1→4)-linked N-acetyl-d-glucosamine residues. The acetylation degree as determined by 13C CP-MAS NMR spectroscopy was over 90 %. The EPS biological activities, such as antioxidant effect and antitumor properties, were evaluated. To the best of our knowledge, this is the first study on the production of a new alternative of extracellular chitin-like polysaccharide with promising bioactive properties from the filamentous fungus M. alpina.

Activity of Estafietin and Analogues on Trypanosoma cruzi and Leishmania braziliensis.

Sesquiterpene lactones are naturally occurring compounds mainly found in the Asteraceae family. These types of plant metabolites display a wide range of biological activities, including antiprotozoal activity and are considered interesting structures for drug discovery. Four derivatives were synthesized from estafietin (1), isolated from Stevia alpina (Asteraceae): 11ßH,13-dihydroestafietin (2), epoxyestafietin (3a and 3b), 11ßH,13-methoxyestafietin, (4) and 11ßH,13-cianoestafietin. The antiprotozoal activity against Trypanosoma cruzi and Leishmania braziliensis of these compounds was evaluated. Epoxyestafietin was the most active compound against T. cruzi trypomastigotes and amastigotes (IC50 values of 18.7 and 2.0 µg/mL, respectively). Estafietin (1) and 11ßH,13-dihydroestafietin (2) were the most active and selective compounds on L. braziliensis promastigotes (IC50 values of 1.0 and 1.3 µg/mL, respectively). The antiparasitic activity demonstrated by estafietin and some of its derivatives make them promising candidates for the development of effective compounds for the treatment of Chagas disease and leihsmaniasis.

Long-term in situ persistence of biodiversity in tropical sky islands revealed by landscape genomics.

Tropical mountains are areas of high species richness and endemism. Two historical phenomena may have contributed to this: (i) fragmentation and isolation of habitats may have promoted the genetic differentiation of populations and increased the possibility of allopatric divergence and speciation and (ii) the mountain areas may have allowed long-term population persistence during global climate fluctuations. These two phenomena have been studied using either species occurrence data or estimating species divergence times. However, only few studies have used intraspecific genetic data to analyse the mechanisms by which endemism may emerge at the microevolutionary scale. Here, we use landscape analysis of genomic SNP data sampled from two high-elevation plant species from an archipelago of tropical sky islands (the Trans-Mexican Volcanic Belt) to test for population genetic differentiation, synchronous demographic changes and habitat persistence. We show that genetic differentiation can be explained by the degree of glacial habitat connectivity among mountains and that mountains have facilitated the persistence of populations throughout glacial/interglacial cycles. Our results support the ongoing role of tropical mountains as cradles for biodiversity by uncovering cryptic differentiation and limits to gene flow.

Pollen and stigma size changes during the transition from tristyly to distyly in Oxalis alpina (Oxalidaceae).

Pollen and stigma size have the potential to influence male fitness of hermaphroditic plants, particularly in species presenting floral polymorphisms characterised by marked differences in these traits among floral morphs. In this study, we take advantage of the evolutionary transition from tristyly to distyly experienced by Oxalis alpina (Oxalidaceae), and examined whether modifications in the ancillary traits (pollen and stigma size) respond to allometric changes in other floral traits. Also, we tested whether these modifications are in accordance with what would be expected under the hypothesis that novel competitive scenarios (as in distylous-derived reproductive system) exert morph- and whorl-specific selective pressures to match the available stigmas. We measure pollen and stigma size in five populations of O. alpina representing the tristyly-distyly transition. A general reduction in pollen and stigma size occurred along the tristyly-distyly transition, and pollen size from the two anther levels within each morph converged to a similar size that was characterised by whorl-specific changes (increases or decreases) in pollen size of different anthers in each floral type. Overall, results from this study show that the evolution of distyly in this species is characterised not only by changes in sexual organ position and flower size, but also by morph-specific changes in pollen and stigma size. This evidence supports the importance of selection on pollen and stigma size, which increase fitness of remaining morphs following the evolution of distyly, and raises questions to explore on the functional value of pollen size in heterostylous systems under pollen competition.

Relative growth of carcass tissues of goat kids from five breed Types finished on pasture or feedlot / Crescimento relativo dos tecidos da carcaça de cabritos de cinco grupos raciais terminados em pastagem ou confinamento

The aim of this study was to evaluate the effect of cross breeding, finishing system, and gender on the relative growth of carcass tissues of dairy kids. Seventy eight kids (39 male and 39 female) from five breed types were used: Alpine; ½ Boer + ½ Alpine (½ BA); ½ Nubian + ½ Alpine (½ ANA); ¾ Boer + » Alpine (¾ BA); and ½ Nubian + » Boer + » Alpine (TC). Kids were distributed into two finishing systems: in pasture with doe (FS1) and weaned in feedlot (FS2). Kids were slaughtered at a mean age of 128.4 ± 7.9 days and mean live weight of 22.07 kg. The mean weight of half carcasses was 5.09 kg. To determine allometric growth, we used the exponential equation Y= aXb. In the half carcass, muscle tissue showed comparatively early growth in group ½ BA, whereas fat tissue of animals in FS1 had relatively late growth. Females exhibited early growth of muscle tissue, while in males this tissue was intermediate. The ½ BA first-cross improved carcass characteristics by enhancing the growth of muscle tissue.(AU)

Este trabalho foi realizado com o objetivo de avaliar a influência dos cruzamentos, sistema de terminação e sexo no crescimento relativo dos tecidos da carcaça de cabritos em crescimento. Foram utilizados 78 cabritos, 39 machos e 39 fêmeas, de cinco grupos raciais: 13 cabritos Alpinos; 14 1/2 Boer + 1/2 Alpino (1/2 BA); 15 1/2 Anglo Nubiano + 1/2 Alpino (1/2 ANA); 18 3/4 Boer + 1/4 Alpino (3/4 BA); e 18 ½ Anglo Nubiano + » Boer + » Alpino (TC). Os grupos foram distribuídos em dois sistemas de terminação, cabrito em pastagem com a mãe (ST1) e cabrito desmamado em confinamento (ST2). Os animais foram abatidos em média aos 22,07 kg de peso vivo e 128,4 ± 7,9 dias. O peso médio das meias carcaças foi de 5,09 kg. Para determinação do crescimento alométrico foi utilizada a equação exponencial Y = aXb. O tecido muscular, em relação ao peso da meia carcaça, apresentou crescimento precoce no grupo ½ BA. O tecido adiposo dos animais do ST1 foi depositado tardiamente. As fêmeas apresentaram crescimento precoce de tecido muscular, em relação ao peso da meia carcaça, enquanto nos machos tal comportamento foi intermediário. O cruzamento 1/2 BA pode melhorar as características da carcaça em função de potencializar o crescimento do tecido muscular.(AU)

Relative growth of carcass tissues of goat kids from five breed Types finished on pasture or feedlot / Crescimento relativo dos tecidos da carcaça de cabritos de cinco grupos raciais terminados em pastagem ou confinamento

The aim of this study was to evaluate the effect of cross breeding, finishing system, and gender on the relative growth of carcass tissues of dairy kids. Seventy eight kids (39 male and 39 female) from five breed types were used: Alpine; ½ Boer + ½ Alpine (½ BA); ½ Nubian + ½ Alpine (½ ANA); ¾ Boer + » Alpine (¾ BA); and ½ Nubian + » Boer + » Alpine (TC). Kids were distributed into two finishing systems: in pasture with doe (FS1) and weaned in feedlot (FS2). Kids were slaughtered at a mean age of 128.4 ± 7.9 days and mean live weight of 22.07 kg. The mean weight of half carcasses was 5.09 kg. To determine allometric growth, we used the exponential equation Y= aXb. In the half carcass, muscle tissue showed comparatively early growth in group ½ BA, whereas fat tissue of animals in FS1 had relatively late growth. Females exhibited early growth of muscle tissue, while in males this tissue was intermediate. The ½ BA first-cross improved carcass characteristics by enhancing the growth of muscle tissue.

Este trabalho foi realizado com o objetivo de avaliar a influência dos cruzamentos, sistema de terminação e sexo no crescimento relativo dos tecidos da carcaça de cabritos em crescimento. Foram utilizados 78 cabritos, 39 machos e 39 fêmeas, de cinco grupos raciais: 13 cabritos Alpinos; 14 1/2 Boer + 1/2 Alpino (1/2 BA); 15 1/2 Anglo Nubiano + 1/2 Alpino (1/2 ANA); 18 3/4 Boer + 1/4 Alpino (3/4 BA); e 18 ½ Anglo Nubiano + » Boer + » Alpino (TC). Os grupos foram distribuídos em dois sistemas de terminação, cabrito em pastagem com a mãe (ST1) e cabrito desmamado em confinamento (ST2). Os animais foram abatidos em média aos 22,07 kg de peso vivo e 128,4 ± 7,9 dias. O peso médio das meias carcaças foi de 5,09 kg. Para determinação do crescimento alométrico foi utilizada a equação exponencial Y = aXb. O tecido muscular, em relação ao peso da meia carcaça, apresentou crescimento precoce no grupo ½ BA. O tecido adiposo dos animais do ST1 foi depositado tardiamente. As fêmeas apresentaram crescimento precoce de tecido muscular, em relação ao peso da meia carcaça, enquanto nos machos tal comportamento foi intermediário. O cruzamento 1/2 BA pode melhorar as características da carcaça em função de potencializar o crescimento do tecido muscular.

Expression analysis for genes involved in arachidonic acid biosynthesis in Mortierella alpina CBS 754.68.

The time courses for production of fungal biomass, lipid, phenolic and arachidonic acid (ARA) as well as expression of the genes involved in biosynthesis of ARA and lipid were examined in Mortierella alpina CBS 754.68. A significant increase in the arachidonic acid content in lipids that coincided with reduced levels of lipid was obtained. Reduced gene expression occurred presumably due to the steady reduction of carbon and nitrogen resources. However, these energy resources were inefficiently compensated by the breakdown of the accumulated lipids that in turn, induced up-regulated expression of the candidate genes. The results further indicated that the expression of the GLELO encoding gene is a rate-limiting step in the biosynthesis of ARA in the early growth phase.

Expression analysis for genes involved in arachidonic acid biosynthesis in Mortierella alpina CBS 754.68

The time courses for production of fungal biomass, lipid, phenolic and arachidonic acid (ARA) as well as expression of the genes involved in biosynthesis of ARA and lipid were examined in Mortierella alpina CBS 754.68. A significant increase in the arachidonic acid content in lipids that coincided with reduced levels of lipid was obtained. Reduced gene expression occurred presumably due to the steady reduction of carbon and nitrogen resources. However, these energy resources were inefficiently compensated by the breakdown of the accumulated lipids that in turn, induced up-regulated expression of the candidate genes. The results further indicated that the expression of the GLELO encoding gene is a rate-limiting step in the biosynthesis of ARA in the early growth phase.

Expression analysis for genes involved in arachidonic acid biosynthesis in Mortierella alpina CBS 754.68

The time courses for production of fungal biomass, lipid, phenolic and arachidonic acid (ARA) as well as expression of the genes involved in biosynthesis of ARA and lipid were examined in Mortierella alpina CBS 754.68. A significant increase in the arachidonic acid content in lipids that coincided with reduced levels of lipid was obtained. Reduced gene expression occurred presumably due to the steady reduction of carbon and nitrogen resources. However, these energy resources were inefficiently compensated by the breakdown of the accumulated lipids that in turn, induced up-regulated expression of the candidate genes. The results further indicated that the expression of the GLELO encoding gene is a rate-limiting step in the biosynthesis of ARA in the early growth phase.

Influence of region and season on kid behavior on pasture / Influência do local e estação do ano sobre o comportamento de cabritos a pasto

The behavior (frequency of animais grazing) and physiological profiles of anglonubians, alpine x anglonubians and saanen x anglonubians kids were studied during the summer (january/ february) and winter (june/july) on two different places with different weather conditions (Itapetininga and Nova Odessa in the State of São Paulo). The breed didnt affect the frequency distribution (FD) taken every 30 minutes of animals grazing, from 7:30am to 6:30pm. Places, season and grass availability were significative (P 0.01) for FD. At 8:00am with the animals at rest, the breed didnt show difference (P > 0.05) for rectal temperature (RT) and respiratory rate (RR). At 2:00pm also at rest, the RT and RR were higher (P 0.01) for the saanen crossbred, when compared to the other breeds and also, higher (P 0.01) for the anglonubians when compared to the alpine crossbred. Places and season affected (P 0.01) RT and RR independently of the time of the day. The hematological data were not influenced by breed, places and season, and showed the species standard profile.

Estudou-se o comportamento (freqüência de animais em pastejo) e os aspectos fisiológicos de cabritas anglonubianas, pardo alpino x anglonubianas (pardos) e saanen x anglonubianas (brancos), em pastagem, durante o verão (janeiro e fevereiro) e inverno (junho e julho), em dois locais de condições climáticas distintas (Itapetininga e Nova Odessa). Observou-se que o tipo racial não influenciou a distribuição da freqüência de animais em pastejo, observada a cada 30 minutos, das 7 às 19h30. Todavia, verificou-se efeito significativo (P 0,01) do local, da estação do ano e notadamente da disponibilidade de forragem no pasto. Às 8 horas, com os animais em repouso, não houve diferença (P > 0,05) da temperatura retal (TR) e freqüência respiratória (FR) em função do tipo racial. Às 14 horas, com os animais também em repouso, os animais brancos apresentaram TR e FR significativamente maiores que os anglonubianos e estes que os pardos. O local e a estação do ano afetaram significativamente a TR e FR em qualquer dos horários. Os dados hematológicos não variaram em função do tipo racial, local ou estação do ano, ficando dentro dos valores normais para a espécie.

Influence of region and season on kid behavior on pasture / Influência do local e estação do ano sobre o comportamento de cabritos a pasto

The behavior (frequency of animais grazing) and physiological profiles of anglonubians, alpine x anglonubians and saanen x anglonubians kids were studied during the summer (january/ february) and winter (june/july) on two different places with different weather conditions (Itapetininga and Nova Odessa in the State of São Paulo). The breed didnt affect the frequency distribution (FD) taken every 30 minutes of animals grazing, from 7:30am to 6:30pm. Places, season and grass availability were significative (P 0.01) for FD. At 8:00am with the animals at rest, the breed didnt show difference (P > 0.05) for rectal temperature (RT) and respiratory rate (RR). At 2:00pm also at rest, the RT and RR were higher (P 0.01) for the saanen crossbred, when compared to the other breeds and also, higher (P 0.01) for the anglonubians when compared to the alpine crossbred. Places and season affected (P 0.01) RT and RR independently of the time of the day. The hematological data were not influenced by breed, places and season, and showed the species standard profile.

Estudou-se o comportamento (freqüência de animais em pastejo) e os aspectos fisiológicos de cabritas anglonubianas, pardo alpino x anglonubianas (pardos) e saanen x anglonubianas (brancos), em pastagem, durante o verão (janeiro e fevereiro) e inverno (junho e julho), em dois locais de condições climáticas distintas (Itapetininga e Nova Odessa). Observou-se que o tipo racial não influenciou a distribuição da freqüência de animais em pastejo, observada a cada 30 minutos, das 7 às 19h30. Todavia, verificou-se efeito significativo (P 0,01) do local, da estação do ano e notadamente da disponibilidade de forragem no pasto. Às 8 horas, com os animais em repouso, não houve diferença (P > 0,05) da temperatura retal (TR) e freqüência respiratória (FR) em função do tipo racial. Às 14 horas, com os animais também em repouso, os animais brancos apresentaram TR e FR significativamente maiores que os anglonubianos e estes que os pardos. O local e a estação do ano afetaram significativamente a TR e FR em qualquer dos horários. Os dados hematológicos não variaram em função do tipo racial, local ou estação do ano, ficando dentro dos valores normais para a espécie.

Pharmacological and toxicological evaluation of Alpinia speciosa

Alpinia speciosa Schum or A. nutans is a plant of the Zingiberanceae family, Known popularly as "colony" (colônia) and used as a diuretic and to control hypertensión. We have determinated the concentration of Na+ and K+ found in the alcoholic extract and in the tea concoction. They contained 51.0mEq Na+, and 132 mEq K+ in the extract, and 0,0 mEq of Na+ and 26 mEq K+ in the tea. Phytochemical analysis of the leaves demonstrated the presence of catecquic tanins, phenols and alkaloids, and also some essential oils. When injected intra-peritoneally the hydroalcoholic extract, in range of 100 a 1400 mg/Kg, (or 2500-18000 mg/Kg orally) produced in mice: writhing, psychomorot excitation, hypokinesis and pruritus. The LD50 by ip was 0.760 + or - 0.126 g/Kg and 10.0 + or - 2.5 g/Kg by oral administration for the hydroalcoholic extract. Subacute toxicity made injecting daily for 30 days the LD10 in rats caused an increase in transaminases and lactate dehydrogenase, whereas other parameters such as nlood glucose, urea and creatinine were normal. A histopathological analysis of liver, spleen, gut, lung and heart showed no alterations. The drug also produced a prolongation of the sleeping time. The hydroalcoholic extract induced int he rat and in the dog a dose-dependent fall in blood pressure in doses of 10 to 30 mg/Kg. In isolated atria the extract induced a reduction of the frequnecy and in the inotropic responses. Neither the extract nor the tea had an effect on the diuresis of the rat.