Iron Uptake Controls Trypanosoma cruzi Metabolic Shift and Cell Proliferation.

(1) Background: Ionic transport in Trypanosoma cruzi is the object of intense studies. T. cruzi expresses a Fe-reductase (TcFR) and a Fe transporter (TcIT). We investigated the effect of Fe depletion and Fe supplementation on different structures and functions of T. cruzi epimastigotes in culture. (2) Methods: We investigated growth and metacyclogenesis, variations of intracellular Fe, endocytosis of transferrin, hemoglobin, and albumin by cell cytometry, structural changes of organelles by transmission electron microscopy, O2 consumption by oximetry, mitochondrial membrane potential measuring JC-1 fluorescence at different wavelengths, intracellular ATP by bioluminescence, succinate-cytochrome c oxidoreductase following reduction of ferricytochrome c, production of H2O2 following oxidation of the Amplex® red probe, superoxide dismutase (SOD) activity following the reduction of nitroblue tetrazolium, expression of SOD, elements of the protein kinase A (PKA) signaling, TcFR and TcIT by quantitative PCR, PKA activity by luminescence, glyceraldehyde-3-phosphate dehydrogenase abundance and activity by Western blotting and NAD+ reduction, and glucokinase activity recording NADP+ reduction. (3) Results: Fe depletion increased oxidative stress, inhibited mitochondrial function and ATP formation, increased lipid accumulation in the reservosomes, and inhibited differentiation toward trypomastigotes, with the simultaneous metabolic shift from respiration to glycolysis. (4) Conclusion: The processes modulated for ionic Fe provide energy for the T. cruzi life cycle and the propagation of Chagas disease.

Molecular phenotypes of bovine blastocyst derived from in vitro-matured oocyte supplemented with PAPP-A.

Insulin-like growth factor-1 (IGF-1) regulates cellular lipid content, whereas pregnancy-associated plasma protein-A (PAPP-A) increases IGF-1 bioavailability. Using in vitro-matured cumulus-oocyte complexes, we aimed to evaluate the impact of PAPP-A on the blastocyst lipid content, embryo cryotolerance and embryonic transcriptional profile. We determined that PAPP-A did not affect the lipid content of oocytes, blastocysts, or blastocyst yield (P > 0.05). However, PAPP-A modulated the embryo transcriptional profiles by downregulating PPARGC1A and AKR1B1, which are related to lipid metabolism; CASP9, a pro-apoptotic gene; and IFN-τ, a marker of embryo quality (P < 0.05). Furthermore, the use of PAPP-A improved blastocyst re-expansion in the first 3 h of culture after vitrification (P < 0.05). Although PAPP-A did not affect the blastocyst lipid content or embryo production, we suggest that embryonic transcriptional modulation could contribute to maintain the balance in embryo lipid metabolism. Furthermore, PAPP-A's approach seems to control key intracellular pathways that improve post-cryopreservation development of blastocysts.

Trace mineral mixture supplemented to in vitro maturation medium improves subsequent embryo development and embryo quality in cattle.

Trace minerals participate in reproductive processes and are crucial for oocyte maturation. The objective of the present study was to investigate the effect of combined supplementation with copper (Cu), manganese (Mn), selenium (Se) and zinc (Zn) during bovine in vitro maturation (IVM) on subsequent embryo development and quality. The IVM medium was supplemented as follows: a) Control (no mineral supplementation); b) MScz (6 ng/mL Mn + 100 ng/mL Se + 200 ng/mL Cu + 400 ng/mL Zn); c) MScZ (6 ng/mL Mn + 100 ng/mL Se + 200 ng/mL Cu + 1200 ng/mL Zn); d) MSCz (6 ng/mL Mn + 100 ng/mL Se + 600 ng/mL Cu + 400 ng/mL Zn). Supplementation with MScz and MSCz produced more blastocysts compared with the control. Total blastocyst cell number was higher when minerals were added at any combination. Day-8 blastocysts derived from oocytes treated with minerals had lower intracellular reactive oxygen species concentration and lipid content than the control. In conclusion, combined supplementation with Cu, Mn, Se and Zn during bovine oocyte IVM increased in vitro production performance, improving embryo developmental ability and quality.

Docosahexaenoic acid and phenazine ethosulfate are not efficient lipid modulators for porcine in vitro maturation systems.

The high lipid content in porcine oocytes impairs in vitro embryo production (IVP). Here, we evaluated the influence of two different lipid modulators during in vitro maturation (IVM) on the embryo development and the lipid content of oocytes and embryos. In Experiment I, oocytes were exposed to 50 µM docosahexaenoic acid (DHA) with (+) or without (-) the presence of porcine follicular fluid (pFF). In Experiment II, phenazine ethosulfate (PES) was added during IVM at two concentrations (0.5 and 0.05 µM). The pFF- with 50 µM DHA treatment impaired nuclear maturation, cleavage and blastocyst rates (p < 0.05). Oocytes in pFF- media accumulated less lipids (p < 0.05). The addition of 0.5 µ M PES reduced all development rates (p < 0.05) and resulted in higher lipid content for oocytes and embryos. Only 0.05 µM PES oocytes matured similarly to the control (p > 0.05), although embryo development and embryo lipid content was similar to 0.5 µM PES oocytes (p > 0.05). Thus, 50 µM DHA supplementation in the IVM medium without pFF impaired oocyte maturation and embryo development rates without interfering in oocyte lipid content even in the presence of pFF. Maturation with PES neither favored porcine embryo development nor reduced their lipid content.

Fatty acid profile of milk from Nordestina donkey breed raised on Caatinga pasture.

In this research communication we describe the composition of fatty acids (FA) present in the milk of the Nordestina donkey breed, and how they differ during lactation. Milk samples were taken from 24 multiparous lactating Nordestina donkeys that grazed the Caatinga, comprising 5 animals at each of around 30, 60 and 90 d in milk (DIM) and a further 9 animals ranging from 120 to 180 DIM. The milk fat content was analysed by mid infrared spectroscopy and the FA profile by gas chromatography. The milk fat percentage ranged from 0.45 to 0.61%. The main FA found in milk were 16:0 and 18:1c9. These did not differ among DIM classes and comprised 23% and 25% of total FA. Notably, the α-Linolenic acid (18:3 n-3) was the third most abundant FA and differed (P < 0.05) with DIM, being lowest in the 30 and 60 DIM samples (around 10.7% of total FA) and highest in the 60 and 90 DIM classes (around 14.6% of total FA). The low-fat content and the FA profile of the donkey milk gives it potential as a functional ingredient, which could help to preserve the commercial viability of the Nordestina donkey breed.

Acute supplementation of growing rats with Brazil nut flour increases hepatic lipid content but prevents oxidative damage in the liver.

Brazil nuts (BN) (Bertholletia excels, Bonpl.), are of great importance because of their nutritional properties and economic value. They can be consumed in natura or as flour (BNF). In this study, we evaluated the effects of BNF and BN intakes (Both 5% and 15%) on metabolic parameters of rats for 15 days. Serum Selenium (Se) levels were higher in BN-15% and BNF-15% groups. Lipid content was reduced in retroperitoneal and epididymal adipose tissues in all groups and in the BN-5% group in the liver. However, liver lipids increased in the BNF-15% group. The levels of carbonylated proteins and lipid peroxidation in the liver were not altered. The data reveal that the increase in hepatic lipids in the BNF-15% group probably occurred due to the high concentration of free fatty acids present in the flour. The Se bioavailability in the diet contributed to the preservation of the liver function in rats. PRACTICAL IMPLICATIONS: The consumption of BN is common in the population. However, changes in eating habits have led to a more frequent consumption of vegetable derivatives, such as drinks and oils. The cake residue generated after processing is still considered of high nutritional value, since it is a source of protein and minerals such as Se. Because of its low cost, the use of pie at the industrial level is becoming increasingly more interesting for the development of new products, and the Brazil nut flour (BNF) is considered a good option. Our study showed that just like BN, BNF can be a source of selenium for the body, although changes in lipid metabolism and physiological parameters can be observed depending on the amount used. We believe that the results of this investigation can be used to guide the development of new technologies and products containing BN.

Lipid profile and fatty acid composition of marine fish species from Northeast coast of Brazil.

Total lipid content, fatty acid composition and nutritional profile of lipid fraction of fourteen fish species from the Brazilian Northeastern coast were evaluated. Lipid content was determined by Bligh and Dyer methodology, whereas the fatty acid composition was determined by gas chromatography with flame ionization detector. The indices of atherogenicity, thrombogenicity and hypocholesterolemic/hypercholesterolemic ratio were used to evaluate the nutritional quality of lipid fraction. The total lipid content ranged from 0.25 to 3.09%, with higher values in L. synagris. A significant difference (p < 0.05) were noted in fatty acids composition between the fourteen fish species. PUFA were the major fatty acids in twelve of fourteen species and C. leiarchus showed the highest content. The n-3 content ranged from 69.83 to 270.62 mg g-1 of total lipids, being EPA and DHA the major fatty acids in most of the species studied. The species L. synagris, M. bonaci and S. brasiliensis exceeded the WHO daily intake recommendation of 250 mg per day from those fatty acids. The quality indexes of the lipid fraction as well as the n-6/n-3 and PUFA/SFA ratios indicated that all fish species can be healthy nutritional options, and can bring many benefits to human health. The principal component analysis resulted in a two-principal-component model that described 79.78% of data variance. Also, it highlighted that despite the differences between the breeding strategies of fish families, the species could be grouped according to their similarity in fatty acids composition.

Stimulation action of mefenpyr-diethyl on soybean, wheat, and signal grass plants.

Mefenpyr-diethyl is a safener used for protection of cereal plants under applications of ACCase and ALS inhibitor herbicides. Current studies are describing safeners using a new approach, relating these products to stimulation action on plants. The objective of this work was to evaluate the stimulation action of mefenpyr-diethyl on soybean, wheat, and signal grass plants. The experiment was conducted in a greenhouse, under a completely randomized design, with four replications, in two seasons. Mefenpyr-diethyl (50 g a.i. ha-1) was applied on soybean plants (at V4 stage), and wheat and signal grass plants (both with 15 cm height). The variables evaluated were plant height, dry matter, and lipid content of the three species, and number of tillers of wheat and signal grass plants. The application of mefenpyr-diethyl in the first season increased the number of tillers of wheat and height of soybean plants. The soybean presented 24 and 14% more dry matter than the control in the first and second season, respectively, and 0.5% more lipid content in plants treated with mefenpyr-diethyl. These results show the stimulation action of mefenpyr-diethyl on wheat and soybean plants, denoting its potential for growth promotion and indicating the need for studies with this approach. No effect was found for the signal grass plants.

Exploratory Study of Fatty Acid Profile in Two Filmy Ferns with Contrasting Desiccation Tolerance Reveal the Production of Very Long Chain Polyunsaturated Omega-3 Fatty Acids.

Lipids are fundamental components of cell membranes and play a significant role in their integrity and fluidity. Alteration in lipid composition of membranes has been reported to be a major response to abiotic environmental stresses. This work was focused on the characterization of frond lipid composition and membrane integrity during a desiccation-rehydration cycle of two filmy fern species with contrasting desiccation tolerance: Hymenophyllum caudiculatum (less tolerant) and Hymenophyllum plicatum (more tolerant). The relative water content decreased without differences between species when both filmy ferns were subjected to desiccation. However, H. plicatum reached a higher relative water content than H. caudiculatum after rehydration. Fatty acids profiles showed the presence of a very long chain polyunsaturated fatty acid during the desiccation-rehydration cycle, with eicosatrienoic acid being the most abundant. Additionally, propidium iodide permeation staining and confocal microscopy demonstrated that, following the desiccation-rehydration cycle, H. plicatum exhibited a greater membrane integrity than H. caudiculatum. The lack of some very long chain fatty acids such as C22:1n9 and C24:1n9 in this species contrasting with H. plicatum may be associated with its lower membrane stability during the desiccation-rehydration cycle. This report provides the first insight into the fatty acid composition and dynamics of the membrane integrity of filmy ferns during a desiccation-rehydration cycle. This could potentially play a role in determining the different levels of desiccation tolerance and microhabitat preferences exhibited by Hymenophyllaceae species.

Influence of cAMP modulator supplementation of in vitro culture medium on Bos taurus indicus embryos.

The effectiveness of the use of natriuretic peptide C (NPPC) in the blocking of meiosis has already been proven in several species. However, there are no reports on the use of NPPC in the activation of metabolic processes in embryos. Whereas modulations of cAMP concentrations alter the lipid metabolism of bovine oocytes, the present study aims to evaluate the effect of NPPC on the development, lipid content and transcript levels of genes related to lipid metabolism of IVP bovine embryos. For this purpose, ovaries were obtained from a slaughterhouse, and oocytes were fertilized in vitro (D0). From D5 of in vitro culture, embryos were treated with 100 nM NPPC (NPPC group) or with no NPPC (Control group) and evaluated in terms of Blastocyst (D7) and hatching rates (D10). For the assessment of the cytoplasmatic lipid amounts, blastocysts were stained with Sudan Black B dye. The embryonic lipid profile was investigated by electrospray ionization desorption-mass spectrometry (DESI-MS). The abundance of nine transcripts related to lipid metabolism were assessed using the Biomark HD system. For statistical analysis, blastocyst and hatching rates, lipid content by the Sudan Black B and variation of gene expression between groups were compared by Student t-test. For lipid profile analysis, principal component analysis (PCA) and fold-change were performed. The embryo lipid content was similar between NPPC (881 ±â€¯3.7) and Control (883 ±â€¯5.2) groups (p > 0.05). However, cholesteryl esters and TAGs were downregulated by NPPC at multiple levels according to the DESI-MS profiles. Of the analyzed genes, ELOVL6 and SREBF1 showed an up-regulation in the control group (p < 0.05), while CPT2 was observed to be up-regulated in the NPPC-treated embryos. There was no significant difference in the blastocyst production rate between NPPC (44.4%) and Control (42.4%), however the hatching rate at D10 was higher (p < 0.05) in the NPPC group (69.77%) when compared to the Control group (48.33%). These findings demonstrate that NPPC alters the mRNA expression of genes related to lipid metabolism and that it exerts a positive effect on the hatching rates of IVP Bos taurus indicus embryos.

Oil quality in sea-level quinoa as determined by cultivar-specific responses to temperature and radiation conditions.

BACKGROUND: There is renewed interest in quinoa as a potential source of vegetable oils; however, there is no information about how environmental conditions affect its fatty acid composition, a critical indicator of its oil quality. The fatty acid concentrations of four cultivars adapted to temperate environments were compared at three sowing dates to evaluate the effect of environmental conditions during the seed-filling period on the variation in oil quality. RESULTS: The interaction between cultivar and sowing date was the main source of variation explaining the changes in the lipid content and fatty acid concentrations in quinoa. Most of the variation in the concentration of unsaturated fatty acids was attributed to the temperature and solar radiation during the seed-filling period; cultivar-specific responses to photo-thermal conditions were observed among the sea-level quinoa cultivars evaluated. CONCLUSION: The lipid content and concentration of fatty acids in quinoa are affected by sowing date. This effect is exerted through changes in temperature and solar radiation conditions. This managing practice can therefore be used to achieve quinoa oil with different qualities. © 2019 Society of Chemical Industry.

Maternal influence on early lipid content in an introduced partially anadromous population of Oncorhynchus mykiss.

This study evaluated how the maternal migratory tactic in a partially anadromous population of Oncorhynchus mykiss may influence the early energetic status of their offspring. Total lipid content variation (% dry mass) of recently emerged fry caught in the Santa Cruz River, Argentina, was evaluated as a function of their maternal origin (anadromous v. resident) and fork length (LF ). Lipid content of fry decreased with LF and was higher for offspring of anadromous mothers.

Contenido lipídico, actividad mitocondrial y desarrollo embrionario temprano de ovocitos provenientes de vacas mestizas (Bos taurus indicus) / Lipid content, mitochondrial activity and early embryo development in oocyte collected from crossbred cows (Bos taurus indicus) / Conteúdo lipídico, atividade mitocondrial e desenvolvimiento embrionário precoce de oócitos coletados de vacas mestiças (Bos taurus indicus)

El objetivo de la presente investigación fue el de valorar el efecto de la predominancia fenotípica sobre el contenido lipídico como indicador de la calidad ovocitaria. Los COC’s fueron recuperados mediante aspiración folicular y sometidos a maduración in vitro (MIV), fecundación in vitro (FIV) y cultivo in vitro (CIV). Se determinó el contenido lipídico y actividad mitocondrial en ovocitos inmaduros y MIV. La tasa de maduración total se ubicó en 75%, con valores de 80,6% y 69,3% para ovocitos predominantemente B. indicus y predominantemente B. taurus, respectivamente. La tasa de fecundación total fue de 27,6%, para ovocitos predominantemente B. indicus y predominantemente B. taurus, esta fue de 26,1% y 29%, respectivamente. Se observó un total de embriones divididos de 52,1% y 58,9% tras 48 y 72 horas post inseminación (hpi), respectivamente. Por otro lado, para el grupo predominantemente B. indicus se apreció un 55,5% y 57,5% de embriones divididos tras 48 y 72 hpi. Mientras que para el grupo predominantemente B. taurus se apreció un 52,1% y 58,9% de embriones divididos tras 48 y 72 hpi. Ovocitos inmaduros presentaron mayor número de gotas lipídicas pequeñas (p <0,0001), en contraste con los ovocitos MIV que presentaron mayor número de gotas lipídicas medianas y grandes (p < 0,0001). Ovocitos predominantemente B. indicus presentaron mayor número de gotas lipídicas pequeñas (p = 0,0005) y medianas (p = 0.005), mientras que para las gotas lipídicas grandes no se apreciaron diferencias significativas. Ovocitos MIV presentaron mayor actividad mitocondrial que el grupo de ovocitos inmaduros (p < 0,05), sin observarse efecto de la predominancia fenotípica sobre este parámetro. La valoración del contenido lipídico no resultó un factor predictivo de la calidad ovocitaria en hembras mestizas.

The objective of this research was to evaluate the effect of phenotypic predominance on lipid content, mitochondrial activity and early developmental competence as indicators of oocyte quality. Cumulus-oocyte complexes (COCs) were recovered through follicular aspiration, and underwent in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of presumptive zygotes. Lipid content and mitochondrial activity in immature and IVM oocytes were determined. A maturation rate of 80.6% and 69.3% was found for oocytes predominantly B. indicus and predominantly B. taurus, respectively. Total fertilization rate was 27.6%; 26.1% for predominantly B. indicus oocytes and 29% for predominantly B. taurus oocytes. A total of 55.5% and 57.5% of cleaved embryos after 48 and 72 h post-insemination (hpi) in predominantly B. indicus group were observed, respectively. As for the predominantly B. taurus group, 48.6% and 60.4% of cleaved embryos were found after 48 and 72 hpi, respectively. In both groups, immature oocytes showed a greater amount of small lipidic droplets (p <0.0001); IVM decreased the number of small lipid droplets (p < 0.0001) and increased the number of medium and large lipid droplets (p < 0.0001). Predominantly B. indicus oocytes had a greater number of small and medium-sized lipid droplets, while there were no significant differences in large lipid droplets. IVM oocytes had higher mitochondrial activity than immature oocytes group (p < 0.05) without any effect of phenotypic predominance on this parameter. Assessment of lipid content was not a predictive factor of oocyte quality in crossbred cows.

O objetivo deste trabalho foi avaliar o efeito da predominância fenotípica no conteúdo lipídico como um indicador da qualidade do oócito. Os COC foram recuperados por aspiração folicular e submetidos a maturação in vitro (IVM), fertilização in vitro (FIV) e cultura in vitro (CIV). Determinou-se o conteúdo lipídico e atividade mitocondrial em oócitos imaturos e IVM. A taxa de maturação total era de 75%, com valores de 80,6% e 69,3% para oócitos predominantemente B. Indicus e predominantemente B. taurus, respectivamente. A taxa de fertilização total foi de 27,6%, para oócitos predominantemente B. indicus e predominantemente B. taurus, este foi de 26,1% e 29%, respectivamente. Um total de 52,1% de embriões divididos e 58,9% foi observada após 48 e 72 horas após a inseminação (hpi), respectivamente. Além disso, para o grupo predominantemente B. indicus 55,5% e 57,5% embrião clivada após 48 e 72 hpi foi observado. Enquanto para o grupo predominantemente B. taurus 52,1% e 58,9% dos embriões divididos após 48 e 72 hpi respectivamente. Oócitos imaturos mostraram gotas lipídicas mais pequenas (p<0,0001), em contraste com oócitos IVM com maior número de gotas lipídicas médias e grandes (p <0,0001). Oócitos predominantemente B. indicus mostraram mais gotas lipídicas pequenas (p = 0,0005) e médias (p = 0,005), enquanto que para as gotas lipídicas grandes não foram observadas diferenças significativas. Oócitos IVM tiveram maior atividade mitocondrial que o grupo de oócitos imaturos (p < 0,05), não houve efeito de predominância fenotípica sobre este parâmetro. A avaliação do conteúdo lipídico não foi um preditor de qualidade dos oócitos nas vacas mestiças.

Contenido lipídico, actividad mitocondrial y desarrollo embrionario temprano de ovocitos provenientes de vacas mestizas (Bos taurus indicus) / Lipid content, mitochondrial activity and early embryo development in oocyte collected from crossbred cows (Bos taurus indicus) / Conteúdo lipídico, atividade mitocondrial e desenvolvimiento embrionário precoce de oócitos coletados de vacas mestiças (Bos taurus indicus)

El objetivo de la presente investigación fue el de valorar el efecto de la predominancia fenotípica sobre el contenido lipídico como indicador de la calidad ovocitaria. Los COCs fueron recuperados mediante aspiración folicular y sometidos a maduración in vitro (MIV), fecundación in vitro (FIV) y cultivo in vitro (CIV). Se determinó el contenido lipídico y actividad mitocondrial en ovocitos inmaduros y MIV. La tasa de maduración total se ubicó en 75%, con valores de 80,6% y 69,3% para ovocitos predominantemente B. indicus y predominantemente B. taurus, respectivamente. La tasa de fecundación total fue de 27,6%, para ovocitos predominantemente B. indicus y predominantemente B. taurus, esta fue de 26,1% y 29%, respectivamente. Se observó un total de embriones divididos de 52,1% y 58,9% tras 48 y 72 horas post inseminación (hpi), respectivamente. Por otro lado, para el grupo predominantemente B. indicus se apreció un 55,5% y 57,5% de embriones divididos tras 48 y 72 hpi. Mientras que para el grupo predominantemente B. taurus se apreció un 52,1% y 58,9% de embriones divididos tras 48 y 72 hpi. Ovocitos inmaduros presentaron mayor número de gotas lipídicas pequeñas (p <0,0001), en contraste con los ovocitos MIV que presentaron mayor número de gotas lipídicas medianas y grandes (p < 0,0001). Ovocitos predominantemente B. indicus presentaron mayor número de gotas lipídicas pequeñas (p = 0,0005) y medianas (p = 0.005), mientras que para las gotas lipídicas grandes no se apreciaron diferencias significativas. Ovocitos MIV presentaron mayor actividad mitocondrial que el grupo de ovocitos inmaduros (p < 0,05), sin observarse efecto de la predominancia fenotípica sobre este parámetro. La valoración del contenido lipídico no resultó un factor predictivo de la calidad ovocitaria en hembras mestizas.(AU)

The objective of this research was to evaluate the effect of phenotypic predominance on lipid content, mitochondrial activity and early developmental competence as indicators of oocyte quality. Cumulus-oocyte complexes (COCs) were recovered through follicular aspiration, and underwent in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of presumptive zygotes. Lipid content and mitochondrial activity in immature and IVM oocytes were determined. A maturation rate of 80.6% and 69.3% was found for oocytes predominantly B. indicus and predominantly B. taurus, respectively. Total fertilization rate was 27.6%; 26.1% for predominantly B. indicus oocytes and 29% for predominantly B. taurus oocytes. A total of 55.5% and 57.5% of cleaved embryos after 48 and 72 h post-insemination (hpi) in predominantly B. indicus group were observed, respectively. As for the predominantly B. taurus group, 48.6% and 60.4% of cleaved embryos were found after 48 and 72 hpi, respectively. In both groups, immature oocytes showed a greater amount of small lipidic droplets (p <0.0001); IVM decreased the number of small lipid droplets (p < 0.0001) and increased the number of medium and large lipid droplets (p < 0.0001). Predominantly B. indicus oocytes had a greater number of small and medium-sized lipid droplets, while there were no significant differences in large lipid droplets. IVM oocytes had higher mitochondrial activity than immature oocytes group (p < 0.05) without any effect of phenotypic predominance on this parameter. Assessment of lipid content was not a predictive factor of oocyte quality in crossbred cows.(AU)

O objetivo deste trabalho foi avaliar o efeito da predominância fenotípica no conteúdo lipídico como um indicador da qualidade do oócito. Os COC foram recuperados por aspiração folicular e submetidos a maturação in vitro (IVM), fertilização in vitro (FIV) e cultura in vitro (CIV). Determinou-se o conteúdo lipídico e atividade mitocondrial em oócitos imaturos e IVM. A taxa de maturação total era de 75%, com valores de 80,6% e 69,3% para oócitos predominantemente B. Indicus e predominantemente B. taurus, respectivamente. A taxa de fertilização total foi de 27,6%, para oócitos predominantemente B. indicus e predominantemente B. taurus, este foi de 26,1% e 29%, respectivamente. Um total de 52,1% de embriões divididos e 58,9% foi observada após 48 e 72 horas após a inseminação (hpi), respectivamente. Além disso, para o grupo predominantemente B. indicus 55,5% e 57,5% embrião clivada após 48 e 72 hpi foi observado. Enquanto para o grupo predominantemente B. taurus 52,1% e 58,9% dos embriões divididos após 48 e 72 hpi respectivamente. Oócitos imaturos mostraram gotas lipídicas mais pequenas (p<0,0001), em contraste com oócitos IVM com maior número de gotas lipídicas médias e grandes (p <0,0001). Oócitos predominantemente B. indicus mostraram mais gotas lipídicas pequenas (p = 0,0005) e médias (p = 0,005), enquanto que para as gotas lipídicas grandes não foram observadas diferenças significativas. Oócitos IVM tiveram maior atividade mitocondrial que o grupo de oócitos imaturos (p < 0,05), não houve efeito de predominância fenotípica sobre este parâmetro. A avaliação do conteúdo lipídico não foi um preditor de qualidade dos oócitos nas vacas mestiças.(AU)

Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification / Acúmulo de lipídios intracitoplasmáticos, desenvolvimento e criotolerância de embriões bovinos produzidos in vitro e tratados com diferentes concentrações de forskolin antes da vitrificação

The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5µM (Forsk 2.5 group), 5.0µM (Forsk 5.0 group) or 10.0µM (Forsk 10.0 group). Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05) in the blastocyst rates between the Control group (44.9%) and the other treatments, the embryo production was lower (P<0.05) in Forsk 10.0 group (38.8%) compared to Forsk 2.5 (50.5%) and Forsk 5.0 (54.7%) groups. The intracytoplasmic lipid content (expressed in arbitrary units of pixels) in blastocysts from the Control group (1.00±0.03) was similar (P>0.05) to that found in Forsk 2.5 (0.92±0.03) and Forsk 10.0 groups (1.06±0.03) groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04) was lower than in the Control group (P<0.05). Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P<0.05) in Forsk 5.0 group (72.2%) compared to the Control group (46.2%). In conclusion, pre-treatment with forskolin at a concentration of 5.0 µM for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.(AU)

Os embriões foram produzidos a partir de ovários obtidos em abatedouro e foram alocados em quatro grupos experimentais. Nos grupos tratados, o forskolin foi adicionado ao meio de cultivo in vitro no dia 6 do cultivo e os embriões foram incubados durante 24 horas com uma das seguintes concentrações: 2,5µM (grupo Forsk 2,5), 5,0µM (grupo Forsk 5,0) ou 10,0µM (grupo Forsk 10,0). Os embriões do grupo controle foram cultivados na ausência de forskolin. No dia 7 do cultivo, os blastocistos expandidos foram corados com o corante lipofílico Sudan Black B para a determinação do teor de lípidos intracitoplasmáticos ou foram criopreservados através do protocolo Vitri-Ingá®. Não foi observada diferença significativa (P>0,05) na taxa de produção de blastocistos entre o grupo Controle (44,9%) e os demais tratamentos, todavia observou-se menor produção de embriões (P<0,05) no grupo Forsk 10,0 (38,8%) em comparação com os grupos Forsk 2,5 (50,5%) e Forsk 5,0 (54,7%). A quantidade de lipídeos intracitoplasmáticos do grupo Controle (1,00±0,03) foi semelhante (P>0,05) a dos grupos Forsk 2,5 (0,92±0,03) e Forsk 10,0 (1,06±0,03); no entanto, o acúmulo de lípidos nos blastocistos do grupo Forsk 5.0 (0,82 ± 0,04) foi menor do que no grupo controle (P<0,05). A partir destes resultados, o grupo Forsk 5,0 foi testado quanto à criotolerância e foi observado que a taxa de re-expansão da blastocele 24 horas após o aquecimento foi maior (P<0,05) no grupo Forsk 5,0 (72,2%) quando comparado ao grupo Controle (46,2%). Em conclusão, o pré-tratamento com forskolin na concentração de 5,0 µM durante 24 horas antes da vitrificação foi eficiente para promover a redução da quantidade de lipídeos intracitoplasmáticos e, consequentemente, melhorou a criotolerância de embriões bovinos produzidos in vitro.(AU)

Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification / Acúmulo de lipídios intracitoplasmáticos, desenvolvimento e criotolerância de embriões bovinos produzidos in vitro e tratados com diferentes concentrações de forskolin antes da vitrificação

The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5µM (Forsk 2.5 group), 5.0µM (Forsk 5.0 group) or 10.0µM (Forsk 10.0 group). Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05) in the blastocyst rates between the Control group (44.9%) and the other treatments, the embryo production was lower (P<0.05) in Forsk 10.0 group (38.8%) compared to Forsk 2.5 (50.5%) and Forsk 5.0 (54.7%) groups. The intracytoplasmic lipid content (expressed in arbitrary units of pixels) in blastocysts from the Control group (1.00±0.03) was similar (P>0.05) to that found in Forsk 2.5 (0.92±0.03) and Forsk 10.0 groups (1.06±0.03) groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04) was lower than in the Control group (P<0.05). Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P<0.05) in Forsk 5.0 group (72.2%) compared to the Control group (46.2%). In conclusion, pre-treatment with forskolin at a concentration of 5.0 µM for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.(AU)

Os embriões foram produzidos a partir de ovários obtidos em abatedouro e foram alocados em quatro grupos experimentais. Nos grupos tratados, o forskolin foi adicionado ao meio de cultivo in vitro no dia 6 do cultivo e os embriões foram incubados durante 24 horas com uma das seguintes concentrações: 2,5µM (grupo Forsk 2,5), 5,0µM (grupo Forsk 5,0) ou 10,0µM (grupo Forsk 10,0). Os embriões do grupo controle foram cultivados na ausência de forskolin. No dia 7 do cultivo, os blastocistos expandidos foram corados com o corante lipofílico Sudan Black B para a determinação do teor de lípidos intracitoplasmáticos ou foram criopreservados através do protocolo Vitri-Ingá®. Não foi observada diferença significativa (P>0,05) na taxa de produção de blastocistos entre o grupo Controle (44,9%) e os demais tratamentos, todavia observou-se menor produção de embriões (P<0,05) no grupo Forsk 10,0 (38,8%) em comparação com os grupos Forsk 2,5 (50,5%) e Forsk 5,0 (54,7%). A quantidade de lipídeos intracitoplasmáticos do grupo Controle (1,00±0,03) foi semelhante (P>0,05) a dos grupos Forsk 2,5 (0,92±0,03) e Forsk 10,0 (1,06±0,03); no entanto, o acúmulo de lípidos nos blastocistos do grupo Forsk 5.0 (0,82 ± 0,04) foi menor do que no grupo controle (P<0,05). A partir destes resultados, o grupo Forsk 5,0 foi testado quanto à criotolerância e foi observado que a taxa de re-expansão da blastocele 24 horas após o aquecimento foi maior (P<0,05) no grupo Forsk 5,0 (72,2%) quando comparado ao grupo Controle (46,2%). Em conclusão, o pré-tratamento com forskolin na concentração de 5,0 µM durante 24 horas antes da vitrificação foi eficiente para promover a redução da quantidade de lipídeos intracitoplasmáticos e, consequentemente, melhorou a criotolerância de embriões bovinos produzidos in vitro.(AU)

Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification

ABSTRACT: The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5M (Forsk 2.5 group), 5.0M (Forsk 5.0 group) or 10.0M (Forsk 10.0 group). Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05) in the blastocyst rates between the Control group (44.9%) and the other treatments, the embryo production was lower (P 0.05) in Forsk 10.0 group (38.8%) compared to Forsk 2.5 (50.5%) and Forsk 5.0 (54.7%) groups. The intracytoplasmic lipid content (expressed in arbitrary units of pixels) in blastocysts from the Control group (1.00±0.03) was similar (P>0.05) to that found in Forsk 2.5 (0.92±0.03) and Forsk 10.0 groups (1.06±0.03) groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04) was lower than in the Control group (P 0.05). Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P 0.05) in Forsk 5.0 group (72.2%) compared to the Control group (46.2%). In conclusion, pre-treatment with forskolin at a concentration of 5.0 M for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.

RESUMO: Os embriões foram produzidos a partir de ovários obtidos em abatedouro e foram alocados em quatro grupos experimentais. Nos grupos tratados, o forskolin foi adicionado ao meio de cultivo in vitro no dia 6 do cultivo e os embriões foram incubados durante 24 horas com uma das seguintes concentrações: 2,5M (grupo Forsk 2,5), 5,0M (grupo Forsk 5,0) ou 10,0M (grupo Forsk 10,0). Os embriões do grupo controle foram cultivados na ausência de forskolin. No dia 7 do cultivo, os blastocistos expandidos foram corados com o corante lipofílico Sudan Black B para a determinação do teor de lípidos intracitoplasmáticos ou foram criopreservados através do protocolo Vitri-Ingá®. Não foi observada diferença significativa (P>0,05) na taxa de produção de blastocistos entre o grupo Controle (44,9%) e os demais tratamentos, todavia observou-se menor produção de embriões (P 0,05) no grupo Forsk 10,0 (38,8%) em comparação com os grupos Forsk 2,5 (50,5%) e Forsk 5,0 (54,7%). A quantidade de lipídeos intracitoplasmáticos do grupo Controle (1,00±0,03) foi semelhante (P>0,05) a dos grupos Forsk 2,5 (0,92±0,03) e Forsk 10,0 (1,06±0,03); no entanto, o acúmulo de lípidos nos blastocistos do grupo Forsk 5.0 (0,82 ± 0,04) foi menor do que no grupo controle (P 0,05). A partir destes resultados, o grupo Forsk 5,0 foi testado quanto à criotolerância e foi observado que a taxa de re-expansão da blastocele 24 horas após o aquecimento foi maior (P 0,05) no grupo Forsk 5,0 (72,2%) quando comparado ao grupo Controle (46,2%). Em conclusão, o pré-tratamento com forskolin na concentração de 5,0 M durante 24 horas antes da vitrificação foi eficiente para promover a redução da quantidade de lipídeos intracitoplasmáticos e, consequentemente, melhorou a criotolerância de embriões bovinos produzidos in vitro.

Chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted by emulsified canola oil / Composição química, propriedades microbiológicas e perfil de ácidos graxos de Salame tipo Italiano com substituição da gordura animal por óleo de canola emulsionado

ABSTRACT: Vegetable oils have been used to substitute pork backfat to improve the fatty acid profile of fermented sausages. The aim of this study was to assess the chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted (15% and 30%) by emulsified canola oil. Fat contents decreased while moisture contents increased in Italian-type salami with emulsified canola oil. The growth of lactic acid bacteria in salami was not affected by canola oil and absence of fecal coliforms, coagulase-positive staphylococci, and Salmonella were reported during processing of fermented sausages. Lower levels of saturated fatty acids (SAFAs), higher levels of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were observed in salami with emulsified canola oil. Together, our results indicated that substituting pork backfat with emulsified canola oil improved the nutritional values of Italian-type salami.(AU)

RESUMO: O uso de óleos vegetais como substitutos da gordura animal em embutidos fermentados tem surgido como uma alternativa de produzir produtos cárneos com um melhor perfil lipídico. Assim, o objetivo deste trabalho foi avaliar a composição química, propriedades microbiológicas e o perfil de ácidos graxos de salame tipo Italiano elaborado com a substituição parcial da gordura suína (15 e 30%) por emulsão com óleo de canola. A incorporação do óleo de canola reduziu o teor de gordura e aumentou o teor de umidade dos embutidos fermentados. Em relação às características microbiológicas, a adição de óleo de canola emulsionado não alterou o crescimento das bactérias láticas durante o processamento. Ainda, não foram detectadas contagens de coliformes fecais, staphylococci coagulase positiva e Salmonella. Em relação ao perfil de ácidos graxos, foi possível observar a redução nos teores totais de ácidos graxos saturados (SAFAs) e uma elevação nos valores de ácidos graxos monoinsaturados (MUFAs) e poliinsaturados (PUFAs) nos tratamentos adicionados de óleo de canola. Dessa forma, a substituição da gordura suína por óleo de canola emulsionado em salame tipo Italiano torna-se uma alternativa interessante na produção de produtos cárneos com melhores características nutricionais.(AU)

Chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted by emulsified canola oil / Composição química, propriedades microbiológicas e perfil de ácidos graxos de Salame tipo Italiano com substituição da gordura animal por óleo de canola emulsionado

ABSTRACT: Vegetable oils have been used to substitute pork backfat to improve the fatty acid profile of fermented sausages. The aim of this study was to assess the chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted (15% and 30%) by emulsified canola oil. Fat contents decreased while moisture contents increased in Italian-type salami with emulsified canola oil. The growth of lactic acid bacteria in salami was not affected by canola oil and absence of fecal coliforms, coagulase-positive staphylococci, and Salmonella were reported during processing of fermented sausages. Lower levels of saturated fatty acids (SAFAs), higher levels of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were observed in salami with emulsified canola oil. Together, our results indicated that substituting pork backfat with emulsified canola oil improved the nutritional values of Italian-type salami.

RESUMO: O uso de óleos vegetais como substitutos da gordura animal em embutidos fermentados tem surgido como uma alternativa de produzir produtos cárneos com um melhor perfil lipídico. Assim, o objetivo deste trabalho foi avaliar a composição química, propriedades microbiológicas e o perfil de ácidos graxos de salame tipo Italiano elaborado com a substituição parcial da gordura suína (15 e 30%) por emulsão com óleo de canola. A incorporação do óleo de canola reduziu o teor de gordura e aumentou o teor de umidade dos embutidos fermentados. Em relação às características microbiológicas, a adição de óleo de canola emulsionado não alterou o crescimento das bactérias láticas durante o processamento. Ainda, não foram detectadas contagens de coliformes fecais, staphylococci coagulase positiva e Salmonella. Em relação ao perfil de ácidos graxos, foi possível observar a redução nos teores totais de ácidos graxos saturados (SAFAs) e uma elevação nos valores de ácidos graxos monoinsaturados (MUFAs) e poliinsaturados (PUFAs) nos tratamentos adicionados de óleo de canola. Dessa forma, a substituição da gordura suína por óleo de canola emulsionado em salame tipo Italiano torna-se uma alternativa interessante na produção de produtos cárneos com melhores características nutricionais.

Chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted by emulsified canola oil / Composição química, propriedades microbiológicas e perfil de ácidos graxos de Salame tipo Italiano com substituição da gordura animal por óleo de canola emulsionado

ABSTRACT: Vegetable oils have been used to substitute pork backfat to improve the fatty acid profile of fermented sausages. The aim of this study was to assess the chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted (15% and 30%) by emulsified canola oil. Fat contents decreased while moisture contents increased in Italian-type salami with emulsified canola oil. The growth of lactic acid bacteria in salami was not affected by canola oil and absence of fecal coliforms, coagulase-positive staphylococci, and Salmonella were reported during processing of fermented sausages. Lower levels of saturated fatty acids (SAFAs), higher levels of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were observed in salami with emulsified canola oil. Together, our results indicated that substituting pork backfat with emulsified canola oil improved the nutritional values of Italian-type salami.

RESUMO: O uso de óleos vegetais como substitutos da gordura animal em embutidos fermentados tem surgido como uma alternativa de produzir produtos cárneos com um melhor perfil lipídico. Assim, o objetivo deste trabalho foi avaliar a composição química, propriedades microbiológicas e o perfil de ácidos graxos de salame tipo Italiano elaborado com a substituição parcial da gordura suína (15 e 30%) por emulsão com óleo de canola. A incorporação do óleo de canola reduziu o teor de gordura e aumentou o teor de umidade dos embutidos fermentados. Em relação às características microbiológicas, a adição de óleo de canola emulsionado não alterou o crescimento das bactérias láticas durante o processamento. Ainda, não foram detectadas contagens de coliformes fecais, staphylococci coagulase positiva e Salmonella. Em relação ao perfil de ácidos graxos, foi possível observar a redução nos teores totais de ácidos graxos saturados (SAFAs) e uma elevação nos valores de ácidos graxos monoinsaturados (MUFAs) e poliinsaturados (PUFAs) nos tratamentos adicionados de óleo de canola. Dessa forma, a substituição da gordura suína por óleo de canola emulsionado em salame tipo Italiano torna-se uma alternativa interessante na produção de produtos cárneos com melhores características nutricionais.