Metabolomics of soybean green stem and foliar retention (GSFR) disease using mass spectrometry and molecular networking.

RATIONALE: The nematode Aphelenchoides besseyi is the causal agent of green stem and foliar retention, a soybean disease recently described in Brazil. This condition can reduce soybean yield by up to 100%. However, little is known about chemical interactions between the plant and pathogen. Therefore, this work aimed to investigate metabolites from healthy soybean roots and from soybean roots that were inoculated with A. besseyi. METHODS: A. besseyi were multiplied in vitro with Fusarium sp. colonies in Petri dishes for 25 days, and were axenically inoculated into hydroponics healthy soybean plants. The metabolites were extracted from the roots of healthy and A. besseyi-infected plants 16 days post-inoculation. These extracts were analyzed using an untargeted metabolomic method with an ultra-high-performance liquid chromatography/electrospray ionization /tandem mass spectrometry (UHPLC/ESI-MS/MS) and molecular networking approach. RESULTS: Roots from infected plants showed morphological alterations such as shrinkage, darkening, and arching. Similarly, they also showed an increased presence of flavonoids, compared with healthy roots. Compounds such as neobavaisoflavone, glycitin, genistin, and genistein were putatively identified and had greater intensity in inoculated roots. These compounds are linked to the defensive mechanisms in plants against nematodes. Moreover, coumaric acid, also exclusively putatively identified in inoculated roots, shows activity related to inhibition of root growth. CONCLUSIONS: Liquid chromatography, mass spectrometry, and molecular networking approaches proved to be a powerful tool for the metabolomic study of GSFR. This study showed metabolomics differences of protective substances in the roots, evidencing a quick response of the plant to the attack of A. besseyi.

Untargeted Metabolomics Analysis by UHPLC-MS/MS of Soybean Plant in a Compatible Response to Phakopsora pachyrhizi Infection.

Phakopsora pachyrhizi is a biotrophic fungus, causer of the disease Asian Soybean Rust, a severe crop disease of soybean and one that demands greater investment from producers. Thus, research efforts to control this disease are still needed. We investigated the expression of metabolites in soybean plants presenting a resistant genotype inoculated with P. pachyrhizi through the untargeted metabolomics approach. The analysis was performed in control and inoculated plants with P. pachyrhizi using UHPLC-MS/MS. Principal component analysis (PCA) and the partial least squares discriminant analysis (PLS-DA), was applied to the data analysis. PCA and PLS-DA resulted in a clear separation and classification of groups between control and inoculated plants. The metabolites were putative classified and identified using the Global Natural Products Social Molecular Networking platform in flavonoids, isoflavonoids, lipids, fatty acyls, terpenes, and carboxylic acids. Flavonoids and isoflavonoids were up-regulation, while terpenes were down-regulated in response to the soybean-P. pachyrhizi interaction. Our data provide insights into the potential role of some metabolites as flavonoids and isoflavonoids in the plant resistance to ASR. This information could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.

Unraveling Asian Soybean Rust metabolomics using mass spectrometry and Molecular Networking approach.

Asian Soybean Rust (ASR), caused by the biotrophic fungus Phakopsora pachyrhizi, is a devastating disease with an estimated crop yield loss of up to 90%. Yet, there is a nerf of information on the metabolic response of soybean plants to the pathogen Untargeted metabolomics and Global Natural Products Social Molecular Networking platform approach was used to explore soybean metabolome modulation to P. pachyrhizi infection. Soybean plants susceptible to ASR was inoculated with P. pachyrhizi spore suspension and non-inoculated plants were used as controls. Leaves from both groups were collected 14 days post-inoculation and extracted using different extractor solvent mixtures. The extracts were analyzed on an ultra-high performance liquid chromatography system coupled to high-definition electrospray ionization-mass spectrometry. There was a significant production of defense secondary metabolites (phenylpropanoids, terpenoids and flavonoids) when P. pachyrhizi infected soybean plants, such as putatively identified liquiritigenin, coumestrol, formononetin, pisatin, medicarpin, biochanin A, glyoceollidin I, glyoceollidin II, glyoceollin I, glyoceolidin II, glyoceolidin III, glyoceolidin IV, glyoceolidin VI. Primary metabolites (amino acids, peptides and lipids) also were putatively identified. This is the first report using untargeted metabolomics and GNPS-Molecular Networking approach to explore ASR in soybean plants. Our data provide insights into the potential role of some metabolites in the plant resistance to ASR, which could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.

Superovulatory response, production and quality of embryos of cows fed on linseed or canola seed supplemented diets - doi: 10.4025/actascianimsci.v34i3.13372

Superovulatory response, production and embryo quality and the effects of seasonal changes on embryo production of Nellore cows fed on either linseed (n-3) or canola seeds (n-6) were evaluated. Sixteen Nellore cows (550 ± 48.0 kg) were fed on three diets: control diet (CON), diet with linseed (LIN) and diet with canola seed (CAN) during four seasons. There was no difference in superovulatory response and in total corpora lutea produced when diets are taken into consideration. The number of cows that responded to superovulatory treatment was higher in the winter (93.8%) than in the summer (62.5%). No difference with regard to the three diets existed on the average number of total structures (6.20, 4.96 and 6.50), unfertilized structures (2.53, 1.17 and 1.60) or congealable embryos (3.40, 1.30 and 3.80). Average degenerated embryos were higher for cows fed on LIN diet (2.48) than for those fed on CON diet (0.32); the CAN diet presented an intermediate response (1.10). Mean total structures produced were lower during the winter (3.57) and summer (3.50) than during the autumn (8.75) and spring (7.25).

Substituição do milho pelo resíduo da fecularia de mandioca no desempenho e nas características de carcaça de bovinos terminados em confinamento / Levels of replacing corn by cassava starch on performance and carcass characteristics of bulls finished in feedlot

Objetivou-se com este trabalho avaliar o efeito do nível de substituição (0; 12,5; 22,8 e 32,7%) do milho pelo resíduo de fecularia de mandioca sobre o desempenho e características de carcaça de bovinos em confinamento. Trinta e dois machos inteiros mestiços (½ Aberdeen Angus vs. ½ Nelore) com 18 meses de idade e 380 + 24 kg de peso vivo foram utilizados em delineamento experimental inteiramente casualizado, com quatro tratamentos e oito repetições. O experimento foi desenvolvido durante 56 dias com 14 dias de adaptação. As rações completas [volumoso (casca de algodão) + concentrado (milho, farelo de soja e resíduo desidratado de fecularia de mandioca)] foram fornecidas às 8 h e 16 h aos animais. Foi analisado o peso inicial (PI), peso final (PF) ganho médio diário (GMD), ingestão de matéria seca (IMS), conversão alimentar da matéria seca (CAMS), peso de carcaça quente (PCQ), rendimento de carcaça quente (RCQ), espessura de gordura de cobertura (EGC), área de olho de lombo (AOL), comprimento de perna (CP) e espessura de coxão (EC). Os resultados observados para PI, PF, GMD, CAMS, RCQ, EGC, AOL, CP e EC não apresentaram diferenças (P > 0,05) entre os tratamentos. Os resultados obtidos para desempenho e características de carcaça foram satisfatórios.

This work was carried out to study the effects of replacing corn by different levels of cassava starch (0; 12.5; 22.8 and 32.7%) on performance and carcass characteristics of bulls finished in feedlot. Thirty-two crossbred bulls (½ Aberdeen Angus vs. ½ Nellore) with 18 months old and 380 + 24 kg live weight were used in a completely randomized experimental design with four treatments and eight replications. The experiment was realized during 56 days with 14 days for adaptation. The complete diets [roughage (cottonseed hulls) + concentrate (corn, soybean meal and cassava starch)] were given at 8 am and at 4 pm to bulls. It was analyzed the initial weight (IW), final weight (FW), average daily gain (ADG), dry matter intake (DMI), feed conversion (FC), hot carcass dressing (HCD), fat thickness (FAT), Longissimus muscle area (LMA), leg length (LL) and cushion thickness (CT). The IW, FW, ADG, FC, HCW, HCD, FT, LMA, LL and CT did not present difference (P > 0.05) among levels of replacing corn by cassava starch. The results obtained on performance and carcass traits using cassava starch by-products as a replacement for corn can be considered satisfactory.

Monensina sódica ou extrato de própolis na dieta de bovinos Nelore terminados em confinamento: composição química e perfil de ácidos graxos do músculo Longissimus / Sodium monensin or propolis extract in the diet of Nellore bulls finished in feedlot: chemical composition and fatty acid profile of Longissimus muscle

Este trabalho foi realizado para avaliar a composição química e a composição de ácidos graxos do músculo Longissimus de 33 machos não castrados da raça Nelore com 27 meses de idade e peso médio inicial de 402 ± 7.58 kg terminados em confinamento. Foram usados três tratamentos: Controle CON, Monensina MON e Extrato de Própolis PRO. Os bovinos foram mantidos confinados durante 84 dias e abatidos com peso vivo médio final de 488 ± 24.9 kg. A adição dos aditivos (monensina ou extrato de própolis) não tiveram influência (P > 0,10) na composição química do músculo Longissimus dos bovinos. A composição de ácidos graxos saturados (AGS) foi semelhante entre os tratamentos (P > 0,10). A composição de ácidos graxos monoinsaturados foi diferente entre os tratamentos (P 0,01). Valor superior foi observado nos bovinos da dieta CON (47,0%). Da mesma forma, houve diferença (P 0,01) entre tratamentos para os ácidos graxos poliinsaturados (AGPI). Bovinos do tratamento (9,92%) apresentaram maior (P 0,01) valor para os AGPI. Os bovinos dos tratamentos CON (6,74%) e PRO (6,93%) apresentaram menores valores (P 0,01) para os AGPI.

This work was carried out to evaluate the chemical composition and fatty acid profile of Longissimus muscle of 33 Nellore bulls with 27 months old and initial average weight of 402 ± 7.58 kg finished in feedlot. Three treatments (Control CON, Monensin MON and Propolis extract PRO) were evaluated. The animals were kept in feedlot during 84 days and slaughtered with final average weight with 488 ± 24.9 kg. The addition of additives (monensin or propolis extract) did not influence (P > 0.10) the chemical composition of Longissimus muscle of bulls. Saturated fatty acid (SFA) did not have difference (P > 0.10) among treatments. Monounsaturated fatty acids (MUFA) have difference (P 0.01) among treatments. The highest value was observed in CON treatment (47.0%). There is also difference (P 0.01) among treatments for polyunsaturated fatty acids (PUFA). Bulls of MON (9.92%) treatment obtained highest (P 0.01) value for PUFA. Bulls of CON (6.74%) and PRO (6.93%) have lowest (P 0.01) values for PUFA.

Superovulatory response, production and quality of embryos of cows fed on linseed or canola seed supplemented diets - doi: 10.4025/actascianimsci.v34i3.13372

Superovulatory response, production and embryo quality and the effects of seasonal changes on embryo production of Nellore cows fed on either linseed (n-3) or canola seeds (n-6) were evaluated. Sixteen Nellore cows (550 ± 48.0 kg) were fed on three diets: control diet (CON), diet with linseed (LIN) and diet with canola seed (CAN) during four seasons. There was no difference in superovulatory response and in total corpora lutea produced when diets are taken into consideration. The number of cows that responded to superovulatory treatment was higher in the winter (93.8%) than in the summer (62.5%). No difference with regard to the three diets existed on the average number of total structures (6.20, 4.96 and 6.50), unfertilized structures (2.53, 1.17 and 1.60) or congealable embryos (3.40, 1.30 and 3.80). Average degenerated embryos were higher for cows fed on LIN diet (2.48) than for those fed on CON diet (0.32); the CAN diet presented an intermediate response (1.10). Mean total structures produced were lower during the winter (3.57) and summer (3.50) than during the autumn (8.75) and spring (7.25).

Chemical composition and fatty acids profile of five carcass cuts of crossbred heifers finished in feedlotChemical composition and fatty acids profile of five carcass cuts of crossbred heifers finished in feedlot / Composição química e perfil de ácidos graxos de cinco diferentes cortes de novilhas mestiças (Nelore vs Charolês)

This work was carried-out to study the chemical composition and fatty acids profile among chuck, rump, striploin, topside and knuckle from crossbred (½ Nellore x ½ Charolais) heifers finished in feedlot. The knuckle, striploin and topside showed higher (P 0.05) moisture (75.19, 73.87 e 74.36%) and ash contents (1.09; 1.08 and 1.13%) than the chuck (71.18 e 0.93%) and rump (71.58 e 1.07%), respectively. On the other hand, lipid contents of chuck (7.40%) and rump (5.19%) were higher (P 0.05) than striploin (2.42%), topside (1.76%) and knuckle (1.69%). The chuck cholesterol content (57.63mg/100 g of muscle) was higher (P 0.05) than striploin (49.71mg/100 g of muscle) and topside (46.55mg/100 g of muscle). The knuckle percentages of C 18:3 n-3 (0.98%), C20:4n-6 (1.88%), C 20:5 n-3 (0.74%), AGPI (9.34%), n-6 (7.61%) and n-3 (1.73%) were higher (P

O objetivo do presente estudo foi comparar a composição química e perfil de ácidos graxos entre acém, alcatra, contrafilé, coxão mole e patinho de novilhas cruzadas (½ Nelore vs ½ Charolês) terminadas em confinamento. O patinho, coxão mole e contrafilé apresentaram maior percentagem (p 0,05) de umidade (75, 19; 73,87 e 74,36%) e cinzas (1, 09; 1,08 e 1,13%) em relação ao acém (71,18 e 0,93%) e alcatra (71,58 e 1,07%), respectivamente. Por outro lado, as percentagens de lipídios totais do acém (7,40%) e alcatra (5,19%) foram superiores (p 0,05) aos observados no contrafilé (2,42%), coxão mole (1,76%) e patinho (1,69%). O colesterol total do acém (57,63 mg/100 g de músculo) foi superior (p 0,05) ao observado no contrafilé (49,71 mg/100 g de músculo) e coxão mole (46,55 mg/100 g de músculo). As percentagens de C 18:3 n-3, C 20:4 n-6, C 20:5 n-3, AGPI, n- e n-3 do patinho (0,98, 1,88, 0,74, 9,34, 7,61 e 1,73%) foram superiores (p 0,05) às observadas no acém (0,38, 1,13; 0,39; 4,22; 3,45 e 0,77%) e contrafilé (0,53; 0,93; 0,26; 5,16; 4,37 e 0,79%), respectivamente. Desta forma, os cortes que apresentaram menores riscos à saúde humana com relação ao perfil de ácidos graxos, foram o contrafilé, o coxão mole e o patinho.

Chemical composition, fatty acid profile and CLA levels in the Longissimus muscle of Caracu and Caracu vs. Charolais cattle / Composição química, perfil de ácidos graxos e níveis de CLA no músculo Longissimus de bovinos Caracu e Caracu vs. Charolês

This work was conducted in order to study the chemical composition, fatty acid profile, as well as the levels of n-3 and n-6 fatty acids and conjugated linoleic acid (CLA) present in the Longissimus muscle of Caracu and Caracu vs. Charolais genetic groups of cattle. This study was carried out at the Experimental Farm of the Agronomic Institute of Paraná, in southern Brazil. Twenty animals (10 Caracu CAR and 10 Caracu vs. Charolais CAC) were used, with an initial average age between 8 and 10 months. The young bulls were slaughtered at 450 kg and 18 months of age. The moisture and crude protein percentages were similar between the two genetic groups. However, the percentage of ash was higher in the CAC group. Conversely, total lipid levels was lower in the CAC group. The fatty acid profiles were similar for CAR and CAC bulls. Percentages of polyunsaturated fatty acids (PUFA) and n-6 were higher in the CAC group. The monounsaturated fatty acid (MUFA), saturated fatty acid, and n-3 percentages, as well n-6/n-3 and PUFA/MUFA ratios, were similar between CAR and CAC bulls. The 18:2 n-6, 18:2 c 9 t 11 and 20:3 n-6 contents (mg/g of total lipids) were higher in the CAR group.

Este trabalho foi realizado para estudar a composição química, perfil de ácidos graxos e a quantificação dos ácidos graxos n-3, n-6 e ácido linoléico conjugado (CLA) no músculo Longissimus de bovinos inteiros dos grupos genéticos Caracu e Caracu vs. Charolês. Este estudo foi realizado na Fazenda Experimental do Instituto Agronômico do Paraná. Foram utilizados vinte animais (10 Caracu CAR e 10 Caracu vs. Charolês CAC) com idade inicial de 8-10 meses. Os animais foram abatidos com peso médio de 450 kg e 18 meses. A percentagem de umidade e proteína total foram similar entre os dois grupos genéticos. Entretanto, a percentagem de cinzas foi maior no grupo CAC. Ao contrário, a percentagem de lipídeos totais foi menor no grupo CAC. O perfil de ácidos graxos foi similar entre os animais CAR e CAC. A percentagem de ácidos graxos poliinsaturados (AGPI) e n-6 foi maior para animais do grupo CAC. As percentagens de ácidos graxos monoinsaturados (AGMI), ácidos graxos saturados (AGS), n-3 e as razões de n-6/n-3 e AGPI/AGS foram similar entre os animais CAR e CAC. A quantificação dos ácidos graxos 18:2 n-6, 18:2 cis 9 trans 11 and 20:3 n-6 (mg/g of total lipids) foi maior em animais do grupo CAR.

Chemical composition and fatty acids profile of five carcass cuts of crossbred heifers finished in feedlotChemical composition and fatty acids profile of five carcass cuts of crossbred heifers finished in feedlot / Composição química e perfil de ácidos graxos de cinco diferentes cortes de novilhas mestiças (Nelore vs Charolês)

This work was carried-out to study the chemical composition and fatty acids profile among chuck, rump, striploin, topside and knuckle from crossbred (½ Nellore x ½ Charolais) heifers finished in feedlot. The knuckle, striploin and topside showed higher (P 0.05) moisture (75.19, 73.87 e 74.36%) and ash contents (1.09; 1.08 and 1.13%) than the chuck (71.18 e 0.93%) and rump (71.58 e 1.07%), respectively. On the other hand, lipid contents of chuck (7.40%) and rump (5.19%) were higher (P 0.05) than striploin (2.42%), topside (1.76%) and knuckle (1.69%). The chuck cholesterol content (57.63mg/100 g of muscle) was higher (P 0.05) than striploin (49.71mg/100 g of muscle) and topside (46.55mg/100 g of muscle). The knuckle percentages of C 18:3 n-3 (0.98%), C20:4n-6 (1.88%), C 20:5 n-3 (0.74%), AGPI (9.34%), n-6 (7.61%) and n-3 (1.73%) were higher (P

O objetivo do presente estudo foi comparar a composição química e perfil de ácidos graxos entre acém, alcatra, contrafilé, coxão mole e patinho de novilhas cruzadas (½ Nelore vs ½ Charolês) terminadas em confinamento. O patinho, coxão mole e contrafilé apresentaram maior percentagem (p 0,05) de umidade (75, 19; 73,87 e 74,36%) e cinzas (1, 09; 1,08 e 1,13%) em relação ao acém (71,18 e 0,93%) e alcatra (71,58 e 1,07%), respectivamente. Por outro lado, as percentagens de lipídios totais do acém (7,40%) e alcatra (5,19%) foram superiores (p 0,05) aos observados no contrafilé (2,42%), coxão mole (1,76%) e patinho (1,69%). O colesterol total do acém (57,63 mg/100 g de músculo) foi superior (p 0,05) ao observado no contrafilé (49,71 mg/100 g de músculo) e coxão mole (46,55 mg/100 g de músculo). As percentagens de C 18:3 n-3, C 20:4 n-6, C 20:5 n-3, AGPI, n- e n-3 do patinho (0,98, 1,88, 0,74, 9,34, 7,61 e 1,73%) foram superiores (p 0,05) às observadas no acém (0,38, 1,13; 0,39; 4,22; 3,45 e 0,77%) e contrafilé (0,53; 0,93; 0,26; 5,16; 4,37 e 0,79%), respectivamente. Desta forma, os cortes que apresentaram menores riscos à saúde humana com relação ao perfil de ácidos graxos, foram o contrafilé, o coxão mole e o patinho.

Chemical composition, fatty acid profile and CLA levels in the Longissimus muscle of Caracu and Caracu vs. Charolais cattle / Composição química, perfil de ácidos graxos e níveis de CLA no músculo Longissimus de bovinos Caracu e Caracu vs. Charolês

This work was conducted in order to study the chemical composition, fatty acid profile, as well as the levels of n-3 and n-6 fatty acids and conjugated linoleic acid (CLA) present in the Longissimus muscle of Caracu and Caracu vs. Charolais genetic groups of cattle. This study was carried out at the Experimental Farm of the Agronomic Institute of Paraná, in southern Brazil. Twenty animals (10 Caracu CAR and 10 Caracu vs. Charolais CAC) were used, with an initial average age between 8 and 10 months. The young bulls were slaughtered at 450 kg and 18 months of age. The moisture and crude protein percentages were similar between the two genetic groups. However, the percentage of ash was higher in the CAC group. Conversely, total lipid levels was lower in the CAC group. The fatty acid profiles were similar for CAR and CAC bulls. Percentages of polyunsaturated fatty acids (PUFA) and n-6 were higher in the CAC group. The monounsaturated fatty acid (MUFA), saturated fatty acid, and n-3 percentages, as well n-6/n-3 and PUFA/MUFA ratios, were similar between CAR and CAC bulls. The 18:2 n-6, 18:2 c 9 t 11 and 20:3 n-6 contents (mg/g of total lipids) were higher in the CAR group.   

Este trabalho foi realizado para estudar a composição química, perfil de ácidos graxos e a quantificação dos ácidos graxos n-3, n-6 e ácido linoléico conjugado (CLA) no músculo Longissimus de bovinos inteiros dos grupos genéticos Caracu e Caracu vs. Charolês. Este estudo foi realizado na Fazenda Experimental do Instituto Agronômico do Paraná. Foram utilizados vinte animais (10 Caracu CAR e 10 Caracu vs. Charolês CAC) com idade inicial de 8-10 meses. Os animais foram abatidos com peso médio de 450 kg e 18 meses. A percentagem de umidade e proteína total foram similar entre os dois grupos genéticos. Entretanto, a percentagem de cinzas foi maior no grupo CAC. Ao contrário, a percentagem de lipídeos totais foi menor no grupo CAC. O perfil de ácidos graxos foi similar entre os animais CAR e CAC. A percentagem de ácidos graxos poliinsaturados (AGPI) e n-6 foi maior para animais do grupo CAC. As percentagens de ácidos graxos monoinsaturados (AGMI), ácidos graxos saturados (AGS), n-3 e as razões de n-6/n-3 e AGPI/AGS foram similar entre os animais CAR e CAC. A quantificação dos ácidos graxos 18:2 n-6, 18:2 cis 9 trans 11 and 20:3 n-6 (mg/g of total lipids) foi maior em animais do grupo CAR.