Evaluation Of Mycotoxin Content In Soybean (Glycine max L.) Grown In Rwanda.

Soybean is a critical food and nutritional security crop in Rwanda. Promoted by the Rwandan National Agricultural Research System for both adults and as an infant weaning food, soybean is grown by approximately 40% of households. Soybean may be susceptible to the growth of mycotoxin-producing moulds; however, data has been contradictory. Mycotoxin contamination is a food and feed safety issue for grains and other field crops. This study aimed to determine the extent of mycotoxin contamination in soybean, and to assess people's awareness on mycotoxins. A farm-level survey was conducted in 2015 within three agro-ecological zones of Rwanda suitable for soybean production. Soybean samples were collected from farmers (n=300) who also completed questionnaires about pre-and post-harvest farm practices, and aflatoxin awareness. The concentration of total aflatoxin in individual soybean samples was tested by enzymelinked immunosorbent assay (ELISA) using a commercially-available kit. Other mycotoxins were analyzed using liquid chromatography-mass spectrometry (LCMS/MS) on 10 selected sub samples. Only 7.3% of the respondents were aware of aflatoxin contamination in foods, but farmers observed good postharvest practices including harvesting the crop when the pods were dry. Using enzyme-linked immunosorbent assay (ELISA), only one sample had a concentration (11 µg/kg) above the most stringent EU maximum permitted limit of 4 µg/kg. Multi-mycotoxins liquid chromatography-mass spectrometry (LC-MS/MS) results confirmed that soybeans had low or undetectable contamination; only one sample contained 13µg/kg of sterigmatocystine. The soybean samples from Rwanda obtained acceptably low mycotoxin levels. Taken together with other studies that showed that soybean is less contaminated by mycotoxins, these results demonstrate that soybean can be promoted as a nutritious and safe food. However, there is a general need for educating farmers on mycotoxin contamination in food and feed to ensure better standards are adhered to safeguard the health of the consumers regarding these fungal secondary metabolites.

The effect of different concentrations of linseed oil or fish oil in the maternal diet on the fatty acid composition and oxidative status of sows and piglets.

N-3 polyunsaturated fatty acids (PUFA) are essential for foetal development. Hence, including n-3 PUFA in the sow diet can be beneficial for reproduction. Both the amount and form (precursor fatty acids vs. long chain PUFA) of supplementation are important in this respect. Furthermore, including n-3 PUFA in the diet can have negative effects, such as decreased arachidonic acid (ARA) concentration and increased oxidative stress. This study aimed to compare the efficacy to increase eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) concentrations in the piglet, when different concentrations of linseed oil (LO, source of precursor α-linolenic acid) or fish oil (FO, source of EPA and DHA) were included in the maternal diet. Sows were fed a palm oil diet or a diet including 0.5% or 2% LO or FO from day 45 of gestation until weaning. Linoleic acid (LA) was kept constant in the diets to prevent a decrease in ARA, and all diets were supplemented with α-tocopherol acetate (150 mg/kg) and organic selenium (0.4 mg/kg) to prevent oxidative stress. Feeding 0.5% LO or 0.5% FO to the sows resulted in comparable EPA concentrations in the 5-day old piglet liver, but both diets resulted in lower EPA concentrations than when 2% LO was fed. The highest EPA concentration was obtained when 2% FO was fed. The DHA level in the piglet liver could only be increased when FO, but not LO, was fed to the sows. The 2% FO diet had no advantage over the 0.5% FO diet to increase DHA in the piglet. Despite the constant LA concentration in the sow diet, a decrease in ARA could not be avoided when LO or FO were included in the diet. Feeding 2% FO to the sows increased the malondialdehyde concentration (marker for lipid peroxidation) in sow plasma, but not in piglets.

Linoleate isomerase activity occurs in lactic acid bacteria strains and is affected by pH and temperature.

AIMS: To investigate the ability of lactic acid bacteria (LAB) to convert linoleic acid (LA) and α-linolenic acid (α-LNA) to conjugated linoleic acid (CLA) and conjugated linolenic acid (CLNA), respectively. To assess pH and temperature influences on CLA and CLNA production by Lactobacillus sakei LMG 13558. METHODS AND RESULTS: A screening of 48 LAB yielded one Lactobacillus curvatus, five Lactobacillus plantarum and four Lact. sakei strains displaying linoleate isomerase (LAI) activity. CLNA conversion percentages varied largely (1-60%). CLA conversion, occurring in three strains, was lower (2-5%). The LAI gene sequences of the ten LAI-positive strains shared 75-99% identity with the LAI gene sequence of a Lact. plantarum AS1.555. At pH 6.2, CLA and CLNA production by Lact. sakei LMG 13558 was higher at 30°C than at 20 and 25°C. At pH 5.5 (30°C) or 37°C (pH 6.2), LA was not converted and α-LNA only slightly converted. CONCLUSIONS: LAB show strain-dependent LAI activity. Production of CLA and CLNA is affected by pH and temperature, as shown for Lact. sakei LMG 13558. SIGNIFICANCE AND IMPACT OF THE STUDY: Several LAB produce CLA and/or CLNA, as shown for Lact. sakei and Lact. curvatus for the first time. These findings offer potential for the manufacturing of fermented functional foods.

Effect of dietary antioxidant supplementation on the oxidative status of plasma in broilers.

In this study, the effect of dietary antioxidants on the plasma oxidative status of growing birds fed a diet rich in polyunsaturated fatty acids was investigated. One-day-old broilers were fed for 42 days a diet containing 4% linseed oil and supplemented with single plant extracts rich in antioxidants (natural tocopherols, rosemary, grape seed, green tea, tomato) or a combination of some of these plant extracts, in two different total doses (100 and 200 mg product/kg feed). A diet with synthetic antioxidants with and without α-tocopheryl acetate (200 mg/kg feed) were also included. The plasma oxidative status was evaluated measuring the ferric reducing ability of plasma (FRAP), the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Lipid peroxidation was measured by thiobarbituric acid-reactive substances (TBARS). No significant effect of the dietary treatments was observed for FRAP as well as for TBARS. However, diet affected GSH-Px activity (p = 0.002) and a trend for an effect on SOD activity was observed (p=0.084). A higher GSH-Px activity was found for 200 mg/kg tomato extract and natural α-tocopherol in relation to the corresponding 100 mg/kg treatment, and the lowest GSH-Px activity was measured for the synthetic antioxidants treatment. The lowest and highest SOD activity were found for the 200. and 100 mg/kg treatment with tomato extract respectively. In conclusion, the oxidative status and lipid oxidation of plasma in broilers was not affected by feeding natural antioxidant extracts at the doses in the present study, but some changes in antioxidant enzyme activities were observed, of which the implication remains to be elucidated.

Effects of diet, age and gender on the polyunsaturated fatty acid composition of broiler anatomical compartments.

1. The objective of this study was to investigate the effect of dietary fatty acid (FA) composition, age, and gender on the FA composition of different broiler anatomical compartments. Four dietary fat sources (palm fat, P; soybean oil, S; linseed oil, L; fish oil, F) were added to a wheat-soybean meal based diet at 30 g/kg in addition to 50 g/kg palm fat. Diets were fed separately to female and male birds from d 1 to either d 21 or d 42 of age. 2. The total FA content (mg/100 g tissue) and the FA composition (g/100 g FAME) was determined in 7 anatomical compartments (skinless thigh muscle, skinless breast muscle, liver, heart, brain, abdominal fat pad, and remainders plus carcase trimmings named Rest compartment). The FA profiles differed greatly among compartments and were strongly affected by diet. 3. The S diet resulted in a 2-3-fold increase of the proportion of C18:2n-6 and C20:4n-6 in all compartments compared to the other diets, except for brain. 4. The response in the proportion of C18:3n-3 following feeding the L diet was much greater, with 4-20-fold increases compared to the other diets, except again for brain. 5. In all compartments except brain, the L diet resulted in approximately a 2-fold increase in the proportion of C22:6n-3 compared to the P and S diets and smaller, but also significant, increases for C20:5n-3 and C22:5n-3. 6. The proportions of C20:5n-3, C22:5n-3 and C22:6n-3 were much higher on the F diet in all compartments, except for brain, compared to the P and S diets. The F diet resulted in higher proportions of C22:6n-3 than the proportions of C20:5n-3 and C22:5n-3 in breast and thigh meat, and liver, in contrast to the other diets. 7. Brain was less responsive than other tissues to changes in the dietary FA composition. The proportion of C22:6n-3 in brain was similar on the L and F diets. 8. The effect of gender on the FA composition of different anatomical compartments was marginal. The long-chain n-3 and n-6 PUFA in breast, liver, and brain decreased with age.

Effect of diet, sex and age on fatty acid metabolism in broiler chickens: SFA and MUFA.

The present study was conducted to evaluate the effect of different dietary lipid sources, age and sex on the SFA and MUFA metabolism in broiler chickens using a whole body fatty acid balance method. Four dietary lipid sources (palm fat, Palm; soyabean oil, Soya; linseed oil, Lin; and fish oil, Fish) were added at 3 % to a basal diet containing 5 % Palm. Diets were fed to female and male chickens from day 1 to either day 21 or day 42 of age. The accumulation (percentage of net intake and ex novo production) of SFA and MUFA was significantly lower in broilers fed on Palm than in broilers fed on the other diets (85.7 v. 97.4 %). Conversely, beta-oxidation was significantly higher in Palm-fed birds than the average of the other dietary treatments (14.3 v. 2.6 %). On average, 33.1 % of total SFA and MUFA accumulated in the body were elongated, and 13.8 % were Delta-9 desaturated to longer chain or more unsaturated metabolites, with lower proportions being elongated and desaturated for the Palm and Fish diets than for the Soya and Lin diets. Total in vivo apparent elongase activity decreased exponentially in relation to the net intake of SFA and MUFA, while it increased with age. Total in vivo apparent Delta-9 desaturase activity was not significantly affected by dietary treatment or age. Total ex novo production and beta-oxidation of SFA and MUFA showed a negative and positive curvilinear relationship with net intake of SFA and MUFA, respectively. Sex had no effect on SFA and MUFA metabolism.

Effect of diet, sex and age on fatty acid metabolism in broiler chickens: n-3 and n-6 PUFA.

The PUFA metabolism in broiler chicken was studied through the whole body fatty acid balance method. Four dietary lipid sources (palm fat, Palm; soyabean oil, Soya; linseed oil, Lin; fish oil, Fish) were added at 3 % to a basal diet containing 5 % palm fat. Diets were fed to female and male birds from day 1 to either day 21 or day 42 of age. Birds fed the Lin diet showed a significantly higher 18 : 2n-6 accumulation compared with the other diets (85.2 v. 73.6 % of net intake), whereas diet did not affect 18 : 3n-3 accumulation (mean 63 % of net intake). Bioconversion of 18 : 2n-6 significantly decreased in the order Palm>Lin>Soya>Fish (4.7, 3.9, 3.4 and 1 % of net intake, respectively). The 18 : 3n-3 bioconversion on the Palm and Soya diets was similar and significantly higher than in broilers on the Lin diet (9.1 v. 5.8 % of net intake). The beta-oxidation of 18 : 2n-6 was significantly lower on the Lin diet than on the other diets (10.8 v. 23.3 % of net intake), whereas beta-oxidation of 18 : 3n-3 was significantly higher on the Fish diet than on the other diets (41.5 v. 27.3 % of net intake). Feeding fish oil suppressed apparent elongase and desaturase activity, whereas a higher dietary supply of 18 : 3n-3 and 18 : 2n-6 enhanced apparent elongation and desaturation activity on the PUFA involved in the n-3 and n-6 pathway, respectively. Accumulation of 18 : 2n-6 and 18 : 3n-3 increased and beta-oxidation decreased with age. Sex had a marginal effect on the PUFA metabolism.

Crystallization behavior of milk fat obtained from linseed-fed cows.

Milk with an increased content of unsaturated fatty acids was obtained by incorporating 60% of extruded linseed into the concentrate of cows. Two groups of Holstein cows (3 animals/group) were fed a concentrate (control or linseed enriched) together with the same roughage diet (ad libitum). After an adaptation period of 3 wk, evening and morning milk samples were collected every 7 d for 3 wk. Milk was decreamed and anhydrous milk fat (AMF) was isolated from the fat fraction by using the Bureau of Dairy Industries method. The objective of this study was to investigate if the crystallization mechanism of milk fat changed when the content of unsaturated fatty acids was increased. Therefore, the crystallization behavior of a milk fat enriched with unsaturated fatty acids was compared with that of a control milk fat. Nonisothermal crystallization was investigated with differential scanning calorimetry, and 1-step and 2-step isothermal crystallization behaviors were investigated using pulsed nuclear magnetic resonance, differential scanning calorimetry, and x-ray diffraction. A higher content of unsaturated fatty acids in AMF resulted in an increased proportion of low melting triglycerides. These triglycerides lowered the solid fat content profile, particularly at refrigerator temperatures. Furthermore, they induced some changes in the crystallization and melting behaviors of milk fat compared with a control AMF, although no fundamental changes in the crystallization mechanism could be revealed. Even though a lower melting point could be observed for milk fat with a higher content of unsaturated fatty acids, a similar degree of supercooling was needed to initiate crystallization, resulting in a shift in onset temperature of crystallization toward lower temperatures. In addition, slower crystallization kinetics were measured, such as a lower nucleation rate and longer induction times, although crystallization occurred in a similar polymorphic crystal lattice. During melting, a shift in offset temperature toward lower temperatures could be observed for the 3 melting fractions of AMF in addition to a higher proportion of low melting triglycerides. These results demonstrate that a higher content of unsaturated fatty acids has some effect on the crystallization behavior of milk fat. This knowledge could be used to produce dairy products of similar or superior quality compared with conventional products by intervening in the production process of dairy products.

Genetic parameters for intramuscular fatty acid composition and metabolism in pigs.

The aim of this study was to estimate genetic parameters for pork intramuscular fatty acid (FA) composition and indices for desaturase and elongase activities involved in n-3 and n-6 PUFA metabolism. The LM of 437 slaughter pigs was analyzed for FA composition (expressed as g/100 g of FA). Indices for enzyme activities were calculated from product to precursor FA ratios. Genetic parameters were estimated with single- and multi-trait animal models. The total FA content, reflecting the intramuscular fat content, was either included or not in the model. Results from the models without total FA content showed relatively high heritability estimates, generally above 0.50, for the proportion of the most important MUFA and PUFA, compared with much smaller values for the SFA. When total FA content was included in the models, heritability values decreased (P < 0.001) for most individual FA and for all sums of FA groups, except for C18:0, C18:3n-6, and C18:3n-3. Heritability estimates for the ratios C20:4n-6/C18:2n-6 and C22:6n-3/C18:3n-3, reflecting the overall conversion in the n-6 and n-3 PUFA pathway, respectively, were 0.29 and 0.35, respectively, with total FA content in the model and increased to 0.38 and 0.49, respectively, if total FA content was not in the model. Heritabilities for other more specific indices were of the same order. Genetic correlations between PUFA proportions and indices for enzyme activities with ADG were mostly negative, whereas the correlations with carcass lean meat percentage were mostly positive. It was concluded that there is meaningful genetic variation for long-chain PUFA metabolism that is only partly dependent on the carcass and muscle fat content. This may allow selection for improved FA composition of pork.

Effect of supplementation of the maternal diet with fish oil or linseed oil on fatty-acid composition and expression of Δ5- and Δ6-desaturase in tissues of female piglets.

The present study investigated whether enrichment of the pig maternal diet with n-3 polyunsaturated fatty acids (PUFA) affects the fatty-acid composition of female piglets via enhancing of expression of the lipogenic enzymes Δ5-desaturase (Δ5d) and Δ6-desaturase (Δ6d). The sows (50% Landrace × 50% Large White) were fed a control diet or one of the experimental diets starting at day 45 in gestation. The experimental diets were supplemented either with linseed oil or fish oil, whereas the control diet contained palm oil. Expression of Δ5d and Δ6d, and fatty-acid composition was determined by Western blotting and gas-liquid chromatography, respectively, in muscle, subcutaneous adipose tissue and liver. The highest Δ5d protein expression was observed in the piglets' muscle, followed by subcutaneous adipose tissue, with the lowest level in the liver. Expression of Δ6d in the piglets' tissues followed an opposite pattern, and was highest in the liver, followed by subcutaneous adipose tissue, with the lowest level in muscle. Supplementation of the maternal diet with fish oil or linseed oil increased the level of n-3 PUFA of the piglets in a tissue-specific manner. The response of Δ6d and Δ5d protein expression in female piglets, with average birth weight 2.4 kg, to the dietary manipulation was also tissue-specific. It is suggested that the increase in n-3 PUFA content in the progeny was related, at least partially, to the activation of Δ6d and Δ5d expression.

Lipid and protein oxidation of broiler meat as influenced by dietary natural antioxidant supplementation.

Natural tocopherols (TC), rosemary (RO), green tea (GT), grape seed, and tomato extracts were supplemented in single and in combinations at total concentrations of 100 and 200 mg.kg(-1) of feed in a 4% linseed oil-containing diet to investigate the oxidative stability of broiler breast muscle. Supplementation with 300 mg.kg(-)1 of synthetic antioxidants alone and synthetic antioxidants with alpha-tocopheryl acetate at a concentration of 200 mg.kg(-1) (100 IU) feed was used as a control. Fresh patties were prepared and stored under light at 4 degrees C. After freezing for 8 mo and overnight thawing, 3 other patties were prepared and similarly stored under light at 4 degrees C. During display, samples were evaluated for oxidative stability measurements. For lipid oxidation, the treatment with synthetic antioxidants and 200 mg.kg(-1) of alpha-tocopheryl acetate yielded the lowest TBA reactive species (TBARS) values. For TC, grape seed, and tomato extracts, TBARS values for 100 mg.kg(-1) were higher (P < 0.05) than 200 mg.kg(-1) treatments, whereas no differences (P > 0.05) in TBARS values were observed for RO between 100 and 200 mg.kg(-1). In contrast, GT showed higher TBARS values at 200 mg.kg(-1). Administration of combinations of TC, RO, and GT did not reveal synergistic effects but confirmed the increase in TBARS values with increasing doses of GT. No differences (P > 0.05) among the different antioxidant treatments were detected for protein oxidation. The muscle alpha-tocopherol content linearly responded to the feed alpha-tocopherol content and thus there were no indications for a sparing effect on alpha-tocopherol from other antioxidant treatments. In summary, dietary natural antioxidant extracts were less effective than the treatment with synthetic antioxidants combined with alpha-tocopheryl acetate for protecting against oxidation, but there were marked differences between different natural antioxidant extracts.

Fatty acid profile and oxidative stability of pork as influenced by duration and time of dietary linseed or fish oil supplementation.

In this experiment, the effect of duration and time of feeding n-3 PUFA sources on the fatty acid composition and oxidative stability of the longissimus thoracis (LT) muscle was investigated. Linseed (L) and fish oil (F), rich in alpha-linolenic acid and eicosapentaenoic and docosahexaenoic acid (EPA and DHA), respectively, were supplied equivalent to a level of 1.2% oil (as fed), either during the whole fattening period or only during the first (P1; 8 wk) or second (P2; 6 to 9 wk until slaughter) fattening phase. All diets were based on barley, wheat, and soybean meal and were fed ad libitum. Crossbred pigs (n = 154; Topigs 40 x Piétrain) were randomly allotted to the 7 feeding groups. In the basal diet (B), only animal fat was used as the supplementary fat source. Three dietary groups were supplied the same fatty acid source during both fattening phases (i.e., group BB, LL, and FF). For the other 4 dietary groups, the fatty acid source was switched after the first phase (groups BL, BF, LF, and FL; the first and second letter indicating the diet in P1 and P2, respectively). Twelve animals per feeding group were selected based on average live BW. The LT was analyzed for fatty acid composition; lipid stability (thiobarbituric acid-reactive substances) and color stability (a* value, % of myoglobin pigments) were determined on the LT after illuminated chill storage for up to 8 d. The alpha-linolenic acid, EPA, and docosapentaenoic acid incorporation was independent of the duration of linseed feeding (1.24, 0.54, and 0.75% of total fatty acids, respectively, for group LL). Supplying fish oil during both phases resulted in the greatest EPA and DHA proportions (1.37 and 1.02% of total fatty acids; P < 0.05), but the content of docosapentaenoic acid was not affected. The proportion of DHA was greater when fish oil was administered during P2 compared with P1 (P < 0.05). There was no effect of diet on meat ultimate pH and drip loss or on lipid or color oxidation.

Effect of dietary rosemary and α-tocopheryl acetate on the oxidative stability of raw and cooked pork following oxidized linseed oil administration.

The effect of a 2% dietary administration to pigs of oxidized linseed oil (targeted level of 150mEq.O(2)/kg oil after heating at 50°C and exposure to air for 3-4 days following addition of 10ppm CuSO(4)), either or not in combination with antioxidants, on the oxidative stability of raw and cooked pork during illuminated chill storage was assessed. The antioxidant treatments were: 40ppm α-tocopheryl acetate, 40ppm rosemary extract, 40ppm rosemary extract+2ppm gallic acid, and 40ppm α-tocopheryl acetate+40ppm rosemary extract. A total of 20ppm of α-tocopheryl acetate (ATA) was added to all diets in order to meet the physiological requirement of the animals. The antioxidant treatments did not exert any effect on colour and protein oxidation. Lipid oxidation was only decreased by dietary ATA when comparing the ATA supplemented groups combined versus a control treatment group for raw but not for cooked meat. This was due to a higher content of α-tocopherol in the meat and subcutaneous fat. The lipid oxidation results suggested a lack of antioxidant effect for the rosemary extract. No evidence for a synergistic effect of the antioxidant combinations was observed.

Influence of dietary selenium and vitamin E on quality of veal.

Three groups of six calves each were fed a milk replacer and a starter concentrate for 15weeks. Calves of the first group received the basal diet containing selenium (Se) and vitamin E at 0.095-0.128mg and 30-33mg per kg of total solids, respectively. Calves of the second group received the basal diet supplemented with Se-enriched yeast to increase dietary Se concentration to 0.50mg/kg. The third group of calves received the latter diet supplemented with vitamin E to increase its concentration to 100mg/kg. There was no effect of diet on growth rate, digestibility of dry matter and Se, chemical composition of meat (M. Longissimus thoracis et lumborum), meat colour and fatty acid profile of meat lipids. The Se supplementation significantly increased Se concentration in muscle from 0.21 to 0.43mg/kg. The activity of glutathione peroxidase (GSH-Px) in muscle and liver tissue of Se-supplemented animals was increased by 56% and 67%, respectively, compared to the control. The combined supplementation of vitamin E and Se significantly improved the lipid stability of meat compared to the control diet, but not compared to the Se-supplemented diet. It can be concluded that dietary Se supplementation increases the concentration of Se and the GSH-Px activity in meat, but has limited potential for improving meat oxidative stability.

Effect of grazing pastures of different botanical composition on antioxidant enzyme activities and oxidative stability of lamb meat.

The aim of this work was to study the influence of different pastures (Intensive ryegrass, Botanically diverse and Leguminosa rich pastures) on the antioxidant status and oxidative stability of meat from lambs that had been exclusively grazing for three months. Lipid, colour and protein oxidation, α-tocopherol content and activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (Cat) and glutathione peroxidase (GSH-Px)) were measured in Longisimus thoracis et lumborum muscle samples taken 1 day after slaughter. Pasture type significantly affected protein oxidation and the activity of GSH-Px, but no significant differences were found for the α-tocopherol content, colour and lipid oxidation, and the activities of SOD and Cat. Grazing a Botanically diverse pasture induced significantly higher protein oxidation in meat, as measured by the free thiol and carbonyl contents, compared to a Leguminosa rich or Intensive ryegrass pasture (P<0.05). The GSH-Px activity was significantly higher in meat from lambs on the Leguminosa rich pasture compared to the other pasture groups (P<0.01).