Trans 10, cis 12-conjugated linoleic acid reduced reproductive ability by disrupting the estrus cycle in female mice.

As a positional and geometrical isomer of linoleic acid, trans 10, cis 12 conjugated linoleic acid (t10c12-CLA) reduces white fat by reducing food intake, modulating lipid metabolism, and stimulating energy expenditure. However, the t10c12-CLA products are mostly mixtures, making it difficult to obtain accurate results. Studies are needed to investigate the effects of pure t10c12-CLA on animals and humans. In this study, we used the biallelic transgenic (tg) mice, which could produce t10c12-CLA itself, to investigate the effects of pure t10c12-CLA on female reproductive ability. The results showed that the body and relative ovary weights had no significant difference between tg and wild-type (wt) littermates at ages 3 or 10 weeks. While the fecundity test found that tg mice had a significantly longer first litter time (32.0 ± 4.70 days vs. 21.3 ± 2.31 days, P<0.05), and a significantly lower number of litters (4.75 ± 2.75 vs. 6.67 ± 0.57, P<0.05) when compared with wt mice during continuous mating within seven months. Hormone profiles showed that serum estradiol levels did not change in tg mice; however, significantly (P<0.05) decreased progesterone and increased prostaglandin E2 levels were observed in tg mice compared with those of wt mice. Hematoxylin-eosin staining showed no pathological characteristics in tg ovaries, except for the increased atresia follicles (P<0.05). Moreover, the tg mice had a significantly more extended diestrus period than the wt mice (48.4 ± 6.38% vs. 39.6 ± 3.81%, P<0.05). In summary, t10c12-CLA could affect serum progesterone and prostaglandin E2 levels, lead to a disordered estrus cycle, and impact the reproductive performance of female mice. This study provided theoretical and biosafety recommendations for applying t10c12-CLA in female mammals.

Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk

Abstract Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 g.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

Resumo O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 g.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk / Purificação parcial da enzima isomerase do ácido linoleico da bactéria Lactobacillus paracasei isolada do leite

Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 µg.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 µg.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid gastric administration during the pregestational and gestational periods does not influence the follicular endowment of the progeny.

Fetal programming suggests that maternal stimulation and nutrition during the period of fetal development can program the progeny. Conjugated linoleic acid (CLA), an isomer of linoleic acid, has been characterized in several aspects, but few studies have been performed on its involvement in reproduction and fetal programming. The aim of this study was to evaluate the F1, F2 and F3 progeny of female mice supplemented with CLA during the pregestational and gestational periods with respect to biometric and reproductive parameters, as well as ovarian morphophysiology. The F1 progeny of mothers supplemented with CLA exhibited stable weight gain, while the F2 progeny showed no effects (P=0.0187 and P=0.0245, respectively). A reduction in Lee's Index was observed in both generations at the second post-weaning evaluation week in the animals treated with CLA (P=0.0100 and P=0.0078, respectively). The F2 generation showed an increase in the anogenital index in both sexes of the animals treated with CLA (P= 0.0114 and P<0.0001, female and male respectively). CLA administration to mothers did not affect any of the following in their progeny: ovarian follicle mobilization (P>0.05), follicle number (P>0.05) and the integrated density of the lipid content of oocytes included in antral follicles (P>0.05). This study evaluated the use of CLA in mothers and found that it did not affect the progeny regarding murine reproductive performance, suggesting that this supplement can be used safely.

Biofortification of cow milk through dietary supplementation with sunflower oil: fatty acid profile, atherogenicity, and thrombogenic index.

The present study aimed to assess the effects of replacing the starchy ingredients of concentrate by increasing the levels of sunflower oil on the production, composition, fatty acid profile, and evaluate the atherogenicity and thrombogenic index of Jersey cow's milk. Eight Jersey cows were arranged in a double Latin square and distributed in treatments consisting of supplementation with increasing levels of sunflower oil replacing the corn grain and wheat bran of concentrate, including the following: T0 (control diet), without sunflower oil and with 38 g ether extract (EE)/kg dry matter (DM); T1 = 65 g EE/kg DM; T2 = 86 g EE/kg DM; and T3 = 110 g EE/kg DM. The daily milk production was measured, and the corrected milk production was calculated. Milk samples were analyzed by infrared spectroscopy to determine fat, protein, lactose, and total solids, whereas the lipid profile was assessed by gas chromatography. Milk production, energy-corrected milk production, fat content, daily fat production, lactose, and total solids were not affected by the treatments. Protein, lactose, and total solids concentrations decreased. Short-, medium-, and odd-chain fatty acids decreased with an increase in sunflower oil levels. Conversely, linear increases in long-chain, monounsaturated, and polyunsaturated fatty acid concentrations were observed. There were significant increases in stearic and elaidic acids and conjugated linoleic acid isomers, especially in vaccenic and rumenic acids. There was a positive effect on the milk atherogenicity, thrombogenicity, and nutraceutical indices. Dietary supplementation with sunflower oil changes the milk FA profile, decreases the atherogenicity and the thrombogenicity indices, and improve the nutraceutical index up to the addition of 86 g EE/kg DM de sunflower oil in the diets of Jersey cows.

Cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid gastric administration during the pregestational and gestational periods does not influence the follicular endowment of the progeny

Abstract Fetal programming suggests that maternal stimulation and nutrition during the period of fetal development can program the progeny. Conjugated linoleic acid (CLA), an isomer of linoleic acid, has been characterized in several aspects, but few studies have been performed on its involvement in reproduction and fetal programming. The aim of this study was to evaluate the F1, F2 and F3 progeny of female mice supplemented with CLA during the pregestational and gestational periods with respect to biometric and reproductive parameters, as well as ovarian morphophysiology. The F1 progeny of mothers supplemented with CLA exhibited stable weight gain, while the F2 progeny showed no effects (P=0.0187 and P=0.0245, respectively). A reduction in Lee's Index was observed in both generations at the second post-weaning evaluation week in the animals treated with CLA (P=0.0100 and P=0.0078, respectively). The F2 generation showed an increase in the anogenital index in both sexes of the animals treated with CLA (P= 0.0114 and P<0.0001, female and male respectively). CLA administration to mothers did not affect any of the following in their progeny: ovarian follicle mobilization (P>0.05), follicle number (P>0.05) and the integrated density of the lipid content of oocytes included in antral follicles (P>0.05). This study evaluated the use of CLA in mothers and found that it did not affect the progeny regarding murine reproductive performance, suggesting that this supplement can be used safely.

Dietary Effect of Palm Kernel Oil Inclusion in Feeding Finishing Lambs on Meat Quality.

This study evaluated the effects of palm kernel oil (PKO) in the diet of lambs on carcass characteristics, quality, and fatty acid profile of the meat. Forty uncastrated male Santa Inês lambs were used and divided among the treatments: PKOzero without inclusion; PKO1.3­added 1.3%; PKO2.6­added 2.6%; PKO3.9­added 3.9%; PKO5.2­added 5.2%. The carcass characteristics, the variables related to meat color, and the chemical composition of the Longissimus lumborum of lambs were not affected by the PKO inclusion. The weight of the carcasses at slaughter, hot and cold, half carcass, loin-eye area, and commercial cuts decreased linearly when PKO was added to the lamb diet (p < 0.01). CCY decreased linearly to the inclusion level of 2.66% PKO (RMSE 2.204). Total conjugated linoleic acid CLA and C18:3 n-3 GA concentrations remained stable until the inclusion levels of 3.44% PKO (RMSE 0.0956) and 2.17% (RMSE 0.0637), decreasing its concentrations as the increased level of PKO. The presence of PKO in the lambs' diet up to the level of 5.2% did not change the meat quality characteristics; thus, from the point of view of lamb meat production and fatty acid profile, the inclusion of PKO is not beneficial.

Lymphocyte and CD62E expression in lichen planus and lichenoid reaction.

BACKGROUND: It is difficult to distinguish the clinical and histopathological aspects of oral lichen planus lesions from those of oral lichenoid reaction. Some criteria were proposed to distinguish them, mainly because they have different biological behaviors. The aim of the present study was to compare the lymphocyte population and the expression of E-selectin between these lesions. METHODS: Participants with a clinical diagnosis of oral lichen planus (GOLP) and oral lichenoid reaction (GOLR) who needed to perform a biopsy were selected. The tissue was frozen and immunostaining was performed for CD3/CD4, CD3/CD8, CD4/CLA, CD8/CLA, and CD62E. The analysis of each immunostaining was accomplished using the ImageJ program. RESULTS: In total, 25 participants with oral lichen planus and 11 with oral lichenoid reaction were seen. In the evaluation of CD3 + CD4+/CD3 + and CD3 + CD8+/CD3 + proportions, there was a higher percentage of these cells in the oral lichen planus group when compared with the oral lichenoid reaction group (p = 0.027 and p = 0.038 respectively). The average number of CLA + lymphocytes for CD4+/CLA + and CD8+/CLA + in both groups was not statistically significant (p = 0.840; d = 0.363). In GOLP, the number of CD4 + CLA+/E-selectin and CD8 + CLA+/E-selectin was not statistically significant (p = 0.951 and p = 0.454 respectively); neither in GOLR (p = 0.454 and p = 0.989 respectively). CONCLUSION: Our results indicate that CD3 + CD4+, CD3 + CD8+, CD4 + CLA+, CD8 + CLA + lymphocytes and E-selectin are present in both lesions. However, the proportion of CD3 + CD4+/CD3 + and CD3 + CD8/CD3 + cells is higher in the oral lichen planus group when compared with the oral lichenoid reaction group, suggesting that these cells may be important for the etiopathogenic mechanism of these lesions.

Effects of cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid supplementation on maternal reproductive parameters.

Conjugated linoleic acid (CLA) is a mixture of positional isomers of linoleic acid found in ruminant products and meat. The diet supplementing with CLA is an emerging area, requiring studies to elucidate its effects on animals and human reproduction, as well as its side effects. Therefore, the aim of this study was to evaluate the effects of CLA gastric administration, during the pregestational and gestational period in biometric and reproductive parameters, as well as in ovarian morphophysiology. Animals were distributed in three groups: (1) control (n = 10); (2) fish oil (n = 10); and (3) CLA (n = 10), that daily received, by gavage, phosphate-buffered saline, fish oil and CLA, respectively, carried out over 50 days (before mating, mating and pregnancy). There was an increment in the nasoanal distance and Lee index of the CLA and fish oil-treated groups during the first weeks (P > 0.05). CLA administration did not affect the ovarian follicle mobilization (P > 0.05), the number of follicles (P > 0.05) and the integrated density of lipid content of oocytes included in antral follicles (P > 0.05). There was no effect of CLA administration on the litter weight (P > 0.05; F2 and F3), however, an increment (P < 0.05) in the number of pups per litter (F2) was observed. Overall, this study demonstrated the absence of side effects of the CLA gastric administration on mice reproductive performance and suggests that this treatment would transgenerationally enhance fertility in this species.

Oilseed Supplementation Improves Milk Composition and Fatty Acid Profile of Cow Milk: A Meta-Analysis and Meta-Regression.

Oilseed supplementation is a strategy to improve milk production and milk composition in dairy cows; however, the response to this approach is inconsistent. Thus, the aim of this study was to evaluate the effect of oilseed supplementation on milk production and milk composition in dairy cows via a meta-analysis and meta-regression. A comprehensive and structured search was performed using the following electronic databases: Google Scholar, Primo-UAEH and PubMed. The response variables were: milk yield (MY), atherogenic index (AI), Σ omega-3 PUFA, Σ omega-6 PUFA, fat, protein, lactose, linoleic acid (LA), linolenic acid (LNA), oleic acid (OA), vaccenic acid (VA), conjugated linoleic acid (CLA), unsaturated fatty acid (UFA) and saturated fatty acid (SFA) contents. The explanatory variables were breed, lactation stage (first, second, and third), oilseed type (linseed, soybean, rapeseed, cottonseed, and sunflower), way (whole, extruded, ground, and roasted), dietary inclusion level, difference of the LA, LNA, OA, forage and NDF of supplemented and control rations, washout period and experimental design. A meta-analysis was performed with the "meta" package of the statistical program R. A meta-regression analysis was applied to explore the sources of heretogeneity. The inclusion of oilseeds in dairy cow rations had a positive effect on CLA (+0.27 g 100 g−1 fatty acids (FA); p < 0.0001), VA (+1.03 g 100 g−1 FA; p < 0.0001), OA (+3.44 g 100 g−1 FA; p < 0.0001), LNA (+0.28 g 100 g−1 FA; p < 0.0001) and UFA (+8.32 g 100 g−1 FA; p < 0.0001), and negative effects on AI (−1.01; p < 0.0001), SFA (−6.51; p < 0.0001), fat milk (−0.11%; p < 0.001) and protein milk (−0.04%; p < 0.007). Fat content was affected by animal breed, lactation stage, type and processing of oilseed and dietary NDF and LA contents. CLA, LA, OA and UFA, desirable FA milk components, were affected by type, processing, and the intake of oilseed; additionally, the concentrations of CLA and VA are affected by washout and design. Oilseed supplementation in dairy cow rations has a positive effect on desirable milk components for human consumption. However, animal response to oilseed supplementation depends on explanatory variables related to experimental design, animal characteristics and the type of oilseed.

Fatty acid profile and beef quality of Nellore and Angus bulls fed whole shelled corn

This study aimed to evaluate the quality traits and fatty acid profile of beef from Nellore and Angus bulls fed whole shelled corn (WSC) and ground corn plus maize silage (GC) diets. Eighteen Nellore and 18 Angus young bulls [381 ± 12 kg initial body weight (BW) and an average age of 20 ± 1.9 months] were used in a completely randomized design using a 2 × 2 factorial arrangement and were slaughtered at a final BW of 451.5 kg and 545.5 kg, respectively. Twenty–four hours after slaughter, samples of longissimus thoracis muscle were collected for the analysis of lipid oxidation, color, fatty acid profile, shear force, and cooking loss. There was no effect of diet × breed interaction on meat color, lipid oxidation, shear force, and cooking loss. Angus beef had lower shear force (p < 0.05) than Nellore beef and had a greater concentration of linoleic acid and polyunsaturated fatty acids (PUFA) (p < 0.01). Beef of bulls fed WSC tended to have greater concentration of CLA C18:2c9t11 (p = 0.09), greater concentration of CLA C18:2t10c12 (p = 0.01), and PUFA (p = 0.05), and consequently, higher oxidation levels. Angus bulls produced beef with greater tenderness and PUFA concentration. The results of fatty acid show a possible change in biohydrogenation when animals are fed the WSC diet, reducing lipogenesis, as this diet increases the C18:2t10c12 content.

Fatty acid profile and beef quality of Nellore and Angus bulls fed whole shelled corn

This study aimed to evaluate the quality traits and fatty acid profile of beef from Nellore and Angus bulls fed whole shelled corn (WSC) and ground corn plus maize silage (GC) diets. Eighteen Nellore and 18 Angus young bulls [381 ± 12 kg initial body weight (BW) and an average age of 20 ± 1.9 months] were used in a completely randomized design using a 2 × 2 factorial arrangement and were slaughtered at a final BW of 451.5 kg and 545.5 kg, respectively. Twenty­four hours after slaughter, samples of longissimus thoracis muscle were collected for the analysis of lipid oxidation, color, fatty acid profile, shear force, and cooking loss. There was no effect of diet × breed interaction on meat color, lipid oxidation, shear force, and cooking loss. Angus beef had lower shear force (p < 0.05) than Nellore beef and had a greater concentration of linoleic acid and polyunsaturated fatty acids (PUFA) (p < 0.01). Beef of bulls fed WSC tended to have greater concentration of CLA C18:2c9t11 (p = 0.09), greater concentration of CLA C18:2t10c12 (p = 0.01), and PUFA (p = 0.05), and consequently, higher oxidation levels. Angus bulls produced beef with greater tenderness and PUFA concentration. The results of fatty acid show a possible change in biohydrogenation when animals are fed the WSC diet, reducing lipogenesis, as this diet increases the C18:2t10c12 content.

Fatty acid profile and beef quality of Nellore and Angus bulls fed whole shelled corn

ABSTRACT: This study aimed to evaluate the quality traits and fatty acid profile of beef from Nellore and Angus bulls fed whole shelled corn (WSC) and ground corn plus maize silage (GC) diets. Eighteen Nellore and 18 Angus young bulls [381 ± 12 kg initial body weight (BW) and an average age of 20 ± 1.9 months] were used in a completely randomized design using a 2 × 2 factorial arrangement and were slaughtered at a final BW of 451.5 kg and 545.5 kg, respectively. Twentyfour hours after slaughter, samples of longissimus thoracis muscle were collected for the analysis of lipid oxidation, color, fatty acid profile, shear force, and cooking loss. There was no effect of diet × breed interaction on meat color, lipid oxidation, shear force, and cooking loss. Angus beef had lower shear force (p 0.05) than Nellore beef and had a greater concentration of linoleic acid and polyunsaturated fatty acids (PUFA) (p 0.01). Beef of bulls fed WSC tended to have greater concentration of CLA C18:2c9t11 (p = 0.09), greater concentration of CLA C18:2t10c12 (p = 0.01), and PUFA (p = 0.05), and consequently, higher oxidation levels. Angus bulls produced beef with greater tenderness and PUFA concentration. The results of fatty acid show a possible change in biohydrogenation when animals are fed the WSC diet, reducing lipogenesis, as this diet increases the C18:2t10c12 content.

Oleate Hydratase in Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230 Catalyzes the Reversible Conversion between Linoleic Acid and Ricinoleic Acid.

Conjugated linoleic acid (CLA) has been the subject of numerous studies in recent decades because of its associated health benefits. CLA is an intermediate product of the biohydrogenation pathway of linoleic acid (LA) in bacteria. Several bacterial species capable of efficiently converting LA into CLA have been widely reported in the literature, among them Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230. Over the last few years, a multicomponent enzymatic system consisting of three enzymes involved in the biohydrogenation process of LA has been proposed. Sequencing the genome of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 revealed only one gene capable of encoding an oleate hydratase (OleH), unlike the presence of multiple genes typically found in similar strains. This study investigated the biological effect of the OleH enzyme of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 on the hydration of LA and dehydration of ricinoleic acid (RA) and its possible role in the production of CLA. The OleH was cloned, expressed, purified, and characterized. Fatty acid measurements were made by an internal standard method using a gas chromatography-coupled flame ionization detector (GC-FID) system. It was found that the enzyme is a hydratase/dehydratase, leading to a reversible transformation between LA and RA. In addition, the results showed that L. delbrueckii subsp. bulgaricus LBP UFSC 2230 OleH protein plays a role in stress tolerance in Escherichia coli. In conclusion, the OleH of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 catalyzes the initial stage of saturation metabolism of LA, although it has not converted the substrates directly into CLA. IMPORTANCE This study provides insight into the enzymatic mechanism of CLA synthesis in L. delbrueckii subsp. bulgaricus and broadens our understanding of the bioconversion of LA and RA by OleH. The impact of OleH on the production of the c9, t11 CLA isomer and stress tolerance by E. coli has been assisted. The results provide an understanding of the factors which influence OleH activity. L. delbrueckii subsp. bulgaricus LBP UFSC 2230 OleH presented two putative fatty acid-binding sites. Recombinant OleH catalyzed both LA hydration and RA dehydration. OleH was shown to play a role in bacterial growth performance in the presence of LA.

Draft genome sequence of Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230: a tool for preliminary identification of enzymes involved in CLA metabolism.

Several Lactobacillus ssp. are recognized as potential conjugated linoleic acid (CLA) producers. We have previously reported the ability of a range of Lactobacillus delbrueckii subsp. bulgaricus strains to produce CLA in fermented milk, being a potential candidate for the fermented dairy food chain. This study reports the draft genome sequence of L. bulgaricus strain LBP UFSC 2230, isolated from Italian Grana Padano cheese. Draft genome sequence originated in a total of 4,310,842 paired-end reads that were quality trimmed and assembled into 135 contigs with a total length of 604,745,873 bp, including 2086 protein coding genes and an average GC content of 49.7%. Draft genome sequence represents an important tool to identify the enzymes involved in this strain's CLA metabolism. We identified a gene encoding an enzyme involved in biohydrogenation of linoleic acid pathway, oleate hydratase.

Effect of conjugated linoleic acid feeding periods on fatty acid profile and nutritional quality of Astyanax altiparanae / Efeito do tempo de alimentação com ácido linoleico conjugado no perfil de ácidos graxos e qualidade nutricional de Astyanax altiparanae

The effects of feeding diets with conjugated linoleic acid (CLA) to Astyanax altiparanae for different periods were investigated on carcasses’ fatty acid profile and nutritional quality. The trial was laid out in a complete randomized design with eight feeding periods (0, 7, 14, 21, 28, 35, 42, and 49 days) and four replicates. A total of 240 fish (3.4 ± 0.3 g) were distributed into four aquariums and fed with the diet containing 32% crude protein, 15 MJ kg-1 gross energy, and 2.5% of CLA. Feeding CLA diet for 49 days resulted in the higher deposition of CLA, CLA isomers, and docosahexaenoic acid (DHA). The isomer c9,t11 was highly deposited than the t10,c12. Polyunsaturated fatty acids (PUFA), n-3, and n-6 increased with feeding period, whereas saturated (SFA), monounsaturated, and medium-chain fatty acids decreased. PUFA/SFA ratio, DHA/EPA, EPA+DHA, and thrombogenicity index increased linearly. Whereas atherogenicity index reduced, hypocholesterolemic/hypercholesterolemic ratio increased in quadratic effects over feeding time. No difference was observed for the n-6/n-3 ratio. Feeding A. altiparanae with 2.5% of CLA for a minimum of 35 days improves the fatty acid profile and provides a commercial product with good nutritional quality and functional benefits.

Os efeitos da alimentação com dietas contendo ácido linoleico conjugado (CLA) por diferentes períodos foram investigados no perfil de ácidos graxos e na qualidade nutricional das carcaças de Astyanax altiparanae. O experimento foi conduzido em delineamento inteiramente causalizado com oito períodos de alimentação (0, 7, 14, 21, 28, 35, 42 e 49 dias) e quatro repetições. Um total de 240 peixes (3,4 ± 0,3 g) foram distribuídos em quatro aquários e alimentados com dieta contendo 32% de proteína bruta, 15 MJ kg-1 de energia bruta e 2.5% de CLA. A alimentação com CLA por 49 dias resultou em maior deposição de CLA, isômeros e ácido docosahexaenóico (DHA). O isômero c9,t11 foi mais depositado do que o t10,c12. Os ácidos graxos n-3 e n-6 aumentaram com o período de alimentação, enquanto os ácidos graxos saturados (SFA), monoinsaturados e de cadeia média diminuíram. A razão PUFA/SFA, DHA/EPA, EPA + DHA e índice de trombogenicidade aumentaram linearmente. Enquanto o índice de aterogenicidade reduziu, a razão hipocolesterolêmica/hipercolesterolêmica aumentou em efeito quadrático com o aumento do tempo de alimentação com CLA. Nenhuma diferença foi observada para a razão n-6/n-3. Assim, a alimentação de A. altiparanae com 2.5% de CLA por um mínimo de 35 dias melhora o perfil de ácidos graxos e fornece um produto comercial com boa qualidade nutricional e benefícios funcionais.

Effect of conjugated linoleic acid feeding periods on fatty acid profile and nutritional quality of Astyanax altiparanae / Efeito do tempo de alimentação com ácido linoleico conjugado no perfil de ácidos graxos e qualidade nutricional de Astyanax altiparanae

The effects of feeding diets with conjugated linoleic acid (CLA) to Astyanax altiparanae for different periods were investigated on carcasses fatty acid profile and nutritional quality. The trial was laid out in a complete randomized design with eight feeding periods (0, 7, 14, 21, 28, 35, 42, and 49 days) and four replicates. A total of 240 fish (3.4 ± 0.3 g) were distributed into four aquariums and fed with the diet containing 32% crude protein, 15 MJ kg-1 gross energy, and 2.5% of CLA. Feeding CLA diet for 49 days resulted in the higher deposition of CLA, CLA isomers, and docosahexaenoic acid (DHA). The isomer c9,t11 was highly deposited than the t10,c12. Polyunsaturated fatty acids (PUFA), n-3, and n-6 increased with feeding period, whereas saturated (SFA), monounsaturated, and medium-chain fatty acids decreased. PUFA/SFA ratio, DHA/EPA, EPA+DHA, and thrombogenicity index increased linearly. Whereas atherogenicity index reduced, hypocholesterolemic/hypercholesterolemic ratio increased in quadratic effects over feeding time. No difference was observed for the n-6/n-3 ratio. Feeding A. altiparanae with 2.5% of CLA for a minimum of 35 days improves the fatty acid profile and provides a commercial product with good nutritional quality and functional benefits.(AU)

Os efeitos da alimentação com dietas contendo ácido linoleico conjugado (CLA) por diferentes períodos foram investigados no perfil de ácidos graxos e na qualidade nutricional das carcaças de Astyanax altiparanae. O experimento foi conduzido em delineamento inteiramente causalizado com oito períodos de alimentação (0, 7, 14, 21, 28, 35, 42 e 49 dias) e quatro repetições. Um total de 240 peixes (3,4 ± 0,3 g) foram distribuídos em quatro aquários e alimentados com dieta contendo 32% de proteína bruta, 15 MJ kg-1 de energia bruta e 2.5% de CLA. A alimentação com CLA por 49 dias resultou em maior deposição de CLA, isômeros e ácido docosahexaenóico (DHA). O isômero c9,t11 foi mais depositado do que o t10,c12. Os ácidos graxos n-3 e n-6 aumentaram com o período de alimentação, enquanto os ácidos graxos saturados (SFA), monoinsaturados e de cadeia média diminuíram. A razão PUFA/SFA, DHA/EPA, EPA + DHA e índice de trombogenicidade aumentaram linearmente. Enquanto o índice de aterogenicidade reduziu, a razão hipocolesterolêmica/hipercolesterolêmica aumentou em efeito quadrático com o aumento do tempo de alimentação com CLA. Nenhuma diferença foi observada para a razão n-6/n-3. Assim, a alimentação de A. altiparanae com 2.5% de CLA por um mínimo de 35 dias melhora o perfil de ácidos graxos e fornece um produto comercial com boa qualidade nutricional e benefícios funcionais.(AU)

Induction of accessory corpus luteum in goats and sheep: From physiological basis to reproductive effects / Indução de corpo lúteo acessório em caprinos e ovinos: Das bases fisiológicas aos efeitos reprodutivos

A identificação das ondas foliculares ovarianas e de seu padrão hormonal revelou que os folículos ovarianos dominantes da onda ovulatória (FDOO) crescem em ambiente hormonal com predominância crescente de estradiol, diferentemente daqueles da primeira (FDPO) e das ondas foliculares intermediárias (FDOI), que crescem sob forte impacto da progesterona (P4). O hormônio luteinizante (LH) é considerado o hormônio gonadotrófico decisivo para direcionar se um folículo dominante ovulará (↑ LH) ou não (↓ LH). Estratégias foram desenvolvidas para aumentar o LH endógeno (administração de GnRH) ou fornecer LH exógeno de origem suína (pLH) ou, ainda, gonadotrofinas semelhantes ao LH, como gonadotrofina coriônica humana (hCG). Estas medidas são capazes de disponibilizar LH para maturação final, ovulação e/ou luteinização de FDPO ou FDOI, formando corpos lúteos acessórios (CLa). Como consequência, a P4 aumenta e favorece o estabelecimento da gestação, sobretudo em condições em que a P4 for o fator limitante para a implantação e manutenção embrionária. Em ovelhas e cabras, em diferentes estudos, a hCG foi administrada de cinco a sete dias após o início do estro e revelou que o FDPO responde positivamente à administração de hCG, formando CLa e/ou promovendo a hipertrofia do CL formado originalmente, aumentando a área luteal. O incremento da P4 normalmente acompanha o aumento de área do tecido luteal. Como efeito final e mais desejável, a gestação e o nascimento de cordeiros/cabritos também aumentam. Esses conceitos serão discutidos na presente revisão sobre indução de CLa em ovinos e caprinos.

The identification of ovarian follicular waves and associated hormonal milieux has revealed that dominant follicles of the ovulatory wave (OWDF) grow in a hormonal environment where there is an increasing predominance of estradiol, unlike first-wave dominant follicles (FWDF) and intermediatewave dominant follicles (IWDF), which grow under increasing progesterone (P4) concentrations. The luteinizing hormone (LH) is considered the decisive gonadotropic hormone to direct whether a dominant follicle will (↑ LH) or will not (↓ LH) ovulate. Based on this, strategies have been developed to either increase endogenous LH (GnRH administration) or provide exogenous LH of porcine origin (pLH) or LH-like gonadotropins, such as human chorionic gonadotropin (hCG). Such strategies are able to provide LH for final maturation, ovulation, and/or luteinization of the FWDF or IWDF, forming accessory corpora lutea (aCL). As a consequence, P4 increases and favors the establishment of pregnancy, particularly when P4 is the limiting factor for the success of the conceptus implantation and maintenance. In sheep and goats, previous studies have administered hCG five to seven days after the onset of estrus and revealed that FWDF positively respond to hCG administration by either forming aCLs and/or promoting hypertrofia of the original CL which, in turn, increases its luteal tissue area. Normally, P4 synthesis increases along with the increase in luteal tissue area. As a final and most desirable outcome, pregnancy and the birth of lambs/kids also increase. These concepts will be discussed in this review, focusing on aCL induction in sheep and goats.

NanH and PknG putative virulence factors as a recombinant subunit immunogen against Corynebacterium pseudotuberculosis infection in mice.

Despite the economic and zoonotic relevance of caseous lymphadenitis, a competent immunoprophylaxis tool is still necessary. Here, we evaluated two putative virulence factors of Corynebacterium pseudotuberculosis, rNanH, and rPknG, as recombinant subunit vaccines in a murine model against the infection by C. pseudotuberculosis. Three groups of ten Balb/c mice each were inoculated with a sterile 0.9% saline solution (G1), rNanH (G2), or rPknG (G3) in formulations containing saponin as an adjuvant. The mice received two vaccine doses intercalated by a 21-day interval and were challenged with 2 × 104 CFU/mL of the C. pseudotuberculosis MIC-6 strain 21 days after the last immunization. The total IgG, IgG1, and IgG2a production levels increased significantly in the experimental groups (G2 and G3) on day 42. The highest levels of IgG2a antibodies in G2 and G3 were observed compared to IgG1 levels. G3 showed a significant (p < 0.05) humoral response through higher production of total IgG at day 42 when compared to G2. A significant increase of mRNA expression levels of interleukin (IL)-17, tumor necrosis factor, and interferon-γ was observed only in G2, while IL-4 was significantly produced only by G3. The levels of IL-10 and IL-12 obtained were not significant in any group. The survival rates after the challenge were 20% for G3 and 60% for G2 (p < 0.05). Our findings suggest that the formulation containing rNanH and saponin (G2) resulted in the best protection against the challenge and was able to elicit a Th1 immune response in mice, and can be considered as a promising antigen in the development of an effective vaccine against caseous lymphadenitis.

Current applications of Colloidal Liquid Aphrons: Predispersed solvent extraction, enzyme immobilization and drug delivery.

Colloidal Liquid Aphrons (CLAs) are micron sized discrete spherical solvent droplets formed by the dispersion of polyaphrons into a bulk aqueous phase at a low phase volume ratio where they can be kept homogenously suspended with only minimal agitation. CLAs have high stability due to the presence of a surfactant 'shell' surrounding the solvent core, and possess large surface areas per unit volume for mass transfer due to their small size. Therefore, CLAs are well suited for applications in pre-dispersed solvent extraction (PSE), enzyme immobilization, and have the potential to be used as a drug delivery system. Using PSE, CLAs have been used to remove metals such as Ni2+, Cu2+, Fe3+, Cr3+ and Mg2+ from dilute streams, separate organic dyes such as Yellow 1 from wastewater, extract succinic and lactic acid, reactively extract phenylalanine, and separate suspensions. CLAs have also been used to immobilize enzymes such as lipase, lysozyme and albumins with cases of superactivity being reported due to the influence of surfactant and solvent interactions with the enzyme. Furthermore, due to their similarity to current drug delivery systems such as microemulsions and hydrogels, and other advantages, CLA systems have the potential to be adapted for drug delivery systems also. This article provides a complete list of the current applications of Colloidal Liquid Aphrons (CLAs) in PSE and enzyme immobilization, and also presents insight into how CLAs can be utilized as a drug delivery method in the future. Finally, this review ends by summarizing potentially interesting research areas to pursue in this field.