scholarly journals Silage quality and biogas production from Spartina pectinata L. fermented with a novel xylan-degrading strain of Lactobacillus buchneri M B/00077

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marta Kupryś-Caruk ◽  
Renata Choińska ◽  
Agnieszka Dekowska ◽  
Katarzyna Piasecka-Jóźwiak
Keyword(s):  
Lignocellulosic Biomass ◽  
Culture Medium ◽  
Biogas Production ◽  
Sole Source ◽  
Spartina Pectinata ◽  
Lactobacillus Buchneri ◽  
Xylanolytic Activity ◽  
Genes Encoding

AbstractThe aim of the current study was to determine the ability of the Lactobacillus buchneri M B/00077 strain to degrade xylan, its impact on the quality of silage made from the lignocellulosic biomass of Spartina pectinata L., as well as the efficiency of biogas production. In the model in vitro conditions the L. buchneri M B/00077 strain was able to grow in a medium using xylan as the sole source of carbon, and xylanolytic activity was detected in the post-culture medium. In the L. buchneri M B/00077 genome, genes encoding endo-1,4-xylanase and β-xylosidase were identified. The silages prepared using L. buchneri M B/00077 were characterized by a higher concentration of acetic and propionic acids compared to the controls or the silages prepared with the addition of commercial xylanase. The addition of bacteria increased the efficiency of biogas production. From the silages treated with L. buchneri M B/00077, 10% and 20% more biogas was obtained than from the controls and the silages treated with commercial xylanase, respectively. The results of the current study indicated the strain L. buchneri M B/00077 as being a promising candidate for further application in the field of pretreatment of lignocellulosic biomass.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

140 EFFECT OF A NOVEL BOVINE EMBRYO CULTURE MEDIUM TO IMPROVE BLASTOCYST DEVELOPMENT

2013 ◽  
Vol 25 (1) ◽  
pp. 217
Author(s):  
R. F. Gonçalves ◽  
C. Figueiredo ◽  
M. A. Achilles
Keyword(s):  
Culture Medium ◽  
Apoptotic Cell ◽  
Cell Number ◽  
Total Cell ◽  
Cell Numbers ◽  
Group 2 ◽  
Group 1 ◽  
Hatching Rates

There are still immense differences in the quality of in vitro-produced embryos compared to their in vivo-generated counterparts. These differences include a higher sensitivity of in vitro-produced embryos towards cryopreservation. The quality of such embryos has been evaluated using various parameters like morphological examination, assessment of total cell numbers, or pregnancy rates after transfer. In the present study, the effects of glycine, alanine, taurine, and glutamine addition to SOF (Achilles Genetics culture medium, Achilles Genetics®, Garça, SP, Brazil) on the in vitro development (cleavage and blastocyst rates) and quality (total cell and apoptotic cell numbers) of bovine embryos were determined. Ovaries of Nelore cows were obtained from a slaughterhouse. Cumulus–oocyte complexes (COC) were collected from follicles ≥4 mm in diameter, matured in TCM-199, and fertilized with frozen–thawed Nelore bull semen (IVF = Day 0). On Day 1, presumptive zygotes were cultured in SOF supplemented with fetal bovine serum (FBS, group 1, n = 550) or in Achilles Genetics culture medium (SOF supplemented with Achilles Mixture and FBS, group 2, n = 557) at 38.5°C and 5% CO2 in air until Day 9. Embryos were evaluated during culture: at Day 3 cleavage rates, at Day 7 blastocyst rates, and on Day 9 hatching rates. Experiments were replicated 5 times, analysed using ANOVA, followed by a comparison of means by Tukey test (P ≤ 0.05). Blastocysts at Day 8 from Group 1 (n = 75) and Group 2 (n = 75) were fixed and permeabilized for TUNEL assay (DeadEndTM Florimetric TUNEL System, Promega, Madison, WI, USA), according to the manufacturer instructions. Total cell number, apoptotic cell number, and apoptotic cell index (calculated by dividing the apoptotic cell number by total cell number) were analyzed by analysis of variance and means were compared by Student Newman Keuls test. The threshold of significance was set at P ≤ 0.05. Cleavage rates were 79.2 ± 2.5 for group 1 and 91.0 ± 2.5 for group 2. Blastocyst and hatching rates (calculated on the total of zygotes) for group 2 (47.4 ± 2.8; 82.1 ± 1.5) were significantly greater than for group 1 (39.8 ± 2.8; 74.3 ± 1.5). The total cell numbers were not different (P > 0.05) between group 1 (112.7 ± 2.9) and group 2 (111.1 ± 2.7). Blastocysts from group 2 showed lower (P < 0.05) number of apoptotic cells (10.7 ± 1.2) than those from group 1 (20.9 ± 1.2). These results indicate that the addition of glycine, alanine, taurine, and glutamine to SOF (Achilles Mixture) may be an important energy source for the bovine blastocyst and could act synergistically to enhance embryo development to the hatching stage and embryo quality. Financial support from CNPq and FAPESP.

scholarly journals One of the trends of oxidative protection in in vitro fertilization programs

2020 ◽  
Vol 14 (4) ◽  
pp. 502-514
Author(s):  
O. S. Vachlova ◽  
T. A. Oboskalova
Keyword(s):  
In Vitro Fertilization ◽  
Culture Medium ◽  
Inhibitory Effect ◽  
Biological Action ◽  
Published Data ◽  
International Studies ◽  
Vitro Fertilization ◽  
Contraceptive Effect

Here we review published data from experimental and clinical international studies examining pathogenetic effects of melatonin upon using programs of In Vitro Fertilization (IVF); highlighting various viewpoints on its biological action as a regulator of circadian rhythms: on the one hand, the inhibitory effect of melatonin on pulsating secretion of gonadotropin-releasing hormone was considered, thereby achieving a contraceptive effect; on the other hand, its ability to induce the secretion of human chorionic gonadotropin ensuring ovulation process, was discussed. We also review the data on melatonin acting as a highly active antioxidant. While using melatonin as a metabolic supplement in IVF programs, its positive effect on oocyte morphology and quality of fertilization, embryo division was observed. Moreover, we also highlight the results of studies examining melatonin-related effects on quality of fertilization and embryo division after adding it to culture medium. Such effects demonstrated dose-depended pattern. Taking into account the data of the analyzed publications, adding exogenous melatonin to culture medium may represent a new strategy for personalized approach to improve outcome of IVF programs. Its effectiveness should be further investigated and considered for introduction within the framework of pregravid preparation.

scholarly journals In vitro maintenance, under slow-growth conditions, of oil palm germplasm obtained by embryo rescue

2015 ◽  
Vol 50 (5) ◽  
pp. 426-429 ◽  
Author(s):  
Julcéia Camillo ◽  
Jonny Everson Scherwinski-Pereira
Keyword(s):  
Plant Growth ◽  
Oil Palm ◽  
Culture Medium ◽  
Slow Growth ◽  
Elaeis Guineensis ◽  
Embryo Rescue ◽  
Growth Conditions ◽  
In Vitro Maintenance

The objective of this work was to evaluate the in vitro maintenance of oil palm (Elaeis guineensis and E. oleifera) accessions under slow-growth conditions. Plants produced by embryo rescue were subject to 1/2MS culture medium supplemented with the carbohydrates sucrose, mannitol, and sorbitol at 1, 2, and 3% under 20 and 25±2ºC. After 12 months, the temperature of 20°C reduced plant growth. Sucrose is the most appropriate carbohydrate for maintaining the quality of the plants, whereas mannitol and sorbitol result in a reduced plant survival.

Supplementation of insulin–transferrin–selenium to embryo culture medium improves the in vitro development of pig embryos

Zygote ◽  
2013 ◽  
Vol 22 (3) ◽  
pp. 411-418 ◽  
Author(s):  
Ziban Chandra Das ◽  
Mukesh Kumar Gupta ◽  
Sang Jun Uhm ◽  
Hoon Taek Lee
Keyword(s):  
Nuclear Transfer ◽  
Culture Medium ◽  
Mammalian Species ◽  
Oxygen Species ◽  
Embryo Culture Medium ◽  
Maturation Medium ◽  
Blastocyst Quality ◽  
Vitro Development

SummaryInsulin, transferrin and selenium (ITS) supplementation to oocyte maturation medium improves the post-fertilization embryonic development in pigs. ITS is also commonly used as a supplement for the in vitro culture (IVC) of embryos and stem cells in several mammalian species. However, its use during IVC of pig embryos has not been explored. This study investigated the effect of ITS supplementation to IVC medium on the in vitro development ability of pig embryos produced by parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT). We observed that ITS had no significant effect on the rate of first cleavage (P > 0.05). However, the rate of blastocyst formation in ITS-treated PA (45.3 ± 1.9 versus 27.1 ± 2.3%), IVF (31.6 ± 0.6 versus 23.5 ± 0.6%) and SCNT (17.6 ± 2.3 versus 10.7 ± 1.4%) embryos was significantly higher (P < 0.05) than those of non-treated controls. Culture of PA embryos in the presence of ITS also enhanced the expansion and hatching ability (29.1 ± 3.0 versus 18.2 ± 3.8%; P < 0.05) of blastocysts and increased the total number of cells per blastocyst (53 ± 2.5 versus 40.9 ± 2.6; P < 0.05). Furthermore, the beneficial effect of ITS on PA embryos was associated with significantly reduced level of intracellular reactive oxygen species (ROS) (20.0 ± 2.6 versus 46.9 ± 3.0). However, in contrast to PA embryos, ITS had no significant effect on the blastocyst quality of IVF and SCNT embryos (P > 0.05). Taken together, these data suggest that supplementation of ITS to the IVC medium exerts a beneficial but differential effect on pig embryos that varies with the method of embryo production in vitro.

scholarly journals Effect of Anti-Mullerian Hormone in Culture Medium on Quality of Mouse Oocytes Matured In Vitro

PLoS ONE ◽  
2014 ◽  
Vol 9 (6) ◽  
pp. e99393 ◽  
Author(s):  
Yihui Zhang ◽  
Li Shao ◽  
Yixin Xu ◽  
Yigui Cui ◽  
Jiayin Liu ◽  
...  
Keyword(s):  
Culture Medium ◽  
Mouse Oocytes

scholarly journals Perkembangan Sel Endometrium Domba setelah Inkubasi dalam Kolagenase dan Dikultur In Vitro dengan Estradiol dan Progesteron

2019 ◽  
Vol 20 (3) ◽  
pp. 298
Author(s):  
Ananda Ananda ◽  
Ekayanti Mulyawati Kaiin ◽  
Ni Wayan Kurniani Karja ◽  
Mohamad Agus Setiadi
Keyword(s):  
Cell Culture ◽  
Cell Viability ◽  
Incubation Time ◽  
Culture Medium ◽  
Proliferation Rate ◽  
Endometrial Cell ◽  
Test Quality ◽  
Endometrial Cells

The aim of the present study was to determine the optimal incubation time of collagenase on concentration, viability, quality of endometrial cell culture, and the role of estradiol and progesterone on ovine endometrial cells proliferation. Endometrium minced was incubated into collagenase with different duration of time: 1 hour, 3 hours, and 6 hours repectively. Cell concentration and viability were calculated after incubation. The quality of cell culture was observed on 1st, 3rd, and 5th day after seeding. The best incubation result was then continued with the addition of the E2 (100 pg/ mL), P4 (100 ng/mL), E2:P4 (100 pg/mL : 10 ng/mL), and E2:P4 (10 pg/mL : 100 ng/mL) into the culture medium. After nine days, cell culture was harvested by trypsinization. Concentration and cell viability were analyzed using analysis of variance, followed by Duncan test. Quality of endometrial cell culture was analyzed descriptively. Results showed that there was a significant decreased in the concentration and cell viability obtained in each treatment of collagenase incubation time and optimum treatment of endometrial cell culture was found at 3 hours. Experiment on culture of endometrial cell revealed that proliferation rate treated by E2 and P4 was better then control (P<0.05). Futhermore, additional of E2 into the culture medium even E2 alone (100 pg/mL or higher E2 combination with P4 (100 pg/mL : 10 ng/mL) showed better proliferation rate than P4 alone (100 ng/mL) or higher P4 combination (10 pg/mL : 100 ng/mL). In conclusion, suplementation of 100 pg/mL of estradiol (E2) could support better proliferation of ovine endometrial cells in vitro.

scholarly journals Chitosan-propolis combination inhibits anthracnose in 'Hass' avocados

2018 ◽  
pp. 681
Author(s):  
A.K. Marino, José S.P. Junior ◽  
Kelly M. Magalhães ◽  
Ben-Hur Mattiuz
Keyword(s):  
Mycelial Growth ◽  
Culture Medium ◽  
Colletotrichum Gloeosporioides ◽  
Control Treatment ◽  
Distilled Water ◽  
Mycelium Growth ◽  
In Vivo Growth

The effect of chitosan-propolis combination on the control of in vitro and in vivo growth of Colletotrichum gloeosporioides, a causal agent of anthracnose, as well as on the quality of avocado of Hass cultivar has been evaluated.  Chitosan and propolis were added to the culture medium separately and in combinations to verify the efficacy of inhibition of C. gloeosporioides mycelium in vitro. Avocados were immersed for 1 minute in the treatments that best inhibited the mycelium growth (in vitro) with the aim of testing the fungus control in vivo. The tested treatments were 1.5% chitosan and combinations of propolis (1.0%, 1.5%, 2.0%) with 1.5% chitosan and then inoculated with C. gloeosporioides. Control fruits were dipped in distilled water only. In the second experiment, avocados were immersed for 1 minute in the treatments that best inhibited the mycelium growth (in vivo) with the aim of testing the fruits quality. The tested treatments were 1.5% chitosan isolated and in the combination of 1.5% chitosan with 1.5% propolis. The control treatment were immersed only in distilled water. Avocados were stored at 22 ± 0.3 °C and 90 ± 3% RH for 7 days. The use of 1.5% chitosan in combination with 2.0% propolis controlled the mycelial growth of Colletotrichum gloeosporioides in vitro. In addition, coating with only 1.5% chitosan provided the best results in avocados, reducing the severity and incidence of anthracnose and maintaining the fruit quality.

Plant protein hydrolysates (plant peptones) as substitutes for animal proteins in embryo culture medium

2009 ◽  
Vol 21 (4) ◽  
pp. 587 ◽  
Author(s):  
F. George ◽  
D. Kerschen ◽  
A. Van Nuffel ◽  
J. F. Rees ◽  
I. Donnay
Keyword(s):  
Culture Medium ◽  
Similar Rate ◽  
Protein Hydrolysates ◽  
Plant Protein ◽  
Fluid Medium ◽  
Anti Oxidant ◽  
The Cost ◽  
Oviduct Fluid

The aim of the present study was to improve the sanitary quality of in vitro-produced bovine embryos by using plant protein hydrolysates (plant peptones) as substitutes for animal proteins. Peptones were compared with bovine serum albumin (BSA) as the protein source in synthetic oviduct fluid medium and the quality of the resulting embryos was determined. Two batches of peptones (wheat and cotton) were selected on the basis of their anti-oxidant properties. When added to the culture medium, both peptones (at 0.56 mg mL–1 for cotton peptone and at 0.18 mg mL–1 for wheat peptone) led to similar developmental and hatching rates compared with 4 mg mL–1 BSA and embryos were equally resistant to freezing and able to elongate after transfer. Surprisingly, a significant decrease in reduced glutathione (GSH) content was observed when embryos were produced with plant peptone instead of BSA. Supplementation of the culture medium with precursors of GSH (cysteine and β-mercaptoethanol) significantly increased the GSH content. A shift of the sex ratio towards male embryos was seen for Day 8 embryos cultured with wheat peptone, whereas no shift was observed for embryos cultured in the presence of BSA or polyvinylpyrrolidone. In conclusion, culture with plant peptones enables embryos to be obtained at a similar rate and of similar quality to that seen following the use of BSA. The use of the plant peptones increased the sanitary quality of the embryos and decreased the cost of embryo production.

Preimplantation development and viability of in vitro cultured rabbit embryos derived from in vivo fertilized gene-microinjected eggs: apoptosis and ultrastructure analyses

Zygote ◽  
2005 ◽  
Vol 13 (2) ◽  
pp. 125-137 ◽  
Author(s):  
A.V. Makarevich ◽  
P. Chrenek ◽  
N. Žilka ◽  
J. Pivko ◽  
J. Bulla
Keyword(s):  
Culture Medium ◽  
Detrimental Effect ◽  
Transgenic Animals ◽  
Cell Number ◽  
High Rate ◽  
Cleavage Stage ◽  
Thin Sections

Microinjection (Mi) of gene constructs into pronuclei of fertilized eggs is a widely used method to generate transgenic animals. However, the efficiency of gene integration and expression is very low because of the low viability of reconstructed embryos resulting from cell fragmentation and cleavage arrest. As a consequence, only a few viable embryos integrate and express transgene. Since cellular fragmentation and cleavage stage arrest in embryos may be associated with apoptosis, we aimed to test the hypothesis that the low viability of Mi-derived eggs is caused by a high rate of apoptosis in embryos, as a result of the detrimental effect of Mi. Pronuclear stage eggs (19–20 hours post-coitum, hpc) were microinjected with several picolitres of DNA construct into the male pronucleus (gene-Mi); the intact eggs (non-Mi) or eggs microinjected with phosphate-buffered saline (PBS-Mi) served as controls. Epidermal growth factor (EGF; 0, 20 and 200 ng/ml) was added to the culture medium and the embryos were cultured up to 94–96 hpc. Apoptosis was detected using the TUNEL assay, and the ultrastructure was analysed using electron microscopy of Durcupan ACM thin sections of the embryo. Gene-Mi embryos had significantly lower (p<0.05) blastocyst yields and a higher percentage of cleavage-arrested embryos than those in the non-Mi group. In gene-Mi groups, approximately 40% of all cleavage-stage-arrested embryos had fragmented blastomeres. Both gene-Mi-and PBS-Mi-derived blastocysts had a significantly higher TUNEL index (p<0.001) and lower total cell number (p<0.05) than the non-Mi embryos. Comparison of the quality of gene-Mi embryos with that of PBS-Mi embryos indicated that the deleterious effect of Mi on the embryo was caused by the Mi procedure itself, rather than DNA. EGF (at 20 ng/ml) had beneficial effects on the quality of gene-Mi-derived embryos, eliminating the influence of the Mi procedure on apoptosis and embryo cell number. Ultrastructural analysis confirmed a higher occurrence of apoptotic signs (nuclear membrane blebbing, areas with electron-dense material, numerous apoptotic bodies) in Mi-derived cleavage-arrested embryos compared with untreated or Mi-derived normal-looking embryos. These findings suggest an association between embryo cleavage arrest and apoptosis in Mi-derived embryos. Inclusion of EGF in the embryo culture medium can eliminate the detrimental effect of Mi on embryo quality.

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