scholarly journals The effect of aqueous plant extracts (Hibiscus sabdariffa and Glycyrrhiza glabra), on the growth of Entamoeba histolytica in vitro

2009 ◽  
Vol 3 (1) ◽  
pp. 50-55
Author(s):  
Zahra’a Abdul-Raheem Ahmed ◽  
Amna N. Jasim ◽  
Ali H. Ad’hiah
Keyword(s):  
Entamoeba Histolytica ◽  
Plant Extracts ◽  
Culture Media ◽  
Agar Medium ◽  
Glycyrrhiza Glabra ◽  
Reproduction Rate ◽  
Hibiscus Sabdariffa ◽  
Significant Difference ◽  
Infusion Agar

Entamoeba histolytica parasite was isolated from a stool sample, cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM). The effect of two aqueous plant extracts (Hibiscus sabdariffa and Glycyrrhiza glabra) on the growth and activity of the parasite in the two culture media was investigated. The aqueous extracts of H. sabdariffa and G. glabra were effective in reducing the parasite size in the LEM medium. With respect to the reproduction rate, the third concentration (19.71 mg/ml) of H. sabdariffa was significantly effective in inhibiting such rate to 57.6 and 83.6% in LEM and LIAM media, respectively, while for G. glabra, no significant difference in the reproduction rate was observed in both culture media.

scholarly journals The effect of calcium antagonists on the growth of Entamoeba histolytica in vitro

2010 ◽  
Vol 4 (1) ◽  
pp. 5-10
Author(s):  
Zahra’a Abdul-Raheem Ahmed ◽  
Ali H. Ad’hiah ◽  
Amna N. Jasim
Keyword(s):  
Entamoeba Histolytica ◽  
Calcium Antagonists ◽  
Culture Media ◽  
Agar Medium ◽  
Reproduction Rate ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Infusion Agar ◽  
Ethylene Diaminetetraacetic Acid

he E. histolytica parasite was maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM). The effect of two calcium antagonists (Nifedipine and Ethylene-diaminetetraacetic acid EDTA) on the growth and activity of the parasite in the two culture media was investigated. The calcium antagonists Nifedipine and EDTA inhibited the reproduction rate of E. histolytica in a concentration-dependent manner. For Nifedipine, a concentration of 41.6 mg/ml inhibited the reproduction rate to 99.7% in both media. The EDTA had an approximate effect (98.2 and 95.8)% at a concentration of 0.83 mg/ml in LEM and LIAM media, respectively. Additionally, some cases of a parasite encystment were observed in LEM medium that was treated with Nifedipine.

scholarly journals The role of some types of erythrocytes on the growth of Entamoeba histolytica trophozoite in vitro

2010 ◽  
Vol 7 (1) ◽  
pp. 200-206
Author(s):  
Baghdad Science Journal
Keyword(s):  
Stool Sample ◽  
Entamoeba Histolytica ◽  
Culture Media ◽  
Agar Medium ◽  
Reproduction Rate ◽  
Sheep Erythrocytes ◽  
Histolytica Trophozoite ◽  
Infusion Agar

The parasite E.histolytica was first isolated from a stool sample, and then cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM) . Then, the effect of some types of erythrocytes (human and sheep), on the growth and activity of the parasite in the two culture media was investigated. The parasite was able to ingest and lysis erythrocytes of human and sheep that were supplemented to the culture media and such manipulation was able to augment the reproduction rate of the cultivated E. histolytica, however, such consequence was media- and concentration-dependent. The reproduction rate was significantly increased (66.0, 57.5 and 58.6%, respectively) in LEM medium containing human erythrocytes types B at 0.11 x 106 cells/ml and O at 0.13 x 106 and 0.15 x 106 cells/ml. The sheep erythrocytes showed a similar enhancement (56.1%) at a concentration of 0.13 x 106 cells/ml. In contrast, adding erythrocytes to LIAM medium did not enhance the reproduction rate of the parasite significantly.

scholarly journals Cultivation of Entamoeba histolytica in vitro and diagnose the bacterial growths in culture media

2009 ◽  
Vol 6 (3) ◽  
pp. 442-447
Author(s):  
Baghdad Science Journal
Keyword(s):  
Escherichia Coli ◽  
Entamoeba Histolytica ◽  
Culture Media ◽  
Agar Medium ◽  
Cyst Formation ◽  
Essential Requirement ◽  
Bacteria Growth ◽  
Bacterial Content ◽  
Infusion Agar

The parasite was isolated from a stool sample, cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM) . The culture was maintained for up to 21 months, and the best time to maintain the parasite was every 48 hours, although the growth in the culture media continued for 13 days without a maintenance. Additionally, no cyst formation was observed during cultivation of parasite in the two culture media. Although, was observe young cyst formed in LEM media were deletion of maintained. The diagnosis of bacteria growth in the culture media, bacterial content (Escherichia coli) was an dominance and essential requirement for a successful cultivation of Entamoeba histolytica in the two culture media.

scholarly journals Impact of L-carnitine supplementation on the in vitro developmental competence and cryotolerance of buffalo embryos

2021 ◽  
pp. 3164-3169
Author(s):  
Mohamed M. M. El-Sokary ◽  
Al-Shimaa Al-H. H. El-Naby ◽  
Amal R. Abd El Hameed ◽  
Karima Gh. M. Mahmoud ◽  
T. H. Scholkamy
Keyword(s):  
Embryo Culture ◽  
In Vitro Maturation ◽  
Culture Media ◽  
Effective Concentration ◽  
Developmental Competence ◽  
Carnitine Supplementation ◽  
Oocyte Developmental Competence ◽  
High Lipid ◽  
Significant Difference

Background and Aim: Despite many trials, buffalo embryos have poor cryosurvivability because of their high lipid content. L-carnitine was found to be a lipid-reducing agent when added to oocyte and embryo culture media. The study aimed to determine the most effective concentration of L-carnitine to improve the oocyte developmental competence and cryotolerance of buffalo embryos. Materials and Methods: In vitro maturation and embryo culture media were supplemented with four concentrations of L-carnitine: 0 (control), 0.25, 0.5, and 1 mM. Good-quality embryos on 7 days were vitrified using mixtures of dimethyl sulfoxide and ethylene glycol at two concentrations (3.5 and 7 M). Results: The result showed that the cleavage and morula rates were significantly (p<0.05) higher in the 0.5 mM group. Blastocyst rates were significantly (p<0.05) higher at both 0.5 and 1 mM. The rates of viable embryos directly after thawing were significantly (p<0.05) increased in the 0.5 mM group. No significant difference was found in embryos cultured for 24 h after warming among all the groups. Conclusion: The addition of L-carnitine at a concentration of 0.5 mM to the culture media improves the oocyte developmental competence and cryotolerance of buffalo embryos directly after warming but not after 24 h of culture. Nevertheless, further studies must identify how L-carnitine exerts its beneficial micromechanisms.

160 COMPARATIVE STUDY OF IN VITRO CULTURE MEDIA AND ASSISTED HATCHING TECHNIQUES ON MOUSE EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 188
Author(s):  
N. C. Negota ◽  
L. P. Nethenzheni ◽  
M. L. Mphaphathi ◽  
D. M. Barry ◽  
T. L. Nedambale
Keyword(s):  
In Vitro Culture ◽  
Chorionic Gonadotropin ◽  
Culture Medium ◽  
Culture Media ◽  
Assisted Hatching ◽  
Hatching Rate ◽  
Significant Difference ◽  
Equine Chorionic Gonadotropin ◽  
F1 Generation

The in vitro culture media and assisted hatching techniques remain challenging obstacles to be utilised widely. Mechanical, chemical, enzymatic thinning, and laser-assisted techniques have been used previously but information is still lacking on its application in livestock. The aim of this study was to compare the effect of 2 in vitro culture media (Hamster F10 and TMC-199) and 4 (mechanical, chemical, enzymatic, and laser) assisted hatching techniques on blastocyst formation and hatching rate using murine embryos as a model. The C57/b and Balb/c breeds were raised until they reached maturity and bred naturally to produce F1 generation. The light in the breeding house was controlled at 14 h light and 10 h dark. Feed and water were provided ad libitum for the mice. Superovulation of females were stimulated using equine chorionic gonadotropin and human chorionic gonadotropin. The F1 generation was used for the collection of the 400 blastocysts and randomly allocated into 4 assisted hatching techniques. Blastocysts were paired into a group of 10 and replicated 4 times for each assisted hatching technique. The general linear model of SAS version 9.4 (SAS Institute Inc., Cary, NC, USA) was used to analyse the data. Assisted hatching techniques of laser, mechanical, enzymatic, and chemical yielded 46.9 ± 37.1, 51.1 ± 40.2, 39.1 ± 35.8, and 33.3 ± 4.5%, respectively, under in vitro culture of Hamster F10. The TCM-199, laser, mechanical, enzymatic, and chemical assisted hatching techniques yielded 56.3 ± 43.3, 52.6 ± 35.5, 49.2 ± 37.5, and 33.9 ± 35.5%, respectively, with a significant difference. There was no significant difference observed in assisted hatching techniques and Hamster F10 culture medium. However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM than cultured in Hamster F10. Hatching rate of blastocysts increased from chemical, enzymatic, mechanical, and laser with response to Hamster F10 and TCM; thus, laser is a suitable assisted hatching technique with TCM-199.

Development of prepubertal goat oocytes after their in vitro maturation and chemical activation

Zygote ◽  
2020 ◽  
Vol 28 (6) ◽  
pp. 447-452
Author(s):  
Seungbum Hong ◽  
Binoy S. Vettical ◽  
Nisar Ahmad Wani
Keyword(s):  
Culture Media ◽  
Chemical Activation ◽  
Control Treatment ◽  
Developmental Potential ◽  
Embryo Culture Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Further Development ◽  
Oviductal Fluid

SummaryExperiments were conducted to study in vitro maturation of prepubertal goat oocytes and their developmental potential after chemical activation. In Experiment 1, cumulus–oocytes complexes collected from the ovaries of prepubertal goats slaughtered at a local abattoir were matured in vitro in TCM-199-based medium supplemented with 10 µg/ml luteinizing hormone (LH) (treatment 1) or 10 µg/ml LH + 0.1 mM l-cysteine (treatment 2). In Experiment 2, mature oocytes were activated with either 5 µM ionomycin or 7% ethanol. After 18 h, some oocytes were randomly fixed and stained to evaluate their chromatin status, while others were cultured in embryo culture medium to study their further development. In Experiment 3, oocytes activated with 5 µM ionomycin were cultured for 7 days in one of the four different culture media [Charles Rosenkrans medium (CR-1), TCM-199, potassium simplex optimization medium (KSOM) and synthetic oviductal fluid (SOF)] to study their developmental potential. The maturation rate in control, treatment 1, and treatment 2 media did not differ from each other (P > 0.05). However, the lowest degeneration of oocytes was observed in treatment 3 (P < 0.05) when compared with the other two groups. The proportion of activated oocytes was higher, while non-activated oocytes were lower in ionomycin group when compared with the group activated with ethanol (P < 0.05). The proportions of oocytes cleaved were 65.7, 56.8, 61.0 and 54.4% in CR-1, TCM-199, KSOM and SOF medium, respectively, with no significant difference. However, further development of cleaved oocytes was better in KSOM followed by SOF.

Expression of mRNA encoding leukaemia inhibitory factor (LIF) and its receptor (LIFRβ) in buffalo preimplantation embryos produced in vitro: markers of successful embryo implantation

Zygote ◽  
2012 ◽  
Vol 21 (2) ◽  
pp. 203-213 ◽  
Author(s):  
S. Eswari ◽  
G. Sai Kumar ◽  
G. Taru Sharma
Keyword(s):  
Culture Media ◽  
Total Cell ◽  
Inhibitory Factor ◽  
Preimplantation Embryos ◽  
Leukaemia Inhibitory Factor ◽  
Hatching Rate ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Significant Difference

SummaryThe objective of this study was to evaluate the effect of supplementation of recombinant leukaemia inhibitory factor (LIF) in culture media on blastocyst development, total cell number and blastocyst hatching rates and the reverse transcription-polymerase chain reaction analysis of preimplantation buffalo embryos to determine whether they contain the LIF-encoding mRNA and its beta receptor (LIFRβ) genes in different stages of preimplantation buffalo embryos. Cumulus–oocyte complexes retrieved from slaughterhouse buffalo ovaries were matured in vitro and fertilized using frozen buffalo semen. After 18 h of co-incubation with sperm, the presumptive zygotes were cultured in modified synthetic oviductal fluid without (control) or with rhLIF (100 ng/ml). There was no significant difference in the overall cleavage rate up to morula stage however the development of blastocysts, hatching rate and total cell numbers were significantly higher in the LIF-treated group than control. Transcripts for LIFRβ were detected from immature, in vitro-matured oocytes and in the embryos up to blastocyst stage, while transcripts for the LIF were detected from 8–16-cell stage up to blastocyst, which indicated that embryo-derived LIF can act in an autocrine manner on differentiation process and blastocyst formation. This study indicated that the addition of LIF to the embryo culture medium improved development of blastocysts, functional (hatching) and morphological (number of cells) quality of the blastocysts produced in vitro. The stage-specific expression pattern of LIF and LIFRβ mRNA transcripts in buffalo embryos indicated that LIF might play an important role in the preimplantation development and subsequent implantation of buffalo embryos.

scholarly journals The difference of the salivary volume before and after drinking the rosella tea (Hibiscus sabdariffa)

2010 ◽  
Vol 22 (3) ◽  
Author(s):  
Mutiara Indah Permata Sari Islami ◽  
Edeh Roletta Haroen ◽  
Sri Tjahajawati
Keyword(s):  
In Vitro Study ◽  
T Test ◽  
Hibiscus Sabdariffa ◽  
Test Analysis ◽  
Community Benefits ◽  
Significant Difference ◽  
Before And After ◽  
Quasi Experimental ◽  
The Difference

Introduction: Roselle plants (Hibiscus sabdariffa) is one of the herbs that began to be used by the community. Benefits of this plant is quite a lot for health. The portion taken for consumption is the red flower petals. oselle tea is one of the sour beverages which can affect the salivary gland secretion. The purpose of this study is to analyzed the difference of salivary volume before and after drinking roselle tea. Methods: This study has been conducted to 40 students of Faculty of Dentistry, Padjadjaran University, ranging from 18-23 years of age with good general condition. This study is quasi-experimental in vitro study using the paired test analysis with α = 0,05 of the data collected from salivary volume. Results: The result of study indicates that the average of salivary volume before drinking roselle tea is 1,90 milliliter. After drinking roselle tea, the average of salivary volume is 4,54 milliliter. The result of paired test analysis shows that t-test is 16,172 and t-table is 2,022. The value of t-test > t-table. Result of statistic shown there is significant difference of salivary volume before and after drinking roselle tea. Conclusion: There is a difference of salivary volume before and after drinking roselle tea.

scholarly journals 146COMPARISON OF EMBRYO CULTURE MEDIA FOR BLASTOCYST DEVELOPMENT AFTER INTRACYTOPLASMIC SPERM INJECTION OF IN VITRO-MATURED EQUINE OOCYTES

2004 ◽  
Vol 16 (2) ◽  
pp. 195
Author(s):  
Y.H. Choi ◽  
D.D. Varner ◽  
K. Hinrichs
Keyword(s):  
In Vitro Culture ◽  
Embryo Culture ◽  
Intracytoplasmic Sperm Injection ◽  
Culture Media ◽  
Polar Body ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Vitro Fertilization ◽  
Significant Difference

Research on in vitro culture of equine embryos has been scant, due to failure of equine in vitro fertilization to be repeatably successful. We have recently obtained high fertilization rates of equine oocytes via intracytoplasmic sperm injection (ICSI) using a piezo drill (Choi et al., 2002 Reproduction 123, 455–465). Culture of presumptive zygotes in G1.2/2.2 medium resulted in 63% cleavage and an average of 15 cells at 4d, but only 2 to 9% blastocyst development at 7 days (Choi et al., 2003 Theriogenology 59, 1219–1229). In the present study, we evaluated the effect of two different culture media, G1.3/G2.3 v. DMEM/F-12, with or without FBS, on blastocyst development after ICSI. Oocytes were collected from slaughterhouse-derived ovaries by follicular scraping and were matured in vitro for 24h in M199 with 10% FBS and 5μUmL−1 FSH. After culture, oocytes having a polar body (198/305; 65%) were fertilized by ICSI with frozen-thawed equine sperm using a piezo drill. Presumptive zygotes were cultured in 1 of 4 media: G1.3/G2.3 (which includes 0.8% BSA) with or without 10% FBS, or in DMEM/F-12 with 0.5% BSA, with or without 10% FBS. Culture was performed in microdroplets at 5μL/zygote under oil at 38.2°C in an atmosphere of 5% CO2, 5% O2 and 90% N2 for 7.5 days. In G1.3/2.3 treatments, G1.3 media were completely refreshed at 48h, zygotes were transferred to G2.3 (with or without FBS as per the first stage) at 96h, and were completely refreshed with the same media at 144h. In DMEM/F-12 treatments, media were completely refreshed every other day. Three to 5 replicates were performed in each treatment, and data were analyzed by chi-square test. There were no significant differences in cleavage rates (59–64%) among treatments. The rate of development to blastocyst, per oocyte injected, in G1.3/G2.3/BSA (1/49, 2%) was significantly lower (P&lt;0.05) than that for the other three treatments: G1.3/2.3/BSA/FBS (9/49, 18%), DMEM/F-12/BSA (9/50, 18%), or DMEM/F-12/BSA/FBS (10/50, 20%). There was no significant difference in blastocyst development among the latter three treatments. These findings indicate that G1.3/2.3 media with BSA only do not adequately support growth of equine embryos. Development of up to 20% of injected oocytes to the blastocyst stage in G media supplemented with FBS, in DMEM/F-12/BSA or in DMEM/F-12/BSA/FBS represents the highest in vitro equine blastocyst rate in medium alone (i.e. without co-culture) yet reported. The success of DMEM/F-12 as an embryo culture medium may provide a relatively simple basis for equine in vitro culture programs. To determine whether this medium was able to support further developmental competence, we cultured equine embryos resulting from nuclear transfer of in vitro-matured oocytes in DMEM/F-12+10% FBS (without BSA). We transferred 4 resulting blastocysts to recipient mares by transcervical transfer; one pregnancy is ongoing at 230d gestation at the time of this writing. This work was supported by the Link Equine Research Endowment Fund, Texas A&amp;M University.

scholarly journals Embryo Development of Cypripedium formosanum in Relation to Seed Germination In Vitro

2005 ◽  
Vol 130 (5) ◽  
pp. 747-753 ◽  
Author(s):  
Yung-I Lee ◽  
Nean Lee ◽  
Edward C. Yeung ◽  
Mei-Chu Chung
Keyword(s):  
Seed Germination ◽  
Embryo Development ◽  
Developmental Stages ◽  
Culture Media ◽  
Nile Red ◽  
Germination Percentage ◽  
Zygotic Embryogenesis ◽  
Seed Collection ◽  
Significant Difference

This investigation documents the key anatomical features in embryo development of Cypripedium formosanum Hayata, in association with the ability of embryos to germinate in vitro, and examines the effects of culture media and seed pretreatments on seed germination. A better understanding of zygotic embryogenesis for the Cypripedium L. species would provide insights into subsequent germination events and aid in the in vitro propagation of these endangered species. In seeds collected at 60 days after pollination (DAP), soon after fertilization, no germination was recorded. The best overall germination was found at 90 DAP (≈70%), at which time early globular to globular embryos with a single-celled suspensors can be observed. After 135 DAP, the seeds germinated poorly. At this time the inner integument shrinks and forms a tight layer, which encloses the embryo, the so-called “carapace.” Using Nile red stain, a cuticular substance was detected in the carapace, which may play a role in the impermeability of the mature seed and may help the seeds survive in the stringent environment. At maturity (after 210 DAP), the embryo proper has an average size of eight cells along its length and six cells across the width. Lipids and proteins are the main storage products within the embryo. To improve seed germination, experiments were conducted to test the suitability of various media and pretreatments of seeds. When different media were used, except for the Harvais medium at 120 DAP, there was no significant difference in seed germination at three different developmental stages tested. Soaking mature seeds in 1% NaOCl or treating them with ultrasound may slightly increase the germination percentage. For seed germination, our results indicate that the timing of seed collection outweighs the composition of medium and the seed pretreatments.

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