In vitro assessment of corpus luteum function in cattle following Ovsynch and CIDR ovulation synchronization protocols

2004 ◽  
Vol 84 (4) ◽  
pp. 721-724 ◽  
Author(s):  
M. Aali ◽  
J. A. Small ◽  
G. Giritharan ◽  
N. Ramakrishnappa ◽  
K. M. Cheng ◽  
...  

Non-lactating beef cows (N = 40) were used to determine in vitro production of progesterone by CLs collected on days 6–8, 13–15 and 19–20, following Ovsynch or CIDR ovulation synchronization protocols. Progesterone released by the CL tissues into the medium was measured after 1 h of incubation (control) and after 6 h of hormone treatments (LH, PGF2α or LH + PGF2α). In vitro progesterone production did not differ (P > 0.05) between Ovsynch and CIDR ovulation synchronization protocols. Pooled data, irrespective of ovulation synchronization treatments, showed interaction (P < 0.05) between hormone treatment and stage of CL. Key words: Corpus luteum, progesterone, cows, ovulation synchronization

1970 ◽  
Vol 18 (1) ◽  
pp. 25-35 ◽  
Author(s):  
Md. Tariqul Islam ◽  
E. R. Joachim Keller ◽  
D. Philibert Dembele

Nodal explants of 12 accessions from four species of yam (Dioscorea spp.) were cultured for six weeks on MS to evaluate the influence of IAA, Kn, NAA and BAP on the production of leaves and microtubers. Four Dioscorea polystachya Turcz., three each of D. bulbifera L. and D. sansibarensis Pax. and two D. japonica Thunb. accessions were used. Five and 10 mg/l of Kn along with IAA and sucrose, and 0.2 and 0.5 mg/l of NAA, sucrose and with or without BAP were used in four treatments. The accessions Yam 23 and Yam 25 of D. sansibarensis failed to initiate any leaf under four treatments. The remaining accessions produced 0.11 to 1.76 leaves per explant. The medium containing IAA with higher concentration of Kn (10 mg/l) and 3% sucrose was found to be best for in vitro production of leaf (0.71/explant) and the most productive species was D. japonica (1.36), followed by D. polystachya (1.19/explant). At the same culture period, Yam 16 of D. bulbifera failed to initiate any microtuber at IAA with Kn, and NAA with or without BAP. The remaining accessions produced 0.09 to 1.15 microtubers per explant. Lower concentration of Kn (5 mg/l) with IAA and sucrose was favourable for producing microtubers (0.61/explant on an average), the best species being D. sansibarensis (1.27) followed by D. japonica (0.59/ explant). Finally, the presence of BAP adversely affected the production of microtuber among Dioscorea species. Key words: Dioscorea spp., Microtuber, Nodal culture, Propagation D.O.I. 10.3329/ptcb.v18i1.3260 Plant Tissue Cult. & Biotech. 18(1): 25-35, 2008 (June)


Author(s):  
Gerrit Stuivenberg ◽  
Brendan Daisley ◽  
Polycronis Akouris ◽  
Gregor Reid

AbstractRecent studies suggest histamine and d-lactate may negatively impact host health. As excess histamine is deleterious to the host, the identification of bacterial producers has contributed to concerns over the consumption of probiotics or live microorganisms in fermented food items. Some probiotic products have been suspected of inducing d-lactic-acidosis; an illness associated with neurocognitive symptoms such as ataxia. The goals of the present study were to test the in vitro production of histamine and d-lactate by a 24-strain daily synbiotic and to outline methods that others can use to test for their production. Using enzymatic based assays, no significant production of histamine was observed compared to controls (P > 0.05), while d-lactate production was comparable to a commercially available probiotic with no associated health risk. These assays provide a means to add to the safety profile of synbiotic and probiotic products.


2011 ◽  
Vol 23 (1) ◽  
pp. 209
Author(s):  
S. Miyashita ◽  
K. Miyata ◽  
C. Tachibana ◽  
Y. Inaba ◽  
H. Koyama ◽  
...  

The objective of this study was to investigate the effect of stage of corpus luteum (CL) development on the in vitro production of bovine embryos. Ovaries were classified according to the expected day of the oestrous cycle based on the morphology of the ovaries. Ovaries with a corpus hemorrhagicum and the remnant of the follicular lumen filled with blood were considered the early luteal stage (Days 2 to 4; Day 0 = day of ovulation, n = 46). Ovaries with a large mass of orange tissue in the CL were classified as the midluteal stage (Days 7 to 10, n = 42). Cumulus–oocyte complexes (COC) were collected by aspiration of 2- to 6-mm follicles. The COC were classified into the following grades: COC with >3 compact layers of cumulus cells and evenly granulated cytoplasm were classified into Grade 1; COC with >3 layers cumulus cells and evenly granulated cytoplasm were classified into Grade 2; COC with partially remaining cumulus cells and abnormal cytoplasm were classified into Grade 3; COC without cumulus cells or those with expanded cumulus cells were classified into Grades 4 and 5, respectively. Grades 1 and 2 COC were in vitro matured for 20 h in TCM-199 supplemented with 5% calf serum and 0.02 mg mL–1 of FSH at 38.5°C under an atmosphere of 5% CO2 in air. Matured COC were inseminated with 5 × 106 sperm for 18 h. Presumptive zygotes were cultured in CR1aa medium supplemented with 5% calf serum at 38.5°C under an atmosphere of 5% O2, 5% CO2, and 90% N2 for 9 days (fertilization = Day 0). The mean number of COC and the proportion of COC classified as Grades 1 and 2 were analysed by ANOVA. Cleavage rates on Day 3 and blastocyst rates on Days 7 to 9 were analysed by a chi-square test. The mean number of recovered oocytes in the early luteal stage (18.7 ± 9.5) was significantly higher (P < 0.05) than the number in the midluteal stage (12.2 ± 5.7). The proportion of Grades 1 and 2 oocytes in the early luteal stage [66.7% (531/789)] was significantly higher (P < 0.01) than that in the midluteal stage [51.6% (252/484)]. The cleavage and blastocyst rates in the early luteal stage [60.9% (181/297) and 32.7% (97/297), respectively] were significantly higher (P < 0.05) than those in the midluteal stage [50.7% (76/150) and 20.7% (31/150) respectively].The present study suggests that the stage of development of the CL in bovine ovaries influences the number of recovered oocytes per ovary and the development of in vitro production of bovine embryos.


2021 ◽  
Vol 42 (3) ◽  
pp. 1147-1158
Author(s):  
Maria Fernanda Zamai ◽  
◽  
Fábio Luiz Bim Cavalieri ◽  
Marcia Aparecida Andreazzi ◽  
Fabio Morotti ◽  
...  

Reproductive biotechnologies are emerging as an important element for livestock; however, some strategies must be modified to adapt to different breeding systems, such as the use of follicular synchronization protocols. This study aimed to evaluate follicular synchronization using estradiol benzoate (EB), in the presence of the corpus luteum (CL) from Wagyu oocyte donors in in vitro embryo production (IVEP). Rounds of IVEP were performed in heifers and cows (n=19) that were classified into three groups: G1/CL - animals with CL, G2/WCL - animals without CL, and G3/CL + EB - animals with CL that were subjected to follicular synchronization with EB at D0. The groups G1/CL and G2/WCL were considered the control and undertook the natural process of follicular dynamics. The results showed that the synchronization of the follicular wave with the application of EB in the presence of CL, presented a smaller number of small (6.05 ± 0.55) and large follicles (0.45 ± 0.15), but increased (P < 0.05) the number of medium-sized follicles (16.20 ± 0.90). However, the results of ovum pick up showed that regardless of whether or not EB was applied, and regardless of the presence or absence of CL in the Wagyu donor, there was no difference among the groups (P > 0.05) concerning the number of viable oocytes and the viability rate. It was concluded that follicular synchronization using EB in Wagyu oocyte donors that presented a CL, increased the number of medium-sized follicles. However, there was no improvement in the efficiency of ovum pick up, in vitro embryo production, and pregnancy rate.


1970 ◽  
Vol 28 (1) ◽  
pp. 31-38
Author(s):  
MNS Talukder ◽  
A Iqbal ◽  
MAMY Khandoker ◽  
MZ Alam

An alternative to superovulation is in-vitro production (IVP) of embryos where the efficient collection and grading of oocytes is important. Ovaries from an abattoir were collected and categorized as type I with no corpus luteum (CL), and type II with CL. The length, width and weight of type I and type II ovary were 1.4 ± 0.03 and 1.5 ± 0.08 cm; 0.8 ± 0.04 and 1.0 ± 0.07 cm; 0.6 ± 0.07 and 0.7 ± 0.04 gm, respectively, each significantly (P<0.05) higher in type II ovaries. A total of 80 and 78 follicles were observed and 60 and 61 follicles aspirated from left and right ovaries, respectively, from each of 25 ovaries. Out of 133 follicles 100 were aspirated from 40 type-I ovaries, and 21 aspirated from 10 type-II ovaries. The differences in the number of normal, abnormal and total cumulus-oocytecomplex (COCs) per ovary between left and right ovaries were not significant (P>0.05). The number of normal (1.9 ± 0.11) and total (2.5 ± 0.14) COCs per ovary were significantly (P<0.05) higher in ovaries without than in those with CL (1.2 ± 0.36 and 2.0 ± 0.30, respectively). But the number (0.80 ± 0.13) of abnormal COCs per ovary was significantly (P<0.05) higher in ovaries with CL than in those without (0.7 ± 0.09). Significantly (P<0.05) higher percentage of COCs expansion was grade A (6.9 ± 2.05) than grade B (53.1 ± 1.27) COCs. It is suggested that type I (without CL) ovaries and follicles of 2-6 mm diameter are suitable to collect good quality COCs for in-vitro maturation (IVM) of oocytes and the culture condition for IVM of sheep COCs are reported. DOI: http://dx.doi.org/10.3329/bvet.v28i1.8811   Bangl. vet. 2011. Vol. 28, No. 1, 31-38


2012 ◽  
Vol 47 (No. 2 - 3) ◽  
pp. 33-37
Author(s):  
S. Čech ◽  
V. Havlíček ◽  
M. Lopatářová ◽  
M. Vyskočil ◽  
R. Doležel

Effectiveness of in vitro production of embryos (IVP) is limited among other factors by the recovery rate of oocytes. Gonadotropin superstimulation can improve the recovery rate of oocytes. The effect of FSH treatment on follicular population and recovery rate of oocytes at ovum pick-up (OPU) in the growing phase of the 1st as well as the 2nd follicular wave after superstimulation was the object of our experiment. Twelve unpregnant milking cows (15&ndash;20 kg milk per day) housed on a dairy farm were used in the experiment. The cows bearing corpus luteum were synchronized by PGF<sub>2 </sub>(day 0) and they were treated by FSH (Folicotropin inj. sicc. ad us vet., Spofa Prague, Czech Republic, single doses 80, 80, 80, 80, 40 and 40 UI) in 12 h intervals on days 12, 13 and 14. Transvaginal ultrasonographic puncture of oocytes in cows bearing a new corpus luteum was performed on day 7 (OPU 1, various phase of the follicular wave, removal of the dominant follicle) and it was repeated on days 10 (OPU 2, growing phase of the follicular wave &ndash; control), 16 (OPU&nbsp;3, growing phase of the first follicular wave after superstimulation) and 20 (OPU 4, growing phase of the second follicular wave after superstimulation). All follicles &gt; 2 mm were punctured. The ovarian follicles (ultrasonographically) and numbers and qualities of obtained oocytes (microscopically) were evaluated during and immediately after OPU. Follicular population was divided to small (FS, 2&ndash;5 mm), medium (FM, 5&ndash;9 mm) and large (FL, &gt; 9 mm) follicles. Oocytes were classified as 1st (intact cumulus, &gt; 3 layers of cumulus cells), 2nd (complete 1&ndash;3 layers of cumulus cells), 3rd (incomplete layers of cumulus cells, expanded cumulus mass) and 4th (absence of corona cells, degenerated oocytes) classes. Although we found the least of FS (x = 1.0) during OPU 3, significantly more FM (x = 24.7) and FL (x = 3.1) follicles were found at this procedure in comparison with others. Likewise a significantly higher number of oocytes (x = 8.1) was obtained at OPU 3 in comparison with OPU 1 and OPU 2. Significantly higher number of FM (x = 6.1) was found and non-significantly higher number of oocytes was obtained at OPU 4 in comparison with OPU 1 and 2. The results show that administration of FSH increases the number of follicles and the number of collected oocytes in the growing phase of the 1st follicular wave after superstimulation, nevertheless a higher number of follicles and a higher recovery rate of oocytes can be expected in the growing phase of the 2nd follicular wave after superstimulation as well.


2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 372-372
Author(s):  
Araksiya Smekalova ◽  
Elena K Montvila ◽  
Olga Konovalova ◽  
Olga Mityashova ◽  
Irina Lebedeva

Abstract Growth hormone (GH) is an endocrine and paracrine/autocrine regulator of the hen ovarian function, with the GH receptor concentration in the granulosa layer (GL) being maximum in the largest preovulatory follicle (Lebedeva et al. 2004, Biol.Reprod. 71:1174–1181). In the present study, GH effects on in vitro production of progesterone by GL from the two largest yellow follicles (F1 and F2) were investigated due to the hen age and the presence of the theca layer (TL). Young hens with long clutch (YLC, 32–33 week-old, &gt;10 eggs per clutch) and old hens with short clutch (OSC, 74–76 week-old, 3–6 eggs per clutch) were used. After isolation, GL from F1 and F2 follicles (n = 8–9) was cultured separately or jointly with the respective TL for 18 h in the presence or absence of chicken GH (25 ng/ml). Concentrations of progesterone in culture media were measured by ELISA. The data were analyzed by repeated measures ANOVA. When GL from F1 follicle cultured alone, GH did not affect progesterone production in YLC hens and decreased it from 30,5±3,4 to 20,5±2,9 ng/mg tissue (P &lt; 0.01) in OSC hens. Conversely, when tested GL from F2 follicle, GH increased progesterone output from 15,8±2,4 to 20,4±2,5 ng/mg tissue (P &lt; 0.05) in YLC birds and had no effect on the output in OSC birds. During co-culture of GL and TL, GH raised 1.4–1.5 times the production of progesterone in the case of F1 follicle and did not change it in the case of F2 follicle in hens of both ages. The findings indicate that the steroidogenic response of GL from the two largest preovulatory follicles to GH differs in young and old hens. However, the interaction with TL modifies the GL response and makes it similar in birds regardless the age and reproductive status. The study was supported by RFBR (19-016-00216).


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