Although the in vivo recovery of Not worried about looks age or race and GC using OPU systems have been ly reported, in most cases, the samples were collected from large or preovulatory follicles Fuck locals Pansey Alabama 18 — 21 ], or were pooled [ 22 ].
Moreover, the amount of FF or GC recovered from each follicle was seldom reported [ 22 — 25 ], and this is a key piece of information that is required to de endocrine and molecular studies on an individual basis.
Whether OPU systems can be used to effectively sample small and medium follicles is particularly important in studies with Zebu breeds. The diameter of the dominant follicle at deviation [ 2627 ] and at ovulation [ 28 ] in Bos indicus is smaller than in Bos taurus.
Thus, studies in Zebu and crossbreed animals would require the sampling follicles of smaller sizes than those ly reported in other breeds. The aim of the present study was to adapt and evaluate the efficiency of an OPU system in recovering individual samples of FF and GC from follicles of different diameters in vivo.
To adapt and evaluate the Girls from Bangor Maine nude use of an OPU system for the recovery of Beautiful older ladies want xxx dating Rock Springs Wyoming and GC samples from follicles of different diameters in vivo, three experimental trials were performed.
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The first trial was deed to adapt the system to collect FF and GC samples from follicles of different diameters. The second and third trials were deed to evaluate the efficiency of the adapted OPU system for the in vivo recovery of samples of FF and GC, Free fuck Waynesburg. All animals were cycling and no reproductive abnormalities were observed in prior gynecological exams.
The crossbred cows were kept in an outdoor grazing system Brachiaria decumbens and the Holstein heifers were confined in free stalls and fed with corn silage and concentrate.
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Water and minerals were available to the animals ad libitum. Trial I: development of the adapted OPU system A preliminary in vitro test was performed to calculate fluid losses using a Lonely wife wants casual sex Troutdale OPU Teflon tubing circuit that was 80 cm long and of 1.
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The known volumes of blood plasma that were expected to correspond to the volume of FF in follicles of 12, 10, 8, 6, and 4 mm in diameter were placed in adult friend finder. Five replicates were prepared for each follicle diameter. Thereafter, these volumes were aspirated using a 20 G needle connected to one tip of the Telfon circuit and were recovered in separate 1.
The conventional OPU system was later adapted to reduce the losses observed for 8, 6, and 4 mm follicles. A sterile 0.
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Using this adapted OPU system, known amounts of blood plasma, expected to correspond to the volumes of FF in follicles of 4 and 6 mm in diameter, were aspirated, recovered in the semen straws, Get girl fuck Digby transferred to 1. The losses were calculated as ly described. Two 0.
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The cotton plug was removed from the first semen straw. Figure 1 An illustration of the adapted OPU. To connect the straw, a small silicone tube 3 was used to Woman seeking nsa East Marion one tip to the circuit, while the other tip was directly connected to the needle 6. For follicles from 7.
The cotton plug of the second straw was removed and another small silicone tube was used to connect the two semen straws. Full size image Trial II: follicular fluid recovery in vivo using the adapted OPU system For the second trial, the adapted system Sex date Gaithersburg I was used to collect FF directly from the ovarian follicles of the cows.
The animals were scanned daily with a transrectal B-mode ultrasonography 7.
The follicles that were Swingers Personals in Flynn in each wave were measured and individually aspirated at specific diameters. They were then grouped into 12specifically, 4.
The main procedures for OPU were carried out as ly described [ 27 ]. After the aspiration of each follicle, the Teflon system and the needle were replaced to avoid cross-contamination.
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Instead of a pump, the differential pressure applied to the system to recover the FF was created with a 10 mL syringe. Immediately after follicle collapse, the needle was removed from the ovary Free nude women in Chestertown Maryland the system was checked for the presence of FF and blood contamination.
The volume of the FF sample was determined as ly described Trial Ibased on the sample weight and taking the density of the FF to be 1. The emergence of the follicular wave was synchronized in 20 Holstein Date married women 10 Gir heifers, using the same protocol described in the section. The heifers were scanned daily by transrectal B-mode ultrasonography 7. Thereafter, the follicular dynamics were monitored every 12 h and the largest follicle was aspirated at 6, 8, 10 Best girls to fuck in Leamington 12 mm in Holstein, or at 4, 6, 8 or 10 mm in Gir heifers.
These diameters were established by taking into consideration the differences in follicle size at deviation between the breeds 8. The follicle aspiration procedures Housewives wants sex tonight WI Evergreen 54840 were as described in the section.
They were also inspected for the presence or absence of the cumulus-oocyte complex COC by stereomicroscope. If present, COC was removed from the fluid. The samples were then centrifuged at X g for 10 min to separate the fluid and the cells.
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The of cells was determined with a hemocytometer. The samples that were visually contaminated with blood or with fewer thanGC were discarded.
Statistical analyses The outcome variables, including volume losses, follicular fluid volume, of GC, and quantity of RNA recovered, were evaluated for normality and homoscedasticity Horny friends in Baltimore Maryland Lilliefors and Cochran and Bartlett tests.
The efficiency of the in vivo recovery of the FF sample was determined by regression analysis.
The frequencies of visible blood contamination and FF samples with less thanGC were evaluated by Chi-squared test. Table 1 Recovery efficiency in vitro using a conventional 80 cm Teflon tubing or an adapted with 0.
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