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F Martinat-Botté

Publications and source records attributed to F Martinat-Botté.

13 recordsLinked to original sources
PubMed2007

The effectiveness of the stereomicroscopic evaluation of embryo quality in vitrified-warmed porcine blastocysts: an ultrastructural and cell death study.

The objective of this study was to analyze the validity of the stereomicroscopic evaluation of vitrified-warmed (V-W) porcine blastocysts. Unhatched blastocysts were obtained from Large-white gilts (n=10). Blastocysts (n=156) were vitrified using the Open Pulled Straw technology. After warming, V-W blastocysts were cultured for 24h (V24). Then, their developmental progression was morphologically assessed by stereomicroscopy and classified as: V24 viable re-expanded blastocysts; V24 viable hatched blastocysts or V24 degenerated. Blastocysts which re-expanded or hatched after warming were considered viable. Some fresh blastocysts were not vitrified and were evaluated after 24h in culture (F24). By stereomicroscopic analysis all the fresh blastocysts were considered viable. Some F24, V24 re-expanded viable, V24 hatched viable and V24 degenerated blastocysts were processed for transmission electron microscopy (n=13, 19, 9 and 9, respectively) or assessed by TUNEL for cell-death evaluation (n=16, 21, 11 and 21, respectively). All V24 hatched blastocysts showed similar ultrastructure to fresh blastocysts. However, some V24 re-expanded blastocysts considered viable (6/19) revealed ultrastructural alterations. Degenerated V24 blastocysts showed ultrastructural disintegration. Hatched V24 blastocysts did not differ (p>0.05) from F24 hatched blastocysts with regard to the ratio of dead cells (2.8+/-0.5% versus 1.9+/-0.3%, respectively). However, V24 expanded blastocysts had higher (p<0.01) cell death levels (4.3+/-3.4%) than those observed in the F24 expanded blastocysts (1.1+/-0.3%). The degenerated blastocysts showed the highest cell-death index (19.4+/-6.3%). In summary, V-W blastocyst hatching during in vitro culture appears to coincide with good ultrastructure and low cell-death index, suggesting that the hatching rate assessed by stereomicroscopy is more appropriate than embryo re-expansion for an evaluation of V-W blastocyst quality.

Animals↗
PubMed2006

[Cryopreservation and transfer of pig in vivo embryos: State of the art].

The present review article describes the development of porcine cryopreservation and transfer of embryos and the sanitary regulations related to these technologies. Rapid progress has been made during the last decade in cryopreservation by using vitrification which involves a very rapid cooling rate. Currently, this technology is suitable for morulae and blastocysts and to date, more than 300 piglets are born after surgical transfers of vitrified embryos. Moreover, farrowings are obtained after non-surgical transfers of fresh or vitrified embryos. However, further improvements are required to permit the dissemination of these technologies in the near future. According to the recommendations of the International Embryo Transfer Society (IETS) regarding international transportation, embryos must be washed and cryopreserved with an intact zona pellucida. Transmission by embryo transfer of virus or bacteria to swine has been studied and rather converging results are now published. The risk of disease transmission by the embryo transfer is reduced. The major advantage of these technologies is the possibility to transport and store genetic material whilst reducing the risks of pathogen transmission.

Animals↗
PubMed2005

Piglets born after non-surgical deep intrauterine transfer of vitrified blastocysts in gilts.

The aims of this study were: (1) to evaluate the effect of the number of previous estrus of recipient gilts on effectiveness of intrauterine insertion of a flexible catheter designed for non-surgical deep intrauterine catheterization during diestrus in pigs; and (2) to determine the farrowing rate and the litter size after non-surgical deep intrauterine embryo transfer (ET) of porcine blastocysts vitrified by the open pulled straw (OPS) method. In experiment 1, 27 large white hyperprolific gilts (LWh) with 2-6 previous estrus were used. Intrauterine insertions of the flexible catheter were carried out at day 5.5-6 of the estrous cycle (D0=onset of estrus). During insertions, no or only moderate reactions were observed in 88.9% of gilts and was not related (P >0.05) to the number of estrus prior to the insertion periods. The number of the estrus had a significant effect (P <0.05) on the difficulties found during the procedure. In the 100% of gilts with two estrus (N=6) it was not possible to insert the flexible catheter through the cervix. In gilts with three or more estrus, it was possible to pass the cervix and to progress along a uterine horn in 80.9% of the cases. In 86.7% of the gilts, the tip of the flexible catheter achieved the second or third quarter of the uterine horn. In experiment 2, following non-surgical deep intrauterine transfer of 20 vitrified/warmed blastocysts, 9 Meishan recipients (42.9%) farrowed an average of 5.4 +/- 0.8 piglets (range 3-9) of which 0.6 +/- 0.3 piglets (range 0-2) were born dead. In conclusion, this study shows that it is possible to obtain birth of piglets following non-surgical deep intrauterine embryo transfer (ET) of vitrified/warmed blastocysts. Non-surgical deep intrauterine ET and OPS vitrification methods are promising procedures to be used together for the introduction of new genetic material in a farm.

Animals↗
PubMed2000

Piglets born after vitrification of embryos using the open pulled straw method.

Morulae and unhatched blastocysts from Large White hyperprolific (LWh) and Meishan (MS) gilts were selected to test an ultrarapid open pulled straw (OPS) vitrification method with two media. The viability of vitrified/warmed embryos was estimated by the percentage of embryos that developed to the hatched blastocyst stage in vitro or by birth after transfer. In Experiment 1, two cryoprotectant dilution media were compared for cryopreservation of MS and LWh blastocysts: TCM was a standard Hepes-buffered TCM199 + 20% NBCS medium and PBS was a PBS + 20% NBCS medium. After a two-step equilibration in ethylene glycol, dimethyl sulfoxide, and sucrose, 2-5 blastocysts were loaded into OPS and plunged into liquid nitrogen. Embryos were warmed; a four-step dilution with decreasing concentrations of sucrose was applied. In PBS, LWh blastocysts (27%) had a lower viability in vitro than MS blastocysts (67%; P = 0.001). In TCM, no significant difference was observed between genotypes (41% for LWh and 43% for MS blastocysts) and both viability rates were lower than that of the control groups. In Experiment 2, morula-stage LWh and MS embryos were vitrified and warmed using PBS. The viability rate was low and did not differ between LWh (11%) and MS (14%). In Experiment 3, 200 MS and 200 LWh blastocysts were vitrified/warmed as described in Experiment 1 (PBS). In each of 20 MS recipients, 20 embryos were transferred. The farrowing rate was 55% and recipients farrowed four and five piglets (median) for MS and LWh blastocysts, respectively. The OPS method is therefore appropriate for cryopreservation of unhatched porcine blastocysts.

Animals↗
PubMed1998

Intrauterine infusion of high doses of pig trophoblast interferons has no antiluteolytic effect in cyclic gilts.

In the pig species, the preimplanting trophoblast is known to synthesize and secrete high amounts of interferon during early development. Previous experiments in cyclic gilts using total conceptus secretory proteins suggested that porcine trophoblastic interferons, unlike those of ruminants, exert no effect on the luteal cycle. In the present experiment, cyclic Meishan gilts were divided into two groups, cannulated on both uterine horns, and given daily injections of either a placebo or increasing doses of a mixture of recombinant interferon-gamma and interferon-delta, on Days 11-14 of the estrous cycle. In treated gilts, the injected doses were much higher than those previously found in uterine perfusates from pregnant gilts. However, no significant differences could be found between the control (n = 4) and the treated (n = 5) group concerning the days of the estrous cycle for mid-decrease of progesterone (control: Day 14.5+/-0.57 [mean+/-SD]; treated: Day 15+/-1.25), the day of estrus (control: Day 19+/-0.96; treated: Day 19.6+/-0.55), and the subsequent ovulation rate (control: 14+/-2.2 corpora lutea; treated: 13.1+/-1.1 corpora lutea). These data confirm that pig trophoblastic interferons, unlike those of ruminants, do not themselves exert an antiluteolytic effect. A possible synergistic effect of embryonic estrogens on the luteal functions of nonpregnant sows remains to be determined.

Animals↗
PubMed1994

Presence of leukaemia inhibitory factor and interleukin 6 in porcine uterine secretions prior to conceptus attachment.

Leukaemia inhibitory factor (LIF) plays an important role in embryo development and implantation. We detected peak LIF activity in porcine uterine luminal fluids (ULF) at day 12 of gestation and during day 7 and 13 of the oestrous cycle. A radio-receptor competition assay showed the presence of a molecule in ULF specifically binding to human LIF receptor (LIF-R). LIF activity was partially neutralized by anti-human LIF antibody. Interleukin-6 (IL-6) activity was detected in ULF throughout the oestrous cycle and pre-implantation period. An anti-murine alpha chain (gp80) of IL-6 receptor (IL-6R) specifically neutralized this activity. LIF and IL-6 mRNA were only detected in day 11 endometrium. The presence of LIF or IL-6 in the uterine cavity has not been previously reported. Our results extend LIF production by endometrium during the oestrous cycle and pre-implantation period to another mammalian species other than mouse.

Animals↗
PubMed1992

Oestrus and LH responses to oestradiol during lactational anoestrus in Chinese Meishan and large white sows.

To investigate endocrine mechanisms associated with the occasional occurrence of fertile oestrus during lactation in the high prolific Chinese Meishan (MS) breed, the incidence of oestrus and changes in plasma luteinizing hormone (LH) levels before and after oestradiol benzoate (OB, 15 micrograms/kg body weight) administration on day 22 was compared in 4 MS and 6 Large White (LW) sows. All sows exhibited oestrus in response to OB. Only 1 sow (MS) ovulated in response to OB, became pregnant and farrowed. Mean plasma LH levels before OB were low (MS: 0.38 +/- 0.06 ng LH/ml, LW: 0.29 +/- 0.04 ng LH/ml, ns). LH levels above 2 ng/ml (surge) occurred in 2/4 MS and 2/6 LW sows at 60 +/- 5 h after OB. The MS sow that ovulated had an LH surge level of 4.5 ng/ml plasma at 40 h after OB. These results indicate minor breed differences in the control of LH secretion during lactational anoestrus.

Anestrus↗
PubMed1990

Interferon-gamma gene and protein are spontaneously expressed by the porcine trophectoderm early in gestation.

The nature and the source of the antiviral activity found in the reproductive tract of pregnant gilts early in gestation were analyzed. Two antigenically distinct antiviral activities were found in uterine flushings and in supernatants of conceptus-conditioned culture medium between days 12 and 20 of gestation, using Madin Darby bovine kidney cells and vesicular stomatitis virus as a challenge in the antiviral bioassay. One component was antigenically identified as interferon-gamma (IFN-gamma). Northern blot analysis of conceptus poly(A)+ RNA with a human IFN-gamma cDNA probe revealed two mRNA of 1.3 and 1.4 kb. In addition, immunoprecipitation of metabolically labeled conceptus secretory proteins with an antiserum raised against purified porcine rIFN-gamma resulted in four bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with molecular mass 18.5 to 24.5 kDa. Pre-electrophoresis incubation of the immunoprecipitate with glycopeptidase F, which removes N-linked carbohydrates, yielded a single band of 16.5 kDa. Finally, staining of ultrathin sections by indirect immunofluorescence using the same antiserum to rIFN-gamma revealed that all cells of extra-embryonic trophectoderm contained intensely fluorescent granules in their apical cytoplasm. Neither endoderm nor embryonic cells stained positive. These results clearly show that IFN-gamma, known so far as a T or NK cell-derived lymphokine, is spontaneously and intensively secreted by the porcine trophectoderm, an embryonic tissue not related to the hematopoietic lineage. They also suggest that the implanting conceptus, at least in the porcine species, could play an active role in immune interactions with the mother.

Animals↗
PubMed1986

Use of ultrasound scanning in swine for detection of pregnancy and some pathological conditions.

The adaptation of ultrasound scanning to the diagnosis of pregnancy and other physiological conditions in swine was examined. The scanner was equipped with a 3.5-MHz transducer. The probe was placed against the skin in the area of the abdominal flank of a standing sow and moved between the first and third posterior teats. The 2,257 sows were all presumed to be pregnant and underwent a single echography examination between 18 and 90 d post-insemination. For pregnant sows, the accuracy rate of diagnosis averaged 98.7 %. The majority of errors were made between 18 and 21 d of pregnancy in sows which subsequently gave birth to fewer than five piglets. This technique allowed us to detect only 54 % of non-pregnant sows. Nevertheless, a high accuracy rate was obtained when examination was performed within 7 d before the return of estrus (80 %). Conditions at the time of the examination were also an important factor. Ultrasound scanning opened up possibilities for observing other conditions such as cystic ovaries and metritis.

Journal Article↗
PubMed1985

Control of pig reproduction in a breeding programme.

Important improvements in the management of sows have been made over the past 20 years in Europe and the U.S.A. Nevertheless, annual productivity varied between 15 and 25 piglets weaned per sow per year in France for 1983. Reasons for such a difference in productivity have been analysed quantitatively. Marked differences exist between herds in the genotype of pig used and in the age of puberty. Transportation acts as a stress stimulus and stimulates puberty attainment. However, there is variability in the occurrence of oestrus amongst different purchased batches of pigs. Recent endocrine results led to four phases in sexual maturation of gilts being distinguished (perinatal, infancy, activation and resting) and permitted a better understanding of the variation in the onset of puberty. After a lactation of 3 weeks or more, about 25% of primiparous sows did not return to oestrus within 1 week of weaning. Fertility of these sows with a delayed oestrus was 10% lower than in sows having a normal oestrus after weaning. The past 3 decades of research on control of ovulation have yielded positive results. However, there is no satisfactory method for the induction of puberty. One method used to regulate the oestrous cycle in gilts is treatment with a progestagen. This method is still available commercially in France. The close synchronization obtained with a progestagen treatment has led to the use of AI at fixed days (Days 6 and 7) in crossbred gilts but not for purebred gilts. Some environmental factors (e.g. male stimulation, age of gilts at treatment, diet and month of AI) have been analysed to evaluate their effects on farrowing rate and litter size. After weaning, a short treatment with a progestagen (20 mg/day) tends to improve the synchronization of oestrus (3-9 days), especially in primiparous sows. Farrowing rate and litter size are similar in treated sows and control sows. However, this technique does not permit fixed-time AI without detection of oestrus. Diagnosis of pregnancy was successful with ultrasound scanning. This technique can be performed 20 days after insemination if the sow is tied up during the examination and at 22 days if the sow is restrained in a retention box. From 22 days after mating, errors made for positive and negative diagnoses were less than 5%. These techniques contribute to a better control of reproduction in different management systems.

Animal Husbandry↗
PubMedDate not supplied

Relationship between peri-oestrus progesterone levels and time of ovulation by echography in pigs and influence of the interval between ovulation and artificial insemination (AI) on litter size.

Two methods for the determination of ovulation were compared to one ultrasonography performed 5 times a day. Time of ovulation by echography was 40 +/- 5.8 h (mean +/- SD) after the onset of oestrus. Preovulatory LH rise (two blood samples per day) began near the onset of oestrus but, in our conditions, this parameter could not be used to predict ovulation. The basal level of progesterone (two blood samples per day) was determined with a non-linear model, the timing when progesterone rose more than one SD (0.3 ng x mL(-1)) coincided with the timing of ovulation determined by echography (R2 = 0.98). This method was efficient and was used in a field trial to measure the consequences of the variability of the interval between Al and ovulation on litter size. The interval between Al and ovulation had an effect on litter size; litter size decreased by one piglet when this interval increased by 10h.

Animals↗
PubMedDate not supplied

Birth of piglets after OPS vitrification and transfer of compacted morula stage embryos with intact zona pellucida.

In swine, five to six days post-insemination, morulae and blastocysts are collected together after uterine flushing. The purpose of this study was to vitrify zona pellucida-intact morulae with Open Pulled Straw (OPS) technology and obtain piglets after transfer. Morulae (200) were vitrified after a two-step equilibration in ethylene glycol, dimethyl sulfoxide and sucrose in Hepes-buffered TCM199 + 20% NBCS medium (TCM). 2-6 morulae were loaded into OPS and plunged into liquid nitrogen. At embryo warming, a three-step dilution with decreasing concentrations of sucrose was applied. In each of 10 recipients, 20 morulae were transferred surgically. Day 25, gestation rate and the farrowing rate were 80% and 70%, respectively. The pregnant recipients farrowed from 1 to 8 piglets and the survival of total transferred embryos was 13%. Although survival rates are still compromised, OPS technology is therefore appropriate to cryopreserve porcine morulae with intact zona pellucida.

Animals↗