Journal of reproduction and fertility. Supplement (J Reprod Fertil Suppl)

Publisher: Society for Reproduction and Fertility

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Other titles Reproduction (Cambridge, England)
ISSN 0449-3087
OCLC 49335849
Material type Series
Document type Journal / Magazine / Newspaper

Publications in this journal

  • [Show abstract] [Hide abstract]
    ABSTRACT: The exact nature of cryo-injury to dog semen and the stage of cryopreservation at which it occurs have not been evaluated fully and were investigated in the present study. Semen samples were examined immediately before and after addition of semen extender, cooling and freeze-thawing in a Tris-based medium. Vital and ultrastructural assessments were made immediately after each treatment and after incubation for 4 h at 39 degrees C after each treatment. Addition of semen extender produced no significant ultrastructural changes in spermatozoa; however, cooling resulted in an immediate increase in the number of acrosomal abnormalities and a subsequent decrease in sperm viability. Freeze-thawing caused both an immediate and a delayed decrease in sperm viability. This effect may have been an exacerbation of the effects of cooling as many acrosomal abnormalities were present or it may have been the result of initially unrecognized damage. Cooling and freezing of dog spermatozoa may have both immediate and delayed effects on the ultrastructure of spermatozoa. The immediate effects of cooling and freezing may either kill spermatozoa or render them incapable of fertilization by damaging the acrosome, whereas delayed effects may reduce sperm longevity by altering plasma membrane structure.
    Journal of reproduction and fertility. Supplement 02/2001; 57:357-63.
  • L Moe ·
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    ABSTRACT: Data from two population-based studies in four Norwegian counties were used to calculate the crude incidence of mammary tumours, and the age- and breed-specific incidence of mammary tumours in female dogs of three different breeds. The largest study comprised 14401 histologically verified tumour cases from four counties covered by the Norwegian Canine Cancer Register. The registry covers about 25% of the total Norwegian dog population. The second study was a census in Norway that was sent to all owners of the following breeds: boxers, bichon frisé and Bernese mountain dogs, to estimate the age distribution of the female dog population at risk of developing mammary tumours. The crude incidence of malignant mammary tumours in female dogs of any breed was 53.3%. The highest relative risk ratio of mammary tumours was found in boxers, cocker spaniels, English springer spaniels and dachshunds. The mean age of histologically diagnosed mammary tumours was 7.9 years in boxers and 7.8 years in springer spaniels, compared with 8.8 years in all other breeds. In the four Norwegian counties from 1992 to 1997, the population-based incidence rates (for all ages) of malignant mammary tumours per 1000 female dogs per year were 35.47 in boxers, 3.87 in Bernese mountain dogs and 17.69 in bichon frisé. Mammary cancer is the most common tumour in female dogs in Norway, and represents a population of almost entirely reproductively intact females. The age-specific incidence rates for mammary cancer vary considerably among the three breeds that were studied in detail.
    Journal of reproduction and fertility. Supplement 02/2001; 57:439-43.
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    ABSTRACT: The aim of this study was to determine whether nuclear transplantation could be used to clone a dog using donor nucleus cells collected from an adult female. Fibroblasts obtained from skin biopsies were fused with enucleated bovine or canine oocytes. The resulting cloned embryos were cultured in vitro to monitor embryonic development. A proportion of the resulting embryos was transferred into surrogate bitches for development to term. When canine oocytes were used as recipient ova for canine fibroblasts, 23% of the resulting embryos cleaved at least once after culture in vitro. Five cloned embryos were transferred into three synchronized recipient bitches, but no pregnancies resulted. When bovine oocytes were used as recipinets for canine fibroblasts, 38% cleaved to the two- to four-cell stage and 43% cleaved to the eight- to 16-cell stage. Forty-seven of these embryos were transferred into four recipient females, resulting in a single conceptus that ceased development at about day 20 of gestation. The desire for cloned dogs is considerable and will undoubtedly incite the development of successful methods for cloning companion animals. However, significant investment into additional research is required, especially in the areas of in vitro maturation of oocytes and control of the oestrous cycle of bitches.
    Journal of reproduction and fertility. Supplement 02/2001; 57:287-93.
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    ABSTRACT: Our understanding of molecular mechanisms of fertilization in mammals has lagged behind the rapid development of reproductive technology over the last decade. Significant advances in knowledge have resulted mainly from studies in laboratory animals in vitro. However, this situation has changed in the last few years as targeted mutagenesis in mice has provided important new information about genes and proteins involved in basic aspects of sperm-egg interactions in vivo. In this brief review the current knowledge about the generic aspects of mammalian fertilization is summarized, together with particular features of reproductive physiology related to cats and dogs.
    Journal of reproduction and fertility. Supplement 02/2001; 57:105-10.
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    ABSTRACT: Taurine and hypotaurine have been found in spermatozoa and seminal plasma of numerous species and are known to have beneficial effects on sperm characteristics in mammals. Taurine is considered an essential dietary constituent in cats. Dietary deficiency has been associated with a range of serious clinical disorders. Quantification of taurine and hypotaurine in the genital tracts of male cats has not been reported. In this study, the concentrations of taurine and its precursors were measured in serum, spermatozoa, epididymal fluid and seminal plasma from cats. The concentrations of taurine measured in serum samples confirmed that the cats were not deficient in taurine. Significant amounts of taurine and hypotaurine were found in spermatozoa, seminal plasma and epididymal flushing fluid. Hypotaurine was not detected in serum samples. These results indicate that hypotaurine may be synthesized in cat testes or epididymides. Cysteamine was not detected in any of the samples.
    Journal of reproduction and fertility. Supplement 02/2001; 57(1):93-5.
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    ABSTRACT: The aims of this study were: (i) to establish a reliable model for the study of cystic endometrial hyperplasia in ovariectomized bitches; and (ii) to assess the roles of oestrogen and progesterone in the pathogenesis of irritant-induced cystic endometrial hyperplasia. Greyhound bitches (n = 15) were ovariectomized and divided into five groups (n = 3 per group). After 3-4 weeks, oestradiol benzoate (0.6-4.8 micrograms kg-1, i.m.) was administered twice a day for 12 days to the bitches in group 1, followed by progesterone (0.2-1.8 mg kg-1, i.m.) twice a day for 30-33 days. These dosages were chosen to mimic the plasma hormone concentrations of a normal oestrous cycle. A silk suture was inserted by laparotomy into the left uterine horn 12 days into the simulated dioestrus (determined by vaginal cytology) and necropsy was performed after a further 12 days. For groups 2-5, the silk suture was positioned at ovariectomy. After a further 3-4 weeks, these bitches were treated with progesterone (group 2: 1.8 mg kg-1 i.m. twice a day), oestradiol benzoate (group 3: 0.6-4.8 micrograms kg-1 i.m. twice a day), oestradiol benzoate and progesterone together (group 4: previous dosages) or vehicle (group 5). Necropsies were performed after 12-13 days of treatment. Cystic endometrial hyperplasia was induced in the suture-containing uterine horns of all bitches in groups 1 and 4, and in two bitches in group 2. Cystic endometrial hyperplasia did not develop in any control (no suture) uterine horns, or in either uterine horn of the bitches treated with either oestradiol only or vehicle. These results indicate that progesterone is necessary for the development of irritant-induced cystic endometrial hyperplasia and that oestradiol potentiates the effects of progesterone. The protocol used for bitches in group 1 would be a suitable model for further studies of the pathogenesis of cystic endometrial hyperplasia.
    Journal of reproduction and fertility. Supplement 02/2001; 57:407-14.
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    ABSTRACT: The aim of this study was to identify and characterize populations of trophoblast cells in canine placenta during different stages of fetal development using lectin histochemistry. Dogs have endotheliochorial placentation and trophoblast cell invasion continues after chorioallantois villous penetration early in pregnancy, leading to formation of a labyrinth. Specialized subpopulations of cells differentiate, such as syncytial trophoblast that invades the maternal epithelium early in placentation and surrounds and forms intimate cuffs around maternal blood vessels. Marginal haematomata, which are lined by specialized phagocytic cytotrophoblast cells, form by mid-gestation. Invasive 'extravillous' cells advance into and remodel maternal endometrial tissues further. Placentas and attached uterine tissues were collected and sampled from six bitches at mid-gestation (days 31-33 of gestation) and 12 females in late gestation (day 42-term) for characterization of these tissues and identification of other populations of trophoblast cells. Uterine tissues from nonpregnant bitches were also collected at oestrus (n = 2) and during the luteal phase (n = 1). In histochemical studies, two of six biotinylated lectins that were tested stained cytotrophoblast and syncytial trophoblast cell populations differentially. Arachis hypogaea agglutinin (PNA) was specific for cytotrophoblasts in placental tissue lining villi and cytotrophoblastic cells with phagocytic or absorptive phenotypes in the necrotic zone at mid-gestation. In late gestation, cytotrophoblast cells with an absorptive phenotype at the interface between the labyrinth and lacunar glandular chambers were stained with PNA. Staining of other cells was minimal, with the exception of deep endometrial glands. Lectin binding using Maclura pomifera agglutinin (MPL) specifically stained the same cells as PNA and the population of invasive syncytial trophoblast cells remodelling maternal blood vessels and small maternal vessels at the materno-fetal interface, as well as trophoblast cells within necrotic zones at mid-gestation. Both lectins were positive for phagocytic cytotrophoblast cells lining the haematophagus organs. The results of this study demonstrate that lectin histochemistry is a useful tool for staining subpopulations of cytotrophoblast and syncytial trophoblast cells.
    Journal of reproduction and fertility. Supplement 02/2001; 57:199-206.
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    ABSTRACT: The aim of this study was to investigate the secretion patterns of FSH: (i) during the canine oestrous cycle with particular attention to the luteal period; and (ii) during 24 h in male and female dogs. Plasma FSH concentrations were measured by a highly specific homologous canine immunoradiometrical (IRMA) assay using monoclonal antibodies specific for canine FSH. In the first study, FSH concentrations were measured daily from +/- 15 days before until 150 days after the LH surge in groups of pregnant (n = 5) and nonpregnant (n = 5) Beagle bitches. Day 0 of the cycle was the day of the preovulatory LH peak as measured by LH radioimmunoassay. In the second study, FSH concentrations were measured at 1 h intervals for 24 h in five bitches at day 120 after the LH surge and in five males. From day 35 to day 40 after the LH peak, FSH concentrations were higher in pregnant than in nonpregnant luteal phases. A significant and abrupt decrease in FSH concentrations was observed at about the time of parturition (day 65) and was followed by lower FSH concentrations during lactation compared with nonpregnant bitches. FSH concentrations in nonpregnant animals were fairly constant from the end of oestrus to mid-anoestrus. In both post-lactation and anoestrous bitches, plasma concentrations of FSH increased consistently as anoestrus progressed. No significant differences in the mean hour-to-hour patterns of FSH secretion were observed over 24 h and no differences were detected between male and female dogs. However, a clear pulsatile pattern of secretion was observed in all individuals, both males and females, with an apparent 4.8 h interval between peaks. The results of this study demonstrate major differences in FSH secretion between pregnant and nonpregnant bitches. This finding indicates that there are differences in the regulation of the hypothalamo-pituitary-ovarian axis during pregnancy, possibly related to changes in activity of the corpus luteum. The results of this study also demonstrate a pulsatile pattern of FSH secretion in both male and female bitches, with a 4.8 h interval between peaks, and confirms the progressive increase in plasma FSH concentrations observed during anoestrus.
    Journal of reproduction and fertility. Supplement 02/2001; 57:15-21.
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    ABSTRACT: There have been few reports of artificial insemination using fresh feline semen because of difficulties in collecting semen. It has been shown previously that the number of spermatozoa required for fertilization of cats by intravaginal insemination and unilateral intrauterine horn insemination are 8.0 x 10(7) and 8.0 x 10(6), respectively. There have been no reports of intratubal insemination of cats. Therefore, a study was designed to determine the number of spermatozoa necessary to fertilize cats by intratubal insemination with fresh semen. Four male and 25 mixed breed female cats were used. Semen was collected using an artificial vagina. Insemination was performed by laparotomy 15-30 h after administration of 100 iu hCG on day 2 or 3 of spontaneous oestrus. Cats in which ovulation had not occurred at insemination received an additional 100 iu hCG after surgery. In intratubal insemination, four groups consisting of three, seven, eight and seven cats received 5.0 x 10(3), 5.0 x 10(5), 2.0 x 10(6) and 4.0 x 10(6) viable spermatozoa, respectively, into both uterine tubes at approximately 2 cm from the fimbriae tubae. The volume inseminated was 10-20 microliters. The semen volume was adjusted with egg yolk-Tris-fructose citrate. As a result of artificial insemination, none of the cats that received 5.0 x 10(3) or 5.0 x 10(5) spermatozoa were fertilized. However, fertilization was successful in two of eight animals (25.0%) that received 2.0 x 10(6) spermatozoa and in three of seven animals (42.9%) that received 4.0 x 10(6) spermatozoa. The number of kits was 1-4 (mean +/- SE: 2.4 x 0.6). These results indicate that although large numbers of spermatozoa were inseminated into the uterine tubes, the conception rate was low. The low rate of conception may have been due to problems in inducing capacitation of spermatozoa in the uterine tubes.
    Journal of reproduction and fertility. Supplement 02/2001; 57:347-51.
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    ABSTRACT: A new kit (ReproCHEK RELAXIN) intended for the diagnosis of pregnancy in bitches is now available for veterinary use. This assay measures relaxin concentrations in plasma and whole blood samples, and the presence of significant amounts of relaxin is indicative of pregnancy. A clinical trial was carried out to evaluate the performance of the test. Serial blood samples were collected on alternate days, and relaxin concentrations were determined from day 15 to day 35 after the LH surge (estimated by progesterone concentrations). Pregnancy was confirmed using ultrasonography. At the end of pregnancy, both the day of whelping and the size of the litter were recorded. Pregnancy was established in 61 bitches. The day that pregnancy was detected using the relaxin assay ranged from day 19 to day 28 after the LH surge and had a mean (+/- SD) of 25.4 +/- 2.5 days. The day of parturition was taken as a reference point, and pregnancy was detected from -46 to -38 days (mean -40.2 +/- 2.4 days) before parturition. False positives were not observed in pseudopregnant bitches (n = 16) or in the control group (30 anoestrous and ten unmated bitches). These results demonstrate that the new assay kit is an inexpensive, user-friendly and reliable technique for determining pregnancy.
    Journal of reproduction and fertility. Supplement 02/2001; 57:187-91.
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    ABSTRACT: Blood samples were collected from nine Beagle bitches every 6 h during pro-oestrus and oestrus to measure plasma concentrations of progesterone, oestradiol and LH. The number of ovarian follicles was estimated once a day using transcutaneous ultrasonography. According to the concentrations of plasma progesterone, the bitches were mated once and subsequently ovariohysterectomized 3-7 days after mating. The number of corpora lutea was counted, and oocytes and embryos were collected by flushing of the oviducts. In ovaries that had more than three follicles, the number of follicles observed using ultrasonography was underestimated, whereas the recovery rate (number of oocytes and embryos flushed/number of corpora lutea counted) was 99%. Embryos were processed for sectioning and the developmental stages were determined by counting the number of blastomeres under bright field microscopy. Potentially fertilized oocytes and a zygote were observed on day 7 after the LH peak and the developmental rate was about one cell cycle in 24 h until day 12 after the LH peak. The timing of embryonic development was significantly correlated with the time of the LH peak.
    Journal of reproduction and fertility. Supplement 02/2001; 57:181-6.
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    ABSTRACT: The results of two studies in which the pulsatile secretion patterns of LH and FSH were investigated in six Beagle bitches during early, mid- and late anoestrus and in six anoestrous Beagle bitches treated with bromocriptine are summarized to improve our knowledge of the endocrine changes that lead to a new follicular phase in bitches. Blood samples for the determination of secretory profiles were obtained via jugular venepuncture at 10 min intervals for 6 h. In untreated bitches, blood samples were collected during early, mid- and late anoestrus. In bromocriptine-treated bitches (20 micrograms kg-1 twice each day, starting 100 days after ovulation until the start of the next oestrous cycle), blood samples were collected before treatment and at 2 week intervals after the start of bromocriptine treatment until the next ovulation. In all bitches, FSH and LH were secreted in a pulsatile manner and FSH pulses coincided with LH pulses. Progression from early to late anoestrus was associated with an increase in FSH secretion without a concomitant increase in LH secretion. The bromocriptine-induced shortening of the interoestrous interval was also associated with an increase in FSH secretion without a concomitant increase in LH secretion. These results indicate that in bitches an increase in circulating FSH should be considered to be a critical event required for the initiation of ovarian folliculogenesis and consequently the termination of anoestrus.
    Journal of reproduction and fertility. Supplement 02/2001; 57:11-4.
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    ABSTRACT: The aim of the present study was to assess the progression of meiotic maturation and fertilization of in vitro matured blue fox (Alopex lagopus) oocytes at different time intervals after in vitro maturation and insemination by the use of confocal laser scanning microscopy. A total of 242 immature oocytes from ovarian follicles 1-2 mm in diameter from seven ovulating blue fox vixens at oestrus were cultured for 48 h in TCM-199 before in vitro insemination with 5.0 x 10(5) frozen-thawed fox spermatozoa. Oocytes were transferred to 50 microliters microdroplets of modified Tyrode's medium without glucose at pH 7.7 under mineral oil, inseminated and cultured at 38 degrees C in a humidified atmosphere with 5% CO2 for 2 (n = 10), 4 (n = 22), 8 (n = 41), 20 (n = 52), 24 (n = 10), 30 (n = 48) and 48 h (n = 59). The oocytes and zygotes were stained with propidium iodide and were analysed by confocal laser scanning microscopy or epifluorescence microscopy. Only 125 of 242 oocytes (52%) could be evaluated: of these germinal vesicle breakdown (GVBD) was observed in five of nine oocytes (56%) at 2 h, eight of 18 oocytes (44%) at 4 h, eight of 18 oocytes (44%) at 8 h, 27 of 42 oocytes (64%) at 20-24 h, 14 of 19 oocytes (74%) at 30 h, and 13 of 19 oocytes (68%) at 48 h after insemination. In total, 75 of 125 oocytes (60%) underwent GVBD. All stages from the germinal vesicle to the four-cell stage embryos were observed, but the rate of cleavage was low (9%). Immature oocytes collected from small subordinate ovarian follicles of oestrous vixens after ovulation of the dominant follicles were able to mature and be fertilized, as well as undergo the first two cleavage divisions in vitro.
    Journal of reproduction and fertility. Supplement 02/2001; 57:161-5.
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    ABSTRACT: Transcervical collection of endometrial tissue specimens is a valuable and well established tool for the diagnosis of reproductive disorders in cows, mares and women, but it is not used currently in bitches. Endometrial biopsy samples were collected by transcervical cannulation from the cranial uterine body at defined stages of the oestrous cycle using biopsy forceps. In study 1, the histology of 45 biopsy specimens taken from 23 Beagle bitches were compared with larger tissue samples collected post mortem. For further evaluation of representative biopsy specimens, the expression of oestrogen and progesterone receptors was determined. Only 31.1% of the biopsy samples taken from bitches could be evaluated and they showed stroma, capillaries, luminal epithelium, glandular ducts and apical glands; however, basal glands and myometrium were not observed. Biopsy findings were in agreement with the diagnosis of the uterine specimens concerning endometrial differentiation, inflammatory and degenerative lesions. The expression of oestrogen and progesterone receptors was slightly lower in the biopsy samples. In study 2, a total of 49 serial biopsy samples were taken from 12 Beagle bitches. In eight bitches, the manipulation resulted in a haemomucometra. Treatment with PGF2 alpha and antibiotics led to a clinical cure in five of these dogs, two of which were mated successfully during the next oestrus. Three animals had to undergo ovariohysterectomy. Owing to the small proportion of biopsy specimens that was suitable for analysis and the high risk of biopsy-related endometritis, transcervical biopsy collection cannot be recommended as a routine technique in bitches.
    Journal of reproduction and fertility. Supplement 02/2001; 57:61-5.
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    ABSTRACT: In vitro capacitated canine spermatozoa can interact with both mature and immature homologous oocytes in vitro, but it is difficult to determine whether spermatozoa have actually penetrated the oocyte or have simply bound to the zona pellucida. The aim of the first part of this study was to determine accurately the location of spermatozoa in relation to the oocyte by comparing observations made using confocal and fluorescence microscopy. The fluorescence technique had a sensitivity of 66%, and correctly identified 98% of the spermatozoa that had penetrated or bound oocytes and could discriminate between penetrated and bound spermatozoa with a sensitivity and specificity of 97% and 100%, respectively. This finding demonstrates that evaluation of the assay by fluorescence microscopy is reliable for detecting the presence and location of spermatozoa in relation to homologous oocytes. The aim of the second part of the study was to attempt to simplify the assay using fluorescence microscopy and immature oocytes. No significant difference was found between sperm interaction with mature and immature oocytes, which demonstrates that culturing oocytes before the assay has no benefit, and that the assay can be performed quickly and easily using non-cultured oocytes to provide rapid evidence of the fertilizing potential of spermatozoa.
    Journal of reproduction and fertility. Supplement 02/2001; 57:127-36.
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    ABSTRACT: The aim of this study was to determine whether the canine endometrium could be examined by hysteroscopy. Eight German shepherd bitches were used. Two bitches were in dioestrus, five were in anoestrus and one was about 8 weeks post partum. The uterus was exposed through a ventral celiotomy. A 3 mm straight laparoscope or a 4 mm laparoscope with a 30 degrees angle, and a catheter for inflation of the uterus were used. After hysteroscopy the bitches were spayed and the uterus of each bitch was examined macroscopically and histologically. Endometrium and uterotubal junctions were visible in all bitches. The cervix and caudal uterine body could not be visualized in three bitches as two bitches had uterine discharge and air escaped through the cervix in the other bitch. In the post-partum bitch, dark-brown sites of implantation were observed. Four bitches (aged 9, 22, 25 and 31 months) had endometrial cysts (0.5-2.0 mm in diameter). Hysteroscopy caused petechiae and ecchymosis in the endometrium of four bitches. Traumatization of the endometrium affected only the epithelium and the most superficial part of the lamina propria. In conclusion, hysteroscopy appears to be a very sensitive tool for evaluation of the canine endometrium. Further research is necessary to determine the effects of hysteroscopy on subsequent fertility.
    Journal of reproduction and fertility. Supplement 02/2001; 57:415-7.