Maternal high-fat diet during pregnancy and lactation affects factors that regulate cell proliferation and apoptosis in the testis of adult progeny.

Context A maternal high-fat diet is thought to pose a risk to spermatogenesis in the progeny. Aims We tested whether a maternal high-fat diet would affect Sertoli cell expression of transcription factors (insulin-like growth factor I (IGF-I); glial-cell line-derived neurotrophic factor (GDNF); Ets variant 5 (ETV5)) and cell proliferation and apoptotic proteins, in the testis of adult offspring. Methods Pregnant rats were fed ad libitum with a standard diet (Control) or a high-fat diet (HFat) throughout pregnancy and lactation. After weaning, male pups were fed the standard diet until postnatal day 160. Males were monitored daily from postnatal day 34 to determine onset of puberty. On postnatal day 160, their testes were processed for morphometry and immunohistochemistry. Key results The HFat diet increased seminiferous-tubule diameter (P P P P P P P P Conclusions A maternal high-fat diet alters the balance between spermatogonia proliferation and spermatid apoptosis. Implications A maternal high-fat diet seems to 'program' adult male fertility.

Equine chorionic gonadotropin administered on day 5 of a 7-days fixed-time artificial insemination program improves ovulation synchrony and corpus luteum function in anestrous beef cows.

In order to assess the effect of equine chorionic gonadotropin (eCG) administered on Day 5 or 7 of a fixed-time artificial insemination protocol (FTAI) in anestrous suckled beef cows, two experiments were performed to determine the following endpoints: Experiment 1 (n = 22), preovulatory follicle (POF) diameter, ovulation time, corpus luteum (CL) area, estradiol (E2) and progesterone (P4) concentrations; and Experiment 2 (n = 676), a field trial to evaluate conception rate using the same experimental design. In both experiments, a synchronization protocol using estradiol benzoate (EB) (Day 0), intravaginal progestin device (IVD) (Days 0 through 7), prostaglandin (PGF) (Day 7), eCG (Day 5 or 7), and GnRH (Day 9). Treatment consisted of administering 400 IU of eCG on Day 5 (T5) or Day 7 (T7 or control) concomitant with treatment with PGF2α. In experiment 1, all cows of T5 ovulated by 16 h after GnRH administration. The POF tended (P = 0.06; P = 0.07) to be larger at 1 and 2 d before ovulation in T5. The day before ovulation, E2 tended to be lower (P = 0.06) in T5 compared with T7. The CL area and the P4 concentrations were greater (P = 0.04) on day 9 in T5 compared with T7. In experiment 2, the conception rate was greater (P = 0.04) in T5 (72.2%) compared with T7 (61.0%) group. Therefore, administration of eCG on Day 5 of the designed protocol hastened ovulation of a greater follicle, which produced a larger CL and greater concentrations of progesterone by Day 9 after ovulation, resulting in 11.2% increase in cows pregnant.

Maternal undernutrition during pregnancy and lactation increases transcription factors, ETV5 and GDNF, and alters regulation of apoptosis and heat shock proteins in the testis of adult offspring in the rat.

We tested whether changes in Sertoli cell transcription factors and germ cell heat shock proteins (HSPs) are linked to the effects of maternal undernutrition on male offspring fertility. Rats were fed ad libitum with a standard diet (CONTROL) throughout pregnancy and lactation or with 50% of CONTROL intake throughout pregnancy (UNP) or lactation (UNL) or both periods (UNPL). After postnatal Day 21, 10 male pups per group were fed a standard diet ad libitum until postnatal Day 160 when testes were processed for histological, mRNA and immunohistochemical analyses. Compared with CONTROL: caspase-3 was increased in UNP and UNPL (P=0.001); Bax was increased in UNL (P=0.002); Bcl-2 (P<0.0001) was increased in all underfed groups; glial cell line-derived neurotrophic factor (P=0.002) was increased in UNP and UNL; E twenty-six transformation variant gene 5 and HSP70 were increased, and HSP90 was diminished in all underfed groups (P<0.0001). It appears that maternal undernutrition during pregnancy and lactation disrupts the balance between proliferation and apoptosis in germ cells, increasing germ cell production and perhaps exceeding the support capacity of the Sertoli cells. Moreover, fertility could be further compromised by changes in meiosis and spermiogenesis mediated by germ cell HSP90 and HSP70.

Equine early pregnancy endocrine profiles and ipsilateral endometrial immune cell, gene expression and protein localisation response.

We investigated the early effects of the equine embryo on maternal serum concentrations of insulin-like growth factor 1 (IGF1), leptin and adiponectin, uterine immune cells and genes and proteins related to embryo development and the maintenance of pregnancy. Ipsilateral endometrial expression was assessed on Days 7 and 13 after ovulation for the following transcripts: oestrogen receptor ERα (ESR1), progesterone receptor (PGR), progestin and adipoQ receptor family member 5 (PAQR5), oxytocin receptor (OXTR), prostaglandin-endoperoxide synthase 2 (PTGS2), raf-1 proto-oncogene serine/threonine kinase (RAF1), p21-activated kinase 6 (PAK6), fibroblast growth factor family member 9 (FGF9), IGF1 and its receptor (IGF1R), mucin 1 (MUC1), osteopontin (OPN), leptin receptor (LEPR) and adiponectin receptors 1 and 2 (ADIPOR1 and ADIPOR2). Ipsilateral endometrial immunological cell infiltration and immunohistochemical protein localisation were evaluated on Days 7, 10 and 13 after ovulation for ERα, PGR, OXTR, PTGS2, IGF1, IGF1R, IGF2 and MUC1. Serum hormone concentrations were not affected by reproductive status. Pregnancy downregulated ESR1 and PGR mRNA levels, upregulated the expression of all other genes and affected the expression of all genes, except PGR, on Day 7 (compared with eight genes affected at Day 13). Proteins were affected by pregnancy or by its interaction with other variables (day of extraction and endometrial compartment). Pregnant mares had a higher lymphocyte count, which decreased towards Day 13. The effect of pregnancy on leucocytes and proteins was more evident in superficial endometrial compartments. The results of this study suggest that the equine embryo exerts prompt paracrine regulation of critical biological processes.

Heat shock protein HSP90 immunoexpression in equine endometrium during oestrus, dioestrus and anoestrus.

Heat shock proteins play a crucial role in cellular development, proliferation, differentiation and apoptosis. Heat shock protein 90 (HSP90) has been localised in the human endometrium, where its immunoexpression changes during the menstrual cycle. Similar studies have not been done for the equid species, so the present study aimed to describe endometrial HSP90 immunoexpression in mare endometrium. Endometrial biopsies were formalin-fixed and paraffin-embedded, and sections were stained with haematoxylin-eosin in preparation for HSP90 immunohistochemistry. Immunostaining and morphometric analyses were performed on the epithelial lining, endometrial glands and connective stroma during oestrus, dioestrus phase and anoestrus period (n = 7 per phase or period). Immunoexpression was localised in the basal region of the epithelial cells lining the lumen. Immunoexpression was greater during oestrus than during either dioestrus or anoestrus. During anoestrus, there was little immunostaining in the endometrium, suggesting that HSP90 is involved in the functional modulation of sex steroid receptors in cyclic mares. Indeed, the function of HSP90 as a chaperone in the folding of proteins, such as steroid receptors, might explain the greater intensity of immunostaining during the oestrus and dioestrus phases, compared the anoestrus period. We conclude that, in the mare, HSP90 plays a role in endometrial function and that further studies are needed to test whether it is important in pathological conditions as endometritis.

Maternal undernutrition during pregnancy and lactation affects testicular morphology, the stages of spermatogenic cycle, and the testicular IGF-I system in adult offspring.

Maternal undernutrition decreases sperm production in male offspring, possibly through insulin-like growth factor (IGF-I). To test this hypothesis, we fed pregnant Wistar rats ad libitum with a standard diet (CONTROL) or fed 50% of CONTROL intake, either throughout pregnancy (UNP), lactation (UNL, or both (UNPL). After weaning, male offspring (n = 10 per treatment) were fed a standard diet until postnatal day 160, when testes process for histological and molecular analyses. IGF-I immunostaining area and intensity in the testis were greater (P = 0.003) in the UNPL group compared to CONTROL, but lower in the UNP group (P < 0.0001). Levels of IGF-I receptor transcript were lower in the UNPL and UNL groups, compared to CONTROL. There were more Ki-67-positive germ and Sertoli cells, in all underfed groups than in CONTROL. Compared to CONTROL, frequency of spermatogenic cycle stage VII was lower in all underfed groups, and seminiferous tubule diameter was smaller in UNP and UNPL. Plasma FSH concentrations were greater in UNP male offspring compared to all groups (P = 0.05), whereas inhibin B concentrations were greater in UNP (P = 0.01) and UNL (P = 0.003) than in CONTROL or UNPL. Thus, prenatal undernutrition leads to a decrease in testicular IGF-I levels, whereas of pre- and postnatal undernutrition increased testicular IGF-I levels and decreased amounts of IGF-I receptor mRNA in adult offspring. We conclude that maternal undernutrition during pregnancy and lactation leads to long-lasting effects on adult male offspring testicular morphology, spermatogenesis, and IGF-I testicular system.

Lying Postures of Dairy Cows in Cubicles and on Pasture.

Cows housed indoors with cubicles are probably more restricted in their choice of lying posture and orientation compared with cows housed on pasture. We therefore compared lying postures on pasture in Uruguay and the Netherlands with lying postures in cubicles in the Netherlands, also recording orientation on pasture in Uruguay and divider and bedding type in Dutch cubicles. We visited one farm with four herds in Uruguay, doing live observations, and 25 Dutch farms, taking pictures of cows. Observations of 205 cows on pasture in Uruguay showed more long postures, lying on their belly with their neck stretched. Two herds preferred lying towards north and south, while one herd preferred west and east. Pictures of 217 cows on pasture in the Netherlands showed more wide postures (lying on the side with three or four legs stretched out). Pictures of 527 cows in cubicles in the Netherlands showed more narrow postures (lying on the side with hind legs folded). More long postures (lying on the belly with a stretched neck) and less short postures (lying with the head folded back) were seen in cubicles with soft floors and English dividers; more narrow postures were seen in cubicles with concrete floors. Wide postures were seen more in cubicles with mattresses and free-hanging dividers. We conclude that since cows in cubicles show more narrow postures than on pasture and cannot choose their orientation, their choice in showing preferred behavior is restricted. More research is needed to study the consequences of restricted choice in lying behavior on the health and welfare of dairy cows.

Effect of side of the corpus luteum and pregnancy on estrogen and progesterone receptor expression and localization in the endometrium of mares.

The effect of side of corpus luteum on uterine gene expression and protein localization of estrogen receptor α (ERα) and progesterone receptor (PR) in healthy cyclic and pregnant mares 13 days after ovulation (day 0) was investigated. Transcervical biopsies were performed to collect endometrium ipsilateral and contralateral regarding the side of corpus luteum on day 13 post-ovulation in cyclic (n = 6) and pregnant (n = 6) mares. Blood samples were collected daily from day 0 until the day of biopsy for 17ß-estradiol (E2) and progesterone (P4) determinations. Receptor expression was determined by immunohistochemistry and transcript expression by real time RT-PCR. Serum E2 and P4 concentrations were not affected by reproductive status. The contralateral horn presented higher percentage of positive cells for ERα than the ipsilateral horn (P < .05), but side did not affect PR. ERα showed low staining and no main effect of pregnancy was found, but pregnant mares had lower protein expression of PR (19.8 vs. 40.4 ±â€¯5.3%, P < .01). The contralateral horn tended to present higher expression of ERα mRNA (1.33 vs. 0.97 ±â€¯0.17, P < .10) and PR mRNA (1.96 vs. 1.57 ±â€¯0.52, P < .09). ERα mRNA relative expression was lower in the pregnant group (0.88 vs. 1.44 ±â€¯0.19, P < .05). The interaction of reproductive status and side of corpus luteum tended to affect PR mRNA expression as pregnant mares had a lower PR mRNA content in the ipsilateral horn than cyclic mares. To our knowledge, this is the first study that describes the behavior of steroid receptors in the endometrium of mares regarding side of corpus luteum.

Effect of cow parity and synchronization method with PGF2α on conception rates of Bos indicus cows in Cameroon.

The objective of this work was to evaluate the effect of two synchronization methods with prostaglandins F2α (PGF2α) on heifers and multiparous cows. Fourty-three Bos indicus cows (white and Red Fulani) were divided into four groups in a two-by-two factorial structure, parity x method of synchronization. The synchronization methods consisted of a two-dose regime which involved injection of animals on day 0 with PGF2α (Lutalyse) at 5 ml per cow intramuscularly. On day 11, the injection was repeated at the same dosage. On day 14 (72 h after the second injection), a fixed-time artificial insemination (AI) was done. On day 15 (96 h after the second injection), a second insemination was done. The one-and-a-half-dose regime consisted of an injection similar to the first treatment mentioned above on day 0. Thereafter, cows were observed for heat, and anyone showing heat was inseminated. A second dose was given on day 11 to all animals not having shown any heat. A fixed-time AI was done on days 14 and 15. Blood samples were collected on the day 0 of insemination for each cow while day 11 and day 21 after insemination. Progesterone was analysed by means of standard ELISA progesterone kits to determine its profiles after insemination. Results show no evidence of the effect of treatments on conception rates (P > 0.05). Similarly, heifers and multiparous cows had similar conception rates (P > 0.05). Between 3 weeks and 3 months of pregnancy, there was a loss of embryos of 28% in heifers and 20% in multiparous cows, but the difference between the two groups was not significant (P > 0.05). It recommended that farmers do not synchronize animals with poor body condition score (BCS). They should also monitor weight gains of heifers, remove them from the herd when they have been mixed with young growing bulls and put them in a breeding herd. The two-dose regime is better to be used in areas where the inseminator cannot easily be available.

Effect of pre-partum feed supplementation on post-partum ovarian activity, milk production and calf growth of small holder dairy Cattle in Cameroon.

Seventy-two cows were selected for an on-farm study on the effect of feed supplementation before calving on milk production, ovarian activity and calf growth of Holstein, indigenous Red Fulani cows and their crosses. Pre-partum feed supplementation was done using cotton seed cake (80%), maize (18%), bone meal (1%) and kitchen salt (1% NaCl). Supplementation levels consisted of a low supplementation fed at 1 kg per animal per day and high supplementation fed at 2 kg per animal per day. In addition, Red Fulani cows received the supplements in two different ways namely a pre-partum supplementation consisting of 1 kg per cow per day and pre- and post-partum supplementation consisting of 1 kg per cow per day before calving and 1 kg per cow per day post-partum up to 30 days after calving. Blood samples were analysed using ELISA Progesterone kits to determine the length of post-partum anoestrus. Results show that pre-partum levels of feeding did not have any effect (P > 0.05) on body condition score (BCS) at 12 weeks after calving, calf birth weight, average daily weight gain of calves, milk production and post-partum anoestrus. High BCS at calving was shown to influence BCS at 12 weeks of lactation. Holstein cows had bigger calves (P < 0.01) at birth (45 kg) compared to traditional cows (36 kg) and crosses (34 kg). There was little benefit of pre-partum supplementation on the parameters investigated in this study. Consequently, low income farmers are advised to concentrate their efforts of supplementation early in lactation.

Comparison between two progesterone sources and two oestradiol formulations in a Heatsynch protocol for postpartum cycling dairy cows in pasture.

To compare an injectable progesterone (MAD-4) with an intravaginal device (IPD), and natural O17 with synthetic oestradiol (OB) in a synchronisation protocol, 51 cows were divided into four groups. Each group was treated with one of the two sources of progesterone and one of the two oestradiol formulations. Oestrus behaviour, follicle diameter, and pregnancy rates were evaluated. Oestrus behaviour (p = 0.902), numbers of cows in oestrus (p = 0.917), follicle diameter (p = 0.416), and pregnancy rates (p = 0.873) were similar among the four groups. More cows in the group treated with the IPD and OB scored > 200 oestrus behaviour points compared to the other groups (p = 0.038). A longer interval between the end of treatment and oestrus was observed among cows treated with MAD-4 than cows given the IPD (p = 0.030), but no differences were found between animals receiving the two oestradiol formulations (OB and O17). While the use of MAD-4 requires further testing, similar responses to natural oestradiol observed in the present study could allow the use of this formulation in reproductive protocols because it is not associated with the potential human health risks of OB.