Vascular Endothelial (VEGF) and Epithelial Growth Factor (EGF) as Well as Platelet‐Activating Factor (PAF) and Receptors are Expressed in the Early Pregnant Canine Uterus

The aim of this study was to investigate the course of expression of platelet‐activating factor (PAF), PAF‐receptor (PAF‐R), epidermal growth factor (EGF), EGF‐R, vascular endothelial growth factor (VEGF), VEGF‐R1 and VEGF‐R2 in uterine tissue during canine pregnancy. For this purpose, 20 bitches were ovariohysterectomized at days 10–12 (n = 10), 18–25 (n = 5) and 28–45 (n = 5) days after mating, respectively. The pre‐implantation group was proven pregnant by embryo flushing of the uterus after the operation, the others by sonography. Five embryo negative, that is, non‐pregnant, bitches in diestrus (day 10–12) served as controls. Tissue samples from the uterus (placentation sites and horn width, respectively) were excised and snap‐frozen in liquid nitrogen after embedding in Tissue Tec^®. Extraction of mRNA for RT‐PCR was performed with Tri‐Reagent. In the embryos, mRNA from all factors except VEGF was detected. In the course of pregnancy, significantly higher expression of PAF and PAFR as well as VEGF and VEGFR2 during the pre‐implantation stage than in all other stages and a strong upregulation of EGF during implantation were characteristic. The course of EGF was in diametrical opposition to the course of the receptor. These results point towards an increased demand for VEGF, EGF and PAF during the earliest stages of canine pregnancy.     

Evaluation of the sustainability of participatory management of forest plantations: the case study of Wari-Maro Forest Reserve,Republic of Benin§

This study assessed ecological and socio-economic impacts of a participatory forest management project in the Republic of Benin. The study focused on the Wari-Maro Forest Reserve and the ‘Projet d’Aménagement des Massifs Forestiers’ five years after its completion. A forest inventory was carried out using 37 square plots of 729 m² each to characterise the population structure of two types of plantations established: plantations with exotic species and plantations with native species. In addition, individual surveys were conducted with local households, organs of joint forest management and forestry officers to evaluate their perceptions about the participatory management of the plantations. Finally, the sustainability of the participatory management was assessed with an established rating system. Results showed that plantations with exotic species were more successful than plantations with native species. Local communities argued that they have not been involved in the plantations design but only in the implementation step and that their standards of living have decreased after the project completion. The rating system used showed that the participatory management of plantations had a short-term sustainability. The findings suggest that future projects should be designed and implemented with better participation of local communities as full partners.     

In vitro differentiation of bovine bone marrow‐derived mesenchymal stem cells into male germ cells by exposure to exogenous bioactive factors

Mesenchymal stem cells (MSC) are multipotent progenitor cells defined by their ability to self‐renew and give rise to differentiated progeny. Previous studies have reported that MSC may be induced in vitro to develop into different types of specialized cells including male gametes. In vitro gamete derivation technology has potential applications as an alternative method for dissemination of elite animal genetics, production of transgenic animals and conservation of endangered species. This study aimed at investigating the in vitro effect of BMP4, TGFβ1 and RA on the potential for germ cell (GC) differentiation of bovine foetal MSC (bfMSC) derived from bone marrow (BM). The effect of BMP4, TGFβ1 and RA was analysed on the expression of pluripotent, GC and male GC markers on bfMSC during a 21‐day culture period. bfMSC cultured under in vitro conditions expressed OCT4, NANOG and DAZL, but lacked expression of mRNA of VASA, STELLA, FRAGILIS, STRA8 and PIWIL2. Treatment with exogenous BMP4 and TGFβ1 induced a transient increase (p < .05) in DAZL and NANOG mRNA levels, respectively. However, exposure to RA was more effective in increasing (p < .05) expression of DAZL and regulating expression of OCT4 and mRNA levels of NANOG. These data suggest that bfMSC may possess potential for early GC differentiation, where OCT4, NANOG and specially DAZL may play significant roles in controlling progression along the GC lineage.     

Intake of trichostrongylid larvae by goats and sheep grazing together

SUMMARY Cashmere goats and Merino sheep were grazed together at 7.5 animals per ha on annual rye grass and clover pasture in southern Victoria, a winter rainfall area. Intake of parasitic larvae was measured in oesophageal extrusa samples collected from 2 animals of each species, 4 times in one week, on 7 occasions between mid-March (autumn) and mid-June (winter). Pasture contamination with larvae was measured at the same times. The number of larvae per kg of green grass was lower than on green clover; the most heavily contaminated portion of the pasture was the mat of dead herbage on the ground. The diet selected by goats contained more green grass and dead herbage and less clover that that of sheep (P < 0.01). Goats ingested 643 infective trichostrongylid larvae per kg dry matter intake (DMI) versus 274 per kg DMI for sheep in autumn, increasing to 1892 versus 1143 in early winter. The heavier trichostrongylid burdens of goats compared with sheep, when grazed together, are due in part to greater rates of infection consequent on different grazing patterns as well as greater susceptibility to infection.     

Effects of increased dietary energy and protein during late gestation on mammary development in gilts

Thirty-two gilts were used to evaluate the effects of increased dietary energy and CP during late gestation on mammary development. On d 75 of gestation, gilts were assigned randomly in a 2 x 2 factorial arrangement to adequate (5.76 Mcal ME/d) or increased (10.5 Mcal ME/d) energy and adequate (216 g CP/d) or increased (330 g CP/d) protein. On d 105 of gestation, gilts were slaughtered and total mastectomies were performed. Mammary tissue was separated into mammary parenchymal and mammary extraparenchymal stromal tissue and analyzed for DNA, RNA, protein and lipid. No interactions between dietary energy and protein level were detected (P greater than .20). When adjusted for number of mammary glands and maternal BW (weight of the sow less the weight of the fetuses), mammary parenchymal weight was 27% greater (P less than .03) in gilts fed adequate energy than in gilts fed increased energy, but mammary extraparenchymal stroma weight was unaffected by dietary energy level. Total mammary parenchymal DNA was 30% greater in gilts fed adequate energy than in gilts fed increased energy (P less than .03). Total mammary parenchymal RNA (P less than .02) and total mammary parenchymal protein (P less than .02) also were greater in gilts fed adequate energy than in gilts fed increased energy. Dietary protein level did not affect mammary variables measured, except that increased dietary protein tended to reduce mammary extraparenchymal stromal weight (P less than .09). Increased dietary protein between d 75 and d 105 of gestation did not benefit mammary development, but increased dietary energy was detrimental to development of mammary secretory tissue.     

Effects of immunizing gilts against zearalenone on height of vaginal epithelium and urinary excretion of zearalenone

Two experiments were conducted to develop a vaginal epithelium bioassay for zearalenone (Z) and to determine whether immunization against Z would prevent Z mycotoxicosis. Eleven gilts were ovariectomized and allotted by weight to dietary doses of 50, 150 or 350 micrograms Z/kg BW daily for 3 d. All doses of Z increased height of the vaginal epithelium. Height of the vaginal epithelium in gilts fed 150 or 350 micrograms Z/kg BW increased more than that in gilts fed 50 micrograms Z/kg BW. Twenty-four gilts then were ovariectomized and allotted to be immunized or not immunized. A Z-bovine serum albumin conjugate was injected into gilts to achieve immunization. Ten weeks after initial immunization, antibodies to Z were detected after a 1:10(7) dilution at greater than .1 absorbance units using an enzyme-linked immunosorbent assay, and gilts were allotted by weight to diets with no Z or 150 micrograms Z/kg BW daily for 3 d. Immunization alone had no effect on height of vaginal epithelium, but after 3 and 10 d, immunized gilts fed Z had higher vaginal epithelium than did nonimmunized gilts fed Z. Immunized gilts excreted a larger percentage of ingested Z than nonimmunized gilts did. Therefore, immunizing gilts against Z potentiated both the estrogenic effects of Z and urinary excretion of Z equivalents.     

Expression of Genes in the Canine Pre‐implantation Uterus and Embryo: Implications for an Active Role of the Embryo Before and During Invasion

The aim of the present study was to assess genes expressed in maternal uterine tissue and pre‐implantation embryos which are presumably involved in maternal recognition and establishment of canine pregnancy. For this purpose, 10 pregnant bitches were ovariohysterectomized between days 10 and 12 after mating. Four non‐pregnant bitches served as controls. Early pregnancy was verified by flushing the uterine horns with PBS solution. The collected embryos (n = 60) were stored deep‐frozen (?80°C). Uterine tissue was excised, snaps frozen in liquid nitrogen and homogenized using TRI Reagent. All embryos from one litter were thawed together and also homogenized in TRI Reagent. RT‐PCR was performed to prove mRNA expression of progesterone receptor, key enzymes of the prostaglandin synthesis pathway, selected growth factors, cytokines, immune cell receptors, major histocompatibility complex (MHC) and matrix‐metalloproteinases (MMP). Only pregnant uteri revealed the presence of mRNA for interferon (IFN)‐γ, IL‐4 and CD‐8, which resembles the milieu in humans and other mammalians. Similarly, in day 10 embryos, mRNA for transforming growth factor‐β, insulin‐like growth factor‐1,‐2, hepatocyte growth factor, leukaemia inhibitor factor, tumour necrosis factor‐α, interleukin‐1β,‐6,‐8, cyclooxygenase‐2, CD4⁺ cells, and MMP‐2 and ‐9 were detected, but not MHC‐I or ‐II. We therefore suppose that the canine embryo, like its human counterpart, actively initiates measures to prevent attacks from the maternal immune system to prepare its own adhesion, nidation, growth and further development.     

Lactoferrin increases sperm membrane functionality of frozen equine semen

During cryopreservation, sperm was submitted to an increase in reactive oxygen species generation. This work aimed to improve the quality of frozen equine sperm after the addition of antioxidants lactoferrin (Lf) and catalase (Cat) to a freezing extender. Semen from six stallions was frozen with the extenders: F1) control, INRA 82 freezing extender, F2) F1 + 500 μg/ml Lf and F3) F1 + 200 IU/ml Cat. After thawing, sperm motility parameters, membrane functionality and integrity, and acrosome integrity and spontaneous acrosome‐reacted sperm were evaluated with a computer‐assisted sperm analysis, a hypoosmotic swelling test and epifluorescent microscopy, respectively. Nitrite, hydroperoxide and iron concentrations of frozen semen were measured with spectrophotometry. The percentage of functional membrane sperm treated with Lf was higher (50.7% ± 11.6%) compared to that of the control (37.6% ± 15.6%), while the iron (61.4 ± 11.6 vs 73.3 ± 13.8 mg/dl) and nitrite concentrations (16.3 ± 7.1 vs 25.9 ± 4.2 μM/μg protein) were lower, respectively (p < .05). Thus, it can be suggested that Lf protect stallion spermatozoon during freezing as it has increased the percentage of sperm with functional membrane and decreased the lipid oxidant agents.