Expression Pattern of Vascular Endothelial Growth Factor in Canine Folliculogenesis and its Effect on the Growth and Development of Follicles after Ovarian Organ Culture

In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen‐thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs.     

Impact of Food Restriction on Ovarian Development,RFamide‐Related Peptide‐3 and the Hypothalamic–Pituitary–Ovarian Axis in Pre‐Pubertal Ewes

RFamide‐related peptide‐3 (RFRP‐3), the mammalian ortholog of gonadotropin‐inhibiting hormone, has been implicated as a mediator between reproduction and energy balance. This study aimed to investigate the physiological effects of RFRP‐3 on the process of ovarian development in food‐restricted pre‐pubertal ewes. The results showed that food restriction significantly inhibited the ovarian development and follicular growth. The data of qPCR in the hypothalamic–pituitary–ovarian (HPO) axis showed that food restriction not only upregulated RFRP‐3 mRNA expression but also downregulated the mRNA expression of gonadotropin‐releasing‐hormone receptor, follicle‐stimulating hormone receptor and luteinizing hormone receptor (LHR). Immunohistochemistry of RFRP‐3 in the ovaries suggested that RFRP‐3 may regulate the follicular development. These results suggested that the changes of RFRP‐3 in response to food restriction might influence the HPO axis and inhibit ovarian development.     

Cryopreservation of Ram Semen in Extenders Containing Soybean Lecithin as Cryoprotectant and Hyaluronic Acid as Antioxidant

A soybean lecithin‐based extender supplemented with hyaluronic acid (HA) was assayed for effectiveness to improve the quality of frozen–thawed ram semen. HA has not been tested yet in an extender containing soybean lecithin for freezing ram semen. Thus, the aim of this study was to analyse the effects of soybean lecithin at 1% or 1.5% along with HA at 0, 0.5 and 1 mg ml^‐1 in a Tris‐based extender on the motion characteristics, membrane integrity (HOST), viability, GSH peroxidase (GSH‐PX) activity, lipid peroxidation and acrosomal status after freezing–thawing. Semen was collected from four Mehraban rams during the breeding season and frozen in the six lecithin×HA extenders. The extender containing 1.5% lecithin supplemented with no HA yielded higher total motility (52.5%±1.6), viability (55.8%±1.6) and membrane integrity (44.5%±1.7), but the effects of the lecithin concentration did not reach signification. Linearity‐related parameters, ALH, BCF, lipid peroxidation, GSH‐PX activity, morphology and acrosomal status were not affected by the extender composition. In general, adding HA significantly decreased sperm velocity (1 mg ml^‐1 HA), total motility (only with 1.5% lecithin), viability (1 mg ml^‐1 HA for 1% lecithin; both concentrations for 1.5% lecithin) and membrane integrity. In conclusion, adding HA to the freezing extender supplemented with soybean lecithin failed to improve quality‐related variables in ram semen. Increasing the lecithin content could have a positive effect, but further studies are needed.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

MALDI‐MS Lipid Profiles of Oocytes Recovered by Ovum Pickup from Bos indicus and 1/2 indicus × taurus with High vs Low Oocyte Yields

The aim of the present study was to compare the lipid profile in oocytes of indicus and 1/2 indicus × taurus cows with high and low antral follicle count (AFC)/oocyte yields. After an OPU procedure (D0), antral follicles ≥3 mm were counted by ultrasonography (D4, 19, 34, 49, 64), and cows were assigned to groups with either high AFC (≥30 follicles; indicus, NH group; 1/2 indicus × taurus, AH group) or low AFC (≤15 antral follicles; indicus, NL group; 1/2 indicus × taurus, AL group). The lipid profiles of the oocytes were determined by MALDI‐MS. For GI, GII and GIII oocytes, the indicus samples tend to cluster separately from the 1/2 indicus × taurus samples. The lipid species [PC (P‐38:5) + H]⁺ and/or [PC (P‐36:2) + Na]⁺, [PC (38:2) + H]⁺, [PC (38:5) + Na]⁺ and [TAG (60:8) + NH⁴]⁺ were more abundant in indicus (NH and NL groups) than 1/2 indicus × taurus. The higher lipid content in the indicus oocytes likely reflects differences in the rate of lipid metabolism and may contribute to oocyte competence and embryo development.     

Effect of bovine colostrum intake on growth,reproductive parameters and survival in red kids

The aim of this study is to evaluate the efficacy of frozen Azawak colostrum supplementation on body weight (BW), average daily gain (ADG), reproductive parameters (mean age at first parturition, fertility, fecundity, prolificacy) and mortality rate among red kids. The study was conducted at the goat farm secondary centre of Maradi in Niger from September 2010 to September 2011. The control animals (n = 20) were left with their mother, while the treatment animals (n = 20) received in addition 50 ml/animal/day of bovine colostrum at birth and 15 ml/animal/day from d2 to d15. Weight was measured weekly from birth to d365. Mortalities were also recorded over the same period. For reproductive parameters, observations began at weaning (d197). Growth rate was higher (p < 0.001) in supplemented animal, and the treatment effects on ADG were observed up to 150 day after the end of supplementation. A similar long‐lasting trend was also observed in relation to the mortality rate (25% for ColG vs. 55% for ConG; p = 0.05). The age at first kidding tended to be lower in the treated group (13.8 ± 0.7 vs. 14.1 ± 0.8 month; p < 0.1). In conclusion, mild bovine colostrum supplementation induces a long‐lasting positive impact on growth rate and to a lower extent on reproduction parameters and survival rate.     

Grazing behaviour,physical activity and metabolic profile of two Holstein strains in an organic grazing system

The challenge for sustainable organic dairy farming is identification of cows that are well adapted to forage‐based production systems. Therefore, the aim of this study was to compare the grazing behaviour, physical activity and metabolic profile of two different Holstein strains kept in an organic grazing system without concentrate supplementation. Twelve Swiss (H_CH; 566 kg body weight (BW) and 12 New Zealand Holstein‐Friesian (H_NZ; 530 kg BW) cows in mid‐lactation were kept in a rotational grazing system. After an adaptation period, the milk yield, nutrient intake, physical activity and grazing behaviour were recorded for each cow for 7 days. On three consecutive days, blood was sampled at 07:00, 12:00 and 17:00 h from each cow by jugular vein puncture. Data were analysed using linear mixed models. No differences were found in milk yield, but milk fat (3.69 vs. 4.05%, P = 0.05) and milk protein percentage (2.92 vs. 3.20%, P < 0.01) were lower in H_CH than in H_NZ cows. Herbage intake did not differ between strains, but organic matter digestibility was greater (P = 0.01) in H_CH compared to H_NZ cows. The H_CH cows spent less (P = 0.04) time ruminating (439 vs. 469 min/day) and had a lower (P = 0.02) number of ruminating boli when compared to the H_NZ cows. The time spent eating and physical activity did not differ between strains. Concentrations of IGF‐1 and T3 were lower (P ≤ 0.05) in H_CH than H_NZ cows. In conclusion, H_CH cows were not able to increase dry matter intake in order to express their full genetic potential for milk production when kept in an organic grazing system without concentrate supplementation. On the other hand, H_NZ cows seem to compensate for the reduced nutrient availability better than H_CH cows but could not use that advantage for increased production efficiency.     

Use of NaHCO3 and MgO as additives for sheep fed only pasture for a restricted period of time per day: effects on intake,digestion and the rumen environment

Effects of NaHCO₃ and MgO buffer addition on intake and digestive utilization of a pasture were studied in wethers allowed a restricted time of access to forage. Twelve wethers housed in metabolic cages and fed fresh forage (predominantly Lotus corniculatus) ad libitum for 6 h/d were randomly assigned to one of the following treatments: a control forage without buffer (C) or a forage plus buffer composed of a mixture of 750 g/kg NaHCO₃ and 250 g/kg MgO at 20 g/kg dry matter intake (B). Feeding behaviour, feed and water intake and digestibility, urine output, Na urine elimination, kinetics of passage, ruminal pH and ammonia concentration, N balance and ruminal microbial N synthesis were determined in vivo, and the ruminal liquor activity was evaluated in vitro by fermentation of wheat straw. Addition of buffer increased total water intake (p = 0.05), Na urinary output (p = 0.01), purine derivative excretion in urine (p = 0.05) and tended to decrease mean total retention time in the digestive tract (p = 0.09). However, buffer addition increased ruminal pH (p < 0.001) and tended to decrease the ammonia concentration (p = 0.05). That use of buffer decreased ruminal activity was evidenced by a lower volume of gas produced in vitro (p = 0.01) possibly due to a lower microbial concentration in rumen liquor. The higher rumen dilution rate, likely due to a higher water intake, seems to have been the key driver of the actions of buffer supplementation on the rumen environment. Moreover, addition of NaHCO₃ led to an increased urinary Na excretion, which should be considered due to its likely negative environmental impacts.     

Effect of feeding level on ileal and total tract digestibility of nutrients and energy from soybean meal‐based diets for piglets

A total of 36 piglets with an initial body weight (BW) of 5.6 ± 0.7 kg, fitted with simple T‐cannulas at the distal ileum, were used to evaluate the effect of three graded feeding levels (50, 75 or 100 g/kg BW^0.75 day) on apparent ileal digestibility (AID) and total tract digestibility (ATTD) of dry matter (DM), nitrogen (N) and energy, and on ATTD of organic matter (OM), ether extracts (EE), neutral detergent fibre (NDF), acid detergent fibre (ADF) and digestible (DE), metabolisable (ME) and net energy (NE) content in soybean meal (SBM)–casein–cornstarch‐based diets. The AID of DM, N and energy and ATTD of NDF, ADF and EE in the diets were not affected (p > 0.05) by the feed intake (FI) level. There was a small decrease in ATTD of DM, N (CP), OM, ash and energy, and in DE, ME and NE content in the diets (p < 0.05) with increasing FI level. The net disappearance in the large intestine (in % of ileal recovery) decreased for DM, N and energy (p < 0.05) with increasing FI level. The design of the study allowed for estimating ileal endogenous loss of N and total tract endogenous loss of ash, N and EE, for estimating corresponding true ileal and total tract digestibility values, and for estimating urinary endogenous N loss. High variability in estimates of ileal endogenous N loss and total tract endogenous losses of N, EE and ash reflects great variation in individual endogenous losses between animals. Estimation of true total tract digestibility of N, EE and ash by regression analysis was affected by their decrease in ATTD with increasing FI level, as estimates for true digestibility were lower compared to their apparent values. The present results suggest that FI level can affect both apparent and true total tract nutrient digestibility in piglets.     

Effects of lactulose supplementation on performance,blood profiles,excreta microbial shedding of Lactobacillus and Escherichia coli,relative organ weight and excreta noxious gas contents in broilers

This study was to evaluate the effects of lactulose supplementation on performance, blood profiles, excreta microbial shedding of Lactobacillus and Escherichia coli, relative organ weight and excreta noxious gas contents in broilers. A total of 720 ROSS 308 broilers with a body weight of 46 ± 0.1 g (1 day of age) were used in a 28‐d experiment. Broilers were randomly allotted to 4 experiment diets with 12 replicate pens and 15 birds per pen. Dietary treatments were as follows: NC, negative control (without antibiotic); PC, NC + 0.1% tiamulin; L1, NC + 0.1% lactulose; and L2, NC + 0.2% lactulose. Broilers were fed with phase 1 (1–8 day), phase 2 (9–18 day) and phase 3 (19–28 day) diets in the form of mash. During day 1–8, broilers fed the PC and L2 diets had higher (p < 0.05) body weight gain than those fed the NC diet. During day 19–28, broilers fed the L1 and L2 diets had lower (p < 0.05) feed intake than those fed the NC diet. The feed conversion ratio (FCR) was decreased (p < 0.05) in L1 treatment compared with NC treatment. Overall, the FCR was improved (p < 0.05) in all supplementation treatments compared with NC treatment. The apparently metabolizable nitrogen in L1 treatment was higher (p < 0.05) than that in NC treatment at day 28. The excreta Lactobacillus was increased and E. coli was decreased in PC and L2 treatments compared with NC treatment at day 28 (p < 0.05). The excreta NH₃, H₂S and acetic acid contents were decreased (p < 0.05) in L1 and L2 treatments compared with NC treatment. The relative weight of abdominal fat of broilers fed the PC diet was lowest (p < 0.05) compared with other treatments. In conclusion, this study indicated that dietary supplementation of 0.1% or 0.2% lactulose could improve growth performance, decrease excreta E. coli and excreta NH₃ and H₂S contents.