Effect of Estradiol and Progesterone on Metabolic Biomarkers in Healthy Bitches

The aim of this study was to evaluate the possible association between hormonal changes that occur during oestrus and biomarkers related with glucose metabolism (glucose and insulin), lipid metabolism (lipidic profile and BChE) and adipokines (adiponectin and ghrelin) in healthy bitches. For this purpose, we measured these analytes in serum of bitches, at two times: before (T1) and after (T2) the LH peak that were established according to progesterone concentrations. Increased levels of total cholesterol (p < 0.01), high density lipoprotein cholesterol (HDL‐C) (p < 0.01), low density lipoprotein cholesterol (LDL‐C) (p < 0.01), adiponectin (p < 0.01) and ghrelin (p < 0.05) were observed at T2 in comparison with T1. No statistically significant changes were observed in serum glucose, insulin, homoeostasis model assessment for insulin sensitivity (HOMA), triglycerides and BChE. When all data of T1 and T2 were pooled, serum adiponectin showed positive correlation with progesterone (r = 0.353; p = 0.022) and HDL‐C (r = 0.307; p = 0.048), and negative with insulin (r = ?0.429; p = 0.005), HOMA (r = ?0.446; p = 0.003) and BChE (r = ?0.522; p < 0.001). Ghrelin showed negative correlation with estradiol (r = ?0.701; p = 0.004). BChE was negatively correlated with estradiol (r = ?0.441; p = 0.018) and glucose (r = ?0.343; p = 0.028), and positively with insulin (r = 0.460; p = 0.003) and HOMA (r = 0.505; p < 0.001). In conclusion, changes in metabolic biomarkers occur in bitches after LH peak, characterized by increased lipids (total cholesterol, HDL cholesterol and LDL cholesterol) without changes in BChE activity, and increased adiponectin and ghrelin concentrations, without significant changes in glucose and insulin.     

Colour‐Doppler ultrasound imaging as a laparoscopy substitute to count corpora lutea in superovulated sheep

This study evaluated colour‐Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty‐five Santa Ines ewes were superovulated three times at 21‐day intervals. Corpora lutea were counted by colour‐Doppler UI (CL_DOPPLER) 6 days after each superovulation and confirmed by laparoscopy (CL_LAP) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CL_DOPPLER and CL_LAP, respectively) with a significant positive correlation (r = .94; r²=.89). Colour‐Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation.     

Pre‐Selection Test to Identify High Responder Donor Goats

The aim of the study was to evaluate the feasibility of pre‐selection of high or low responder does prior to the superovulatory protocols. Twenty Saanen does received 800 IU of equine chorionic gonadotropin (eCG) at the end of long‐term progestogen treatment. Fourteen days later, a second progestogen protocol associated with a multiple‐dose follicle stimulation hormone (FSH) treatment (5 IU/kg of FSH, in six decreasing doses between days 4 to 6 of the protocol) was administered. Transrectal ultrasound was used to assess the follicular status at the beginning of superovulatory treatments, at the oestrous onset and on the seventh day of the oestrous cycle for counting corpora lutea (CL). A significant lower number of CL was obtained in eCG‐treated in comparision with FSH‐treated does (p < 0.05). A quartic regression was able to explain the relationship between the number of CL in response to both treatments (r²=0.50; p < 0.05). Seventy per cent (14 of 20) of does maintained the same ovulatory response (high or low) after treatments. The Kappa (κ = 0.40; p < 0.05) and Spearman (rs = 0.39; p = 0.08) coefficients were able to show a relationship between treatments. Regarding the follicular status, there is a significant relationship between the number of small follicles (r = 0.71; r²=0.47; p < 0.01) and total follicles (r = 0.60; p < 0.01) at eCG and first FSH dose with the number of CL. Moreover, it was found a negative relationship between the presence of large follicles and the number of CL in response to eCG treatment (r = ?0.44; p < 0.05), but not from FSH (p > 0.05). In conclusion, the screening test with eCG has the potential to identify Saanen does that will better respond to the superovulatory protocol with FSH. In addition, it highlighted the importance of an ultrasound evaluation prior to the beginning of superovulatory treatments with FSH to characterize the follicular status and identify the potential donors of high ovulatory response in MOET programmes in goats.     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Effect of feeding a high‐carbohydrate or a high‐fat diet on subsequent food intake and blood concentration of satiety‐related hormones in dogs

Although studies in rodents and humans have evidenced a weaker effect of fat in comparison to carbohydrates on the suppression of food intake, very few studies have been carried out in this field in dogs. This study investigates the effects of a high‐carbohydrate (HC ) and a high‐fat (HF ) diets on subsequent food intake and blood satiety‐related hormones in dogs. Diets differed mainly in their starch (442 vs. 271 g/kg dry matter) and fat (99.3 vs. 214 g/kg dry matter) contents. Twelve Beagle dogs received the experimental diets at maintenance energy requirements in two experimental periods, following a cross‐over arrangement. In week 7 of each period, blood concentrations of active ghrelin, glucagon‐like peptide (GLP ‐1), peptide YY , insulin, and glucose were determined before and at 30, 60, 120, 180, and 360 min post‐feeding. The following week, intake of a challenge food offered 180 min after the HC and HF diets was recorded over two days. In comparison to the dogs on the HC diet, those on the HF diet had a higher basal concentration of GLP ‐1 (p  = .010) and a higher total area under the curve over 180 min post‐prandial (tAUC _0–180) (p  = .031). Dogs on the HC diet showed a higher elevation of ghrelin at 180 min (p  = .033) and of insulin at 360 min (p  = .041), although ghrelin and insulin tAUC _0–180 did not differ between the two diets (p  ? .10). Diet had no effect on challenge food intake (p  ? .10), which correlated with the tAUC _0–180 of ghrelin (r = .514, p  = .010), insulin (r = ?.595, p  = .002), and glucose (r = ?.516, p  = .010). Feeding a diet high in carbohydrate or fat at these inclusion levels does not affect the feeding response at 180 min post‐prandial, suggesting a similar short‐term satiating capacity.     

A Field Study to Unravel Factors that are Significantly Associated with the Secretory Activity of the Corpus Luteum During the First Three Postpartum Cycles in High Yielding Dairy Cows,Based on the Amount of Steroidogenic and Endothelial Cells Present in the Luteal Tissue

Fourteen multi‐ and eight primiparous high‐yielding dairy cows were followed from the first till the fourth ovulation postpartum. Cows were randomly divided into two groups and supplemented with soybean (group I; n = 11) or rapeseed meal (group II; n = 11). Both groups were subjected to a biopsy sampling of the corpus luteum (CL) at cycle day 9. The luteal capillary network (visualized by Bandeiraea simplicifolia) was denser in cycles 2 and 3 (p = 0.0005). The same was seen for the surface occupied by steroidogenic cells (visualized by 3β‐hydroxysteroiddehydrogenase) (p = 0.0001). The peripheral blood progesterone concentration showed an increasing trend with increasing cycle number and was higher in primiparous cows (p = 0.013), which had also larger glands on cycle day 9. The area occupied by endothelial cells was positively correlated with the area occupied by steroidogenic cells (r = 0.59; p < 0.0001). Both the areas occupied by endothelial and by steroidogenic cells were negatively correlated with the blood concentration of nonesterified fatty acids (NEFAs) (respectively, r = ?0.377; p = 0.004 and r = ?0.355; p = 0.007). We can conclude that primiparous cows generally have higher peripheral progesterone levels during the first three cycles after calving which is associated with a larger CL. In comparison with those of the first post‐partum cycle, corpora lutea of cycles 2 and 3 have a denser capillary network and a larger area of steroidogenic cells, while these are only associated with a trend of higher peripheral progesterone concentrations.     

Influence of the FecXR Allele in Heterozygous Ewes on Follicular Population and Outcomes of IVP and ET using LOPU‐Derived Oocytes

Ewes heterozygous for the FecX^R allele (R+) in the bone morphogenetic protein 15 (BMP15) gene display increased ovulation rate and prolificacy. Besides this phenotypic advantage, the influence of the FecX^R allele on follicle number and size, oocyte competence and in vitro production (IVP) remains undefined. With these aims, 8 R+ and 8 wild‐type (++) ewes were subjected to 2 laparoscopic ovum pick‐up (LOPU) trials (four sessions per trial; two with and two without FSH) and subsequent IVP and fresh embryo transfer. All follicles >3 mm were punctured (n = 1673). Genotype did not significantly affect the number of punctured follicles per ewe and session (10.4 and 10.2 in R+ and ++ untreated ewes, 17.4 and 14.3 in R+ and ++ FSH‐treated ewes, respectively), but follicular diameter of R+ ewes was significantly reduced compared with ++ ewes (?0.2 mm in untreated and ?0.8 mm in FSH‐treated ewes; p < 0.01). R+ ewes showed higher recovery rate and increased numbers of total and suitable cumulus–oocyte complexes for in vitro maturation (IVM). Similar rates of day 8 blastocysts were observed in R+ (36.1%, 147/407) and ++ (32.6%, 100/307) ewes, but the final output of day 8 blastocysts per ewe and session was higher in R+ ewes (+0.75; p < 0.005), without differences in survival rate at birth of the transferred embryos (40.4%, 21/52 vs 36.4%, 16/44, respectively). In conclusion, a higher number of oocytes proven to be competent for in vitro development and embryo survival after transfer are recovered from R+ ewes, despite the lower mean size of their follicles at puncture.     

Effect of Progesterone Prior to GnRH-PGF2α Treatment on Induction of Oestrus and Pregnancy in Anoestrous Awassi Ewes

An experiment was conducted to examine the effect of progesterone prior to a GnRH‐PGF_2α treatment on oestrus and pregnancy in seasonally anoestrous Awassi ewes. Twenty‐four ewes were randomly assigned to three groups to be pre‐treated with 60 mg medroxyprogesterone acetate sponges (group A), 600 mg progesterone sponges (group B) or blank sponges (group C) for 4 days. All ewes were injected with 100 μg of GnRH 24 h after sponge removal followed, 5 days later, by 20 mg PGF_2α injection. Ewes were exposed to three fertile rams at the time of PGF_2α injection (day 0, 0 h) and were checked for breeding marks at 6‐h intervals for 5 days. Blood samples were collected from all ewes 1 day (day ?10) prior to sponge insertion, at the time of sponge removal (day ?6), 1 day following sponge removal (day ?5, at the time of GnRH injection) and at the time of PGF_2α injection (day 0) for analysis of progesterone. Progesterone concentrations on days ?10 and ?5 were basal and averaged 0.2 ± 0.04 and 0.2 ± 0.2 ng/ml, respectively. Progesterone concentrations on day ?6 were elevated only in group B ewes and were higher (p < 0.0001) than those of groups A and C. Progesterone concentrations on day 0 were higher (p = 0.002) in groups A and B than group C. Oestrous responses occurred only in ewes of groups A and B (p > 0.05). Induced oestrus conception rate was greater (p < 0.01) in group A than groups B and C. Ewes returned to oestrus 17–20 days following day 0 were two of eight, six of eight and three of eight of groups A, B and C, respectively, all of which eventually lambed. The overall lambing rate was 82% in progesterone‐primed ewes compared with only 38% non‐progesterone‐primed ewes (p < 0.05). Progesterone priming apparently sensitizes GnRH‐PGF_2α‐treated seasonally anoestrous ewes and increases their response in oestrus and pregnancy rates.     

Two single nucleotide polymorphisms in the promoter of the ovine myostatin gene (MSTN) and their effect on growth and carcass muscle traits in New Zealand Romney sheep

Myostatin is a negative regulator of muscle growth and development in mammals, and variation in ovine myostatin gene (MSTN) has been demonstrated to be associated with variation in the muscularity of sheep. Polymerase chain reaction–single‐stranded conformational polymorphism (PCR–SSCP) was used to look for single nucleotide polymorphisms (SNPs) in a 304‐bp amplicon from the promoter region of ovine MSTN. Sequence analyses revealed two previously identified SNPs (c.?2449G/C and c. ?2379T/C) that resulted in three haplotypes (H₁ (c.[?2449G; ?2379C]), H₂ (c.[?2449C; ?2379C]) and H₃ (c.[?2449G; ?2379T]). The effect of these SNPs on growth and carcass traits was investigated in 357 NZ Romney lambs. General linear mixed‐effect models revealed that sheep with the genotype c.?2449GC had a higher loin meat yield (p = 0.032) and proportion loin yield (p = 0.028), than those with the genotype c.?2449GG. The genotype c.?2379CC was associated with an increase in three weight traits: birthweight (p = 0.003), tailing weight (p = 0.009) and weaning weight (p = 0.028), when compared with the genotype c.?2379TC, but it was not found to have an association with growth rate. This suggests that c.?2379T/C has an effect that originates at, or before birth. Haplotype H₃ was associated with a decrease in birthweight (p = 0.002), tailing weight (p = 0.003) and weaning weight (p = 0.011). Haplotype H₂ was associated with increased loin yield (p = 0.012) and proportion loin yield (p = 0.002). The SNPs may have value as genetic markers for improved Romney breeding.     

High‐milking sheep have a lower ovulation rate and tend to yield fewer embryos in response to superovulation and intrauterine artificial insemination

Antagonistic relationship between milk yield and reproduction is reported in several livestock species. This study aimed to investigate whether genetic merit for milk production in dairy sheep affects responses to superovulation, embryo yield and quality. A total of 21 cross‐bred Sarda x Lacaune ewes homogeneous for age, parity and stage of lactation were included. The ewes were stratified as high‐producing or low‐producing based on their genetic merit for milk production estimated by a pentatrait repeatability animal model. Oestrus was synchronized using an intravaginal progesterone pessary inserted on Day 0 and removed on Day 14. Superovulatory treatment consisted of 350 I.U. of porcine FSH administered in eight decreasing intramuscular doses every 12 hr with a total dose of 10 ml of solution starting 12 days after insertion of sponges. Laparoscopic artificial insemination (AI) was performed 48 hr after pessary removal. Surgical embryo recovery was performed at Day 8 after pessary removal. Correlation between breeding value for milk production and the number of corpora lutea (CL) was significantly different from zero (?0.49). High‐producing ewes had a lower number of CL than low‐producing counterparts (7.6 ± 2.50 vs 12.1 ± 5.16 respectively; p < .02). Furthermore, there was a tendency for high‐producing ewes to yield fewer embryos than low‐producing females (5.3 ± 3.46 vs 9.18 ± 5.11; p = .09). No differences were observed between ewes in both genetic groups with regard to the number of embryos of grades 1, 2 and 3. To our knowledge, this is the first report highlighting an antagonism between genetic merit for milk production and the ability to produce embryos in sheep. These results deserve to be considered in sheep breeding programmes.     

Effect of added dietary threonine on growth performance,health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to Escherichia coli infection and challenged with E. coli K88ac

Threonine (Thr) is important for mucin and immunoglobulin production. We studied the effect of added dietary Thr on growth performance, health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to E. coli K88ac (ETEC) infection and challenged with ETEC. Forty‐eight 24‐day‐old weaned pigs were divided into two groups by their ETEC susceptibility using mucin 4 (MUC4) gene as a marker (2 MUC4^?/?, not‐susceptible, and 2 MUC4^+/+, susceptible, pigs per litter). Within genotype, pigs were fed two different diets: 8.5 (LThr) or 9.0 (HThr) g Thr/kg. Pigs were orally challenged on day 7 after weaning and slaughtered on day 12 or 13 after weaning. Before ETEC challenge, HThr pigs ate more (p < 0.05). The diet did not affect post‐challenge growth, but HThr tended to increase post‐challenge feed efficiency (p = 0.087) and overall growth (p = 0.087) and feed efficiency (p = 0.055). Before challenge, HThr pigs excreted less E. coli (p < 0.05), while after challenge, diet did not affect the number of days with diarrhoea and ETEC excretion. MUC4^+/+ pigs responded to the challenge with more diarrhoea, ETEC excretion and anti‐K88 IgA in blood and jejunal secretion (p < 0.001). HThr pigs had a higher increase of anti‐K88 IgA values in jejunal secretion (p = 0.089) and in blood (p = 0.089, in MUC4^+/+ pigs only). Thr did not affect total IgA and IgM values, morphometry of jejunum, goblet cells count in colon, total mucin from jejunum and colon, but varied jejunal goblet cells counts (p < 0.05). In the first two post‐weaning weeks, 8.5 g Thr/kg diet may be not sufficient to optimize initial feed intake, overall feed efficiency and intestinal IgA secretion and to control the gut microbiota in the first post‐weaning week, irrespective of the pig genetic susceptibility to ETEC infection.     

Does the Microbial Flora in the Ejaculate Affect the Freezeability of Stallion Sperm?

In an attempt to evaluate the possible relationship between the microbial flora in the stallion ejaculate and its ability to freeze, three ejaculates from five stallions were frozen using a standard protocol. Before freezing, an aliquot was removed for bacteriological analysis. Bacterial growth was observed in all the ejaculates studied. The isolated microorganisms were: Staphylococcus spp. and Micrococcus spp. (in all the stallions), β-haemolytic Streptococcus (in stallions 3 and 4), Corynebacterium spp. (in stallions 1, 3–5), Rhodococcus spp. (in stallion number 2), Pseudomonas spp. (in stallion number 1) and Klebsiella spp. (in stallions 1, 3 and 5). The presence and richness of Klebsiella and β-haemolytic Streptococcus in the ejaculate were related to two sperm variables post-thaw, namely the proportion of dead spermatozoa (ethidium+ cells; r = 0.55, p < 0.05) and the amplitude of lateral displacement of the sperm head (ALH, μm; r = −0.56, p < 0.05), respectively. The degree of growth of Corynebacterium spp. in the ejaculate was positively correlated with the percentage of spermatozoa showing high caspase activity post-thaw (r = 0.62, p < 0.05). The presence and number of colonies of β-haemolytic Streptococcus were negatively correlated (r = −0.55, p < 0.05) with low sperm caspase activity. It is concluded that the microbial flora of the equine ejaculate may be responsible for some of the sublethal damage experimented by the spermatozoa during cryopreservation.     

Effect on Rendement Napole genotype on metabolic markers in Ossabaw pigs fed different levels of fat

We investigated effects of Rendement Napole (RN ) genotype on metabolic markers in Ossabaw pigs fed diets with different levels of dietary fat. Thirty‐two pigs, belonging to either the wild‐type (WT , rn⁺/rn⁺) or carrier (CAR , RN ^?/rn⁺) genotypes (n  = 16/genotype), were divided into two dietary groups, (high fat [HF ] or low fat [LF ]) diets, for 12 weeks (n  = 8 pigs/genotype/diet) after which pigs were killed for gene expression analysis by RT ‐PCR . Feeding HF diet caused increased daily gain (ADG , p  <  .05) and final body weight (BW ) (p  <  .05) in comparison with the LF diet (p  <  .05). Feed efficiency (gain:feed) was higher (p  <  .05) in pigs on the HF and was higher (p  <  .05) in CAR pigs compared to WT . There was genotype × diet interaction (p  =  .05) on final BW such that CAR animals on LF diet had the same final BW as animals of both genotypes on HF diet. Carrier pigs on LF diet had higher (p  <  .05) average daily gain and gain:feed than WT pigs. There was a trend (p  <  .08) for a higher feed consumption in pigs on the LF diet. Backfat thickness was higher (p  <  .01) in pigs on the HF diet. Serum triglyceride was higher (0.62 vs. 0.33 mg/dl, p  <  .01) in pigs on HF diet. Serum insulin was higher (p  <  .05) in CAR versus WT pigs (0.40 vs. 0.015 μg/ml). Pigs on the HF diet had a higher (p  <  .05) serum insulin compared to those on the LF diet (0.032 vs. 0.023 μg/ml). Carnitine palmitoyl transferase 1‐alpha was higher (p  <  .05) in the longissimus dorsi and semitendinosus muscles of pigs on HF diet. Acyl‐CoA oxidase I was elevated (p  <  .05) in the liver of pigs on HF diet. Fatty acid synthase was lower in the longissimus dorsi muscle, liver and mesenteric fat (p  <  .05) of carrier pigs. The RN gene regulates specific metabolic markers in the Ossabaw pigs.     

Accurate Ultrasonographic Prediction of Progesterone Concentrations Greater than 1 ng/ml in Holstein lactating dairy cows

To develop an ultrasonographic assay for determining plasma progesterone concentration (P₄) as < 1 ng/ml or ≥ 1 ng/ml, the corpus luteum (CL) area and P₄ were measured in 1094 multiparous Holstein cows. The area‐measuring function and frozen images were used to outline and measure CL imaged via ultrasonography, and CL area was estimated as a polygon of a continuation straight line. A significant correlation was found between CL area and P₄ (p < 0.001), and this analysis resulted in the following correlation equation: y = ?0.35 + 1.02x (r = 0.81). According to the correlation equation, a CL area of 1.3 cm² indicated a P₄ of 1 ng/ml. Based on this relationship, each animal was categorized into one of six groups, groups differed based on CL area, and the area ranges were as follows: < 1.3 cm² (Group A), 1.3–2.2 cm² (Group B), 2.3–3.2 cm² (Group C), 3.3–4.2 cm² (Group D), 4.3–5.2 cm² (Group E) and > 5.2 cm² (Group F). For each group, the proportion of cows whose P₄ was 1 ng/ml or more was 1.5% in Group A, 83.3% in Group B, 76.6% in Group C, 96.6% in Group D, 99.2% in Group E and 100% in Group F. There was a significant difference between Group A and the other five groups, and between Groups B or C and Groups D, E or F (p < 0.005). These results indicate that a functional CL does not exist when the CL area is less than 1.3 cm² and that it exists when the CL area is 3.3 cm² or more.     

Systemic Concentrations of Endogenous and Exogenous FSH in Anoestrous Ewes Superstimulated with Folltropin®-V

The synchronization of follicular waves with medroxyprogesterone acetate (MAP) and oestradiol‐17β (E₂‐17β) prior to ovarian superstimilation in anoestrous ewes reduces the variability in superovulatory responses by an unknown mechanism. Follicle stimulating hormone (FSH) is a primary promoter of antral follicular development, but the relevance of circulating FSH concentrations to the superovulation performance in ewes has not been examined. Eighteen anoestrous Rideau Arcott ewes (May–June) were superovulated with Folltropin^®‐V (porcine FSH), with (n = 8; treated ewes) or without (n = 10; control ewes) a single i.m. dose of 350 μg of E₂‐17β, given on the sixth day of a 14‐day treatment with MAP‐releasing intravaginal sponges (60 mg). The superovulatory treatment, begun 6 days after E₂‐17β injection, consisted of six i.m. applications of Folltropin^®‐V given twice daily (at 08:00 and 16:00 h), followed by an i.m. injection of GnRH (50 μg). Blood samples collected every 8 h throughout the 3‐day treatment, were analysed by radioimmunoassays for concentrations of ovine and porcine FSH, using species‐specific standards and primary antibodies. Serum concentrations of oFSH were greater (p < 0.05) in the controls compared to treated ewes at 40, 64 and 72 h and the variability in mean oFSH concentrations was greater (p < 0.05) in control ewes at 40, 48, 64 and 72 h after the 1^st Folltropin^®‐V injection. There were no differences (p > 0.05) between the two groups in serum concentrations of pFSH. Significant correlations were recorded between the number of corpora lutea (CL) and oFSH concentrations at 8 h (r = 0.72, p < 0.05), 16 h (r=0.63, p < 0.05) and 64 h (r = 0.84, p < 0.01) after the 1^st Folltropin^®‐V injection. The total number of recovered embryos was positively correlated to oFSH concentrations at 56 h (r = 0.69, p < 0.05). We concluded that changes in endogenous FSH concentrations during ovarian superstimulation with pFSH might contribute to the variability in superovulatory responses in ewes.     

Acid–Base Parameters and Steroid Concentrations in Pre‐Ovulatory Follicles and Plasma of Lactating Dairy Cows with Spontaneous and Synchronized Oestrus or Follicular Cyst

The study aimed to compare the acid–base balance and steroid concentrations between follicular fluids (FF) of pre‐ovulatory follicles derived from a spontaneous oestrus (SO), synchronized or induced oestrus (IO) and follicular cysts (CYS) and between FF and blood in dairy cows. Forty‐two dairy cows were included in this study. The animals were allocated to three groups: SO (n = 23); IO (n = 11) using GnRH at day 0 and day 9 and PGF₂α at day 7; and animals with CYS (n = 10). The follicular fluids (FF) were aspirated from the cyst/pre‐ovulatory follicles (? ≥ 15 mm) after SO and after second GnRH dose in IO by transvaginal ultrasound‐guided ovum pick‐up technique. Blood samples (BL) were collected in heparinized vacutainer tubes. The oxygen tension (pO2) in FF of IO was higher (p < 0.05) than in SO and CYS groups. There were negative correlations (p < 0.001, r = ?0.89) between FF and blood pO2. The carbon dioxide tension (pCO2) and lactate level in FF of CYS group were higher (p < 0.05) than in SO and IO groups. There were negative correlations (p < 0.01, r = ?0.73) between blood and FF pO2. Oestradiol‐17β concentration in pre‐ovulatory follicles and plasma of the SO group was higher (p < 0.001) than in IO and CYS groups. Progesterone concentration in pre‐ovulatory follicles and plasma of the SO and IO groups was lower (p < 0.01) than in CYS group. Plasma androstenedione concentration in SO and IO groups was higher (p < 0.05) than in CYS group. In conclusion, acid–base parameters, E2 and P4 levels in the follicular fluid of both IO and CYS groups were deviated greatly from the physiological level (disturbances of intrafollicular/intracystic environment), which may affect the quality of both the oocyte and the granulosa cells.     

Degradation parameters of amaranth,barley and quinoa in alpacas fed grass hay

This study was conducted to determine the compartment 1 (C1) characteristics of alpacas (fistulated male, 7 ± 1.5 years old, 61 ± 5 kg BW) fed grass hay (GH) supplemented with amaranth (AM), quinoa (Q) and barley (B) grains. Alpacas were provided water ad libitum while housed in metabolism crates. The GH and GH plus treatments were fed at 0700 every day. Treatment periods were for 14 days in which GH or GH plus one of the grain treatments were randomly allocated. On day 14, volatile fatty acids (VFA), pH and ammonia nitrogen (NH₃‐N) were determined at 1, 3, 6, 10, 14, 18 and 24 h post‐feeding. C1 degradation of each feed component was also determined with the alpacas being fed GH only and the samples incubated for 0, 2, 4, 8, 14, 24, 48 and 72 h. Dry matter (DM), neutral detergent fibre (NDF) and crude protein (CP) were determined and were divided into three categories: a = immediately soluble; b = the non‐soluble but degradable; and u = non‐degradable/unavailable, potential extent of degradation (PE), degradation rate (c) and effective degradation (ED). C1 passage rate was determined using acid detergent insoluble ash as a marker and was calculated to be 5.5%?h‐1. Total DM intake was highest (p < 0.05) for B and resulted in a higher (p < 0.05) CP intake. GH and AM were different in mean pH (6.81 and 6.66, respectively). B NH₃‐N was greater (p < 0.05) than the other treatments. Total VFA was greatest (p < 0.05) for AM, with the greatest composition differences being a shift form acetate percentage to butyrate. DM, NDF and CP degradation was different across the treatments, where PE and ED were higher (p < 0.05) for the grain treatments. The pseudo‐grains AM and Q had similar C1 degradation characteristics to B.     

Influence of organic phosphorus on reproductive performance and metabolic profiles of anoestrous Farafra ewes in subtropics at the end of breeding season

The effect of organic phosphorus on metabolic, haematological and hormonal status, restoration of ovarian functions and conception rate in anoestrous Farafra ewes in subtropics were evaluated. Anoestrous Farafra ewes (n  = 24; 34.72 ± 0.52 kg body weight) were allocated into two equal groups: control and phosphorus groups. The ewes of phosphorus group were treated with sodium 4‐dimethylamino‐2‐methyl‐phenyl‐phosphonate as an organic bound phosphorous twice a week for successive 3 weeks. Ovarian follicle development and corpora lutea were checked three times a week till occurrence of oestrus using ultrasonography while pregnancy was confirmed at 30 days post‐service. Plasma metabolites, reproductive hormones, thyroid hormones and minerals were detected at weeks ?2, ?1, 0 (mating day) and + 4 weeks post‐oestrus. Phosphorus group had significantly (p  < .05) short interval to oestrous resumption if compared to control ewes (2.1 ± 0.8 weeks vs . 4.6 ± 1.1 weeks). In addition, phosphorous supplementation significantly (p  < .05) increased the number of antral follicles (developed and their sizes in addition to sizes of corpora lutea (8.72 ± 0.3 mm vs . 7.46 ± 0.9 mm) as well. Number of services per conception (2.6 vs . 1.4; p  < .01) was higher in control group than that of phosphorus group. Pregnancy rate (80 vs . 50%) was significantly (p  < .01) higher in phosphorus group when compared to control. White blood cells in treated ewes (10.8 ± 0.44; p  < .05) and monocytes (2.93 ± 0.13; p  < .01) were higher than that of control group (white blood cells; 9.53 ± 0.50 and monocytes; 2.24 ± 0.14). Metabolic parameters did not differ between phosphorus and control groups during different times of treatment. It could be concluded that phosphorous administration to anoestrous Farafra ewes in subtropics could improve reproductive performance and restore ovarian activity at the end of spring and early summer.     

Effectiveness of controlled internal drug release device treatment to alleviate reproductive seasonality in anestrus lactating or dry Barki and Rahmani ewes during non‐breeding season

This study aimed to evaluate the effectiveness of hormonal treatments on ovarian activity and reproductive performance in Barki and Rahmani ewes during non‐breeding season. Forty‐eight multiparous ewes, 24 Barki and 24 Rahmani ewes were divided into two groups, 12 lactating and 12 dry ewes for each breed. Controlled internal drug release (CIDR) device was inserted in all ewes for 14 days in conjunction with intramuscular 500 IU equine chronic gonadotrophin (eCG) at day of CIDR removal. Data were analysed using PROC MIXED of SAS for repeated measures. Breed, physiological status and days were used as fixed effects and individual ewes as random effects. Barki ewes recorded higher (p < .05) total number of follicles, number of large follicles, serum estradiol concentration and estradiol: progesterone (E₂:P₄) ratio compared to Rahmani ewes. Lactating ewes recorded higher (p < .05) number of small follicles and lower concentration of total antioxidant capacity (TAC) compared to dry ewes. Number and diameter of large follicles recorded the highest (p < .05) values accompanied with disappearance of corpora lutea at day of mating. Serum progesterone concentration recorded lower (p < .05) value at day of mating and the highest (p < .05) value at day 35 after mating. CIDR‐eCG protocol induced 100% oestrous behaviour in both breeds, but Rahmani ewes recorded longer (p < .05) oestrous duration compared to Barki. Conception failure was higher (p < .05) in Barki compared to Rahmani ewes. In conclusion, CIDR‐eCG protocol was more potent in improving ovarian activity in Barki compared to Rahmani ewes, but this protocol seems to induce hormonal imbalance in Barki ewes that resulted in increasing conception failure compared to Rahmani ewes.     

Ovarian Responses, Hormonal Profiles and Embryo Yields in Anoestrous Ewes Superovulated with Folltropin®-V after Pretreatment with Medroxyprogesterone Acetate-releasing Vaginal Sponges and a Single Dose of Oestradiol-17β

In ruminants, superovulatory treatments started at the time of follicular wave emergence result in greater and less variable ovulatory responses and embryo yields compared with the treatments begun in the presence of a large growing antral follicle(s) from the previous waves. The progesterone–oestradiol treatment is routinely used for follicular wave synchronization in cattle. The main objective of this study was to characterize the ovarian responses, hormonal profiles and in vivo embryo production in anoestrous Rideau Arcott ewes (May‐June), which were superovulated after pretreatment with medroxyprogesterone acetate (MAP)‐releasing intravaginal sponges and a single dose of oestradiol‐17β (E₂‐17β). Six days after insertion of MAP sponges, eight ewes were given an i.m. injection of 350 μg of E₂‐17β (E₂‐17β‐treated ewes); 10 ewes were given an i.m. injection of vehicle (control ewes). Multiple‐dose Folltropin®‐V treatment, followed by the bolus injection of GnRH (50 μg i.m.), began 6 days after E₂‐17β/vehicle injection. Transrectal ovarian ultrasonography revealed that: (i) the interval between E₂‐17β/vehicle injection and regression of all follicles ≥5 to 3 mm in diameter was shorter (p < 0.01; 2.6 ± 0.4 vs 4.8 ± 0.6 days respectively); and (ii) the interval between injection and emergence of the next follicular wave was longer (p < 0.05; 5.4 ± 0.3 vs 1.2 ± 0.4 days, respectively) in E₂‐17β‐treated than in control ewes. During the 6 days after injection, the mean FSH peak concentration and basal FSH concentration were lower (p < 0.01) in E₂‐17β‐treated ewes. The mean ovulation rate and the number of recovered embryos did not differ (p > 0.05) between the two groups of ewes. However, the number of luteinized unovulated follicles per ewe, and the variability in the number of luteal structures and overall embryo yield were less (p < 0.05) in E₂‐17β‐treated compared with control ewes. In conclusion, the MAP–E₂‐17β pretreatment significantly reduced the variability in ovarian responses and embryo yields, without affecting the embryo production in superovulated anoestrous ewes.