Profile of Steroid Receptors and Increased Aromatase Immunoexpression in Canine Inflammatory Mammary Cancer as a Potential Therapeutic Target

Canine inflammatory mammary cancer (IMC) has been proposed as a model for the study of human inflammatory breast cancer (IBC). The aims of this study were to compare the immunohistochemical expression of aromatase (Arom) and several hormone receptors [estrogen receptor α (ERα), estrogen receptor β (ERβ), progesterone receptor (PR) and androgen receptor (AR)], in 21 IMC cases vs 19 non‐IMC; and to study the possible effect of letrozole on canine IMC and human inflammatory breast cancer (IBC) in vitro using IPC‐366 and SUM‐149 cell lines. Significant elevations of the means of Arom Total Score (TS), ERβ TS and PR TS were found in the IMC group (p = 0.025, p = 0.038 and p = 0.037, respectively). Secondary IMC tumours expressed higher levels of Arom than primary IMC (p = 0.029). Non‐IMC PR‐ tumours contained higher levels of Arom than non‐IMC PR+ tumours (p = 0.007). After the addition of letrozole, the number of IMC and IBC cells dropped drastically. The overexpression of Arom found and the results obtained in vitro further support canine IMC as a model for the study of IBC and future approaches to the treatment of dogs with mammary cancer, and especially IMC, using Arom inhibitors.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Pregnancy rate after fixed‐time transfer of cryopreserved embryos collected by non‐surgical route in Lacaune sheep

This study investigated the feasibility of applying fixed‐time (cryopreserved) embryo transfer in ewes. Embryos (n = 106) were non‐surgically recovered from superovulated donors (n = 39) on day 6–7 after oestrus. Straws containing one or two embryos (morulae and/or blastocysts) subjected to either slow freezing (SF, n = 62) or vitrification (VT, n = 44) were randomly used within fixed‐time embryo transfer on Day 8.5. Recipient ewes were nulliparous (n = 58) bearing corpora lutea after synchronous oestrous induction protocol. The pregnancy rate was higher (p = .03) in SF (39.4%) than VT (16.9%) and survival rate tended (p = .08) to be higher in SF than in VT (25.8% vs. 15.9%). Lambing rates were similar (p = .13) between SF (20.9%) and VT (15.9%). Embryos recovered by non‐surgical route after cervical dilation treatment and later cryopreserved by either slow freezing or vitrification produced reasonable pregnancy rates after FTET.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Short‐term methionine supplementation during the early post‐partum period in primiparous rabbits improves prolificacy associated with an increase in serum concentrations of IGF‐I

The aim of this study was to evaluate the effects of methionine supplementation on energy metabolism and reproductive performance during the early post‐partum period in primiparous does. Forty nulliparous New Zealand White does were used. Females were randomized in two groups at calving: the control group (n = 20) was fed with the basal diet, and the methionine group (n = 20) was fed the basal diet plus 1 g/animal/day of methionine from the day of calving to 4 days post‐partum. Results showed that methionine supplementation increased (p = 0.032) the concentration of insulin‐like growth factor‐1 with respect to control group 4 days post‐partum. It similarly increased the prolificacy (p = 0.03), the number of kits born alive per litter (p = 0.06) and the body gain weight of the litter during supplementation (p = 0.035). These results were observed despite the does in the methionine group having a deeper negative energy balance than the does in the control group. Finally, methionine supplementation did not affect receptivity (p = 0.23), fertility (p = 0.49), the number of kits born dead per litter (p = 0.86) insulin and metabolites as glucose, non‐esterified fatty acids and triglycerides. In conclusion, our results show that methionine supplementation during the first 4 days of the post‐partum period in rabbits increases total litter size and the corporal weight of kits and is associated with an increase in blood concentration of IGF‐1.     

Gonadotropin‐Releasing Hormone Administration to Dairy Cows without a Corpus Luteum 4 Weeks after Calving Increases Reproductive Performance

This field study investigated whether the administration of a single dose of gonadotropin‐releasing hormone (GnRH) to dairy cows without a corpus luteum (CL) 4 weeks after calving can improve reproductive performance. Holstein dairy cows underwent ultrasonography to assess the presence of ovarian structures at 29.2 ± 5.2 days post‐partum, and cows were divided into two main groups based on the presence (CL group, n = 230) or absence (non‐CL group, n = 460) of a CL. The non‐CL group was further randomly divided into two subgroups based on the administration of GnRH (non‐CL GnRH group, n = 230) or no GnRH (non‐CL control group, n = 230). Subsets of cows from non‐CL control (n = 166) and non‐CL GnRH (n = 175) groups received a second ultrasonography at 44.5 ± 5.4 days post‐partum to assess CL formation. The percentage of cows with CL at the second ultrasonography was greater in the non‐CL GnRH group (70.9%) than in the non‐CL control group (53.0%, p = 0.0006). The hazard of the first post‐partum insemination by 150 days in milk (DIM) was higher in the CL group than in the non‐CL control group (hazard ratio [HR]: 1.36, p = 0.001). The probability of a pregnancy to the first insemination was higher in non‐CL GnRH (odds ratio [OR]: 1.50, p = 0.04) and CL groups (OR: 1.55, p = 0.03) compared to the non‐CL control group. Furthermore, the hazard of pregnancy by 210 DIM was higher in non‐CL GnRH (HR: 1.30, p = 0.01) and CL (HR: 1.51, p = 0.0001) groups than in the non‐CL control group. In conclusion, administration of GnRH to dairy cows without a CL 4 weeks after calving was associated with an increase in ovulation and improved reproductive performance.     

Essential oils improved weight gain,growth and feed efficiency of young dairy calves fed 18 or 20% crude protein starter diets

The objective was to evaluate interactions between starter protein (180 vs. 200 g/kg, DM basis) and a mixture of essential oils (EOs; containing thymol, eugenol, vanillin, limonene and guaiacol) on growth, metabolic and ruminal functions of Holstein dairy calves. In a completely randomized 2 × 2 factorial design, 48 calves, 3 days old (averaging BW 42.7 ± 1.9 kg), were allocated into groups fed the following diets: (i) 180 g/kg CP with no EO (180P‐NEO); (ii) 180 g/kg CP with EO (180P‐EO); (iii) 200 g/kg CP with no EO (200P‐NEO); and (iv) 200 g/kg CP with EO (200P‐EO). The EO was supplemented as 1 g/kg of starter DM. Calves were fed ad libitum starter diet and were weaned at day 59 of age, but diets continued until day 80. There were no interactive effects of CP and EO on intake and growth. Pre‐weaning feed efficiency tended to be increased for 200P‐EO (p = .09). Average daily gain and feed efficiency during pre‐weaning period as well as weaning weight were increased (p < .05) by EO, whereas wither height was increased by EO (p = .03) and tended to be increased for 200P vs. 180P (p = .06). Post‐weaning blood urea nitrogen concentration tended to be lower in 180P vs. 200P (p = .08). Ruminal short‐chain fatty acids concentration was greatest in 200P‐EO. The EO increased both butyrate (p = .02) and propionate proportions (p = .01) and reduced acetate proportional ratio (p < .01). Ruminal ammonia‐N was tended to be lower in calves‐fed EO (p = .05) and was lower in those fed 180P vs. 200P (p < .01). In conclusion, supplementation of the starter diet with essential oil improved weight gain, growth and feed efficiency of dairy calves, irrespective of dietary protein content.     

Associations of Leptin and Pituitary‐Specific Transcription Factor Genes’ Polymorphisms with Reproduction and Production Traits in Dairy Buffalo

This study aimed to detect the genetic variability in Leptin and Pit‐1 genes using polymerase chain reaction–restriction fragment length polymorphism and DNA sequencing also to explore their possible associations with reproductive and productive traits of Egyptian buffaloes. Regarding Leptin gene, three genotypes (AA, AG and GG) were identified with frequency of 0.54, 0.40 and 0.06, respectively, and the genotypes were distributed according to the Hardy–Weinberg equilibrium. Allele A was comparatively higher than G with frequency of 0.74 and 0.26, respectively. For Pit‐1 gene, the association could not be performed due to the monomorphism (BB). The results showed that AA genotypes were found to be superior in most of production and reproduction traits. AA genotypes yielded more milk (2332.34 kg, p = 0.04) with higher fat% (6.10, p = 0.004) and fat yield (155.75 kg, p = 0.06), reach peak milk production at 42.19 days and required 2.19 services for conception in comparison with GG genotypes. Birthweight of animals with AA genotype was lesser than with GG genotype (39.35 and 43.67 kg, p = 0.02, respectively). The days open is numerically better in AA genotype animals (99.35 days), but the difference between the three genotypes was non‐significant. The distinct significant associations reported in this study suggested that Leptin is reputable candidate genetic marker, which might be used to enhance animals’ genetic potential for milk production in conjunction with reproduction.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

The effects of dietary calcium iodate on productive performance,egg quality and iodine accumulation in eggs of laying hens

The aim of this study was to examine the effects of various levels of supplemental calcium iodate (CI) on productive performance, egg quality, blood indices and iodine (I) accumulation in the eggs in commercial laying hens. A total of 240 White Leghorn layers (Hy‐line W36) were divided through a completely randomized design into six treatments with five replicates and eight hens per each at 32 weeks of age. This experiment lasted for 12 weeks. Concentrations of I in the mash diets were 0.74, 3.13, 5.57, 8.11, 10.65 and 12.94 mg I/kg of feed in treatments 1–6 respectively. The added doses of CI were included 0.0 (control), 2.5, 5.0, 7.5, 10.0 and 12.5 mg/kg of diet for treatments 1–6 respectively. There were no significant differences in productive performance among the treatments. The highest eggshell strength was observed in group fed diet containing 3.13 mg I/kg (p = .014). The highest percentage of calcium and lowest percentage of phosphorus in eggshell were observed in group fed diet containing 12.94 mg I/kg (p = .0001). Feeding hens with diet containing 12.94 mg I/kg increased serum triiodothyronine‐to‐thyroxine ratio (p = .0001). Serum alanine aminotransferase activity in hens fed diet containing 12.94 mg I/kg was significantly more than control (p = .041). Blood Serum triglycerides in hens fed diet containing 8.11 mg I/kg were significantly higher than control (p = .0001). Edible fraction of the eggs of birds fed diet containing 12.94 mg I/kg was enriched by I almost 3 times more than those fed diet containing 0.74 mg I/kg. The results suggested that egg production, egg mass, feed intake and feed conversion ratio were not significantly affected by dietary I levels. Iodine accumulation in the eggs were increased by increasing dietary I levels and the level of 10 mg/kg CI could supply I enrichment of the eggs.     

Single layer centrifugation of fresh dromedary camel semen improves sperm quality and in vitro fertilization capacity compared with simple sperm washing

Single layer centrifugation (SLC) through a colloid is a tool for selecting viable mammalian spermatozoa but has not been used previously for fresh dromedary camel sperm. Semen from six camels (2 ejaculates/male) was diluted 1:5 (v:v) or 1:10 (v:v) in a Tris–citrate–fructose buffer for mechanical liquefaction by gentle pipetting. Following liquefaction, semen was processed either by SLC or by centrifugation without a colloid (control). Total and progressive motilities, CASA kinematics, vitality and acrosome integrity (eosin–nigrosin) and plasma membrane integrity (Hypo‐osmotic swelling test; HOST), and fertilizing ability in a heterologous assay (zona‐free goat oocytes) were evaluated. Both total (p = .003) and progressive motilities (p = .003) were higher in SLC‐processed than in control semen samples, irrespective of dilution. Positive HOST values increased when using colloid in 1:5 (p = .001) and 1:10 dilution (p = .010). Colloid‐selected sperm had higher penetration rates than controls (p < .001 and p = .02 for 1:5 and 1:10 dilutions, respectively). However, only the SLC sperm at 1:5 dilution showed higher percentages of pronuclear formation (p = .02) than controls. Dilution effect was only significant for total motility before in vitro fertilization, with higher values for the 1:5 dilution (p = .033). The recovery rates of motile sperm between dilutions were similar (26.1% vs 35.4%; p = .226). In conclusion, SLC is a promising tool for selecting functional dromedary camel sperm and warrants more research.     

Effects of different sources of protein on the growth performance,blood chemistry and polypeptide profiles in the gastrointestinal tract digesta of newly weaned piglets

To determine the effects of different sources of protein on the growth performance of newly weaned piglets, 72 newly weaned piglets were randomly assigned to three groups fed different diets (soya bean, casein and dried distillers' grain with solubles (DDGS) feeds). Casein and DDGS feeds consisted of soya bean feed in which 5% of the CP was replaced with casein‐ or DDGS‐derived CP respectively. Blood and chyme samples were collected from each piglet 2 h post‐feeding on days 0 and 28 of the feeding period. The DDGS feed decreased DMI (p = 0.024) and increased FCR (p = 0.025) due to lower nitrogen utilization (p = 0.078) than those of other feeds. Total amino acid content in chyme demonstrated that casein feed digested rapidly in the duodenum (p = 0.005), whereas DDGS feed was digested primarily in the distal jejunum (p = 0.003) and ileocecum (p = 0.002). However, polypeptide profiles in chyme exhibited a pattern different from those of amino acids. There were no differences in the polypeptide profiles in the stomachs of piglets fed soya bean or casein feeds (p > 0.05), but soya bean group had greater amounts of small polypeptides (mass under m/z 3000 Da) in the duodenum (p = 0.052) than other groups. In contrast, the DDGS feed group had more large polypeptides (m/z 3000–4000 Da) in the stomach than the other groups (p < 0.001). In addition, 10 pairs of polypeptides with matching masses were identified in the plasma and digesta, indicating that polypeptides may have been transported across the intestinal epithelial cells and into the blood. Taken together, substitution of 5% of the CP in soya bean meal‐based feed with DDGS‐derived CP decreased the growth performance of newly weaned piglets due to poor digestibility and N utilization of DDGS feed, as well as untimely digestion of casein feed.     

Effects of body condition score (BCS) on steroid‐ and eicosanoid‐metabolizing enzyme activity in various mare tissues during winter anoestrus

The objective of this study was to determine the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses with varying BCSs. The BCSs of twenty non‐pregnant, anoestrous mares were determined prior to euthanasia, and tissue samples were collected from the liver, kidney, adrenal gland, ovary and endometrium. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A) and uridine 5′‐diphospho‐glucuronosyltransferase (UGT) activities were determined using luminogenic substrates. The MIXED procedure of SAS was used to test the effect of BCS on enzyme activity and differences between tissues. Activity of CYP1A in adrenals was increased (p ≤ .05) in BCS 5 versus BCSs 4 and 6. Activity of CYP1A in the liver was increased (p = .05) in BCS 4 versus BCSs 5 and 6. Activity of CYP1A was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and kidney. Activity of CYP2C was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and endometrium. Activity of CYP3A was only detectable in the liver. Activity of UGT in the kidney was decreased (p = .02) in BCS 4 versus BCSs 5 and 6. Activity of UGT was threefold greater (p < .0001) in the liver than in the kidney, whereas activity of UGT was ninefold greater (p < .0001) in the kidney than in the ovary and endometrium. In general, BCS did not alter the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses. However, tissue differences in these enzymes indicated abundant hepatic metabolism in horses, which is similar to other livestock species.     

Growth performance of piglets during the first two weeks of lactation affects the development of the intestinal microbiota

The aim of this study was to evaluate the effect of newborn piglet weight gain during the first 2 weeks of lactation on the luminal and mucosal microbiota of the ileum and colon. The microbiota from high‐weight‐gain (HWG) and low‐weight‐gain (LWG) 2‐week‐old piglets was characterized by amplicon length heterogeneity PCR (LH‐PCR) and compared using diversity indices and multivariate statistical analyses. At birth, LWG piglets weighted in average 0.26 kg less than HWG piglets (p = .002). The weight difference between LWG and HWG piglets increased with time and reached 2.1 kg after 16 days of lactation (p < .0001). Based on these growth performance differences, estimated colostrum and milk intake was greater in HWG than in LWG piglets (p < .0001). Analysis of the LH‐PCR data of the microbiota using non‐metric multidimensional scaling (NMS) and blocked multiresponse permutation procedure (MRBP) revealed that the microbiota of the HWG and LWG piglets tended to differ in ileal mucosa (p = .097) and differed in colonic lumen (p = .024). The microbiota of HWG piglets had higher levels of Bacteroidetes, Bacteroides and Ruminoccocaceae, and lower proportions of Actinobacillus porcinus and Lactobacillus amylovorus when compared with those of LWG piglets. As the weight gain of nursing piglets is highly correlated with the amount of ingested colostrum and milk, the results strongly suggest that colostrum and milk intake in the first 2 weeks of life influenced the development of the gut microbiota.     

Physiological adaptive indicators in fasted neonate broiler chicks in response to calcium gluconate injection

Four hundred and eighty mixed‐sex broiler chicks aged 3 h after hatching were allotted according to a completely random design in a 6 × 2 × 2 factorial schedule into two groups of 12 replications of 20 chicks each. The main experimental factors were fasting for 0, 6, 12, 24, 36 and 48 h after chick placement and calcium gluconate (Ca‐glu) injection (0 and 0.6 ml). Live body weight (BW) of chicks decreased linearly (Y = 43.36–0.109BW_0 h, r² = 0.876) as neonatal fasting extended. Injection of 0.6 ml Ca‐glu at 3 h post‐hatching did not affect weight loss of chicks. Yolk residuals (YR) utilized linearly (Y = 5.75–0.062YR, r² = 0.956) by 0.062 g/h in neonate fasted chicks up to 48 h, showing no effect of Ca‐glu injection. Neonatal fasting periods longer than 12 h increased liver weight (p < 0.05). The mean absolute and proportional (% of BW_0 h) breast and leg weight were reduced linearly as neonatal fasting extended (p < 0.05). Serum glucose concentration increased up to 6 h and then reduced linearly to 150 mg/dl after 48‐h fasting. The Ca‐glu treatment influenced serum glucose level for a short period up to 6 h of fasting. Serum Ca concentration sharply increased up to threefolds in the birds received Ca‐glu injection resulting in acute hypercalcemia, then decreased to the initial level after 24‐h feed withdrawal (p < 0.05). The mean serum level for creatinine, uric acid, cholesterol, HDL, albumins and total proteins significantly increased during the fasting periods of 6 to 48 h and significantly elevated in the birds receiving 0.6‐ml Ca‐glu injection compared with the non‐treated chicks (p < 0.05). It was concluded that subcutaneous administration of 0.6 ml Ca‐glu in the chick's neck did not suitably support the increased metabolic demands for glucose and calcium in feed‐deprived neonate chicks.     

A longitudinal study on the acceptance and effects of a therapeutic renal food in pet dogs with IRIS‐Stage 1 chronic kidney disease

Currently, nutritional management is recommended when serum creatinine (Cr) exceeds 1.4 mg/dl in dogs with IRIS‐Stage 2 chronic kidney disease (CKD) to slow progressive loss of kidney function, reduce clinical and biochemical consequences of CKD, and maintain adequate nutrition. It is unknown if dietary interventions benefit non‐azotemic dogs at earlier stages. A prospective 12‐month feeding trial was performed in client‐owned dogs with IRIS‐Stage 1 CKD (n = 36; 20 had persistently dilute urine with urine specific gravity (USG) <1.020 without identifiable non‐renal cause; six had persistent proteinuria of renal origin with urine protein creatinine (UPC) ratio >0.5; 10 had both). Ease of transition to therapeutic renal food and effects on renal biomarkers and quality of life attributes were assessed. Dogs were transitioned over 1 week from grocery‐branded foods to renal food. At 0, 3, 6, 9, and 12‐months a questionnaire to assess owner's perception of their pet's acceptance of renal food and quality of life was completed. Renal biomarkers, including serum Cr, blood urea nitrogen (BUN), and symmetric dimethylarginine (SDMA), and USG and UPC ratio were measured. Of 36 dogs initially enrolled, 35 (97%) dogs were transitioned to therapeutic renal food. Dogs moderately or extremely liked the food 88% of the time, ate most or all of the food 84% of the time, and were moderately or extremely enthusiastic while eating 76% of the time. All renal biomarkers (Cr, BUN, and SDMA) were decreased (p ≤ .05) from baseline at 3‐months, and remained decreased from baseline at 12‐months in dogs completing the study (n = 20). Proteinuria was reduced in 12 of 16 dogs (p = .045) with proteinuria. Owners reported improvement in overall health and quality of life attributes, and hair and coat quality (all p < .01). In summary, dogs with IRIS‐Stage 1 CKD readily transition to renal food. Decreasing serum biomarker concentrations and reduction in proteinuria suggest stabilized kidney function.     

The effect of maternal obesity on fatty acid transporter expression and lipid metabolism in the full‐term placenta of lean breed swine

This study was conducted to evaluate the influence of back‐fat thickness (BF), at mating of sows, on the maternal and newborn circulating lipids, expression of placental fatty acids (FA) transporters and lipid accumulation in placenta. Full‐term placentas were obtained by vaginal delivery from BFI (9–14 mm; n = 37), BFII (15–19 mm; n = 43) and BFIII (20–27 mm; n = 38) sows according to BF at mating, and frozen placental sections were analysed for fat accumulation. Blood samples were collected from the sows of day 105 pregnancy and from cord blood at delivery. mRNA and protein expression levels were evaluated with real‐time RT‐PCR and Western blotting. Our results demonstrated that BFII females had significantly increased litter weight and placental efficiency, decreased maternal triglyceride (TG) and non‐esterified fatty acids (NEFA) levels, decreased maternal IL‐6, TNFα and leptin levels compared to BFIII females (p < .05). BFIII sows were associated with significantly decreased newborn TG levels, increased newborn glucose, IL‐6 and TNFα levels compared to BFI or BFII sows (p < .05). BFI and BFII females had significantly decreased placental TG, NEFA and cholesterol (CHOL) contents compared to BFIII females (p < .05). Moreover, decreased CD36, FATP1, FABP4, and FABP1 mRNA and protein and FATP4 protein expression, and increased LPL activity were also observed in BFIII group compared with BFII group (p < .05). PPARγ mRNA and protein and lipogenic genes such as SREBP‐1c, ACSL1, ACCα, FAS and SCD mRNA expression were downregulated or upregulated, respectively, in the placentas of BFIII sows compared to BFI or BFII sows (p < .05). Overall, this study demonstrated that there is no advantage, in terms of litter live size, litter weight and placental FA transport and metabolism, in performing the mating of sows with BF>19 mm.     

Is the Production of Embryos in Small‐Scale Farming an Economically Feasible Enterprise?

The present assay attempts to evaluate the feasibility of using embryo transfer in small community farmers by in vivo study and by modelling the results obtained. From the total of 59 donor cows, 62.7% responded to treatment, with a significant difference (p = 0.002) in the percentage of the response between breeds, being 90.5% (19/21) in Holstein and 47.4% (18/38) in Brahman. A total of 283 embryos were graded as transferable, while 141 as non‐transferable, without difference in the percentage of transferable embryo by breed (p = 0.18). The mean of transferable embryos graded as class I and II was not different between Holstein and Brahman (p = 0.96 and p = 0.92, respectively); besides, no differences were observed in the other grades (non‐transferable). The highest difference in costs, regardless of its quality by breed, was seen in the lower levels of probable fertility of the embryo transferred, even reaching several hundred dollars. When modelling the expected costs for embryo produced and transferred, values can reach nearly $2000.00 when the probable fertility is only 10%. However, when the probable fertility was 60%, embryo cost was close to $300.00. This technology seems to be viable on average or high‐scale systems, having a superovulatory response between 60 and 80% with 4–6 transferrable embryos. Yet, in small‐scale farming, due to the reduced number of donors and/or recipients, the costs surpass the economical feasibility of the technique.     

Nasal immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis for improving ovarian responses and fertility in cross‐bred buffaloes

This study was conducted to determine the effect of immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis on ovarian responses and fertility in cross‐bred buffaloes. A total of 134 cross‐bred buffaloes were divided into four groups: groups T1 (n = 34), T2 (n = 35) and T3 (n = 31) were nasal immunized twice a day with 10 ml of 1 × 10¹⁰ CFU/ml of the C501 (pVAX‐asd‐IS) vaccine for 5, 3 and 1 day, respectively. Group C (n = 34) was nasal immunized with 10 ml PBS for 5 days. All animals were immunized twice with an interval of 14 days and administered with 200 μg of a GnRH analogue on day 28, 0.5 mg PGF_2α on day 35 and 200 μg of the same GnRH analogue on day 37. TAI was performed at 18 and 24 hr after the second GnRH treatment. Fourteen days after primary immunization, C501 (pVAX‐asd‐IS) elicited significant immune responses, and anti‐inhibin IgG antibody titres in group T1 were significantly higher (p < .01) than groups T3 and C. After the second GnRH treatment, the growth speed of the dominant follicles in group T1 was significantly faster (p < .05) than groups T3 and C. The number and diameter of large follicles (≥10 mm) as well as ovulatory follicles in group T1 were the greatest in all groups, resulting in a greater conception rate in buffaloes with positive anti‐inhibin antibodies. These results demonstrate that immunization with the C501 (pVAX‐asd‐IS) vaccine, coupled with the Ovsynch protocol, could be used as an alternative approach to improve reproductive performance in cross‐bred buffaloes.     

Methyl‐β‐Cyclodextrin Improves Sperm Capacitation Status Assessed by Flow Cytometry Analysis and Zona Pellucida‐Binding Ability of Frozen/Thawed Bovine Spermatozoa

Mammalian sperm undergo a series of biochemical transformations in the female reproductive tract that are collectively known as capacitation. Cyclodextrins added to the sperm culture medium have been described to induce in vitro sperm capacitation, enabling its use in protein‐free media. However, the additive capacitating effect of methyl‐β‐cyclodextrin (MβCD) in the medium containing bovine serum albumin (BSA) is unknown in the bovine species. In this study, we evaluated the effects of incubating frozen–thawed bovine spermatozoa in a BSA‐containing medium supplemented with MβCD on different sperm quality and functional parameters. Sperm viability decreased with the addition of MβCD in a dose‐dependent manner (p < 0.05), and DNA damage could be observed but only with the highest concentration of MβCD. However, pre‐incubation of spermatozoa in MβCD‐supplemented medium improved the capacitation status as assessed by the increase in plasma membrane fluidity, intracellular calcium concentration, induced acrosome reactivity and zona pellucida (ZP)‐binding ability (p < 0.05). Thus, we conclude that MβCD supplementation is able to enhance the capacitation status of frozen–thawed bovine spermatozoa cultured in capacitation medium containing BSA and could result in a valid strategy for its application on artificial reproductive technologies such as in vitro fertilization or intracytoplasmic sperm injection.