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.     

Intestinal development of bovine foetuses during gestation is affected by foetal sex and maternal nutrition

We aimed to evaluate the effects of maternal nutrition (MN) and foetal sex on the intestinal development of bovine foetuses throughout different days of gestation (DG). Forty‐four multiparous, dry Holstein × Gyr cows with average initial body weight of 480 ± 10 kg were fed the same diet of either restricted feeding at 1.15% of body weight (CO, n = 24) or fed ad libitum (overnourished, ON, n = 20). Six cows from CO group and five cows from ON group were slaughtered at 139, 199, 241 and 268 DG, and foetuses were necropsied to evaluate the intestinal development. The mass, length and density of foetal intestines were not affected by MN (p ≥ 0.260). An interaction between MN and DG was observed for the villi length of jejunum (p = 0.006) and ileum (p < 0.001). Villi length of jejunum and ileum was higher (p < 0.10) in foetuses from ON‐fed cows than in foetuses from CO‐fed cows at 139 DG. However, at 199 DG, the villi length of jejunum and ileum of foetuses from CO‐fed cows was higher than in foetuses from ON‐fed cows. Despite these differences, MN did not affect the villi length of jejunum and ileum at 268 DG (p > 0.10). Female foetuses had greater small intestine mass (p = 0.093), large intestine mass (p = 0.022), small intestine mass in proportion to body mass (p = 0.017) and large intestine mass in proportion to body mass (p < 0.001) than male foetuses. Female foetuses had also longer small intestine (p = 0.077) and greater small intestine density (p = 0.021) and villi length of jejunum (p = 0.001) and ileum (p = 0.010) than males. We conclude that MN affects the pathway for the development of foetal villi length throughout the gestation in bovine foetuses without changing the final villi length. Female foetuses had higher intestinal mass, density and villi length than males during the foetal phase in bovines.     

Development of maternal and foetal immune responses in cattle following experimental challenge with Neospora caninum at day 210 of gestation

This study examined the immunological responses of pregnant cattle and their foetuses following an experimental challenge with live Neospora caninum tachyzoites at day 210 of gestation. Animals were bled prior to and weekly throughout the experiment and sacrificed at 14, 28, 42 and 56 days post inoculation (dpi). At post mortem examination, samples of lymph nodes and spleen were collected from both dam and foetus for immunological analysis. Subcutaneous (sc) inoculation over the left prefemoral (LPF) lymph node of pregnant cattle at day 210 of gestation, led to the vertical transmission of parasites by 14 dpi, however no foetal deaths were observed in the infected animals. Foetuses from infected dams mounted Neospora-specific humoral and cell-mediated immune (CMI) responses by 14 dpi. These responses involved anti-Neospora IgG, antigen-specific lymphocyte proliferation, and the production of the cytokines IFN–γ, interleukin (IL)-4 and IL-10. There was also evidence of innate immunity during the response against Neospora from infected dams, with statistically significant (p < 0.05) increases in mean expression of toll like receptors (TLR)-2 on 56 dpi in maternal spleen, LPF, right prefemoral (RPF), left uterine (LUL) and right uterine (RUL) lymph nodes and TLR-9 in retropharyngeal (RLN), LPF and RPF lymph nodes from 28 dpi. Statistically significant (p < 0.05) increases in mean TLR-9 were detected in spleen samples from foetuses of infected dams, compared to the foetuses from control animals. Our results show that vertical transmission of the parasite occurred in all infected dams, with their foetuses showing effective Neospora-specific cell mediated, humoral and innate immune responses.     

Effects of Supplementing Holstein Heifers with Dietary Melatonin during Late Gestation on Growth and Cardiovascular Measurements of their Offspring

The objective was to examine the effects of supplementing dams with dietary melatonin during late gestation on offspring growth and cardiovascular measurements. On day 190 of gestation, heifers (n = 20) were blocked by body weight and randomly assigned to one of two dietary treatments consisting of 20 mg of dietary melatonin per day [melatonin (MEL)] or no melatonin supplementation [control (CON)]. Dietary treatments were terminated on day 262 of gestation. At birth, calves were separated from their dams with no further treatments. Calf (n = 18) blood pressure, cortisol, nitrites and total antioxidant capacity were collected on weeks 0, 1, 2, 3 and 4 of age. Calf hepatic portal blood flow and concentrations of insulin‐like growth factor 1 were determined on weeks 0 and 4 of age. Calf body weight, abdominal girth, hip height and wither height increased (p < 0.05) with age. An age by treatment interaction (p < 0.01) was observed for calf body weight, which was increased at weeks 8 and 9 of age in calves born to MEL heifers compared to calves born to CON heifers. Pulse pressure, mean arterial pressure, absolute hepatic portal blood flow and blood flow relative to calf body weight were not different (p > 0.05) between treatments. A main effect of calf age (p < 0.05) was observed for concentrations of insulin‐like growth factor 1, which was decreased at week 4 compared to week 0. An age by treatment interaction (p < 0.05) was observed for cortisol, which was decreased at week 2 in calves from MEL‐treated dams compared to calves from CON‐treated dams. Early post‐natal growth was altered in offspring born to dams supplemented with dietary melatonin. This could lead to further foetal programming implications in conjunction with post‐natal development.     

Effects of maternal treatment with β‐hydroxy‐β‐metylbutyrate and 2‐oxoglutaric acid on femur development in offspring of minks of the standard dark brown type

The aim of the study was to evaluate the effect of the diet, mother type and sex of the offspring on the mechanical and geometric parameters of long bones as well as bone tissue density in minks. Primiparous and multiparous dams were supplemented with β‐hydroxy β‐methylbutyrate (a metabolite of leucine, at the daily dosage of 0.02 g/kg of body weight) and/or 2‐oxoglutaric acid (a precursor of glutamine, at the daily dosage of 0.4 g/kg of body weight) during gestation. The diet did not influence bone tissue density and the length of the humerus. An increase in the length of the femur was noted in male offspring delivered by multiparous dams. The diet resulted in an increase in the weight of the humerus in males from multiparous dams and a decrease in offspring from primiparous dams. Heavier femora were noted in male offspring delivered by both types of dams. The maximum elastic strength of the humerus was higher in the offspring delivered by multiparous than primiparous dams, irrespective of the offspring sex. The diet resulted in reduction in the ultimate strength of the femur in the male offspring delivered by primiparous dams. Only females born by multiparous dams, irrespective of the diet, showed a significant increase in the cross‐sectional area of the humerus, while a significant decline was noted in males delivered by multiparous dams and in all the offspring delivered by primiparous dams. An increase in the cross‐sectional area of the femur was noted in the offspring delivered by multiparous dams, while reduction was observed in the offspring delivered by primiparous dams. These results have shown for the first time that the presence of β‐hydroxy‐β‐methylbutyrate or 2‐oxoglutaric acid in the diet of pregnant primiparous or multiparous dams unambiguously affects the geometry and mechanical properties of offspring's long bones.     

The effects of late gestation nutrient restriction of dams on beef heifer intake,metabolites and hormones during an ad libitum feeding trial

This study's objective was to determine if nutrient restriction during late gestation affected beef heifer feed intake, body weight (BW) gain and endocrine regulation during a 10‐week feeding trial. During the last 100 days of gestation, control (CON) dams were fed to increase body condition score (BCS). Whereas, nutrient‐restricted dams (NR) and NR dams protein supplemented 3 days/week (NRS) were fed to decrease BCS by 1.2. After parturition, all cow‐calf pairs were moved to a common pasture and fed in excess of requirements until weaning. At 15 months of age, heifers were randomly sorted into two pens and adjusted to a commercial total mixed ration over a 2‐week period. Blood samples and BW were taken at the initiation of feeding and on a biweekly basis for the duration of the feeding trial. Feed intake was monitored for 10 weeks using a GrowSafe System. After 10 weeks, an intravenous glucose tolerance test (IVGTT) was performed on 21 randomly subsampled heifers. During the feeding trial, NR heifers consumed more feed than CON and NRS heifers. Heifers from NR dams tended to increase BW compared to NRS and CON heifers when adjusted for initial BW. Heifers from NR and NRS dams had a greater increase in BCS compared to heifers from CON dams. Plasma glucose and insulin concentrations during the feeding trial increased in NR heifers compared to the other groups beginning at 2 and 4 weeks respectively. Plasma leptin concentrations were increased in the NR and NRS heifers compared to the CON heifers beginning at week 4 of feeding. During the IVGTT at the conclusion of the feeding challenge, plasma glucose and insulin were increased in NR heifers compared to other treatment groups. These results show that nutrient restriction during late gestation alters appetite and endocrine regulation in heifer offspring.     

Organogenesis and foetal haemodynamics during the normal gestation of healthy black‐rumped agoutis (Dasyprocta prymnolopha,Wagler, 1831) bred in captivity

The objective of this study was to define the patterns of organogenesis and foetal haemodynamics during the normal gestation of healthy agoutis (Dasyprocta prymnolopha) kept in captivity. Thirty pregnant agoutis that ranged in size from small to medium and weighed between 2.5 and 3 kg underwent B‐mode and Doppler ultrasonography for the biometric evaluation of the foetal organs. The foetal aortic blood flow proved to be predominantly systolic, and the measured flow velocity was 78.89 ± 2.95 cm/s, with a maximum pressure gradient of 2.12 ± 0.27 mmHg. The liver was characterized by its large volume, occupying the entire cranial aspect of the abdominal cavity, and it was associated cranially with the diaphragm and caudally with the stomach. The flow velocity in the portal vein was estimated to equal 12.17 ± 2.37 cm/s, with a resistivity index of 0.82 ± 0.05. The gallbladder was centrally located and protruded cranially towards the diaphragm. The spleen was visualized as an elongated structure with tapered cranial and caudal extremities, and the foetal kidneys were visualized bilaterally in the retroperitoneal region, with the right kidney positioned slightly more cranially than the left. The morphological characterization and hemodynamic analysis of the foetal organs of black‐rumped agoutis via B‐mode and Doppler ultrasonography allow determination of the vascular network and of reference values for the blood flow required for perfusing the anatomical elements essential for maintaining the viability of foetuses at different gestational ages.     

Foetal development of skeletal muscle in bovines as a function of maternal nutrition,foetal sex and gestational age

To determine the effects of maternal nutrition on modifications of foetal development of the skeletal muscle and possible increase in the potential of skeletal muscle growth in cattle, gestating cows were either fed 190% NRC recommendations (overnourished; ON) or 100% NRC recommendation (control; CO). Interaction between maternal nutrition (MN) and the foetal sex (FS) was also investigated. Foetuses were necropsied at four different time points throughout gestation (139, 199, 241 and 268 days of gestation) to assess the mRNA expression of myogenic, adipogenic and fibrogenic markers in skeletal muscle. Phenotypic indicators of the development of skeletal muscle fibres, intramuscular lipogenesis and collagen development were also evaluated. Modifications in mRNA expression of skeletal muscle of foetuses were observed in function of MN and FS despite the lack of effect of MN and FS on foetal weight at necropsy. Maternal ON increased the mRNA expression of the myogenic marker Cadherin‐associated protein, beta 1 (CTNNB1) and adipogenic markers Peroxissome proliferator‐activated receptor gamma (PPARG) and Zinc finger protein 423 (ZNF423) at midgestation. However, no differences on foetal skeletal muscle development were observed between treatments at late gestation indicating that a compensatory development may have occurred on CO foetuses making the effect of MN on skeletal muscle development not significant at late gestation. Moreover, our data have shown an evidence of sexual dimorphism during foetal stage with a greater skeletal muscle development in male than in female foetuses. In conclusion, providing a higher nutritional level to pregnant cows changes the trajectory of the development of skeletal muscle during midgestation, but apparently does not change the potential of post‐natal growth of muscle mass of the offspring, as no differences in skeletal muscle development were observed in late gestation.     

Insulin resistance: Relationship between indices during late gestation in dairy cows and effects on newborn metabolism

To investigate the relationship between insulin resistance indices [“Revised quantitative insulin sensitivity check index” (RQUICKI; RQ), “Revised quantitative insulin sensitivity check index ‐ β‐hydroxybutyrate” (RQUICKI_BHB; RQ_BHB), and “Homeostasis model assessment‐insulin resistance” (HOMA‐IR; HR)], and metabolic parameters in dams during late gestation, and their newborn calves. Blood was sampled twice weekly during the experimental period in 30 dry Holstein cows. In calves, blood sampling and body weight measurements were performed immediately after birth, and in 1‐week‐old male calves, liver and muscle biopsy samples were obtained for determining metabolic factor mRNA levels. RQ and RQ_BHB were negatively correlated with insulin, nonesterified fatty acid, BHB, and albumin and were positively correlated with leptin levels in blood during late gestation (p < .05). RQ, rather than RQ_BHB, reflected metabolism of dams, while stronger positive correlations were present between HR and blood insulin concentrations than other parameters, and calves of dams with high HR had low body weight, and high liver and muscle expression of growth hormone and insulin receptor mRNA (p < .05). RQ and HR of dams during late gestation could serve as indicators of dam metabolism and predictors of metabolism in newborn calves respectively.     

Akabane virus infection in the pregnant ewe. 1. Growth of virus in the foetus and the development of the foetal immune response

Three different pools of the CSIRO 16 strain of Akabane virus differing in their laboratory passage histories were used to inoculate 39 ewes between 32 and 36 days pregnant; 22 pregnant ewes received inocula containing no virus. There was no difference in the development, duration and titre of the viraemia and neutralising antibody response between the three infected groups of ewes. Both infected and control ewes had 141% foetuses when autopsied at 69 to 105 days gestation. Of the 55 foetuses from infected ewes 44 (80%) had gross developmental abnormalities.At autopsy of the dams Akabane virus was isolated only from the uterine caruncle. From foetal samples virus was isolated from a wide range of tissues, from one foetus at 69 days and from the blood of four foetuses at 95 to 106 days gestation. Virus was also isolated from 24 of the choriolllantoic fluid samples and from 37 placentomes of the 44 foetuses with developmental defects, in concentrations ranging from 10² to 10^5.5 TCID₅₀/ml or/g. No virus was isolated from the tissues of the control ewes or their foetuses.Neutralising antibody to Akabane virus was detected in 78% of the foetal sera from the infected group, titres ranging from 2 to 64. IgM and IgG₁ and neutralising antibody were detected in sera of 40 foetuses with developmental abnormalities including three that were of 76 to 78 days gestation. Neutralising antibody was detected only in serum that contained IgG₁ but may also have been associated with IgM in infected foetuses. IgM was detected in the serum of most foetuses including the non-infected controls, but sera from the control foetuses did not contain IgG₁ or neutralising antibody to Akabane virus. No IgG₂ or IgA were detected in any foetal serum.     

Effects of nutrient restriction and melatonin supplementation on maternal and foetal hepatic and small intestinal energy utilization

To determine how nutrient restriction and melatonin supplementation influence ewe and foetal hepatic and small intestinal energy use, 32 primiparous ewes on d 50 of gestation were fed 60% (RES) or 100% (ADQ) of NRC recommendations with 0 (CON) or 5 mg/d (MEL) of dietary melatonin. On d 130 of gestation, small intestine and liver were weighed and collected. Data were analysed as a completely randomized design with a 2 × 2 factorial arrangement of treatments. Liver weight (g/kg EBW) decreased (p = 0.02) in RES ewes. Jejunum weight (g/kg BW) increased (interaction p = 0.04) in ADQ‐MEL ewes compared with all other treatments. Total in vitro O₂ consumption (mol/min/tissue) and total citrate synthase activity (mol/min/tissue and mol/min/kg EBW) in liver decreased (p ≤ 0.03) in RES ewes. Oxygen consumption (mol/min/kg EBW) increased (interaction p = 0.02) in jejunum of ADQ‐CON versus RES‐MEL and ADQ‐CON. Citrate synthase activity (mol/min/kg of EBW) increased (interaction p = 0.03) in jejunum of ADQ‐MEL compared with RES‐MEL and ADQ‐CON. Foetal liver weight (g/kg BW) decreased (p = 0.02) in RES versus ADQ. Foetal small intestine weight (g/kg BW) decreased (interaction p = 0.05) in RES‐MEL versus ADQ‐MEL. Total O₂ consumption (mol/min/tissue) and total citrate synthase activity (mol/min/kg of BW) in foetal liver decreased (p ≤ 0.05) in RES versus ADQ. Foetal small intestinal O₂ consumption (mol/min/kg of BW) was greater (interaction p = 0.03) in RES‐CON and ADQ‐MEL than RES‐MEL and ADQ‐CON. Maternal nutrient restriction had a greater effect than melatonin supplementation on liver and jejunum mass and energy utilization in dams and foetuses. Because intestinal mass and energy utilization were more responsive to melatonin supplementation in ewes fed adequate nutrition compared with restricted ewes, melatonin may have limited use as a therapeutic supplement to help overcome potential negative effects of nutrient restriction.     

Effect of streptozotocin-induced diabetes in primiparous swine on subsequent reproductive performance

A follow-up study was conducted to determine the effects of streptozotocin-diabetes during the first parity on subsequent reproductive performance of sows. Only in the first parity, two doses of streptozotocin (50 and 100 mg/kg body weight) were administered to two groups of pregnant gilts at 80 d of gestation; a third group of gilts served as a control. Second-parity reproductive performance showed that gestation length, placental weight, mean birth weight of the litter, litter size and number of pigs born alive were not affected (P greater than .05) by maternal diabetes. Maternal serum glucose and fructose were greater (P less than .01) in high-dose sows than in the low-dose and the control dams. Serum free fatty acids (FFA) were higher (P less than .05) in high-dose dams than in control dams at d-1 and d 112 of gestation; no differences were observed between the high-dose and the low-dose during the same period. Liver and kidney weight, as well as DNA and RNA content, were greater (P less than .01) in pigs from high-dose dams than in those of the other treatments. Liver protein was elevated (P less than .01) in the progeny of high-dose dams. Dry matter and percent lipid were higher (P less than .05 and P less than .01, respectively), in pigs from high-dose sows than those from other treatment. Serum glucose, fructose and FFA of piglets were not affected by previous treatment of the dam.(ABSTRACT TRUNCATED AT 250 WORDS)     

Flutamide Influences Placental Aldo–Keto Reductase Family 1 Member C1 (AKR1C1) Expression in Pigs

Aldo–keto reductase family 1 member C1 (AKR1C1) catalyses the conversion of progesterone into inactive 20α‐dihydroxyprogesterone. It is suggested that AKR1C1 expression in the placenta prevents from the cytotoxic effect of progesterone on foetuses during late pregnancy. The aim of the study was to determine whether the anti‐androgen flutamide administered during late pregnancy (83–89 days of gestation) or before parturition (101–107 days of gestation) influences AKR1C1 expression in the porcine placenta. AKR1C1 mRNA and protein levels were measured using real‐time PCR and western blotting, respectively. Immunolocalization of AKR1C1 within placentas was also performed. Flutamide significantly increased AKR1C1 mRNA (p = 0.008) and protein (p = 0.019) expression only during the pre‐parturient period in pigs. AKR1C1 protein was immunolocalized in the epithelial and stromal cells of foetal and maternal part of placenta at both stages of gestation. Following flutamide treatment, the intensity of staining was higher (p = 0.045) on day 108 of gestation. In conclusion, porcine placental AKR1C1 expression seems to be regulated by an androgen signalling pathway and may be involved in foetal survival by preventing the detrimental effect of progesterone.     

Maternal and foetal immune responses of cattle following an experimental challenge with Neospora caninum at day 70 of gestation

The immune responses of pregnant cattle and their foetuses were examined following inoculation on day 70 of gestation either intravenously (iv) (group 1) or subcutaneously (sc) (group 2) with live NC1 strain tachyzoites or with Vero cells (control) (group 3). Peripheral blood mononuclear cell (PBMC) responses to Neospora antigen and foetal viability were assessed throughout the experiment. Two animals from each group were sacrificed at 14, 28, 42 and 56 days post inoculation (pi). At post mortem, maternal lymph nodes, spleen and PBMC and when possible foetal spleen, thymus and PBMC samples were collected for analysis. Inoculation with NC1 (iv and sc) lead to foetal deaths in all group 1 dams (6/6) and in 3/6 group 2 dams from day 28pi; statistically significant (p ≤ 0.05) increases in cell-mediated immune (CMI) responses including antigen-specific cell proliferation and IFN-γ production as well as increased levels of IL-4, IL-10 and IL-12 were observed in challenged dams compared to the group 3 animals. Lymph node samples from the group 2 animals carrying live foetuses showed greater levels of cellular proliferation as well as significantly (p ≤ 0.05) higher levels of IFN-γ compared to the dams in group 2 carrying dead foetuses. Foetal spleen, thymus and PBMC samples demonstrated cellular proliferation as well as IFN-γ, IL-4, IL-10 and IL-12 production following mitogenic stimulation with Con A from day 14pi (day 84 gestation) onwards. This study shows that the generation of robust peripheral and local maternal CMI responses (lymphoproliferation, IFN-γ) may inhibit the vertical transmission of the parasite.     

Reproductive Performance and Reconception of Nellore Cows According to Their Pure‐ or Cross‐Bred Calves

The objective of the present research was to evaluate the effect of a calf's genetic group on the productive and reproductive efficiency of its Nellore dam. Fixed‐time artificial insemination was applied to 800 cows using semen extracted from Nellore, Simmental and Angus Red bulls. Four hundred eleven cows calved, producing 119 Nellore, 103 ½Simmental–½Nellore and 189 ½Nellore–½Angus Red calves. The second mating period, which paired Nellore cows with Simmental bulls, was initiated 10 days after parturitions began and lasted for 5 months. Based on the two successive parturitions, the cumulative parturition rate for calving periods of 3, 4 and 5 months was calculated. Although no significant difference was observed for birth weight among the genetic groups, cross‐bred calves weighed, on average, 10% more than did pure‐bred calves at the age of 205 days. Nellore dams experienced a gestation period that was 7 days longer than did the cross‐bred dams, and the former showed a higher parturition rate at 90 and 120 days of the calving season, but not at 150 days (calving rates of 80.6, 76.4 and 76.2% for mothers of Nellore, ½Nellore–½Angus Red and ½Nellore–½Simmental, respectively, p > 0.05). At 90 and 120 days, Nellore dams produced more kg of calf per mated dam. In conclusion, in a short breeding season, Nellore dams nursing pure‐bred Nellore calves were found to have a higher biological efficiency compared with Nellore dams nursing cross‐bred calves.     

Proteomic analysis of amniotic and allantoic fluid from buffaloes during foetal development

The objective of this study was to describe the dynamic changes in protein composition and protein abundance in amniotic and allantoic fluids from buffaloes during gestation. Amniotic and allantoic fluids were collected during the first, second and third trimesters of gestation. The foetuses were measured and weighed. Fluid samples were centrifuged at 800 g for 10 min and then at 10,000 g for 60 min at 4°C. The supernatant was collected to determine the total protein concentration. Based on total protein concentration, an aliquot (50 μg) was used for in‐solution tryptic digestion, and mass spectrometry analysis (nano‐LC‐MS/MS) was performed. A multivariate statistical analysis of the proteomic data was conducted. Across the different stages of buffalo gestation, fifty‐one proteins were found in the amniotic fluid, and twenty‐one were found in the allantoic fluid. A total of twelve proteins were common among the stages, and four presented significant differences (VIP score α > 1). Fibronectin and alpha‐1‐antiproteinase were more abundant in the amniotic fluid than in the allantoic fluid. Alpha‐2‐macroglobulin and alpha‐2‐HS‐glycoprotein were more abundant in the allantoic fluid than in the amniotic fluid. Alpha‐2‐macroglobulin participates in remodelling and growth of the uterus at beginning of the gestation (first trimester), and these findings indicate that can serve as a potential tool for the early diagnosis of pregnancy in buffaloes.     

Effect of Pregnancy and Foetal Number on Diameter of Corpus Luteum,Maternal Progesterone Concentration and Oxidant/Antioxidant Balance in Ewes

The aim of this study was to determine the changes in diameter of corpus luteum (CL), maternal progesterone (P) concentration, lipid peroxidation and non‐enzymatic antioxidant levels along with enzymatic antioxidant activities in pregnant ewes bearing single and twin foetuses. The ewes were selected from healthy animals that were brought to the abattoir for slaughtering. The ewes were divided into three groups: Group 1 (non‐pregnant, non‐oestrous, n = 30), Group 2 (pregnant bearing a single foetus, n = 30) and Group 3 (pregnant bearing twin foetuses, n = 12) after they were slaughtered. Pregnant ewes were in the first half of the pregnancy. The diameter of CL and P concentration of pregnant ewes bearing a single foetus or twin foetuses were found higher than that found in non‐pregnant ewes. Similarly, the P concentration of pregnant ewes bearing twin foetuses was higher than that found in pregnant ewes bearing a single foetus. Malondialdehyde (MDA) level in pregnant ewes bearing twin foetuses was higher than that found in both non‐pregnant and pregnant ewes bearing a single foetus. The serum glutathione (GSH) level and glutathione‐peroxidase (GSH‐Px) activity of pregnant ewes bearing twin foetuses were found lower than that found in non‐pregnant ewes. Additionally, the GSH‐Px activity of pregnant ewes bearing twin foetuses was found lower than that found in pregnant ewes bearing a single foetus. No significant difference was found between pregnant ewes bearing female and male foetus with respect to diameter of CL, P concentration and oxidative stress parameters. There were significant positive correlations between foetal number (0, 1, 2) and diameter of CL, P concentration, MDA level, and between P concentration and diameter of CL, MDA level. However, significant negative correlations were found between foetal number (0, 1, 2) and GSH level, GSH‐Px activity, and between P concentration and GSH‐Px activity. In conclusion, the diameter of CL enlarges, P production increases and oxidant/antioxidant balance impairs because of the gestation stress in ewes during pregnancy.     

Physiological changes in the adenosine deaminase activity,antioxidant and inflammatory parameters in pregnant cows and at post‐partum

The aim of this study was to evaluate the behaviour of adenosine deaminase (ADA) activity, as well as its participation in the immunomodulation of pregnant cows. Thus, sixteen cows were divided into two groups (A and B): the group A was composed by cows not pregnant (n = 8), while the group B was composed by pregnant cows (n = 8). Serum levels of interleukin‐10 (IL‐10), IL‐6, tumour necrosis factor alpha (TNF‐α), reactive oxygen species (ROS) and C‐reactive protein (CRP), as well as ADA and glutathione S‐transferase (GST) activities, were measured on five sampling times (3, 5, 7 and 8 months of gestation, and soon after calving). Serum ADA activity was similar throughout the experiment in the cows belonging to the group A, but its activity increased during the experiment in cows from the group B, that is it was lower in the third and fifth months of pregnancy, and higher on months 7, 8 and after calving when compared to the group A. TNF‐α and IL‐6 serum levels were lower in pregnant cows compared to non‐pregnant animals; however, they significantly increased after calving. Serum levels of IL‐10 increased after 8 months of gestation, but it reduced after calving when compared to the group A, while CRP increased on month 8 of gestation and after calving compared to the group A. Pregnant cows showed lower serum ROS levels on months 3, 5 and 7 of gestation, and higher levels at the post‐partum. Serum GST activity was higher on month 5 of gestation in pregnant cows, but it was lower on months 7, 8 and in the post‐partum compared to the group A. Based on these evidence, we concluded that ADA activity and the others mediators or inflammatory modulators have important role in the maintenance of cow′s gestation due to their immunomodulatory effects.     

Altrenogest Treatment Associated with a Farrowing Induction Protocol to Avoid Early Parturition in Sows

This study investigated the effect of altrenogest treatment on the farrowing development of sows, and birth weight (BW) and piglet survival until the third day of life. Three control groups were used: (i) sows that farrowed spontaneously before 114 day of gestation (CONT <114); (ii) sows that spontaneously farrowed at ≥114 day of gestation (CONT ≥114); (iii) sows that farrowed at ≥114 day with cloprostenol treatment (CONTCLOPR). Other sows were treated with altrenogest (Regumate^®) for 3 days (days 111, 112 and 113 of gestation): one group gave birth spontaneously (ALT) and the other group received altrenogest until day 113 and cloprostenol on day 114 (ALTCLOPR). There were no differences (p > 0.05) in farrowing duration, BW, coefficient of variation (CV) of BW, stillborn piglets, mummified foetuses, percentage of light piglets and survival until Day 3 between sows with and without cloprostenol treatment, in both control (CONT ≥114 vs CONTCLOPR) and altrenogest‐treated sows (ALT vs ALTCLOPR). Further comparisons were performed taking into account three groups: sows with early delivery (CONT <114 – farrowing before 114 days of gestation; n = 56), sows with longer gestation (CONT ≥114 – with and without cloprostenol treatment sows; n = 103) and ALT sows (with and without cloprostenol treatment; n = 105). Gestation length of CONT ≥114 and ALT sows was similar (p > 0.05), but higher than in CONT <114 sows. There were no differences (p > 0.05) between groups in farrowing duration, CV of BW, and percentages of stillborn piglets and mummified foetuses. Sows of CONT <114 group had a larger litter size and a lower BW than sows of the other two groups (p < 0.05). Sows of CONT <114 group had a higher percentage of lighter piglets and a lower piglet survival rate (p < 0.05) than ALT sows. In conclusion, altrenogest treatment proved to be an efficient method to avoid early parturition in 3–5 parity sows resulting in heavier piglets at birth.