Pregnant Sheep Research Articles

Vaccine properties of Brucella melitensis 16MΔwzm and reactivation of placental infection in pregnant sheep.

Brucellosis, a worldwide zoonotic disease, is endemic in many developing countries. Besides causing significant economic losses for the livestock industry, it has severe consequences for human health. In endemic regions, small ruminants infected by Brucella melitensis are the main source of human brucellosis. Rev1, the only vaccine currently recommended to control the disease in sheep and goats, has several drawbacks. Rough lipopolysaccharide (R-LPS) mutants have been tested as alternatives, but most lack efficacy. Those in the Wzm/Wzt system responsible for O-polysaccharide export to the periplasm have been proposed as promising vaccine candidates, although to date they have been scarcely investigated in the natural host. In the present work, we studied the biological properties of a 16MΔwzm in-frame deletion mutant, including its safety in pregnant mice and sheep. In mice, 16MΔwzm prevented placental and fetal infections before parturition and protected against B. melitensis and Brucella ovis infections. In sheep, 16MΔwzm was equally safe in lambs, rams, and non-pregnant ewes, inducing some transient Rose Bengal reactions (<7 weeks). The serological reactions occurred earlier and more strongly in pregnant than in non-pregnant ewes and were significantly reduced when conjunctival rather than subcutaneous vaccination was used. In ewes vaccinated at mid-pregnancy, 16MΔwzm was not shed in vaginal discharges during the pregnancy and did not induce abortions/stillbirths. However, some ewes showed a transitory reactivation of infection in placentas and/or milk at parturition, accompanied by a seroconversion in smooth LPS (S-LPS) and/or R-LPS tests. Overall, 16MΔwzm can be considered as a safe vaccine for lambs, rams, and non-pregnant ewes, but its use at mid-pregnancy should be avoided to prevent vaccine dissemination at parturition. If the efficacy results against B. melitensis and B. ovis observed in mice are confirmed by further studies in the natural host, 16MΔwzm could constitute a useful vaccine.

Phosphate, calcium, and vitamin D signaling, transport, and metabolism in the endometria of cyclic ewes

BackgroundRecent evidence suggests important roles for progesterone (P4) and interferon tau in the regulation of calcium, phosphate, and vitamin D signaling in the uteri of pregnant sheep. However, the effects of P4 and estradiol (E2), with respect to the expression of their receptors PGR and ESR1, respectively, in uterine epithelia on mineral signaling during the estrous cycle has not been investigated. Estrous cycles of mature Suffolk ewes were synchronized, prostaglandin F2α was administered, and ewes were observed for estrus (designated as Day 0) in the presence of vasectomized rams. On Days 1, 9, or 14 of the estrous cycle, hysterectomies were performed.Results25-hydroxyvitamin D was more abundant in plasma from ewes on Day 14 than Day 1 (P < 0.05). Expression of fibroblast growth factor receptor 2 (FGFR2), a disintegrin and metalloprotease 17 (ADAM17), and parathyroid hormone-related protein (PTHrP) mRNAs was greater in endometria on Day 9 compared to Days 1 and 14 (P < 0.01). Similarly, expression of transient receptor potential cation channel subfamily V member 6 (TRPV6) mRNA was greater in endometria on Day 9 than Day 1 (P < 0.05). ATPase plasma membrane Ca2+ transporting 4 (ATP2B4) and S100 calcium binding protein G (S100G) mRNA expression was greater in endometria on Day 14 than on Days 1 and 9 (P < 0.01). In contrast, endometrial expression of vitamin D receptor (VDR) mRNA was lower on Days 9 and 14 than Day 1 (P < 0.01). Expression of klotho (KL) (P < 0.05) and cytochrome P450 family 24 subfamily A member 1 (CYP24) (P < 0.01) mRNAs was lower on Day 14 than Days 1 and 9. ADAM17, FGF23, CYP2R1, CYP27B1, KL, and VDR proteins immunolocalized to the uterine myometrium, blood vessels, and uterine luminal (LE), superficial glandular (sGE), and glandular (GE) epithelia. S100A9 protein was weakly expressed in the uterine myometrium, LE, sGE, and GE. Immunoreactivity of CYP2R1 and KL proteins in uterine LE and sGE was less on Day 1 than on Days 9 and 14. In contrast, S100G protein was expressed exclusively by GE, and immunoreactive S100G protein was less on Day 9. S100A12 protein localized to stromal cells of the uterine stratum spongiosum and blood vessels, but not by uterine epithelial cells.ConclusionCollectively, these results implicate E2, P4, and PGR in the regulation of phosphate, calcium, and vitamin D signaling in cyclic ewes.

Changes in expression levels of Nod-like receptors in the spleen of ewes.

Nucleotide-binding oligomerization domain receptors (NOD-like receptors, NLRs) have critical effects on interfaces of the immune and reproductive systems, and the spleen plays a key role in both innate and adaptive immune functions. It is hypothesized that NLR family participates in maternal splenic immune regulation during early pregnancy in sheep. In this study, maternal spleens were collected on day 16 of the estrous cycle, and days 13, 16 and 25 of gestation (n = 6 for each group) in ewes. Expression of NLR family, including NOD1, NOD2, class II transactivator (CIITA), NLR family apoptosis inhibitory protein (NAIP), nucleotide-binding oligomerization domain, Leucine rich repeat and Pyrin domain containing 1 (NLRP1), NLRP3 and NLRP7, was analyzed using quantitative real-time PCR, Western blot and immunohistochemistry analysis. The results revealed that expression levels of NOD1, NOD2, CIITA and NLRP3 were downregulated at days 13 and 16 of pregnancy, but expression of NLRP3 was increased at day 25 of pregnancy. In addition, expression values of NAIP and NLRP7 mRNA and proteins were improved at days 16 and 25 of pregnancy, and NLRP1 was peaked at days 13 and 16 of pregnancy in the maternal spleen. Furthermore, NOD2 and NLRP7 proteins were limited to the capsule, trabeculae and splenic cords. In summary, early pregnancy changes expression of NLR family in the maternal spleen, which may be related with the maternal splenic immunomodulation during early pregnancy in sheep.

Doppler Sonography of Umbilical Artery throughout Pregnancy in Sheep

The Color Doppler study on 6 pregnant ewes was conducted twice a week from Days 20 to 60 post-mating, then at weekly intervals till the end of experiment by the transrectal and transabdominal ultrasonography. In order to evaluate the waveforms and velocity blood flow patterns, peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistive index (RI), were measured. Data were nalysed by employing SPSS 21. The PSV and EDV increased from 38 to 139 days of gestation. The PI and RI of blood flow showed a non-significant decrease throughout gestation.

Expression pattern of calcium influx receptors in small intestine of pregnant and non-pregnant sheep

Expression pattern of calcium influx receptors in small intestine of pregnant and non-pregnant sheep

Current Energy and Lipid Metabolism Biomarkers in Sheep with Subclinical And Clinical Pregnancy Toxemia

The aim of this study was to evaluate the relationship between existing metabolic biomarkers (asprosin, FABP1, PPARα and FGF21) and clinical parameters and to determine their role in the early diagnosis of sheep pregnancy toxemia. In the study, 70 Akkaraman sheep aged three to five years in the last three weeks of pregnancy were divided into three groups: healthy group (n = 20), subclinical group (n = 30), and clinical group (n = 20). Clinical examination (body temperature, respiratory rate, and heart rate), blood serum biochemistry, and ELISA analyses were performed. In sheep with clinical and subclinical pregnancy toxemia, serum PPARα, β-HBA, NEFA, HbA1c, HDL, triglyceride, creatinine, and phosphorus levels were statistically higher, whereas serum glucose and LDL levels were lower than in healthy sheep. However, serum FGF21, AST, ALT, and VLDL values were not statistically different between the three groups. In addition, serum asprosin and FABP1 levels were higher in the subclinical group than in the clinical group. Thus it was concluded that serum asprosin, FABP1, and PPARα findings could be useful in evaluating lipid and energy metabolism in subclinical and clinical forms of pregnancy toxemia. Since this study was based on blood samples from individually reared sheep herds, many environmental factors (e.g., housing, nutrition, and population density) could not be considered. For this reason, it is thought that there is a need for experimental studies in which environmental variables can be controlled.

Maternal Acceptance of Pregnancy and Implantation Process in Sheep

Maternal Acceptance of Pregnancy and Implantation Process in Sheep

Modulation of Nod-like Receptor Expression in the Thymus during Early Pregnancy in Ewes.

Nucleotide-binding oligomerization domain receptors (NOD-like receptors, NLRs) are involved in modulating the innate immune responses of the trophoblast and the placenta in normal pregnancy. The thymus participates in regulation of innate and adaptive immune responses. However, it is unclear whether expression of NLR is modulated in the maternal thymus during early pregnancy. In this study, thymuses were sampled at day 16 of the estrous cycle, and at days 13, 16 and 25 of gestation (n = 6 for each group) from ewes after slaughter. Different stages were chosen because the maternal thymus was under the different effects of interferon-tau and/or progesterone or not. RT-qPCR, Western blot and immunohistochemistry analysis were used to analyze the expression of the NLR family, including NOD1; NOD2; major histocompatibility complex class II transactivator (CIITA); NLR family apoptosis inhibitory protein (NAIP); nucleotide-binding oligomerization domain and Leucine-rich repeat and Pyrin domain containing protein 1 (NLRP1), NLRP3 and NLRP7. The results showed that expression level of NOD1 was changed with the pregnancy stages, and expression levels of NOD2, CIITA, NAIP, NLRP1, NLRP3 and NLRP7 mRNA and proteins were peaked at day 13 of pregnancy. The levels of NOD2 and CIITA were increased during early pregnancy. The stainings for NOD2 and NLRP7 proteins were located in epithelial reticular cells, capillaries and thymic corpuscles. In summary, pregnancy stages changed expression of NLR family in the maternal thymus, which may be related to the modulation of maternal thymic immune responses, and beneficial for normal pregnancy in sheep.

Effects of tocolytic drugs (isoxsuprine hydrochloride) during the implantation period in pregnant sheep.

In this study, the effects of isoxsuprine hydrochloride applied 14 and 15 days after insemination in Anatolian Merino Sheep on lamb yield and some blood parameters were investigated. The research was conducted during the breeding season and 54 ewes inseminated on the same day were used. The ewes were assigned to three groups. Group I: For the placebo effect, physiological saline was injected on the 14th day into half of the control group and on the 15th day into the other half after insemination (n=18). Group II: Tocolytic drug was injected on the 14th day after insemination (n=18). Grop III: Tocolytic drug was injected on the 15th day after insemination (n=18). As the tocolytic drug, isoxsuprine hydrochloride (HCl) 3 ml (Utelax, Sanovel, Türki̇ye) was used intramuscularly. The number of pregnant and viviparous ewes, single and multiple birth ewes, lambs per viviparous ewes were determined as reproductive parameters. Blood progesterone, cortisol and calcium concentration were determined. As a result, it was determined that the single application of isoxsuprine hydrochloride (3 ml) in the implantation period did not have a positive or negative effect on reproductive parameters, and did not change the blood progesterone, calcium and cortisol concentration in ewes.

LBSUN310 Developmental Programming: Gestational Bpa Treatment Leads To Pancreatic Beta Cell Compromise

Abstract Developmental exposure to endocrine disrupting chemicals (EDC) such as bisphenol A (BPA) have been shown to be associated with many metabolic defects in humans. Studies in animals have documented detrimental effects of fetal exposure to BPA on metabolic variables such as insulin resistance, adiposity, and cardiac function. For instance, developmental exposure of sheep to bisphenol A (BPA), a ubiquitously present EDC, causes insulin resistance and compensatory hyperinsulinemia but the underlying mechanism of it is not fully understood. To determine if pancreatic hyperplasia or hypertrophy contributes to the hyperinsulinemic status and the ontogeny of these effects, pregnant Suffolk sheep were administered daily subcutaneous injections of BPA (0. 05, 0 .5 or 5 mg/Kg BW) in corn oil from days (D) 30-90 of gestation (term 147 days). Controls received corn oil during the same period. Pancreas were taken at Day 65 and 90 of gestation from control and 0.5mg/kg BPA (environmental exposure level)-treated female fetuses, and control and prenatal BPA-treated female offspring at 2 years of age (all 3 doses) during synchronized follicular phase of the cycle. Immunohistochemistry was used to determine beta cell number, size, and islet insulin content. In addition, to determine if the pancreas was fibrotic, islet collagen content was determined by Picro Sirius red staining and whole pancreas collagen content was measured with scroll collagen assay. Data were analyzed by ANOVA (Y2) and Students's t-test (D65 and D90), and magnitude of response by Cohen's Effect Size analysis. Findings showed large magnitude reduction of islet insulin content at day 65, significant decrease in islet insulin and beta cell count at D90 along with large magnitude decrease in pancreas/fetal weight. In contrast, in the Y2 sheep prenatal BPA treatment induced a large magnitude decrease in beta cell count (0. 05mg/kg dose group), a large magnitude increase in islet insulin content (5mg/kg dose group) and large magnitude increase in islet collagen content (all 3 BPA doses). The decrease in insulin in the D65 and D90 sheep along with the decrease in beta cells and pancreas/fetal weight at D90 indicates adverse developmental impact of BPA exposure on pancreatic beta cell differentiation. Increase in collagen accumulation in Y2 sheep likely contributes to the beta cell loss reflected as decrease in beta cell number seen in 0. 05 mg/kg dose of BPA. Absence of beta cell loss in 0.5 and 5mg/kg dose in the face of increased collagen is consistent with the nonmonotonic responses these EDCs manifest. Taken together, these results highlight the risks posed by developmental exposure to BPA on beta cell function and the corresponding risk for beta cell loss as animals age. Supported by NIH R01 grants ES016541 and ES030374 Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m.

Selenium supplementation and pregnancy outcomes

In vertebrates and invertebrates, selenium (Se) is an essential micronutrient, and Se deficiency or excess is associated with gonadal insufficiency and gamete dysfunction in both males and females, leading to implantation failure, altered embryonic development and, ultimately, infertility. During pregnancy, Se excess or deficiency is associated with miscarriage, pre-eclampsia (hypertension of pregnancy), gestational diabetes, fetal growth restriction and preterm birth. None of this is surprising, as Se is present in high concentrations in the ovary and testes, and work in animal models has shown that addition of Se to culture media improves embryo development and survival in vitro in association with reduced reactive oxygen species and less DNA damage. Selenium also affects uterine function and conceptus growth and gene expression, again in association with its antioxidant properties. Similarly, Se improves testicular function including sperm count, morphology and motility, and fertility. In animal models, supplementation of Se in the maternal diet during early pregnancy improves fetal substrate supply and alters fetal somatic and organ growth. Supplementation of Se throughout pregnancy in cows and sheep that are receiving an inadequate or excess dietary intake affected maternal whole-body and organ growth and vascular development, and also affected expression of angiogenic factors in maternal and fetal organs. Supplemental Se throughout pregnancy also affected placental growth, which may partly explain its effects on fetal growth and development, and also affected mammary gland development, colostrum yield and composition as well as postnatal development of the offspring. In conclusion, Se supplementation in nutritionally compromised pregnancies can potentially improve fertility and pregnancy outcomes, and thereby improve postnatal growth and development. Future research efforts should examine in more detail and more species the potential benefits of Se supplementation to reproductive processes in mammals.

The prediction of lambing rate in Awassi sheep using a rapid visual PAG ELISA test

The accurrate diagnosis of pregnancy and prediction of lambing rate are important tools that ensure the proper nutrition of pregnant ewes, optimize the birth weight of offspring and decrease the occurrence of pregnancy toxemia in sheep production. In this study, it was aimed to evaluate the reliability of the rapid visual PAG ELISA test in predicting the lambing rate, when performed in two different stages of pregnancy in Awassi sheep. A total of 210 ewes, belonging to a commercial flock, were assigned to two groups according to their mating date. Group 1 comprised 120 ewes, which were in the period from 28 to 38 days post-mating, and Group 2 comprised 90 ewes, which were in the period from 40 to 59 days post-mating. The rapid visual PAG ELISA test was performed on the serum samples of the ewes, according to the manufacturer’s instructions. The diagnostic test characteristics of the rapid visual PAG ELISA test were assessed for the prediction of lambing rate, using the lambing records as a gold standard. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the rapid visual PAG ELISA test were 95.79%, 52.38%, 90.10%, 73.33%, and 87.93%, respectively, in G1 and 92.75%, 70.00%, 91.43%, 73.68%, and 87.64%, respectively, in G2. The McNemar analysis showed that there was no difference between the rapid visual PAG ELISA test and lambing records in the two groups. Eighty per cent all of the ewes exposed to rams lambed. While the rapid visual PAG ELISA test predicted that 83.4% of the ewes exposed to rams would lamb, in reality it achieved that 75.6% of them lambed. The differences between the predicted and achieved lambing rates of rapid visual PAG ELISA (7.8%) indicate embriyonic/fetal mortality. The results of this study indicated that the lambing rate can be predicted with a high sensitivity from the 28th day of pregnancy, using the rapid visual PAG ELISA test.

20 Young Scholar Talk PHD: Implications of Histone Toxicity and Complement System Activity in Sheep and Cattle During Immunomodulatory States Including Respiratory Disease and Pregnancy

Abstract The immunostimulatory and toxic effects of extracellular histones have been demonstrated in humans and rodents suffering from acute respiratory distress syndrome and acute lung injury. Accumulating evidence suggests intricate interactions between extracellular histones and immune complement system proteins occur as a regulatory mechanism. However, limited information is available on the interactions of complement system activity and histone toxicity during immunomodulatory events in livestock. Therefore, our current research is aimed at investigating the contributions of histone toxicity and complement system activity in relation to respiratory disease and pregnancy in wild and domestic sheep and cattle. In an initial study, feedlot heifers were evaluated for protective capacity against extracellular histones and complement activity related to subsequent disease outcomes. At feedlot arrival, serum samples were collected from 118 heifer calves (BW 229 ± 2.4 kg) and subjected to a histone toxicity assay and a 50% hemolytic complement activity assay. Heifers were retrospectively assigned as either never receiving treatment with antibiotics for bovine respiratory disease (NT; n = 80) or calves that died from bovine respiratory disease within 1 wk of entering the feedlot (DA; n = 9). Serum from DA animals was less (P &amp;lt; 0.001) protective against histone toxicity compared to NT heifers. Likewise, complement activity of DA animals was reduced (P &amp;lt; 0.05) compared to NT animals. These results suggest cattle predisposed to severe cases of respiratory disease have impaired complement system activity presumably leading to reduced protection against histone toxicity. In a second study, wild Alaskan Dall sheep (Ovis dalli dalli) and domestic sheep (Ovis aries) were evaluated for susceptibility to histone toxicity. Serum was collected from Debouillet sheep (n = 150) with no clinical signs of illness and serum of wild Alaskan Dall sheep (n = 135) was collected from live capture and release animals. Dall sheep serum was less (P &amp;lt; 0.05) protective against histone toxicity compared to Debouillet sheep. Low protection against histone toxicity may be related to higher reported disease states for wild sheep. Furthermore, recognizing that disease modulates immune function, it is also recognized pregnancy relies on immune system activation. Therefore, pregnant sheep serum was evaluated to determine the effect of histone toxicity and complement system activity. Serum was collected from Debouillet ewes (n = 8) and analyzed for histone toxicity protectiveness and complement activity. Serum was collected from ewes prior to breeding (d -0) and at d 1 and 17 of the estrous cycle. Serum protective capacity against histones and complement activity was not significantly different on d -0 compared to d 1 and d 17. Collectively, these data implicate the toxic effect of extracellular histones and complement system activity during respiratory disease states with less obvious observations during pregnancy.

Changes in expression of nuclear factor kappa B subunits in the ovine thymus during early pregnancy

There is a pregnant maternal immunological tolerance that protects the fetus and promotes its growth, and nuclear factor kappa B (NF-κB) family participates in the regulation of innate immune and adaptive immune responses. The thymus is related to establishing central tolerance, and early pregnancy has effects on expression of a good number of genes and proteins in the maternal thymus in sheep. However, it is unclear whether early pregnancy changes expression of NF-κB subunits in the ovine thymus. In this study, the thymic samples were collected from day 16 of non-pregnant ewes, and days 13, 16 and 25 of pregnant ewes, and the expression of NF-κB members (NF-κB1, NF-κB2, RelA, RelB and c-Rel) was analyzed through real-time quantitative PCR, Western blot and immunohistochemical analysis. The results showed that c-Rel mRNA and protein upregulated at day 25 of pregnancy, and NF-κB1 mRNA and proteins increased at days 16 and 25 of pregnancy, and RelB mRNA and proteins enhanced during early pregnancy. However, expression levels of NF-κB2 and RelA were decreased during early pregnancy, but upregulated from day 13 to 25 of pregnancy. In addition, the RelA protein was located in the epithelial reticular cells, capillaries and thymic corpuscles. This paper reported for the first time that early pregnancy induced expression of NF-κB1, RelB and c-Rel, but inhibited expression of NF-κB2 and RelA in the maternal thymus during early pregnancy, which is involved in the central immune tolerance, and helpful for successful pregnancy in sheep.

Single nucleotide polymorphisms in surfactant protein A1 are not associated with a lack of responsiveness to antenatal steroid therapy in a pregnant sheep model.

Treatment with antenatal steroids (ANS) is standard practice for reducing the risk of respiratory distress in the preterm infant. Despite clear overall benefits when appropriately administered, many fetuses fail to derive benefit from ANS therapies. In standardized experiments using a pregnant sheep model, we have demonstrated that around 40% of ANS‐exposed lambs did not have functional lung maturation significantly different from that of saline‐treated controls. Surfactant protein A is known to play an important role in lung function. In this genotyping study, we investigated the potential correlation between polymorphisms in SFTPA1, messenger RNA and protein levels, and ventilation outcomes in animals treated with ANS. 45 preterm lambs were delivered 48 h after initial ANS therapy and 44 lambs were delivered 8 days after initial ANS therapy. The lambs were ventilated for 30 min after delivery. SFTPA1 mRNA expression in lung tissue was not correlated with arterial blood PaCO2 values at 30 min of ventilation in lambs delivered 48 h after treatment. SFTPA1 protein in lung tissue was significantly correlated with PaCO2 at 30 min of ventilation in lambs ventilated both 48 h and 8 days after ANS treatment. Six different single nucleotide polymorphisms (SNPs) in the Ovis aries SFTPA1 sequence were detected by Sanger Sequencing. No individual SNPs or SNP haplotypes correlated with alterations in PaCO2 at 30 min of ventilation or SFTPA1 protein levels in the lung. For the subset of animals analyzed in the present study, variable lung maturation responses to ANS therapy were not associated with mutations in SFTPA1.

Trace Elements in Pregnant and Non-pregnant Sheep and Goats and Their Relation to Steroid Hormones

Background: One of the major factors affecting the physiology of pregnancy and reproductive cycle is the trace elements intake and their levels in serum of brood ewes and does. The main objective of this investigation was to study the association between serum concentration of trace elements, ovarian findings, stage of pregnancy and steroid hormones in sheep and goats. Methods: Total of 129 ewes (68 pregnant and 61 non-pregnant) and 54 does (32 pregnant and 22 non-pregnant) were examined clinically and ultrasonographically for pregnancy diagnosis and monitoring the ovarian findings. Pregnant animals were categorized according to the gestational stage into 1 to 2, greater than 2 to 3, greater than 3 to 4 and greater than 4 months. According to the ovarian findings, non-pregnant animals were categorized as having mature corpus luteum (CL), large follicle ( greater than 5 mm) and corpus luteum (CL+F), growing follicles (GF, less than 5.mm) and mature follicle (MF, greater than 5 mm). Animals were bled and serum was harvested and assayed for estrogen (E2), progesterone (P4), manganese (Mn), selenium (Se), iron (Fe) and zinc (Zn) levels. Result: Mn (P=0.01), Fe (P=0.05) and Zn (P=0.001) were lower in pregnant compared with non-pregnant ewes. Fe was significantly higher (P=0.03) in non-pregnant compared with pregnant does at any gestational stage. Ewes with MF had higher Se (P=0.02) and Fe (P=0.001) compared with other ewes. Fe (P=0.001) and Zn (P=0.000) were higher in does having MF compared with other does. A positive association between E2 and serum Fe in non-pregnant ewes (P=0.03) and does (P=0.05) was found. It can be concluded that pregnancy associated with a decrease in serum trace elements in sheep and goats.

The Hematological and Biochemical Profiles of Wonosobo Sheep Blood in Various Physiological Conditions

The study aimed at examining the hematological and biochemical profiles of Wonosobo sheep blood in various physiological conditions. There were 32 Wonosobo sheep in the various physiological conditions, including 17 ewe lamb of 1-2 months of age, 5 pregnant female sheep of 12-48 months of age at 3 months of pregnancy, 5 ewe of 12-48 months of age in second lactation period of 1-2 months and 5 ram of 12-48 months of age. Each of the sheep was put in individual sheepfold and given Pakchong-1 grass, dried kangkong, and concentrate. Drinking water was given ad libitum. Blood sample was drawn from jugular vein and put into tubes for blood hematological and biochemical analyses. The results of the study showed that total leukocyte and lymphocyte were high in young sheep, while neutrophils was high in pregnant female sheep (P&lt;0.05). There was not any significant physiological impact on hemoglobin, MCV, MCH, MCHC, RDW, hematocrit, platelet, monocyte, eusinophil, and basophil (P&gt;0.05). The highest ALP activity and Pi levels were found in the ewe lamb, while low creatinine was found in the ram (P&lt;0.05). The parameter of the activities of SGPT, SGOT, LDH, calcium, kalium, chloride, sodium, magnesium, TIBC, UIBC, TS, globulin, albumin, total protein, BUN, creatinine, glucose, cholesterol, triglyceride, HDL and LDL was not affected by physiological conditions (P&gt;0.05). It was concluded that the impact of the physiological conditions of the Wonosobo sheep caused high total leukocyte and lymphocyte in ewe lamb and high neutrophil in pregnant female sheep, the increase in the Pi level and ALP activity of the ewe lamb, and high creatinine in the ram

Investigation of the effectiveness of ultrasonography in determining pregnancy and the number of fetuses on the 35th day of pregnancy in Awassi sheep

In this study, it was aimed to determine the effectiveness of transabdominal ultrasonographic imaging on the 35th day of pregnancy in determining the pregnancy status and number of fetuses in sheep. 69 Awassi sheep were used in the study. Sheep were mated after oestrus synchronization during the breeding season. On the 35th and 50th days after mating, a pregnancy examination was performed twice transabdominally in each animal by ultrasonography. Sheep with single/multiple offspring were noted according to examination performing on day 35 and lambing record. According to ultrasonography results, early foetal death rate was determined as 6.1%. Sensitivity, specificity, positive predictive value, negative predictive value ratios for pregnancy examination findings and single/multiple offspring data by transabdominal ultrasonography on day 35 were determined as 91.30%, 100%, 100%, 83.3% and 38.46%, 81.25%, 76.92%, 44.82%, respectively. While the consistency of transabdominal ultrasonographic imaging for pregnancy on day 35 and 50 were high agreement (Kappa=0.864, p&amp;lt;0,001), consistency of single/multiple pregnancy findings on day 35 and at birth were low (Kappa=0,170, p&amp;gt;0.05). The rates of transabdominal ultrasonographic examination on day 35 for correct diagnosis of pregnancy status and the number of fetuses were found as 93.93% and 54.76%, respectively. Finally, it can be concluded that transabdominal ultrasonographic examination on day 35 for early pregnancy diagnose in sheep is highly effective in determination of pregnant sheep, and recurrent examination on days 35 and 50 may be useful for detecting of early foetal deaths. In order to determine the number of fetuses, repeated examinations should be performed in the following days of pregnancy.

Ultrasound parameters of early pregnancy and Doppler indices of blood vessels in the placenta and umbilical cord throughout the pregnancy period in sheep

BackgroundUltrasonography is one of the most important techniques that enable the detection and monitoring of pregnancy. One such study using this technique is the assessment of the hemodynamics of fetal and umbilical blood vessels.However, there is little data on blood flow in the placentomes, which is the basic structural unit of the sheep’s placenta. Therefore, the aim of this study was to determine the Doppler parameters in the arterial vessels of the caruncles, cotyledons and the umbilical cord as well as measuring venous flow rates during the entire gestation period of the sheep. Additionally, the usefulness of various other ultrasound parameters in the early diagnosis of pregnancy in sheep was analyzed.ResultsMost of the Doppler parameters in umbilical, cotyledonary and caruncular arteries were significantly correlated with the day of pregnancy (p < 0.01). In the early stages of pregnancy, the peak systolic velocity (PSV), regardless of the location of the artery, was significantly lower than that in the later stages of pregnancy (p < 0.01). PSV was also found to be significantly higher in the umbilical artery than in the cotyledonary and caruncular arteries (p < 0.01).Until the 50th day of pregnancy, the end diastolic velocity (EDV) was not found in the umbilical and cotyledonary arteries. EDV was significantly higher in the caruncular arteries than in the cotyledonary and umbilical arteries (p < 0.01). The resistance index (RI) and pulsatility index (PI) in the early stages of pregnancy were found to be significantly higher than that in the later stages of pregnancy (p < 0.01). The RI and PI were significantly lower in the caruncular arteries than in the arteries of the cotyledons and umbilical cord (p < 0.01). In the umbilical vein, all Doppler parameters were observed to be significantly higher than those in the placentomal veins (p < 0.01 or p < 0.05). Using transrectal ultrasound, pregnancy was detected between 20 and 28 days after mating. The ovaries were observed to have corpora lutea, the diameter of which was fairly consistent from the 17th to the 56th day of pregnancy.ConclusionsIt has been demonstrated that both the location of the arterial vessel in the placental-umbilical circulation and the gestational age have a significant impact on hemodynamic parameters. The results also provide new insights about the blood flow in caruncular and cotyledonary arteries, which could contribute to a more holistic understanding of hemodynamic changes in the placentas of sheep. Analyzing haemodynamic parameters in the umbilical and placental veins are preliminary studies in sheep, but it could inspire further research in this field. Furthermore, the research conducted confirms the practicality and convenience of transrectal ultrasonography in the early diagnosis of pregnancy in sheep and also indicates that the identification and imaging of the corpus luteum using B-mode ultrasonography can be a very early and simple method of confirming effective mating in sheep.

Central nervous system distribution of buprenorphine in pregnant sheep, fetuses and newborn lambs after continuous transdermal and single subcutaneous extended-release dosing

Buprenorphine is used during pregnancy for the treatment of opioid use disorder. Limited data exist on the central nervous system (CNS) permeation and distribution, and on the fetal exposure to buprenorphine. The aim of our study was to determine the extent of buprenorphine distribution to CNS in the pregnant sheep, and their fetus at steady-state, and their newborn lambs postdelivery, using three different dosing regimens. Twenty-eight pregnant ewes in late gestation received buprenorphine via 7-day transdermal patch releasing buprenorphine 20 µg/h (n=9) or 40 µg/h (n=11), or an extended-release 8 mg/week subcutaneous injection (n=8). Plasma, cerebrospinal fluid, and CNS tissue samples were collected at steady-state from ewes and fetuses, and from lambs 0.33 – 45 hours after delivery. High accumulation of buprenorphine was observed in all CNS tissues. The median CNS/plasma concentration -ratios of buprenorphine in different CNS areas ranged between 13 and 50 in the ewes, and between 26 and 198 in the fetuses. In the ewes the CNS/plasma -ratios were similar after the three dosing regimens, but higher in the fetuses in the 40 µg/h dosing group, medians 65 – 122, than in the 20 µg/h group, medians 26 – 54. The subcutaneous injection (theoretical release rate 47.6 µg/h) produced higher concentrations than observed after 40 µg/h transdermal patch dosing. The median fetal/maternal concentration -ratios in different dosing groups ranged between 0.21 and 0.54 in plasma, and between 0.38 and 1.3 in CNS tissues, respectively, with the highest ratios observed in the spinal cord. Buprenorphine concentrations in the cerebrospinal fluid were 8 – 13 % of the concurrent plasma concentration in the ewes and 28 % in the fetuses. Buprenorphine was quantifiable in the newborn lambs’ plasma and CNS tissues two days postdelivery. Norbuprenorphine was analyzed from all plasma, cerebrospinal fluid, and CNS tissue samples but was nondetectable or below the LLOQ in most. The current study demonstrates that buprenorphine accumulates into CNS tissues at much higher concentrations than in plasma in pregnant sheep, fetuses, and their newborn lambs even 45 hours after delivery.