Sheep Placenta Research Articles

Preparation, purification and identification of antioxidant peptides from the placenta of Tibetan sheep

Preparation, purification and identification of antioxidant peptides from the placenta of Tibetan sheep

108 Early colostrum uptake by the newborn requires prepartum initiation of lactogenesis in ungulate species - Sponsored by EAAP

Abstract Due to anatomical characteristics of ungulate placenta types, immunoglobulins (IG) cannot be transferred via the placenta from the maternal to the fetal circulation during pregnancy. Therefore, passive immunization of the newborn ungulate immediately after birth is crucial for its survival. Colostrum, the first milk produced contains immunoglobulins (IG) as well as greater concentrations of several nutrients and bioactive components to provide specific immunity for the first weeks of life as well as an activation of the digestive tract and metabolism through colostral hormones, growth factors, vitamins, and special nutrients such as oligosaccharides. The uptake of colostrum by newborn calves, piglets, or foals within the first few hours after birth requires the availability of colostrum in a sufficiently liquid form. The formation of pre-colostrum starts days or even weeks before parturition, which allows concentrations of IG and other factors far beyond the concentrations in maternal blood circulation. Pre-colostrum components are partially secreted into the alveolar lumen and partially stored in the lactocytes of the mammary gland. These secretions are characterized by increased viscosity and density, which are difficult to harvest by the newborn. Through lactogenesis the secretion of milk components such as lactose is initiated, which causes a considerable water influx into the mammary secretion through an osmotic gradient between maternal blood and mammary secretion. The typical high concentrations of gestagens, mostly progesterone, inhibit the initiation of lactogenesis during pregnancy. A declining gestagen activity is the key signal to initiate a lactogenic prolactin increase and hence milk secretion. Mechanisms and timing of gestagen withdrawal differ considerably among mammalian species. Most species initiate both parturition and lactation through luteolysis (cattle, goat, pig, cat, dog, rabbit, mouse rat). In some species the placenta takes over the main portion of gestagen production (human, sheep, horse). In human, IG are transferred to the fetus during pregnancy, and the start of colostrum secretion only several days after parturition is no problem. In sheep and horses, gestagen secretion during late pregnancy is provided by the placenta. Both of these species are ungulates, and a delayed start of lactogenesis would be critical for the survival of the newborn. Therefore, the placental steroid secretion is subject to characteristic changes in both species towards the end of pregnancy to allow a timely availability of colostrum immediately after birth. At the end of pregnancy, the placenta of sheep produces estrogens at the expense of progesterone, and in horses the less potent dihydroprogesterone is produced instead of progesterone. A considerable variation in the resulting colostrum composition is likely due to the existing wide range of the interval between the commencement of lactogenesis and parturition or due to local regulatory effects at the level of individual glands.

Vaccine strains of Rift Valley fever virus exhibit attenuation at the maternal-fetal placental interface.

Rift Valley fever virus (RVFV) infection causes abortions in ruminant livestock and is associated with an increased likelihood of miscarriages in women. Using sheep and human placenta explant cultures, we sought to identify tissues at the maternal-fetal interface targeted by RVFV. Sheep villi and fetal membranes were highly permissive to RVFV infection resulting in markedly higher virus titers than human cultures. Sheep cultures were most permissive to wild-type RVFV and ΔNSm infection, while live-attenuated RVFV vaccines (LAVs; MP-12, ΔNSs, and ΔNSs/ΔNSm) exhibited reduced replication. The human fetal membrane restricted wild-type and LAV replication, and when infection occurred, it was prominent on the maternal-facing side. Type I and type III interferons were induced in human villi exposed to LAVs lacking the NSs protein. This study supports the use of sheep and human placenta explants to understand vertical transmission of RVFV in mammals and whether LAVs are attenuated at the maternal-fetal interface.IMPORTANCEA direct comparison of replication of Rift Valley fever virus (RVFV) in sheep and human placental explants reveals comparative efficiencies and permissivity to infection and replication. Vaccine strains of RVFV demonstrated reduced infection and replication capacity in the mammalian placenta. This study represents the first direct cross-host comparison of the vertical transmission capacity of this high-priority emerging mosquito-transmitted virus.

Interspecies pregnancy between Ovis canadensis mexicana and Ovis aries with reduced placental P450scc expression and intrauterine growth restriction

The present study aimed to assess the steroidogenic process in the placenta of hybrids Ovis canadensis mexicana × Ovis aries, particularly, to determine the plasma lipid profile and the relative abundance of proteins related to the steroidogenesis, to elucidate in part the mechanism of progesterone deficiency in hybrids. Plasma lipoproteins and placental cholesterol were determined by dry chemical colorimetric assays. Steroidogenic acute regulatory protein (STARD1), StAR-related lipid transfer domain protein 3 (STARD3), cholesterol side-chain cleavage enzyme (P450ssc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) were evaluated by Western blot. In addition, STARD3 was also investigated by immunohistochemical experiments in sheep placenta. Results indicated that cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and very low-density lipoprotein (VLDL) plasma levels were similar between hybrid and control pregnancy groups. Conversely, intracellular cholesterol was increased (P ˂ 0.05) in the placenta tissue of hybrids. No significant differences in the relative abundance of both STARD3, STARD1, and 3β-HSD enzyme were observed in the placenta of hybrids compared to control animals, while the relative abundance of cytochrome P450scc was lower in hybrids than in control animals (P = 0.0279). The presence of STARD3 was observed in the cytoplasmic and perinuclear regions of mononucleate and binucleate cells of the trophoblast in the sheep placenta. The significant decrease in the abundance of P450scc in hybrids’ placenta may explain the decrease in progesterone biosynthesis, which has been related to intrauterine growth restriction (IUGR) in these animals previously.

Bioactive Hydrogel Based on Collagen and Hyaluronic Acid Enriched with Freeze-Dried Sheep Placenta for Wound Healing Support.

In an increasingly aging society, there is a growing demand for the development of technology related to tissue regeneration. It involves the development of the appropriate biomaterials whose properties will allow the desired biological response to be obtained. Bioactivity is strongly affected by the proper selection of active ingredients. The aim of this study was to produce bioactive hydrogel materials based on hyaluronic acid and collagen modified by the addition of placenta. These materials were intended for use as dressings, and their physicochemical properties were investigated under simulated biological environmental conditions. The materials were incubated in vitro in different fluids simulating the environment of the human body (e.g., simulated body fluid) and then stored at a temperature close to body temperature. Using an FT-IR spectrophotometer, the functional groups present in the composites were identified. The materials with the added placenta showed an increase in the swelling factor of more than 300%. The results obtained confirmed the potential of using this material as an absorbent dressing. This was indicated by pH and conductometric measurements, sorption, degradation, and surface analysis under an optical microscope. The results of the in vitro biological evaluation confirmed the cytosafety of the tested biomaterials. The tested composites activate monocytes, which may indicate their beneficial properties in the first phases of wound healing. The material proved to be nontoxic and has potential for medical use.

Toxicity studies of condensed fuzheng extract in mice and rats

Nutritional supplements have been used to improve immune function. Condensed fuzheng extract (CFE) is a well-known traditional Chinese medicine (TCM) formula that is predominantly made from sheep placenta, Astragalus mongholicus Bunge, and Polygonatum kingianum Collett & Hemsl. However, the toxicological profile of CFE has not been determined. In this study, we investigated the acute (14 days) and sub-chronic (90 days) oral toxicities of CFE in mice and rats and the phytochemical composition of CFE. Materials and methodsFor the assessment of acute toxicity, 80 ICR mice of both sexes were randomly divided into four groups. Three groups were treated with 4500, 2250 and 1125 mg/kg/d bw CFE daily (n = 10/group per sex) for 14 days; a separate group was used as control. To test the sub-chronic toxicity, male and female Sprague Dawley rats were orally administered 8150, 4075 or 2037 mg/kg bw of CFE for 90 days; a control group was included. Hematological, biochemical, and histopathological markers were tested at the end of the experiment. The chemical composition of CFE was determined by UPLC-HRMS method. ResultsIn both acute and sub-chronic toxicity studies, no mortalities, indications of abnormality, or treatment-related adverse effects were observed. The LD50 of CFE was higher than 4500 mg/kg. There were no significant changes in the hematological and biochemical data in the treatment group compared with the control group (p > 0.05). Histopathological analyses of the heart, liver, spleen, lungs, kidneys, thymus, testes (male rats) and ovaries (female rats) revealed no anatomical changes of each organ. Phytochemical analysis of CFE revealed the presence of flavonoids (highest abundance), phenols and alkaloids. In conclusion, our results showed that CFE is a safe and non-toxic formula. We also reported phytochemicals in CFE that may possess important pharmacological effects.

Enhancing Ovarian Reserve and Dermatological Health with NanoFiltration-Enhanced Sheep Placenta Supplement: A Clinical Evaluation of Biowell Placenta & DCI 40

Enhancing Ovarian Reserve and Dermatological Health with NanoFiltration-Enhanced Sheep Placenta Supplement: A Clinical Evaluation of Biowell Placenta & DCI 40

The Immune Response in the Uteri and Placentae of Chlamydia abortus-Infected Ewes and Its Association with Pregnancy Outcomes.

The enzootic abortion of ewes, caused by the bacterium Chlamydia abortus (C. abortus), is one of the main causes of abortion in sheep. There are multiple contributory factors, including chlamydial growth, host immune response, and hormonal balance, that result in different pregnancy outcomes, such as abortion, the birth of weak lambs that may die, or healthy lambs. This study aimed to determine the relationship between phenotypical patterns of immune cell infiltration and different pregnancy outcomes in twin-bearing sheep (both lambs born dead; one alive and one dead; both alive) when experimentally infected with C. abortus. Both the sheep uteri and placentae were collected after parturition. All samples were analysed for specific immune cell features, including cell surface antigens and the T-regulatory (Treg) cell-associated transcription factor and cytokines, by immunohistochemistry and in situ hybridisation. Some of these immunological antigens were evaluated in ovine reproductive tissues for the first time. Differential patterns of T helper/Treg cells revealed significant group effects in the placentae. It suggests the potential role that the balance of lymphocyte subsets may play in affecting different pregnancy outcomes in C. abortus-infected sheep. The present study provides novel detailed information about the immune responses observed at the maternofoetal interface in sheep at the time of pre-term abortion or lambing.

Modulatory role of mTOR in trophoblast adaptive response in the early stage of placentation in sheep.

Fibroblast growth factor-2 (FGF2) is essential for early placenta development in sheep. This study shows that the mechanistic target of rapamycin is the key modulator of trophoblast adaptive response under FGF2 modulation. During the early stage of placentation in sheep, normal conceptus development is affected by trophoblast cell functionality, whose dysregulation results in early pregnancy loss. Trophoblast metabolism is supported mainly by histotrophic factors, including fibroblast growth factor-2 (FGF2), which are involved in cell differentiation and function through the modulation of specific cellular mechanisms. The mechanistic target of rapamycin (mTOR) is known as a cellular 'nutrient sensor', but its downstream regulation remains poorly understood. The hypothesis was that during trophoblast development, the FGF2 effect is mediated by mTOR signalling pathway modulation. Primary trophoblast cells from 21-day-old sheep placenta were characterised and subjected to FGF2 and rapamycin treatment to study the effects on cell functionality and gene and protein expression profiles. The model showed mainly mononuclear cells with epithelial cell-like growth and placental morphological properties, expressing typical trophoblast markers. FGF2 promoted cell proliferation and migration under normal culture conditions, whereas mTOR inhibition reversed this effect. When the mTOR signalling pathway was activated, FGF2 failed to influence invasion activity. mTOR inhibition significantly reduced cell motility, but FGF2 supplementation restored motility even when mTOR was inhibited. Interestingly, mTOR inhibition influenced endocrine trophoblast marker regulation. Although FGF2 supplementation did not affect ovine placenta lactogen expression, as observed in the control, interferon-tau was drastically reduced. This study provides new insights into the mechanism underlying mTOR inhibitory effects on trophoblast cell functionality. In addition, as mTOR is involved in the expression of hormonal trophoblast markers, it may play a crucial role in early placenta growth and fetal-maternal crosstalk.

Characterisation of cytochrome P450 isoenzyme activity in sheep liver and placental microsomes

IntroductionDrug metabolism during pregnancy is a complex process that involves maternal, placental and fetal sites of metabolism. Indeed, there is a lack of clarity provided from drug metabolism in human pregnancy due to ethical limitations. Large animal models of human pregnancy provide an opportunity to quantify activity of phase 1 drug metabolism mediated by cytochrome P450 (CYP) enzymes in the maternal, placental, and fetal compartments. Herein, we have validated a comprehensive assay to quantify maternal, placental, and fetal CYP activity. MethodsIsolated microsomes from sheep maternal liver, placenta, and fetal liver (140d gestation, term = 150d) were incubated with CYP-specific probe drugs to quantify the activity of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A. Inhibition studies were performed to validate specificity of probe drugs. The validated assay was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). ResultsCYP1A2, CYP2B6, CYP2C8, CYP2C19, CYP2D6, CYP2E1 and CYP3A were active in maternal liver. In contrast, only CYP1A2, CYP2C8 and CYP2D6 were active in the placenta, whereas CYP2B6, CYP2C8 and CYP2D6 were active in the fetal liver. Of the placental-specific CYPs validated, CYP1A2 increased in type A compared with type D placentomes, whereas CYP2C8 activity increased in type B compared with type A and C. DiscussionThis study has established conditions for compartment-specific CYP activity in the sheep maternal-placental-fetal unit using a validated and standardised experimental workflow. Compartment- and placentome type-specific CYP activity are important considerations when examining drug metabolism in the maternal-placental-fetal unit and in determining the impact of pregnancy complications.

Evaluation of Antiaging Effect of Sheep Placenta Extract Using SAMP8 Mice

Widely used in traditional medicine, sheep placenta extract (SPE) is known for its physiological effects such as wound healing, antioxidant, and anti-inflammatory properties. However, the effect of SPE on antiaging is still unclear. In this study, we investigated the effect of SPE on aging through the senescence-accelerated mouse prone 8 (SAMP8) strain. We designed an experiment using both male and female mice randomly divided into 4 groups (n = 10) as follows: Group A—control group; Group B—low-dose SPE (61.5 mg/kg BW/day); Group C—medium-dose SPE (123 mg/kg BW/day); and Group D—high-dose SPE (184.5 mg/kg BW/day). As a result of measuring the aging index parameters such as skin glossiness, spine lordosis, and kyphosis, it was found that the treatment of SPE lowered the aging index. In addition, we found that biochemical parameters such as lactic acid, glucose, ketone bodies, free fatty acids, tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6) were not changed in the experimental group treated with SPE for 13 weeks. Finally, we found that lipid peroxidation (LPO) was decreased, while the activities of catalase and superoxide dismutase (SOD) were significantly increased in the brain tissues of SPE-treated male and female mice. Supplementation of SPE lowered the oxidative stress caused by the aging process in mice without toxicity and decreased the aging index, suggesting the value of SPE as an effective antiaging treatment.

Endogenous Jaagsiekte sheep retrovirus envelope protein promotes sheep trophoblast cell fusion by activating PKA/MEK/ERK1/2 signaling

BackgroundEndogenous Jaagsiekte sheep retrovirus envelope protein (enJSRV-Env) plays an important role in trophoblast cell fusion in sheep. However, the underlying mechanism remains unclear. Methods: Primary endometrial luminal epithelial cells (LECs) were isolated from the sheep uterus and cocultured with sheep trophoblast cells (STCs). Giemsa staining was conducted to count multinucleated cells in the coculture system. Gain- and loss-of-function assays were performed to explore the role of enJSRV-Env in trophoblast cell fusion in the coculture system. Co-immunoprecipitation and mass spectrometry were carried out to identify the interacting partner of enJSRV-Env in the cocultures. Western blot analysis were conducted to determine the activation of protein kinase A (PKA)/mitogen-activated extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase 1/2 (ERK1/2) signaling. Results: Primary LECs were identified by the expression of epithelial marker cytokeratin 18. Overexpression of enJSRV-Env promoted the formation of multinucleated cells in the coculture system. enJSRV-Env activated and physically interacted with PKA, along with the activation of MEK/ERK1/2 signaling. PKA inhibition completely reversed enJSRV-Env-induced MEK/ERK1/2 activation, and ERK1/2 inhibition abolished enJSRV-Env-induced formation of multinucleated cells in the coculture system. Conclusion: enJSRV-Env promotes trophoblast cell fusion in the sheep placenta by activating PKA/MEK/ERK1/2 signaling. This finding reveals a novel mechanism underlying the contribution of enJSRV-Env to trophoblast cell fusion during placental morphogenesis.

OC01.10: Short‐term fetal cardiovascular adaptation to an artificial placenta in sheep

OC01.10: Short‐term fetal cardiovascular adaptation to an artificial placenta in sheep

In utero deposition of trace elements and metals in tissues

IntroductionAll animals, including humans, are exposed to heavy metals which are known to accumulate in different tissues, especially in bone. During pregnancy, the maternal bone turnover is increased and the metals in the mother’s body can be mobilized into the bloodstream. Heavy metals in maternal blood are known to pass through the placenta to the fetal blood and finally, deposited to bone tissue. However, there are no studies on the concentration of metals in the fetal solid tissues and until now, the rate of metal transfer from mother to fetus is not exactly known. Materials and methodsSamples of the blood, liver, placenta, and three different bones were collected from 17 pregnant ewes and their 27 fetuses. The animals had no known exposure to heavy metals. The concentrations of Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, P, Pb, Rb, Sb, Sn, Sr, Te, Ti, Tl, V, and Zn were analyzed using ICP-MS. Results and discussionThe concentration of Sb, Sn, Te, and Tl were under the detection limit in all the samples. The other metals were found in all maternal and fetal tissues, suggesting that all detectable metals cross the placenta. Blood concentrations were low compared to solid tissue concentrations. The concentrations of essential elements varied between maternal and fetal tissues, which could be explained by biological differences. The differences in concentrations of non-essential elements between the ewe and fetuses were smaller. The most significant differences were between maternal and fetal concentrations of Ba and Sr, which is at least partly explained by the mineralization degree of the bone. ConclusionHeavy metals accumulate in fetal solid tissues in sheep that are not directly exposed to heavy metals. Because of the differences in anatomy between human and sheep placenta, the accumulation in the tissue of human fetuses should be extrapolated cautiously. However, there might be some clinical relevance for fertile aged women who are exposed to heavy metals, such as women who work in the metal industry or who have undergone joint replacement surgery.

Analysis of Coxiella burnetii genomes in context of epidemic Q fever

Q fever is a zoonotic infection caused by Coxiella burnetii. Infected goats, sheep and cattle, materials of animal origin, meat and dairy products are the main sources of C. burnetii for humans. Dust C. burnetii infection is of particular importance due to the high resistance of this pathogen. C. burnetii is considered as a biological weapon and its use as an aerosol causes acute diseases with further disability. Chronic C. burnetii infection can cause endocarditis, leading to a chronic fatigue syndrome or even death. In the Netherlands, a pronounced increase in the incidence of Q fever in humans was detected from 2007 to 2010 (≥ 4,000 acute cases), mostly associated with goat farms. Analysis of 10 C. burnetii genomes allowed molecular epidemiology screening of coding and non-coding structures for studding the possible origin of the outbreak strains. Strains Z3055 (sheep placenta, Germany) and NL3262 (goat placenta, Netherlands) are the most closely related (84.9% homologous sequence). Formal order analysis distinguished 6 groups of C. burnetii strains. Adaptation to a new host emerged as a driving force for the reorganization of C. burnetii genome. The largest epidemic of Q fever could occur because of an inadequate veterinary supervision that led to the epizootic and further epidemic in the farmers producing the goat cheese. The transition of the «sheep» (Z3055-like) strain to the «goat» (NL3262) strains, characterised by a change of the host during the epizootic, could signifcantly contribute to the severity of that Q fever epidemic.

In vivo investigation of ruminant placenta function and physiology-a review.

The placenta facilitates the transport of nutrients to the fetus, removal of waste products from the fetus, immune protection of the fetus and functions as an endocrine organ, thereby determining the environment for fetal growth and development. Additionally, the placenta is a highly metabolic organ in itself, utilizing a majority of the oxygen and glucose derived from maternal circulation. Consequently, optimal placental function is required for the offspring to reach its genetic potential in utero. Among ruminants, pregnant sheep have been used extensively for investigating pregnancy physiology, in part due to the ability to place indwelling catheters within both maternal and fetal vessels, allowing for steady-state investigation of blood flow, nutrient uptakes and utilization, and hormone secretion, under non-stressed and non-anesthetized conditions. This methodology has been applied to both normal and compromised pregnancies. As such, our understanding of the in vivo physiology of pregnancy in sheep is unrivalled by any other species. However, until recently, a significant deficit existed in determining the specific function or significance of individual genes expressed by the placenta in ruminants. To that end, we developed and have been using in vivo RNA interference (RNAi) within the sheep placenta to examine the function and relative importance of genes involved in conceptus development (PRR15 and LIN28), placental nutrient transport (SLC2A1 and SLC2A3), and placenta-derived hormones (CSH). A lentiviral vector is used to generate virus that is stably integrated into the infected cell's genome, thereby expressing a short-hairpin RNA (shRNA), that when processed within the cell, combines with the RNA Induced Silencing Complex (RISC) resulting in specific mRNA degradation or translational blockage. To accomplish in vivo RNAi, day 9 hatched and fully expanded blastocysts are infected with the lentivirus for 4 to 5h, and then surgically transferred to synchronized recipient uteri. Only the trophectoderm cells are infected by the replication deficient virus, leaving the inner cell mass unaltered, and we often obtain ~70% pregnancy rates following transfer of a single blastocyst. In vivo RNAi coupled with steady-state study of blood flow and nutrient uptake, transfer and utilization can now provide new insight into the physiological consequences of modifying the translation of specific genes expressed within the ruminant placenta.

Effects of novel Fufang Biejia Ruangan Tablets with sheep placenta as substitute for Hominis Placenta on CCl4-induced liver fibrosis

ObjectiveFufang Biejia Ruangan Tablet (FBRT) is widely used for the treatment of liver fibrosis. However, Hominis Placenta (HP), as an important adjuvant of FBRT, has been restricted for medicinal using due to the limited availability, ethical controversy and safety issues. The present study aimed to investigate the therapeutic effects of novel FBRT (N-FBRT) with sheep placenta (SP) as substitute for HP on liver fibrosis and explore its possible mechanisms. Different dosages of SP in N-FBRT were also evaluated. MethodsRats were subcutaneously injected with CCl4 to induce liver fibrosis and then treated with N-FBRT and FBRT. The anti-hepatic fibrosis effect was determined based on biomarkers analysis of liver function and hepatic fibrosis, and the liver pathology was visualized by H&E staining and Masson staining. The oxidative stress and inflammatory cytokines were also detected. Immunohistochemical staining of α-SMA, real time PCR and Western blotting were performed to evaluate hepatic stellate cells (HSCs) activation and TGF-β1/Smad signaling pathway. ResultsN-FBRT and FBRT could ameliorate CCl4-induced liver fibrosis and improve liver function, as evidenced by lowering serum biomarkers levels of liver function and hepatic fibrosis, and decreasing hepatic Hyp content and collagen deposition, and improving the hepatic morphology and architecture changes. Moreover, the anti-liver fibrosis effect was better when the dosage of SP used in N-FBRT was 1/2 of HP in FBRT. Administration of N-FBRT markedly alleviated oxidative stress and inflammatory cytokines, and inhibited α-SMA expression. Furthermore, the mRNA expression of Col I, Col III, α-SMA and TGF-β1, and proteins expression of α-SMA, TGF-β1, Smad2/3 and p-Smad2/3 were significantly down-regulated by N-FBRT treatment. ConclusionSP can be used as substitute for HP to prepare N-FBRT for the treatment of liver fibrosis and the anti-liver fibrosis effect of N-FBRT is achieved by eliminating oxidative stress and inflammation, and inhibiting HSCs activation and ECM production by blocking TGF-β1/Smad signaling pathway.

264 Using in vivo RNA Interference to Investigate Ruminant Placental Function

Abstract Pregnant sheep have been used extensively for investigating pregnancy physiology, providing valuable information about the progression of ruminant pregnancy. The ability to place indwelling catheters, within both maternal and fetal vessels, allows for steady-state investigation of blood flow, nutrient uptakes and utilization, and hormone secretion, under non-stressed and non-anesthetized conditions. As such, our understanding of the in vivo physiology of pregnancy in sheep is unrivalled by any other species. However, until recently, a significant deficit existed in determining the specific function or significance of individual genes expressed by the placenta in livestock. To that end, we developed and have been using in vivo RNA interference (RNAi) within the sheep placenta to examine the function and relative importance of genes involved in conceptus development (PRR15 and LIN28), placental nutrient transport (SLC2A1 and SLC2A3), and placenta derived hormones (CSH). The lentiviral vector LL3.7 is used to generate virus that is stably integrated into the infected cell’s genome, thereby expressing a short-hairpin RNA (shRNA), that when processed within the cell, combines with the RNA Induced Silencing Complex (RISC) resulting in specific mRNA degradation or translational blockage. To accomplish in vivo RNAi, day 9 hatched and fully expanded blastocysts are infected with the lentivirus for 4–5 hours, and then surgically transferred to synchronized recipient uteri. Only the trophectoderm cells are infected by the replication deficient virus, leaving the inner cell mass unaltered, and we typically obtain 70–80% pregnancy rates following transfer of a single blastocyst. Data will be presented from two projects. One is focused on generating a deficiency in placental glucose transporters at mid-gestation, and the other on the impact of CSH RNAi during late gestation, demonstrating the utility of this experimental approach for examining gene function within the placenta of livestock. Supported by NIH-NICHD grants HD093701 and HD094952.

Off-Flavor Removal from Sheep Placenta via Fermentation with Novel Yeast Strain Brettanomyces deamine kh3 Isolated from Traditional Apple Vinegar.

Animal placentae can be used as health-promoting food ingredients with various therapeutic efficacies, but their use is limited by their unpleasant odor and taste. This study aimed to investigate the possibility of deodorization of sheep placenta via yeast fermentation. A yeast strain was successfully isolated and identified as a novel Brettanomyces strain (Brettanomyces deamine kh3). The deodorizing efficacy of fermentation of the sheep placenta with B. deamine kh3 was evaluated by 42 panels, based on evaluation of preference, ranking, and aroma profiles, and compared with normal placenta and placenta fermented with B. bruxellensis. The results of the sensory evaluation indicated that fermentation of the sheep placenta with B. deamine kh3 may improve its palatability by increasing flavors such as that of grass (tree), rubber, and burnt, and by decreasing the odor and soy sauce flavor. Solid-phase microextraction-gas chromatography (SPME-GC) showed that major off-flavors in sheep placenta, such as ammonia, dimethyl disulfide, and 1,3-dioxolane, were completely diminished in the sheep placenta fermented with B. deamine kh3. This study presents those major volatile compounds, including 2-isobutyl\\-4,4-dimethyl-1,3-dioxane, and 3-methyl-1-butanol, could be crucial in improving the palatability of the sheep placentae fermented with B. deamine kh3. This study provides a good starting point for the industrial application of a new deodorization method.

Off-Flavor Removal from Sheep Placenta via Fermentation with Novel Yeast Strain Brettanomyces deamine kh3 isolated from Traditional Apple Vinegar

Off-Flavor Removal from Sheep Placenta via Fermentation with Novel Yeast Strain Brettanomyces deamine kh3 isolated from Traditional Apple Vinegar