Vitamin and mineral supplementation and rate of gain during the first trimester of gestation affect concentrations of amino acids in maternal serum and allantoic fluid of beef heifers

The objective of this study was to evaluate the effects of feeding vitamin and mineral (VTM) supplement and (or) rate of gain (GAIN) during early gestation on amino acid (AA) concentrations in allantoic fluid (ALF) and amniotic fluid (AMF) and maternal serum. Seventy-two crossbred Angus heifers (initial BW = 359.5 ± 7.1 kg) were randomly assigned to one of four treatments in a 2 × 2 factorial arrangement with main effects of VTM supplement (VTM or NoVTM) and rate of gain (GAIN; low gain [LG], 0.28 kg/d, vs. moderate gain [MG], 0.79 kg/d). The VTM treatment (113 g•heifer⁻¹•d⁻¹, provided macro and trace minerals and vitamins A, D, and E to meet 110% of the requirements specified by the NASEM in Nutrient requirements of beef cattle. Washington, DC: The National Academies Press. doi:10.17226/19014, 2016) was initiated 71 to 148 d before artificial insemination (AI). To complete the factorial arrangement of treatments, at breeding heifers were either maintained on the basal diet (LG), or received MG diet which was implemented by adding a protein/energy supplement to the LG diet. Thirty-five gestating heifers with female fetuses were ovariohysterectomized on d 83 of gestation and maternal serum, ALF, and AMF were collected. Samples were analyzed for concentrations of neutral AA: Ala, Asn, Cys, Gln, Gly, Ile, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val; cationic AA: Arg, His, and Lys; and anionic AA: Asp and Glu. In serum, a VTM × GAIN interaction (P = 0.02) was observed for Glu, with greater concentrations for VTM-LG than VTM-MG. Concentrations of serum Cys, Met, and Trp were greater (P ≤ 0.03) for MG than LG. In ALF, concentrations of Glu were affected by a VTM × GAIN interaction, where VTM-MG was greater (P < 0.01) than all other treatments. Further, ALF from VTM had increased (P ≤ 0.05) concentrations of His, Asp, and 12 of the 14 neutral AA; whereas GAIN affected concentrations of Arg, Cys, and Asp, with greater concentrations (P ≤ 0.05) in MG heifers. In AMF, AA concentrations were not affected (P ≥ 0.10) by VTM, GAIN, or their interaction. In conclusion, increased concentrations of AA in maternal serum and ALF of beef heifers were observed at d 83 of gestation in response to VTM supplementation and rate of gain of 0.79 kg/d, which raises important questions regarding the mechanisms responsible for AA uptake and balance between the maternal circulation and fetal fluid compartments.     

The effects of maternal nutrition during the first 50 d of gestation on the location and abundance of hexose and cationic amino acid transporters in beef heifer uteroplacental tissues

We hypothesized that maternal nutrition during the first 50 d of gestation would influence the abundance of hexose transporters, SLC2A1, SLC2A3, and SLC2A5, and cationic amino acid transporters, SLC7A1 and SLC7A2, in heifer uteroplacental tissues. Angus-cross heifers (n = 43) were estrus synchronized, bred via artificial insemination, and assigned at breeding to 1 of 2 dietary intake groups (CON = 100% of requirements to achieve 0.45 kg/d of BW gain or RES = 60% of CON intake) and ovariohysterectomized on day 16, 34, or 50 of gestation (n = 6 to 9/d) in a completely randomized design with a 2 × 3 factorial arrangement of treatments. Uterine cross-sections were collected from the horn ipsilateral to the corpus luteum, fixed in 10% neutral buffered formalin, sectioned at 5 µm, and stained via immunofluorescence for transporters. For each image, areas of fetal membrane (FM; chorioallantois), luminal epithelium (ENDO), superficial glands (SG), deep glands (DG), and myometrium (MYO) were analyzed separately for relative intensity of fluorescence as an indicator of transporter abundance. Analysis of FM was only conducted for days 34 and 50. No transporters in target areas were influenced by a day × treatment interaction (P ≥ 0.06). In ENDO, all transporters were differentially abundant from days 16 to 50 of gestation (P ≤ 0.04), and SLC7A2 was greater (P = 0.05) for RES vs. CON. In SG, SLC7A1 and SLC7A2 were greater (P ≤ 0.04) at day 34 vs. day 16. In DG, SLC2A3 and SLC7A1 were greater (P ≤ 0.05) for CON vs. RES heifers; furthermore, SLC7A1 was greater (P < 0.01) at day 50 vs. days 16 and 34 of gestation. In MYO, SLC7A1 was greater (P < 0.01) for CON vs. RES and was greater (P = 0.02) at days 34 and 50 vs. day 16. There were no differences in FM (P ≥ 0.06). Analysis of all uterine tissues at day 16 determined that SLC2A1, SLC2A3, and SLC7A2 were all differentially abundant across uterine tissue type (P < 0.01), and SLC7A1 was greater (P = 0.02) for CON vs. RES. Analysis of all uteroplacental tissues at days 34 and 50 demonstrated that all transporters differed (P < 0.01) across uteroplacental tissues, and SLC7A1 was greater (P < 0.01) for CON vs. RES. These data are interpreted to imply that transporters are differentially affected by day of gestation, and that hexose and cationic amino acid transporters are differentially abundant across utero-placental tissue types, and that SLC7A1 is responsive to maternal nutritional treatment.     

Small phytoplankton and carbon export from the surface ocean

Autotrophic picoplankton dominate primary production over large oceanic regions but are believed to contribute relatively little to carbon export from surface layers. Using analyses of data from the equatorial Pacific Ocean and Arabian Sea, we show that the relative direct and indirect contribution of picoplankton to export is proportional to their total net primary production, despite their small size. We suggest that all primary producers, not just the large cells, can contribute to export from the surface layer of the ocean at rates proportional to their production rates.     

Mammary gland growth and vascularity at parturition and during lactation in primiparous ewes fed differing levels of selenium and nutritional plane during gestation

Background： Objectives were to examine the effects of selenium （Se） supply and maternal nutritional plane during gestation on mammary gland growth, cellular proliferation, and vascularity at parturition and d 20 of lactation. Rambouillet primiparous ewes （n = 84） were allocated to treatments in a 2 x 3 factorial. Factors were dietary Se （adequate Se [ASe, 11.5 μg/kg BW] or high Se [HSe, 77.0 μg/kg BVV]） and nutritional plane （60% IRES], 100% [CON], or 140% [EXC]）. At parturition, lambs were removed and 42 ewes （7/treatment） were necropsied. Remaining ewes were fed a common diet meeting requirements for lactation and mechanically milked twice daily until necropsy on d 20. At both necropsy periods, mammary glands were dissected and tissues harvested. Samples were analyzed for RNA, DNA, and protein content, cell proliferation, and vascularity. Where interactions were present （P 〈 0.05）, least squares means from the highest-order interaction are presented. Results： Final body weight of ewes was least （P 〈 0.002） in RES, intermediate for CON, and greatest for EXC, regardless of stage of the ewe at necropsy （parturition or d 20 of lactation）. In ewes necropsied at parturition, mammary glands were heavier （P = 0.02） in EXC compared to RES, with CON intermediate. Concentration of RNA （rag/g） was decreased （P= 0.01） in EXC compared to CON at parturition. There was a tendency （P= 0.07） for a Se by nutrition interaction in percentage of cells proliferating where ASe-EXC ewes had greater （P_〈 0.02） number of proliferating cells then all other treatments. Mammary vascular area tended （P = 0.08） to be affected by a Se by nutrition interaction where ASe-CON had less （P= 0.007） vascular area than HSe-CON ewes. In ewes necropsied at d 20 of lactation, the number of alveoli per area was decreased （P 〈- 0.05） in RES compared to CON and EXC-fed ewes. Conclusions： Results of this study indicate that proper maternal nutritional plane during gestation is important for mammary gland development, even out to d 20 of lactation.     

The Effectiveness of Immunotherapy in Treating Exercise-Induced Pulmonary Hemorrhage

Inflammatory airway disease has been linked to exercise-induced pulmonary hemorrhage (EIPH), and consequently, we hypothesized that immunomodulation via concentrated equine serum (CES) treatment would reduce EIPH as evidenced by red blood cell (RBC) concentrations in bronchoalveolar lavage fluid (BALF). Separate trials were conducted on Thoroughbred horses treated with either CES (n = 6) or placebo (0.9% saline; n = 4). All horses completed pre-treatment and post-treatment (2 and 4 weeks after initiating treatment) maximal exercise tests on a 10% inclined treadmill (1 m/s/min increments to fatigue) over a 10-week period (2−3 weeks between tests), with bronchoalveolar lavage (BAL) performed 30 minutes after exercise. Treatment ensued 10 days after the pre-treatment exercise test, with horses receiving a series of five CES or placebo injections 24 hours apart (20 mL intratracheal and 10 mL intravenously), with subsequent weekly injections for 5 weeks thereafter. After CES treatment, both EIPH (RBC in BALF) and inflammation (white blood cell concentration [WBC] in BALF) were significantly diminished by the 4-week posttreatment run, demonstrating 46 ± 12% and 24 ± 11% decreases, respectively (P < 0.05). In contrast, EIPH was elevated significantly at the 4-week time point, and inflammation remained constant in the placebo trial. In conclusion, these preliminary data suggest that therapeutic intervention involving immunomodulation may represent a viable approach to reducing the severity of EIPH.     

Omega-3 Fatty Acid Supplementation Reduces Basal TNFα but Not Toll-Like Receptor-Stimulated TNFα in Full-Sized and Miniature Mares

It has been well documented that omega-3 long-chain polyunsaturated fatty acids (n-3 PUFA) confer a wide variety of health benefits to humans and animals. The current study was designed to evaluate the ability of n-3 PUFA to modulate the innate immune response in two diverse breeds of horses. Ten Quarter Horse and 10 American Miniature Horse mares were assigned to either an n-3-PUFA-supplemented or a control diet (five full-sized and five miniature mares/treatment) for 56 days. The treatment diet was designed to deliver 64.4 mg/kg body weight of combined eicosapentaenoic acid and docosahexaenoic acid daily. Blood was collected through jugular venipuncture into heparinized tubes on days 0, 28, and 56. Serum PUFA analysis was conducted by gas chromatography. Peripheral blood mononuclear cell production of tumor necrosis factor-α (TNFα) in response to toll-like receptor (TLR) ligands lipopolysaccharide, flagellin, and lipoteichoic acid was estimated using an equine-specific enyme-linked immunosorbent assay. Peripheral blood samples from day 56 were also analyzed for total and differential leukocyte counts and subjected to flow cytometric analysis. Body type did not affect basal or TLR-stimulated TNFα production. Serum PUFA analysis revealed a decrease in α-linolenic acid and substantial increases in arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, and docosapentaenoic acid at both day 28 and day 56 in horses fed n-3 PUFA (P < .0001 for all). Dietary n-3 PUFA supplementation reduced (P < .05) unstimulated basal, but not TLR-stimulated, TNFα production by peripheral blood mononuclear cells. Supplementation with n-3 PUFA neither affected total or differential leukocyte counts nor selected cell surface markers. These results suggest that n-3 PUFA supplementation in the horse can modify circulating PUFA and alter the inflammatory response by reducing basal TNFα production. Furthermore, under conditions of the current study and considering the end points evaluated, the American Miniature Horse could potentially be used as a model for full-sized horse breeds.     

Omega-3 Fatty Acid Supplementation Affects Selected Phospholipids in Peripheral White Blood Cells and in Plasma of Full-Sized and Miniature Mares

Dietary omega-3 fatty acids (n-3 PUFA), notably eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), impart health benefits in humans and animals. In horses, dietary n-3 PUFAs elevate EPA and DHA and may promote anti-inflammatory effects. No reports document effects of dietary n-3 PUFA on fatty acyl components of circulating and cellular phospholipids in horses nor whether responses to dietary n-3 PUFA are similar among horse breeds. Ten Quarter Horse and 10 American Miniature Horse mares were assigned to n-3 PUFA (64.4 mg· kg body weight [BW]⁻¹·d⁻¹) or control diet for 56 days. Blood was sampled at 0, 28, and 56 days. Apparent phospholipid molecular species from several classes (phosphatidylcholine [PC]; “ether-linked” phosphatidylcholine [i.e., alk(en)yl, acyl glycerophosphocholine] [ePC]; phosphatidylethanolamine [PE]; phosphatidylinositol [PI]; and phosphatidylserine [PS]) were determined in plasma and peripheral blood mononuclear cells (PBMC) by mass spectrometry. Statistical analysis showed that six phospholipid species had diet × day interactions (P < .05) for both plasma and PBMC. Further evaluation of these species demonstrated that the mole percentage of PC(38:6), PC(40:7), PC(42:10), PE(38:5), PE(40:6), and PE(40:7) (where x:y represents total acyl carbon:total carbon-carbon double bonds) in both plasma and PBMC phospholipids was elevated in horses fed n-3 PUFA (P < .001 for all). Analysis of the acyl product ions revealed that these contained an acyl chain of mass consistent with an n-3 PUFA. Thus, supplementation increased n-3 PUFA in selected plasma and PBMC phospholipids. The absence of breed effects suggests that miniature and full-size horses responded similarly to dietary treatment.     

Mammary gland growth and vascularity at parturition and during lactation in primiparous ewes fed differing levels of selenium and nutritional plane during gestation

Background

Objectives were to examine the effects of selenium (Se) supply and maternal nutritional plane during gestation on mammary gland growth, cellular proliferation, and vascularity at parturition and d 20 of lactation. Rambouillet primiparous ewes (n = 84) were allocated to treatments in a 2 x 3 factorial. Factors were dietary Se (adequate Se [ASe, 11.5 μg/kg BW] or high Se [HSe, 77.0 μg/kg BW]) and nutritional plane (60% [RES], 100% [CON], or 140% [EXC]). At parturition, lambs were removed and 42 ewes (7/treatment) were necropsied. Remaining ewes were fed a common diet meeting requirements for lactation and mechanically milked twice daily until necropsy on d 20. At both necropsy periods, mammary glands were dissected and tissues harvested. Samples were analyzed for RNA, DNA, and protein content, cell proliferation, and vascularity. Where interactions were present (P ≤ 0.05), least squares means from the highest-order interaction are presented.

Results

Final body weight of ewes was least (P ≤ 0.002) in RES, intermediate for CON, and greatest for EXC, regardless of stage of the ewe at necropsy (parturition or d 20 of lactation). In ewes necropsied at parturition, mammary glands were heavier (P = 0.02) in EXC compared to RES, with CON intermediate. Concentration of RNA (mg/g) was decreased (P = 0.01) in EXC compared to CON at parturition. There was a tendency (P = 0.07) for a Se by nutrition interaction in percentage of cells proliferating where ASe-EXC ewes had greater (P ≤ 0.02) number of proliferating cells then all other treatments. Mammary vascular area tended (P = 0.08) to be affected by a Se by nutrition interaction where ASe-CON had less (P = 0.007) vascular area than HSe-CON ewes. In ewes necropsied at d 20 of lactation, the number of alveoli per area was decreased (P ≤ 0.05) in RES compared to CON and EXC-fed ewes.

Conclusions

Results of this study indicate that proper maternal nutritional plane during gestation is important for mammary gland development, even out to d 20 of lactation.