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.     

Improved identification of Pseudomonas savastanoi pv. phaseolicola at the molecular level

Polymerase chain reaction (PCR) technology was used to identify Pseudomonas savastanoi pv. phaseolicola at the DNA level. Oligonucleotide primers were designed on the basis of the DNA sequence of the phaseolotoxin gene cluster of P. s . phaseolicola . Identification of the pathogen was significantly improved and appearance of unspecific bands was greatly diminished by experimentally selecting the most suitable annealing temperature (Tm) value. We obtained a single, strong DNA band of 1.4 kb length, specific for the identification of P. s . phaseolicola , using a PCR programme based on a Tm value of 80 °C.     

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.     

Effect of early gestational undernutrition on angiogenic factor expression and vascularity in the bovine placentome

The effect of early gestation maternal undernutrition followed by realimentation on placentomal vascular growth and angiogenic factor expression was determined in multiparous beef cows bred to the same bull. Cows gestating only female fetuses (n = 30) were fed in equal numbers to meet the NRC requirements (control) or were fed below the NRC requirements to lose BW (nutrient restricted; NR) from d 30 to 125 of gestation. After slaughter on d 125 of gestation, 10 control and 10 NR cows were necropsied. The remaining NR cows (n = 5) were then fed to achieve a BCS equal to their control group contemporaries (n = 5) by d 220 of gestation. All cows were fed the control diet from d 220 until 250 of gestation, when the remaining control and NR cows were slaughtered and necropsied. At necropsy, placentomes were fixed via perfusion of the caruncular and cotyledonary arteries to determine capillary vascular density. Cotyledonary (fetal placental) and caruncular (maternal placental) tissues also were snap-frozen in liquid nitrogen, and mRNA concentrations of vascular endothelial growth factor and its 2 specific receptors, fms-like tyrosine kinase and kinase insert domain containing receptor, as well as placental growth factor, were determined. There was no effect of diet or day of gestation on the percentage of proliferating caruncular cells. Although diet did not impact cotyledonary cellular proliferation, there was an increase (P < 0.05) in the percentage of proliferating cells on d 250 compared with d 125 of gestation. Nutrient restriction from d 30 to 125 increased (P < or = 0.10) placental mRNA concentrations of placental growth factor and fms-like tyrosine kinase; however, there was no alteration in vascularity. By d 250 of gestation, NR cows had increased (P < 0.05) caruncular capillary surface density and decreased (P < 0.05) cotyledonary capillary area density, capillary number density, and capillary surface density compared with control cows. Although nutrient restriction had little effect on placental vascularity by d 125, upon realimentation, alterations in vascularity became apparent by d 250 of gestation, suggesting a placental programming effect.     

Effects of maternal nutrition and stage of gestation on body weight, visceral organ mass, and indices of jejunal cellularity, proliferation, and vascularity in pregnant ewe lambs

Peripubertal ewe lambs (44.3 +/- 1.1 kg of initial BW) were used in a 2 x 3 factorial design to test the effects of plane of nutrition (diet) and stage of gestation on maternal visceral tissue mass, intestinal cellularity, crypt cell proliferation, and jejunal mucosal vascularity. Singleton pregnancies to a single sire were established by embryo transfer, and thereafter ewes were offered a control (Control) or high (High) amount of a complete diet (2.84 Mcal/kg and 15.9% CP; DM basis) to promote slow or rapid maternal growth rates. After d 90 of gestation, feed intake of the Control group was adjusted weekly to maintain BCS and meet the increasing nutrient demands of the gravid uterus. Ewes were slaughtered at 50 d (n = 6 Control; n = 5 High), 90 d (n = 8 Control; n = 6 High), or 130 d (n = 8 Control; n = 6 High) of gestation. Ewes were eviscerated and masses of individual organs were recorded. The jejunum was sampled and processed for subsequent analyses. Final ewe BW for Control-fed ewes was similar at d 50 and 90 and increased (P = 0.10) from d 90 to 130 (46.0, 48.9, and 58.2 +/- 1.6 kg, respectively), whereas final BW increased (P 相似文献   

Effects of level and source of dietary selenium on maternal and fetal body weight, visceral organ mass, cellularity estimates, and jejunal vascularity in pregnant ewe lambs

Pregnant Targhee ewe lambs (n = 32; BW = 45.6 +/- 2.2 kg) were allotted randomly to 1 of 4 treatments in a completely randomized design to examine the effects of level and source of dietary Se on maternal and fetal visceral organ mass, cellularity estimates, and maternal jejunal crypt cell proliferation and vascularity. Diets contained (DM basis) either no added Se (control) or supranutritional Se from high-Se wheat at 3.0 ppm Se (SW) or from sodium selenate at 3 (S3) or 15 (S15) ppm Se. Diets were similar in CP (15.5%) and ME (2.68 Mcal/kg of DM) and were fed to meet or exceed requirements. Treatments were initiated at 50 +/- 5 d of gestation. The control, SW, S3, and S15 treatment diets provided 2.5, 75, 75, and 375 microg of Se/kg of BW, respectively. On d 134 +/- 10 of gestation, ewes were necropsied, and tissues were harvested. Contrasts, including control vs. Se treatments (SW, S3, and S15), SW vs. S3, and S3 vs. S15, were used to evaluate differences among Se levels and sources. There were no differences in ewe initial and final BW. Full viscera and liver mass (g/kg of empty BW and g/kg of maternal BW) and maternal liver protein concentration (mg/g) and content (g) were greater (P < 0.04) in Se-treated compared with control ewes. Maternal liver protein concentration was greater (P = 0.01) in SW vs. S3 ewes, and content was greater (P = 0.01) in S15 compared with S3 ewes. Maternal jejunal mucosal DNA concentration (mg/g) was greater (P = 0.08) in SW compared with S3 ewes. Total number of proliferating cells in maternal jejunal mucosa was greater (P = 0.02) in Se-fed compared with control ewes. Capillary number density within maternal jejunal tissue was greater (P = 0.08) in S3 compared with SW ewes. Selenium treatment resulted in reduced fetal heart girth (P = 0.08). Fetal kidney RNA (P = 0.04) and protein concentrations (mg/g; P = 0.03) were greater in Se-treated compared with control ewes. These results indicate that supranutritional dietary Se increases cell numbers in maternal jejunal mucosa through increased crypt cell proliferation. No indications of toxicity were observed in any of the Se treatments.     

Use of the DNA sequence of the intergenic spacer region between the 16S and 23S rRNA genes for the identification of Clavibacter michiganensis subsp. insidiosus at the molecular level

A study has been performed to identify Clavibacter michiganensis subsp. insidiosus at the molecular level, using the polymerase chain reaction (PCR) technique with oligonucleotide primers based on specific sequence recognition of the intergenic spacer region between the 16S and 23S rRNA genes. The pair of primers was designed on the basis of available DNA sequence data for that region in C. m. insidiosus and other bacteria. Using this pair of primers, a large amount of an amplified DNA fragment of 218 bp in length was obtained from C. m. insidiosus. The specificity of this amplification was proved by PCR analysis, using the above-mentioned pair of primers and templates from different bacteria, some related to C. m. insidiosus. The PCR products were analysed using agarose gel electrophoresis.