bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid synthesis of ultralong Fe(OH)3:Cu(OH)2 core–shell nanowires self-supported on copper foam as a highly efficient 3D electrode for water oxidation
Authors: Hou Chun-Chao , Wang Chuan-Jun , Chen Qian-Qian , Lv Xiao-Jun , Fu Wen-Fu , Chen Yong
Published: December 2016

Ultralong Fe(OH)3:Cu(OH)2 core–shell nanowires were facilely grown on an open 3D Cu electrode to function as a highly efficient electrocatalyst for water oxidation.
Atmospheric oxidation of halogenated aromatics: comparative analysis of reaction mechanisms and reaction kinetics
Authors: Kovacevic Goran , Sabljic Aleksandar
Published: December 2016

This study provides valuable insight into the mechanism of tropospheric degradation and fate of halogenated aromatic systems.
Trophic transfer of TiO2 nanoparticles from marine microalga (Nitzschia closterium) to scallop (Chlamys farreri) and related toxicity
Authors: Wang Zhenyu , Xia Bin , Chen Bijuan , Sun Xuemei , Zhu Lin , Zhao Jian , Du Peng , Xing Baoshan
Published: December 2016

In the present study, the trophic transfer of TiO2 NPs from marine microalga Nitzschia closterium to scallop Chlamys farreri was investigated. over the potential risk and environmental impact of manufactured nanoparticles (NPs) have increased in marine environments.
Addressing a bottle neck for regulation of nanomaterials: quantitative read-across (Nano-QRA) algorithm for cases when only limited data is available
Authors: Gajewicz A. , Jagiello K. , Cronin M. T. D. , Leszczynski J. , Puzyn T.
Published: December 2016

The development ofin silicomethods that support human health and environmental risk assessment of engineered nanomaterials is nowadays of high interest, because the application of those methods enables to fill the existing experimental data gaps.
Hydrothermal disruption of algae cells for astaxanthin extraction
Authors: Cheng Xiang , Riordon Jason , Nguyen Brian , Ooms Matthew D. , Sinton David
Published: December 2016

Hydrothermal disruption of Haematococcus pluvialis cysts at high temperature and pressure is demonstrated on a microfluidic platform for the extraction of astaxanthin.
Synthetically tuned structural variations in CePdxGe2−x(x = 0.21, 0.32, 0.69) towards diverse physical properties
Authors: Sarkar Sumanta , Banerjee Swastika , Halappa Pramod , Kalsi Deepti , Mumbaraddi Dundappa , Ghara Somnath , Pati Swapan K. , Sundaresan A. , da Silva Ivan , Rayaprol Sudhindra , Joseph Boby , Peter Sebastian C.
Published: December 2016

Three structural variations of CePdxGe2−xwith versatile properties were synthesized by varying the Pd : Ge ratio.
Mechanochemical catalytic oxidations in the solid state with in situ-generated modified IBX from 3,5-di-tert-butyl-2-iodobenzoic acid (DTB-IA)/Oxone
Authors: Mishra Abhaya Kumar , Moorthy Jarugu Narasimha
Published: December 2016

Highly reactive IBX – generated from o-iodobenzoic acid and Oxone – permits a variety of catalytic oxidations in the solid state.
Furanmonogones A and B: two rearranged acylphloroglucinols with a 4,5-seco-3(2H)-furanone core from the flowers of Hypericum monogynum
Authors: Xu Wen-Jun , Luo Jun , Li Rui-Jun , Yang Ming-Hua , Kong Ling-Yi
Published: December 2016

Rearranged acylphloroglucinols with an unprecedented 4,5-seco-3(2H)-furanone skeleton, furanmonogones A (1) and B (2), were isolated from the flowers of Hypericum monogynum.
Correction: Titanium aminophosphates: synthesis, characterization and crystal violet dye degradation studies
Authors: Rajini Anumula , Adepu Ajay Kumar , Chirra Suman , Siliveri Suresh , Venkatathri Narayanan
Published: December 2016

Correction for ‘Titanium aminophosphates: synthesis, characterization and crystal violet dye degradation studies’ by Anumula Rajini et al., RSC Adv., 2016, 6, 507–514.
Dysphagia and Long-Term Feeding Difficulties in the Pediatric Brain Tumor Population
Authors: Kilcommons Antonia , Rawlinson Deborah
Published: December 2016

This paper sets out the issues encountered by speech-language pathologists (SLPs) when working with children with dysphagia in neuro-oncology. There is little evidence documented on the likely presentation in children with brain tumors and the long-term outcomes for children's eating and drinking. Therefore, the authors in this paper outline their experiences of supporting children post-surgery, during chemotherapy and radiotherapy, and in end of life with regard to their feeding. This is a discussion piece based on clinician experience in the field and supported by referenced evidence from relevant texts. It focuses on neuro-oncology; posterior fossa tumors, complications that arise post-surgery, pontine tumors, feeding patterns in survivorship, and the impact of treatment regimens on oral intake. The article documents the type of difficulties seen in this population including the holistic management of dysphagia and aspiration risk, including the wider psycho-social and sensory-based feeding issues that can arise as a result of a child's cancer treatment.
Considerations in Dysphagia Management Following Esophagectomy
Authors: Evangelista Lisa M. , Coyle James L.
Published: December 2016

Esophageal cancer is the sixth leading cause of death from cancer worldwide. Esophageal resection is the mainstay treatment for cancers of the esophagus. While curative, surgical resection may result in swallowing difficulties that require intervention from speech-language pathologists (SLPs). Minimally invasive surgical procedures for esophageal resection have aimed to reduce morbidity and mortality associated with more invasive techniques. Both intra-operative and post-operative complications, regardless of the surgical approach, can result in dysphagia. This article will review the epidemiological impact of esophageal cancers, operative complications resulting in dysphagia, and clinical assessment and management of dysphagia pertinent to esophageal resection.
Formula of parallel straight planar conductors for minimum ohmic resistance
Authors: Kim D.‐H. , Park Y.‐J.
Published: December 2016

A formula for the optimal relation between the width of a planar conductor (w0), the total width of parallel straight planar conductors (W), and the number of the parallel conductors (N) for the minimum ohmic resistance is presented. The formula is verified for the frequency range between 2 and 27.12 MHz according to N by using the finite element method. By experimentation, it is also shown that the proposed formula can be applied to design low‐loss circular planar coils.
Optimal rate profile for multi‐user multi‐rate transmission systems by bivariate fixed‐point analysis
Authors: He Meilin , Song Guanghui , Cheng Jun
Published: December 2016

A K‐user multi‐rate code is proposed for a Gaussian multiple access channel with binary inputs, equal‐power, and symbol synchronisation. In this multi‐rate transmission, K users are equally divided into M groups. For each user in the mth group, a rate‐ regular repeat‐accumulate code serially concatenated with a length‐ spreading is employed. The transmitted rate of each user in the mth group is . At the receiver, iterative joint decoding (IJD) and hybrid interference cancellation (HIC) schemes are considered. For each decoding scheme, a bivariate fixed‐point analysis is applied to explicitly represent as a function of mutual information outputs. On the basis of these basic explicit representations, a united unreliable region is given, where users in at least one group are undecodable. The complementary set of the united unreliable region gives an optimal rate profile that achieves the maximum sum rate. Numerical results show that, for the IJD scheme with M increments, the maximum sum rate increases, approaches the Shannon limit, and exceeds that in conventional equal rate transmission. The maximum sum rate of the HIC scheme, which provides much lower decoding complexity than the IJD scheme, is superior to the conventional successive interference cancellation scheme.
Phase diagram of crushed powders
Authors: Bodard Sébastien , Jalbaud Olivier , Saurel Richard , Burtschell Yves , Lapebie Emmanuel
Published: December 2016

Compression of monodisperse powder samples in quasistatic conditions is addressed in a pressure range such that particles fragmentation occurs while the solid remains incompressible (typical pressure range of 1–300 MPa for glass powders). For a granular bed made of particles of given size, the existence of three stages is observed during compression and crush up. First, classical compression occurs and the pressure of the granular bed increases along a characteristic curve as the volume decreases. Then, a critical pressure is reached for which fragmentation begins. During the fragmentation process, the granular pressure stays constant in a given volume range. At the end of this second stage, 20%–50% of initial grains are reduced to finer particles, depending on the initial size. Then the compression undergoes the third stage and the pressure increases along another characteristic curve, in the absence of extra fragmentation. The present paper analyses the analogies between the phase transition in liquid-vapour systems and powder compression with crush-up. Fragmentation diagram for a soda lime glass is determined by experimental means. The analogues of the saturation pressure and latent heat of phase change are determined. Two thermodynamic models are then examined to represent the crush-up diagram. The first one uses piecewise functions while the second one is of van der Waals type. Both equations of state relate granular pressure, solid volume fraction, and initial particle diameter. The piecewise functions approach provides reasonable representations of the phase diagram while the van der Waals one fails.
On the measurement of the threshold electric field for runaway electron generation in the Frascati Tokamak Upgrade
Authors: Popovic Z. , Esposito B. , Martín-Solís J. R. , Bin W. , Buratti P. , Carnevale D. , Causa F. , Gospodarczyk M. , Marocco D. , Ramogida G. , Riva M.
Published: December 2016

Experiments have been carried out to evaluate the threshold electric field for runaway generation during the flat-top phase of ohmic discharges in the Frascati Tokamak Upgrade tokamak. An investigation of the conditions for runaway electron generation and suppression has been performed for a wide range of plasma parameter values. The measured threshold electric field is found to be significantly larger (∼2−5 times) than predicted by the relativistic collissional theory of runaway generation, ER=ne e3 lnΛ/4π ε02 me c2, and can be explained to a great extent by an increase of the critical electric field due to the effect of the electron synchrotron radiation losses. These findings are consistent with the results of an ITPA joint experiment to study the onset, growth, and decay of relativistic runaway electrons [Granetz et al., Phys. Plasmas 21, 072506 (2014)]. Confirmation of these results for disruptions with high electric field might imply significantly lower requirements on electron densities for suppression and prevention of runaway formation in ITER.
Foundational theories in topological physics garner Nobel Prize
Authors: Chang Sung
Published: December 2016

The laureates bucked conventional wisdom to redefine the meaning of phases and phase transitions in one- and two-dimensional materials.
Exploring early differentiation and pluripotency in domestic animals
Authors: Roberts R. Michael , Yuan Ye , Ezashi Toshihiko
Published: December 2016

This short review describes some general features of the origins of the pluripotent inner cell mass and epiblast during the early development of eutherian mammals and the two kinds of embryonic stem cell (ESC), naïve and primed type, that have been produced from these structures. We point out that the derivation of pluripotent stem cells from domesticated species continues to be fraught with difficulties, most likely because the culture requirements of these cells are distinct from those of mouse and human ESCs. Generation of induced pluripotent stem cells (iPSCs) from the domesticated species has been more straightforward, although the majority of the iPSC lines remain dependent on the continued expression of one or more integrated reprogramming genes. Although hope for the potential usefulness of these cells in genetic modification of livestock and other domestic species has dimmed, ESCs and iPSCs remain our best source of self-renewing populations of pluripotent cells, with potential usefulness in preserving and propagating valuable animal breeds and making contributions to fields such as regenerative medicine, toxicology and even laboratory meat production.
Logistics of large scale commercial IVF embryo production
Authors: Blondin P.
Published: December 2016

The use of IVF in agriculture is growing worldwide. This can be explained by the development of better IVF media and techniques, development of sexed semen and the recent introduction of bovine genomics on farms. Being able to perform IVF on a large scale, with multiple on-farm experts to perform ovum pick-up and IVF laboratories capable of handling large volumes in a consistent and sustainable way, remains a huge challenge. To be successful, there has to be a partnership between veterinarians on farms, embryologists in the laboratory and animal owners. Farmers must understand the limits of what IVF can or cannot do under different conditions; veterinarians must manage expectations of farmers once strategies have been developed regarding potential donors; and embryologists must maintain fluent communication with both groups to make sure that objectives are met within predetermined budgets. The logistics of such operations can be very overwhelming, but the return can be considerable if done right. The present mini review describes how such operations can become a reality, with an emphasis on the different aspects that must be considered by all parties.
Caprine arthritis encephalitis: an example of risk assessment for embryo trading
Authors: Fieni Francis , Lamara Ali , Ali Al Ahmad Mohamad Zuher , Cortez-Romero Cesar , Pellerin Jean- Louis
Published: December 2016

The risk of transmission of caprine arthritis encephalitis virus (CAEV) during embryo transfer has been demonstrated in vivo through the detection of CAEV proviral DNA in: (1) flushing media for embryo collection; (2) cells of the cumulus oophorus surrounding the oocytes, ovarian follicle, oviduct and uterine tissues; and (3) testis, epididymis, vas deferens and vesicular glands. Experimentally infected embryos without a zona pellucida (ZP), washed 10 times with Minimum Essential Media (MEM) and 5% Fetal Calf Serum (FCS) solution, were capable of transmitting CAEV. In vitro we demonstrated that granulosa, oviductal, epididymal and embryo cells are fully susceptible to CAEV infection and allow active replication. However, AI with in vitro-infected semen can result in the production, after ten washing, of CAEV-free embryos, and ten washing in vitro- or in vivo-infected embryos with an intact ZP, or ten washing oocytes with an intact ZP, resulted in the production of virus-free female gametes or embryos that can be used for IVF or embryo transfer. Therefore, we have demonstrated that: (1) that CAEV-free embryos can be produced by IVF using spermatozoa infected in vitro by CAEV; and (2) embryo transfer can be used under field conditions to produce CAEV-free kids from CAEV-infected biological mothers.
10 PREDICTION OF CALVING TIME USING BODY-SURFACE TEMPERATURE SENSORS AND PEDOMETERS IN BEEF CATTLE
Authors: Nishimura Y. , Mise M. , Imai K. , Dochi O.
Published: December 2016

Predicting the onset of calving is important for the prevention of stillbirths. This study aimed to investigate whether using body-surface temperature and number of steps at the time of calving could be used to predict calving time. Three heifers and 4 cows of the beef cattle variety were used in this study. All animals were housed in calving pens measuring 4.2 × 3.6 m each. Body-surface temperature was recorded using a sensor that was fixed ventrally to the body of the tail head (NIAH, NARO, Tsukuba, Japan). The number of steps the beef cattle took was measured using pedometers (Gyuho, Comtec, Miyazaki, Japan), where each 10 steps resulted in 1 count. The temperature sensors and pedometers were placed on the animals for at least 10 days before the estimated date of delivery. Body-surface temperature and pedometer readings were automatically collected every 2 min and every hour. The data used to predict calving were the means of the body-surface temperature and steps taken, which were recorded from the day the sensors were mounted until 1 day after calving. All data were statistically analysed using t-tests. The mean body-surface temperature before calving was 35.4°C (10 h), which was significantly lower than that recorded 3 days before calving (36.2°C; P < 0.05). The mean body-surface temperature of beef cattle 10 to 6 h before calving (36.0°C) was not significantly different from the mean temperature 3 days before calving. However, from 5 to 1 h before calving, the mean body-surface temperature (34.8°C) was significantly lower than that 3 days before calving (P < 0.05). The mean step number of 101.4 counts, taken 10 h before calving, was significantly increased compared with the mean step number of 44.7 counts 3 days before calving (P < 0.05). The mean step number of 59.7 counts from 10 to 6 h before calving was not significantly different from readings taken 3 days before calving. The mean step number count from 5 to 1 h before calving was 143.1 counts, which increased significantly from readings taken 3 days before calving (P < 0.05). In conclusion, ventral tailhead body-temperature sensors and pedometers can be used to predict the onset of calving in beef cattle. Their body-surface temperature, along with the number of steps taken before calving, decreased and increased, respectively, in the 5 h leading up to calving.
This study was supported by the Council for Science, Technology and Innovation (CSTI), Cross-ministerial Strategic Innovation Promotion Program (SIP), and the Technologies for creating next-generation agriculture, forestry and fisheries (funding agency: Bio-oriented Technology Research Advancement Institution, NARO).
108 IS THE BINDING OF COXIELLA BURNETII TO THE ZONA PELLUCIDAE FOLLOWING IN VITRO INFECTION OF IN VITRO-PRODUCED GOAT EMBRYOS CONCENTRATION DEPENDENT?
Authors: Fieni F. , Alsaleh A. , de Souza-Fabjan J. M. , Mermillod P. , Corbin E. , Nascimento P. , Bruyas J. F. , Pellerin J. L.
Published: December 2016

Previous experiments using in vitro infection have shown that at concentrations of 109 bacteria/mL, Coxiella burnetii strongly adheres to the zona pellucidae (ZP) of caprine embryos produced in vitro or in vivo (Alsaleh et al., 2013). However, spontaneous infection results in bacterial concentrations of between 106 and 107 bacteria/mL (Rodolakis, 2006; Alsaleh et al., 2011). The aim of this study was to determine whether the concentration of Coxiella burnetii affected its ability to bind to the ZP in vitro. A total of 120 ZP-intact 8- to 16-cell embryos, produced in vitro from ovaries collected at slaughter, were infected with Coxiella burnetti (strain CbC1) produced via ovoculture at 109 mL−1 (3 batches of 10 embryos), 107/mL (5 batches of 10 embryos), 105 mL−1 (3 batches of 8 embryos). After overnight incubation at 37°C in 5% CO2, the embryos were recovered and washed in batches, in 10 successive baths of PBS with 5% FCS, in accordance with International Embryo Technology Society guidelines. The 10 wash baths were collected separately and centrifuged for 1 h at 13,000 × g. The presence of C. burnetii was determined by conventional PCR in each batch of embryos and in the pellets of the 10 wash baths (Table 1). As demonstrated previously, Coxiella DNA was detected in embryo batches after 10 washes when a concentration of 109 bacteria/mL was used for in vitro infection, but this binding property did not occur at lower bacterial concentrations. We can conclude that the attachment of Coxiella burnetii to the zona pellucida of in vitro-produced embryos is concentration dependent. This finding illustrates the limitations of in vitro experiments to study the risk of pathogen transmission via embryo transfer.

Table 1.
Detection of Coxiella burnetii (CB) in successive embryo washing baths and batches of 8 to 10 infected ZP-intact 8- to 16-cell embryos after 10 wash cycles, using conventional PCR (C-PCR), as a function of the concentration of CB used for in vitro infection and determined by quantitative PCR (Q-PCR)
111 BIOCHEMICAL COMPOSITION OF FOLLICULAR FLUID IN RELATION TO THE STIMULUS TO INDUCE OVULATION IN ALPACAS (VICUGNA PACOS)
Authors: Huanca W. , Castro A. , Gomez N. , Cordero A.
Published: December 2016

Alpacas, like other camelids, are induced ovulators. A study was designed to determine the effect of the ovulation-inducing stimulus on the biochemical composition of follicular fluid. Adult female alpacas (n = 18) were examined daily for 3 days by transrectal ultrasonography using a 5-MHz linear-array transducer (Aloka SSD-500, Tokyo, Japan). When the largest growing ovarian follicle was ≥7 mm, alpacas were given 1.0 mL of seminal plasma intramuscularly (SP, n = 9) or 40 µg of busereline acetate intramuscularly (GnRH, n = 9). A transvaginal transducer with an attached needle guide (Aloka UST-945BP-5) was used for collection of follicular fluid 22 h post-induction. Follicular contents were then centrifuged at 800 × g for 20 min to separate the fluid from the cells. The follicular fluid was collected and stored at –20°C until analysis with a semi-automatic biochemical analyzer (SINOWA, China). The results were glucose 49.17 and 47.95 (mg/dL; P > 0.05), total protein 1.85 and 1.15 (g/dL; P < 0.05), albumin 1.11 and 1.13 (g/dL; P > 0.05), triglycerides 3.94 and 3.16 (mg/dL; P > 0.05), cholesterol 39.01 and 42.5 (mg/dL; P > 0.05), phosphatase 32.68 and 21.36 (IU/L; P < 0.05), alanine aminotransferase 3.66 and 5.07 (IU/L; P > 0.05), and lactate dehydrogenase 42.17 and 27.27 (IU/L; P > 0.05) for SP or GnRH treatments, respectively. Results suggest the need to continue research to explain the effect of possible differences in total protein, cholesterol, and phosphatase on oocyte-expressed genes and follicular development.
Research was supported by the project no. 405-PNICP-PIAP-2014-UNMSM.
119 ASSESSMENT OF OVARIAN FOLLICULAR DYSPLASIA UTILIZING ULTRASOUND AND HISTOLOGIC EXAMINATION
Authors: Gard J. A. , Roberts J. , Braden T. , Mansour M. , Yelich J. , Irsik K. , Rae O. , Wenzel J. G.
Published: December 2016

A slaughterhouse study commissioned by Florida Cattleman’s Association in 2007 identified ovarian follicular dysplasia (OFD) as a primary cause of infertility in Florida beef cows. Ovaries with OFD have progressive bilateral development of solid clustered follicles containing multiple Call-Exner bodies that originate in the rete ovarii and the hilar region, and progress into the cortex to eventually form bilateral Sertoli-type granulosa theca cell tumours (GTCT). The objectives of this study were to assess the distribution of OFD in cull animals and to evaluate utilisation of ultrasound for diagnosis of OFD in cattle. Ultrasound images of the right and left ovaries from 390 cull cows and heifers representing 4 Florida ranches were made with 5-MHz linear probes (Aloka, Ibex). Then, 10 to 12 females per ranch were followed to slaughter the proceeding day for collection of reproductive tracts. The fixed ovaries were measured, sectioned para-sagittally through the hilus, photographed, and arranged in histology cassettes for complete examination of the cut surface. Large ovarian structures including corpus luteum, Graafian follicles, atretic follicles, dysplastic follicles, rete ovarii, dysplastic follicles, and tumours were counted and measured for each ovary. Ovaries with OFD were graded I to IV. Grade I OFD contained small individual dysplastic follicles with diameter less than 200 µm mostly limited to the rete ovarii and medulla. Grade II OFD possessed dysplastic follicles greater than 200 µm diameter that were present in the medulla and cortex. Grade III OFD had extensive multi-sized dysplastic follicles scattered throughout the entire cortex of the ovary and Grade IV OFD had Sertoli-type GTCT. Grade II–IV often had dystrophic mineralization of dysplastic follicles. Gross morphology of fixed sagittal sections and ultrasound images were blindly compared against OFD grade in 40 individual ovaries. The OFD was identified at slaughter in 29/41 cows and in 1/5 of heifers. The distribution of OFD for 30 affected females was Gr I 16/30, Gr II 9/30, Gr III 4/30, and Gr IV 1/30. Characteristics that could be detected by routine ultrasound included increased size and length, increased hyperechogenicity and decreased number of fluid-filled follicles. Hyperechogenic shadows were evident in higher grade OFD. The study demonstrated that Grade III and IV OFD can be observed by routine ultrasound but Grade I and II may require higher resolution ultrasound probes, imaging analysis software, or Doppler ultrasound.
132 OVERVIEW OF FERTILITY TRAITS IN RUSSIAN HOLSTEIN BULLS USING GENOME-WIDE ASSOCIATION
Authors: Sermyagin A. A. , Naryshkina E. N. , Gladyr E. A. , Yanchukov I. N. , Brem G. , Zinovieva N. A.
Published: December 2016

Holsteins are the most widely distributed cattle breed in Russia. To achieve improvement of both milk production and fertility traits, principles were elaborated to use genomic approaches together with breeding strategies to develop a common breeding index. Combining semen-quality measures for sires with the cows’ reproduction records makes it possible to resolve the problems with decreased conception rates and semen production. Our objective was to search the genomic regions and associated markers for bulls’ fertility traits in Russian Holstein cattle. For the first step, 282 bulls were genotyped using the Bovine SNP50 BeadChip (Illumina Inc., San Diego, CA, USA). The data set originated from the Moscow AI station including records for semen volume (SV), sperm concentration (SC), average SV for multiple ejaculates collected from one sire (ASV), motility (Mt), and number of spermatozoa (NS). The data set consisted of 14,490 records, which varied between 10 and 328 per animal, with 104 sires in the data set. The average age of sires was 80.1 ± 0.3 months. For sires with own records, the deregressed estimated breeding values were calculated using BLUP AM, whereas for the sires without records direct genomic values (DGV) were calculated through GBLUP. The DGV values were used as the animal’s pseudo-phenotypes for a common data set. After quality check in Plink 1.07, 41,435 SNP were taken. Bonferroni correction test for detection of significant associations was applied as P < 1.21 × 10−6. The values for the heritability were 0.322, 0.202, 0.366, 0.014, and 0.169 for SV, SC, ASV, Mt, and NS, respectively. The SNP with a direct significant effect on SV and ASV were identified: ARS-BFGL-NGS-100206 (P = 1.5 × 10−7), BTA-89872-no-rs (P = 6.0 × 10−8), ARS-BFGL-NGS-33119 (P = 2.7 × 10−7), Hapmap26947-BTA-133787 (P = 5.0 × 10−7), BTA-30092-no-rs (P = 9.8 × 10−7), ARS-BFGL-BAC-6515 (P = 1.0 × 10−7), and BTB-01625220 (P = 4.2 × 10−8). For SC and NS traits, the polymorphisms ARS-BFGL-NGS-57433 (P = 7.4 × 10−7) and ARS-BFGL-NGS-12662 (P = 5.1 × 10−7) were found. Two QTL for Mt including 11 SNP were detected on BTA1 (146.3–146.9 Mb) and BTA9 (76.9–77.1 Mb). The DGV effect of SNP (R2) ranged from 8.0 to 10.4% of additive genetic variance. The genes SOX5 (BTA12), PERP (BTA9), and EPT1 (BTA11) were associated with embryonic development, cell death, or apoptosis and synthesis of selenoproteins. Our results may be used to select donor cows for oocytes or embryo collection in the focus of genomic selection.
Supported by the Federal Agency for Scientific Organizations #0600–2014–0007.6 and by the Russian Ministry of Education and Science #RFMEFI60414X0062.
136 IMPACT OF SELECTION SYSTEM BY KINETICS ON THE EARLY EMBRYONIC DEVELOPMENT IN BOVINE OVUM PICKUP-IVF EMBRYOS
Authors: Takayama M. , Moriyoshi M. , Dochi O. , Imai K.
Published: December 2016

Recently, in vitro-produced (IVP) embryos have been increasingly produced using ovum pickup (OPU) and IVF in cows worldwide. However, the conception rate of IVP embryos is lower than that of in vivo-derived embryos. This study was conducted to determine the proportion of embryos that led to a high conception rate when the embryos were selected according to the 4 predicting factors. A total of 30 Holstein and 20 Japanese Black cows were used, and 81 OPU-IVF sessions were performed from October 2014 to May 2016. The collected cumulus-oocyte complexes (COC) were cultured for 22 h. Capacitated sperm (at a final concentration of 5 × 106 spermatozoa/mL) were incubated with COC for 6 h. After insemination, presumptive zygotes were separated from cumulus cells and sperm by pipetting. Then, the presumptive zygotes were cultured for 9 days in CR1aa supplemented with 5% calf serum by using a micro-well culture dish (Dai Nippon Printing, Tokyo, Japan). The kinetics of embryo development was observed at 27, 31, and 55 h post-insemination (hpi). The 4 factors used to select embryos were as follows: (1) time at which first cleavage occurred (less than 27 hpi, or less than 31 hpi, in case any of the zygotes did not cleave at 27 hpi in each culture dish); (2) 2 blastomeres after first cleavage at 31 hpi; (3) absence of fragments after first cleavage at 31 hpi; and (4) 8 or more blastomeres at 55 hpi. The number of blastocysts was analysed at 7, 8, and 9 days post IVF. Additionally, the number of produced embryos that could be used for embryo transfer (ET) was determined. The data were analysed using the chi-square test. The total numbers of blastocysts and produced embryos were 615 and 503, respectively. The numbers of blastocysts and produced embryos selected using the combination of factors 1 to 4 were 200 (32.5%) and 169 (27.5%), respectively. The numbers of blastocysts and produced embryos selected using factor 1 were 397 (64.6%) and 340 (67.6%), using factor 2 were 445 (71.3%) and 378 (75.1%), using factor 3 were 364 (81.8%) and 307 (81.2%), and using factor 4 were 374 (60.8%) and 308 (61.2%), respectively. The numbers of blastocysts and produced embryos that were rejected using a combination of factors 1 to 4 were 123 (27.5%) and 90 (17.9%), respectively. The conception rate of fresh embryo transfer was 46.6% (n = 73). We found that the conception rate of the embryos selected using factors 1 to 4 was significantly (P < 0.05) higher than that of embryos without one factor or more [60.0% (n = 35) v. 29.4% (n = 34)]. These results show the applicability and efficiency of the 4 factors for producing embryos with a high competence for conception.
137 EFFECT OF THE ADDITION OF FOLIC ACID TO MATURATION AND CULTURE MEDIA ON DEVELOPMENT OF THE BOVINE BLASTOCYST AND ITS SURVIVAL RATE AFTER FREEZE-THAWING
Authors: Sato S. , Dochi O. , Imai K.
Published: December 2016

Reactive oxygen species (ROS) are the main causes of cell damage in bovine embryos in vitro. Folic acid (FA) is an antioxidant that protects cells from ROS. We studied the effect of the addition of FA to maturation and culture media on development of bovine blastocysts and their survival rate after freeze-thawing. Cell-oocyte complexes (COC) were allowed to mature in HEPES (25 mM)-buffered TCM199 (TCM199) supplemented with 5% calf serum (CS), 0.02 AU mL−1 of FSH, and FA (0, 2.5, 25, and 50 µM) for 20 hours (20–25 COC/100-µL droplet of the medium). After 6 hours of gamete co-culture (5 × 106 sperm/mL), presumptive zygotes were cultured in CR1aa medium supplemented with 5% CS and FA (0, 2.5, 25, and 50 μM) for 9 days (day of fertilization = Day 0). Expanded blastocysts that developed from Day 7 to 9 were frozen for further study. Each embryo was frozen in Dulbecco’s PBS (D-PBS) supplemented with 20% CS, 1.5 M ethylene glycol (EG), and 0.1 M sucrose (SUC). Embryos were equilibrated with their freezing medium for 15 min and loaded individually into a 0.25-mL straw. These straws were put into the cooling chamber of a programmable freezer precooled at −7°C. After 2 min, straws were seeded and held for 13 min at −7°C. Next, straws were cooled to −30°C at −0.3°C/min before being plunged into liquid nitrogen. Frozen embryos were thawed by allowing straws to stand in air for 7 s and warming them in a 30°C water bath for 20 s. Thawed embryos were washed twice with D-PBS supplemented with 20% fetal calf serum (FCS), which was warmed to 38°C. They were immersed into the same medium at 38°C for 10 min, and each embryo was cultured in a 20-μL droplet of TCM199 supplemented with 10% FCS and 0.1 mM β-mercaptoethanol (TCM-199-βME) for 72 h. Embryo cleavage rate was observed at 55 h post-insemination. Blastocyst rates were analysed at 9 days post-insemination. Rates of embryos developing into reexpanded, hatching, and hatched blastocyst stages were determined after 72 h of thawing. All data were analysed by the chi-square test and Fisher’s exact test. Cleavage and blastocyst rates after insemination at 55 hours and 9 days, respectively, were not significantly different among media containing 0 μM (n = 278; 74.1% and 39.9%), 2.5 μM (n = 260; 74.2% and 45.8%), 25 μM (n = 258; 75.6% and 45.7%), and 50 μM (n = 253; 76.3% and 42.7%) FA. Survival and hatching rates of frozen and thawed expanded blastocysts after 72 h in culture were 62.5% and 56.3%, respectively, in 0 μM FA (n = 16); 85.2% and 74.1% in 2.5 μM FA (n = 27); 66.7% and 62.5% in 25 μM FA (n = 24); and 68.0% and 64.0% in 50 μM FA (n = 25). Blastocysts cultured in media containing 2.5 μM FA tended to have a higher survival rate than those cultured in media containing 0 μM FA, although this difference was not significant (P = 0.09). Inclusion of FA did not appear to influence development or post-thaw survival of bovine blastocysts produced in vitro.
141 BOVINE EMBRYO DEVELOPMENT RATES ARE AFFECTED WHEN OOCYTES ARE MATURED IN DIFFERENT VIALS CONTAINING HEPES/BICARBONATE BUFFERED MEDIUM
Authors: Hashem N. , Secher J. O. , Pryor J. H. , Long C. R. , Looney C. R. , Avery B. , Hyttel P. , Stroebech L.
Published: December 2016

Laboratory ware for the in vitro-produced embryos is generally made from embryo-tested plastic instead of glass. The quality of the plastic is crucial for the outcome because plastic is often toxic to gametes (Nijs et al. 2009 Fertil. Steril. 92, 527–535). In addition, gas molecules permeate through the plastic at a rate that depends on a variety of factors, such as diffusion coefficient and thickness of the plastic. In an incubator with appropriate concentration of CO2 and vented culture vessels, the gas permeability of the plastic is not important. When oocytes are transported outside a controlled atmosphere, gas permeability, toxicity, and oocyte cumulus cell CO2 metabolism could perturb the outcome. Medium containing bicarbonate buffer increases pH outside of a controlled atmosphere within minutes, whereas medium buffered with HEPES maintains suitable pH for hours. Previously, we tested that gas permeability differs among plastic vials and glass vials with no cumulus-oocyte complexes (COC) by measuring pH after 2, 5, and 24 h at the same temperature. The objective of this study was to compare pH post-maturation, blastocyst development rates on Day 8 post-IVF (Day 0 = IVF) between 2 different 1.2-mL polypropylene cryovials (A: VWR DK, 479-1219; B: Sigma-Aldrich, St. Louis, MO, USA, CLS430289), glass vial (VWR DK, NSCAC4015-96), and 4-well plate (4WP) as control (Thermo Fisher Scientific, Waltham, MA, USA, 144444). A total of 1135 abattoir-derived COC in Exp. 1 and 133 in Exp. 2 were divided equally between the treatments (20–25 COC per vessel). Vials/4WP contained 0.8/0.5 mL of BO-IVM HEPES, a HEPES/bicarbonate medium (IVF Bioscience; BO-HEPES-IVM, UK). Maturation lasted 22 to 24 h at 38.8°C in an incubator with either a humidified atmosphere of 5.5% CO2 in air (Exp. 1) or with no CO2 contact (Exp. 2). In Exp. 1, oocyte vials were matured without a vial lid while in Exp. 2 vial lids were closed. Statistical analysis was performed with chi-square and mean ± SD. In Exp. 1, Day 8 blastocyst rates were evaluated as percentage of inseminated oocytes, with 4WP and glass vials significantly higher than cryovials A or B (38 ± 8.9%, 35 ± 7.5% v. 26 ± 3.2%, 26 ± 3.5%; respectively; P < 0.05). In Exp. 2, pH was measured for the 3 vials immediately post-maturation. Day 8 blastocyst rates were significantly higher in the glass vials as compared with cryovials A and B (pH 7.26, 31%; pH 7.60, 20% and pH 7.72, 22%, respectively; P < 0.05). In conclusion, blastocyst rates are affected by type of vial, as well as different gas permeability among other factors influencing pH. Further studies are necessary to optimize the maturation of the oocytes in HEPES-buffered media.
147 THE DEVELOPMENTAL CHARACTERISTICS OF IN VITRO-PRODUCED CATTLE-WISENT (BOS TAURUS-BISON BONASUS) HYBRID EMBRYOS
Authors: Shedova E. N. , Singina G. N. , Bagirov V. A. , Zinovieva N. A.
Published: December 2016

Interspecies hybrids are important resources for research and agriculture. Therefore, the aim of this study was to evaluate development, quality, and viability of embryos produced in vitro using cattle (Bos taurus) oocytes and European bison (Bison bonasus) epididymal sperm. The epididymes were obtained following a forced slaughter of one bull aged 7 years. The sperm was collected by scraping the inner surface of the epididymes, diluted with the cryopreservation medium, and equilibrated for 4 h at 4°C. Thereafter, sperm aliquots (0.2 mL) were frozen in liquid nitrogen vapor for 5 min and then plunged into liquid nitrogen for storage. Prior to fertilization, frozen semen was thawed in pre-warmed medium for 1 min at 37°C and prepared by the swim-up method. The frozen-thawed ejaculated sperm from the Russian Black Pied bulls was used as a positive control. Slaughterhouse-derived cumulus-oocyte complexes were matured for 24 h in TCM-199 supplemented with 10% FCS, 0.2 mM sodium pyruvate, 10 μg mL−1 porcine FSH, and 10 μg mL−1 ovine LH. Matured oocytes (35–40 oocytes per group) were co-incubated for 18 h with homologous (n = 266 oocytes) or heterologous (n = 292 oocytes) sperm (spermatozoa/mL) in 500 µL of TALP containing 10 μg mL−1 heparin, 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine, and 0.1% minimal essential medium nonessential amino acids. After IVF, the oocytes were cultured in CR1aa medium (Rosenkrans 1994 J. Anim. Sci. 72, 434–437) to the blastocyst stage. All the cultures were performed at 38.5°C and 5% CO2 in humidified air. At Days 2 and 7 after insemination, the cleavage and blastocyst rates were determined. In addition, a part of obtained blastocysts was fixed with 4% paraformaldehyde, and the total cell number and apoptotic cell ratio were determined by 4’,6-diamidino-2-phenylindole and TUNEL staining. The remaining blastocysts were cultured up to Day 10, and the hatching rates were assessed. The data (3–5 replicates) were analysed by ANOVA. The cleavage rates did not differ among both male species (72.4 and 77.1%). Furthermore, no significant effects of interspecies fertilization on the blastocyst rate or total cell number per blastocyst were found (27.4 ± 1.6% and 77.0 ± 5.7 for cattle embryos and 26.2 ± 1.9% and 83.1 ± 8.9 for cattle-wisent hybrid embryos). On the other hand, the significant differences between homologous and heterologous fertilization were detected in the rate of hatched blastocysts (60.3 ± 5.1 v. 38 ± 2.9, P < 0.05) and apoptotic cell ratio 7.3 ± 0.8 v. 11.6 ± 1.04, P < 0.05). Our findings demonstrate that hybrid embryos produced by IVF of bovine oocytes with the epididymal sperm of European bison can be developed up to advanced blastocyst stages. However, the hybrid embryos have a lower quality and viability than cattle embryos.
Research was supported by the Program of Presidium of the Russian Academy of Science, project no. IV.13.3.
15 QUANTIFICATION OF BULL SPERM TRAITS AS ASSESSED BY COMPUTER-ASSISTED SEMEN ANALYSIS AND THE RELATIONSHIP TO PREGNANCY RATE FOLLOWING CONTROLLED BREEDING
Authors: Mphaphathi M. L. , Seshoka M. M. , Ramukhithi F. V. , Raphalalani Z. C. , Netshirovha T. R. , Maqhashu A. , Kanuya N. L. , Raito M. B. , Greyling J. P. C. , Nedambale T. L.
Published: December 2016

The bull’s contribution through artificial insemination to reproductive efficiency is of great biological importance. The objectives were (1) to compare the oestrous synchronization response of Bonsmara and Nguni cows; and (2) to find the relationship between cow’s conception rate (in vivo and in vitro fertilization) and bull sperm motility rate assessed by computer-assisted semen analysis (CASA) following AI. For the in vivo sperm fertility test, 100 Bonsmara and 482 Nguni cows were randomly selected and subjected to oestrous synchronization protocol and AI with frozen–thawed assessed semen by CASA before AI. Briefly at Day 0, cows were inserted with an intravaginal CIDR® (1.9 g), which was removed on Day 7. Prostaglandin was then administered (2 mL) on Day 8 and a heatmount detector was placed on the hindquarter of each cow. For the in vitro sperm fertility test, collected oocytes from slaughterhouse were in vitro matured (n = 360) and in vitro fertilized (sperm/mL) in 100-µL droplets (final volume) of BO-IVF medium per treatment bulls (Bonsmara or Nguni bull). The frozen/thawed semen straws of Bonsmara and Nguni bulls were randomly selected and used under the same IVF conditions. The thawed bull’s sperm characteristics were examined by CASA before in vitro fertilization. Data were analysed using ANOVA. Treatment means were compared using the Fisher’s protected least significant difference t-test. There was no significant difference in oestrous response for the Bonsmara (83.0%) and Nguni (90.8%) cows, respectively. The Bonsmara cows recorded a significantly higher pregnancy rate (59.0%) compared with the Nguni (37.1%) cows (P < 0.05). Sperm traits such as total motility (TM), progressive motility and rapid were found to be positively correlated with conception rate (r = 0.06, 0.03, and 0.08, respectively; P < 0.01), although correlations were low. There was no difference in the average frozen–thawed sperm TM rate of Nguni (92.2%) and Bonsmara (81.0%). There was a lower fertilization rate following IVF with Bonsmara and Nguni bull sperm. In conclusion, Nguni cows had similar oestrous response as Bonsmara cows. The sperm traits from Bonsmara and Nguni bulls were found to be related to in vivo conception and in vitro fertilization rate when sperm cells were assessed by CASA technology. However, the pregnancy rate was lower in Nguni cows.
150 PROTEOME OF BOVINE CUMULUS CELLS AS RELATED TO OOCYTE MORPHOLOGY AND IN VITRO EMBRYO PRODUCTION
Authors: Velez I. C. , Ramirez M. M. , Chica A. I. , Urrego R. , Moura A. A. , Vasconcelos F. R. , Nagano C. S. , Rodriguez-Villamil P. , Jimenez-Escobar C. , Zambrano-Varon J.
Published: December 2016

The present study was conducted to study the effect of cumulus-oocyte complex (COC) morphology on subsequent in vitro embryo development and to assess the proteome of their corresponding cumulus cells (CC). Cow ovaries were obtained at an abattoir and COC aspirated from 3–8 mm follicles. The COC were defined as type I (TI): homogeneous ooplasma and ≥4 layers of compact CC; type II (TII): granular ooplasma and ≥4 layers of slight expanded CC. Fifty COC had ~500,000 CC. Cumulus cells were frozen in ammonium bicarbonate and immediately lyophilized for proteome analysis. Other selected COC were matured in vitro in TCM-199-supplemented media for 24 h. After maturation, CC were collected (T24) and processed as described above. The remaining COC were fertilized with sperm from a fertile bull and zygotes, cultured in vitro until Day 7. Ten blastocysts per group were stained (Hoechst 33342) and blastomeres, counted for assessment of embryo quality. The CC proteins were obtained from the following groups: immature type I (TIT0) and type II (TIIT0), and in vitro matured type I (TIT24) and type II (TIIT24). For protein extraction, we used sonication (30 min, 4°C), freezing and unfreezing in liquid nitrogen, and maceration. The CC proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by ESI-MS/MS. Differences in cleavage, embryo rates, and blastomere numbers were analysed by t-test. Protein expression difference was set at 2.5-fold (P < 0.05). In silico protein interactions were investigated using STRING v. 10.0. There were no differences in cleavage (88 ± 4 v. 89 ± 8%) and embryo rates (36 ± 7 v. 33 ± 8%) between COC of TI (n = 220) and TII (n = 161), respectively. Blastomeres were also similar in TI (101) and TII (104) groups. Major proteins expressed in all CC were α-enolase, β-actin, oestradiol 17-β-dehydrogenase 1, glutathione S-transferase, glyceraldehyde 3-phosphate dehydrogenase, heat shock protein β-1, histone H2B type 1-N, histone H4, mitochondrial malate dehydrogenase 2, protein disulfide isomarase A6, triosephosphate isomerase, tubulin α-1C chain, and vimentin. Glyceraldehyde 3-phosphate dehydrogenase appeared to be more expressed in TIT0, whereas tubulin α-1C chain and vimentin had greater expression in TIIT24. As evidenced by in silico analysis, most CC proteins interact among themselves, participating in complex networks involving intracellular signalling and other events. In conclusion, there are no difference in embryo development when using compact and early-expanded COC, indicating that both types can be selected for IVP. Protein profile of cumulus cell may serve as a marker for in vitro embryo competence.
163 EVALUATION OF VIABILITY OF BULL SEMEN COLLECTED BY ELECTRO-EJACULATION USING COMMERCIAL SEMEN EXTENDER AND 2 CULTURE MEDIA AT CONTROLLED ROOM TEMPERATURE
Authors: Raseona A. M. , Ajao O. A. , Nethengwe L. D. , Madzhie L. R. , Nedambale T. L. , Barry D. M.
Published: December 2016

Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.