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Objectives were to evaluate the effects of follicular wave and progesterone concentration on growth of the ovulatory follicle, conceptus elongation, uterine IFN-tau concentration, and transcriptome of conceptus and endometrium of pregnant cows on d 17 of gestation. Nonlactating nonpregnant Holstein cows were assigned randomly to one of 3 treatments: ovulation of a first-wave follicle (FW, n = 15); ovulation of a first-wave follicle and progesterone supplementation (FWP4, n = 12); and ovulation of a second-wave follicle (SW, n = 19). Ovulation of a first- or second-wave follicle was achieved by initiating the Ovsynch protocol (d -9 GnRH, d -2 and -1 PGF(2 alpha), d 0 GnRH and artificial insemination, d 0.7 artificial insemination) on d 0 or 6 of a presynchronized estrous cycle, respectively. Cows in FWP4 received 3 intravaginal inserts containing progesterone at 12, 24, and 48 h after the first GnRH injection that were removed on d -2. Cows were killed on d 17 for collection of the reproductive tract. Transcriptome was evaluated by microarray using the Affymetrix Bovine Array. Orthogonal contrasts were built to assess the effects of progesterone concentration during follicle growth (FW vs. FWP4 + SW) and follicular wave (FWP4 vs. SW). Progesterone concentrations (LSM +/- SEM) from d -9 to -2 were greater for SW, followed by FWP4 and FW (5.38 +/- 0.24, 4.26 +/- 0.28, and 1.17 +/- 0.27 ng/mL). Diameter of the ovulatory follicle (FW = 19.6 +/- 0.6; FWP4 = 15.6 +/- 0.6; SW = 15.2 +/- 0.5 mm) and concentrations of estradiol from d -2 to 1 (FW = 4.05 +/- 0.33; FWP4 = 2.73 +/- 0.35; SW = 2.48 +/- 0.30 pg/mL) were greater for FW compared with FWP4 and SW. Progesterone concentrations from d 3 to 16 were greater for FW compared with FWP4 and SW. A total of 28 singleton conceptuses were collected (FW, n = 8; FWP4, n = 8; SW, n = 12) and only intact conceptuses were included in the analyses of length (FW, n = 8; FWP4, n = 6; SW, n = 12). Although conceptuses were longer for FW compared with FWP4 and SW (FW = 16.6 +/- 2.3; FWP4 = 9.8 +/- 2.2; SW = 9.6 +/- 2.0 cm), treatment did not affect the amount of IFN-tau in uterine flushing. Transcriptome of conceptuses and endometrium of pregnant cows was not extensively affected by follicular wave (8 and 1 differentially expressed transcripts) or concentration of progesterone during follicle growth (0 and 3 differentially expressed transcripts), showing that these factors did not affect conceptuses and endometrium transcriptome in pregnancies that are maintained to d 17. USDA-NIFA [2015-67015-23313]; Southeast Milk Check-Off Dairy Research and Education Projects Published version The authors thank William Rembert and the Central Beef Industries LLC administration and staff for their assistance for collection of reproductive tracts; manag-ers and staff at University of Florida Dairy Unit for their assistance with animal handling and feeding; and Mariana R. Carvalho, JoAo Vitor S. Queiroz, Ronaldo L. A. Cerri, Sha Tao, Milerky C. Perdomo, Izabella M. Thompson, Luiz Siqueira, Paula Tribulo, Gabriel C. Gomes, Mauricio G. Favoreto, Ana P. Monteiro, and Ana L. Sevarolli (University of Florida, Gainesville) for their assistance with sample processing. Funding for this project was provided by grants from the USDA-NIFA #2015-67015-23313 and the Southeast Milk Check-Off Dairy Research and Education Projects to J.E.P. Santos. The authors have not stated any con-flicts of interest.

Made available in DSpace on 2020-12-12T01:44:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-01 The present study attempted to verify the prevalence of and risk factors for diarrhea-causing agents in dairy calves from Brazil. Additionally, ages with a higher risk of occurrence for each agent were verified by means of the receiver operating characteristic (ROC) curve. The collections were performed on 39 farms, belonging to 29 municipalities located in eight states of Brazil. It was possible to conclude that the prevalence of Coronavirus, Rotavirus, Cryptosporidium spp., Eimeria spp., and nematodes was 7.20% (95% CI 4.54–9.78), 6.37% (95% CI 3.85–8.89), 51.52% (95% CI 45.26–55.57), 3.46% (95% CI 2.24–4.67), and 3.46% (95% CI 2.24–4.67), respectively. Ages with higher probabilities of occurrence of these diseases in calves were < 10, > 8, > 6, > 37, and > 36 days, respectively. Diarrhea occurred more significantly (P < 0.0001) in animals less than 21 days old and mainly on those receiving milk through automatic feeders (P < 0.001). Cryptosporidium spp. were a risk factor for the occurrence of Rotavirus, and vice versa (P = 0.0039) and presented a positive correlation with Coronavirus (P = 0.0089). Calves that drink water from rivers, streams, and ponds had a higher chance of being infected by Eimeria spp. (P < 0.0001), as well as developing infection by nematodes (P < 0.0001). The results found in this study highlight the importance of studying the agents of diarrhea together, once they act as coinfection where the losses triggered for the owners will involve some of these agents simultaneously. Centro de Parasitologia Veterinária - CPV Escola de Veterinária e Zootecnia Universidade Federal de Goiás (UFG) MSD Saúde Animal Faculdade de Ciências Agrárias e Veterinárias UNESP/CPPAR Universidade Brasil – Campus de Descalvado Universidade Estadual de Londrina Instituto de Patologia Tropical e Saúde Pública Universidade Federal de Goiás Faculdade de Ciências Agrárias e Veterinárias UNESP/CPPAR

Highlights • RVA vaccination program reduces the frequency and intensity of diarrhea in dairy calves. • The vaccination immune pressure can select specific genotypes in RVA field strains. • RVA genotype G10P[11] in fecal samples of calves from G6P[5] vaccinated dairy cattle herds., Worldwide, neonatal diarrhea is one of the most important health issues affecting dairy calves, and rotavirus A (RVA) is one of its primary causes. Among the measures to mitigate the risk of diarrhea outbreaks, cow vaccination stands out as one of the most important. However, the immune pressure resulting from routine vaccination may be able to select specific G and P genotypes in RVA field strains. This study aimed to determine the frequency and intensity of neonatal diarrhea and the incidence of RVA and attempted to monitor the G and P genotypes present in the RVA strains circulating in a high milk yield cattle herd vaccinated with RVA G6P[5] strain. Fecal samples (n = 1220) from 122 Holstein heifer calves between 0–30 days old that were born from RVA-vaccinated cows were collected at 10 different time points, regardless of the presence or absence of diarrhea. The presence of RVA in fecal samples was determined by the polyacrylamide gel electrophoresis (PAGE) technique and confirmed by reverse transcription polymerase chain reaction (RT-PCR). G and P amplicons from 10 RVA-positive fecal samples from calves of different ages and collections were subjected to nucleotide sequencing. The proportion of the calves and fecal samples that were positive for RVA were 62.3% (76/122) and 8.1% (99/1220), respectively. Using sequence analysis, all 10 RVA field strains presented genotype G10P[11]. The protection of G6P[5] vaccination is clear, as this genotype was not detected in this study, and it is known that vaccination against RVA reduces the incidence of diarrhea independent of genotype involved. This result demonstrates the importance of epidemiological monitoring of RVA genotypes circulating in vaccinated dairy cattle herds to the early detection of new potential pathogenic RVA strains.

Neonatal diarrhea is the main cause of morbidity and mortality in calves up to 30 days old, and rotavirus A (RVA) is the main viral etiology. RVA vaccines are one of the main tools for diarrhea control in neonates. The aim of this cross-sectional study was to monitor by RT-PCR the G and P genotypes of RVA strains identified in dairy cattle herds regularly vaccinated with the RVA UK strain (G6P[5]). Of the 14 randomly selected herds, two were excluded because no calf was diagnosed with diarrhea on the day of fecal collection. Another six herds were also excluded from the study because all 20 diarrheic fecal samples evaluated were RT-PCR-negative. In the remaining six herds, 17 (25.4%) of the 67 diarrheic samples were RVA-positive. One G and P amplicon from each herd were selected for nucleotide sequencing. In the phylogenetic analysis, five RVA strains presented the G6P[11] genotype, and one presented the G10P[11] genotype. The G6 genotype present in all RVA field strains clustered into a distinct phylogenetic arrangement (lineage III) of the UK vaccine strain (lineage IV), characterizing the emergence of a phylogenetically distant G6 strain. In addition, we observed the emergence of strains with G10 and P[11] genotypes characterizing failure in heterologous immune protection. These results show the epidemiological importance of constant monitoring of RVA strains in vaccinated cattle herds and the low frequencies of diarrhea and diagnosis of RVA suggest that a regular vaccination program reduces the frequency and severity of RVA diarrhea in suckling calves.

Made available in DSpace on 2018-11-26T17:10:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Jordan University of Science and Technology This study was conducted to document the pattern of antral follicular wave emergence throughout the 14-day, progesterone (P-4)-based estrous synchronization protocol in ewes that were maintained in subtropical conditions, during the period of increasing day lengths (ID), decreasing day lengths (DD), and the transitional period (TP). In addition, the influence of ovarian status (i.e., size of ovarian antral follicles and the presence of corpora lutea) at the outset of P-4 treatment on ensuing ovarian follicular wave development was examined. Sexually mature Santa Ines ewes (n =70) were subjected to one of the two estrous synchronization protocols in the three seasons. On Day 0, the ewes received an i.m. injection of prostaglandin F-2 alpha and an intravaginal P4-releasing device that remained in place for 14days (G-1CIDR) or was replaced on Day 7 (G-2CIDR). Daily ultrasonography of ovaries was conducted from Days 0 to 15. Mean (+/- SEM) numbers of follicular waves per ewe were 3.7 +/- 0.1 and 3.6 +/- 0.1 for G-1CIDR and G-2CIDR (P > 0.05). The number of emerging follicular waves was greater (P

Made available in DSpace on 2018-11-26T17:20:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Zoetis Saude Animal (Campinas -SP, Brazil) The effects of addition of gonadotrophin-releasing hormone (GnRH) to a progesterone plus oestradiol-based protocol and timing of insemination in Holstein cows treated for timed AI (TAI) were evaluated. Cows (n=481) received a progesterone device and 2mg oestradiol benzoate. After 8 days, the device was removed and 25mg dinoprost was administered. Cows were allocated to one of three (Study 1; n=57) or four (Study 2; n=424) groups, accordingly to ovulation inducer alone (Study 1; oestradiol cypionate (EC), GnRH or both) or ovulation inducer (EC alone or combined with GnRH) and timing of insemination (48 or 54h after device removal; Study 2). In Study 1, the diameter of the ovulatory follicle was greater for GnRH than EC. Oestrus and ovulation rates were similar regardless of ovulatory stimuli. However, time to ovulation was delayed when GnRH only was used. In Study 2, cows treated with GnRH or not had similar pregnancy per AI (P/AI) 30 days (41.5% vs 37.3%; P=0.28) and 60 days (35.9% vs 33.0%; P=0.61) after TAI. TAI 48 and 54h after device removal resulted similar P/AI at 30 days (40.3% vs 38.5%; P=0.63) and 60 days (33.8% vs 35.1%; P=0.72). Thus, adding GnRH at TAI does not improve pregnancy rates in dairy cows receiving EC. The flexibility of time to insemination enables TAI of a large number of cows using the same protocol and splitting the time of AI. Univ Sao Paulo, Dept Reprod Anim, Fac Med Vet & Zootecnia, Ave Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP, Brazil Univ Sao Paulo, Dept Med Vet, Fac Zootecnia & Engn Alimentos, Ave Duque de Caxias Norte 225,Campus USP, BR-13635900 Pirassununga, SP, Brazil MSD Saude Anim, Ave Nacoes Unidas 14171,Torre C,Crystal Tower, BR-04794000 Sao Paulo, SP, Brazil Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil Cent Pioneiros, Ave Pioneiros, BR-84145000 Carambei, PR, Brazil Univ Estadual Paulista, Dept Prod Anim, Fac Med Vet & Zootecnia, Dist Rubiao Jr S-N,Caixa Postal 560, BR-18618970 Botucatu, SP, Brazil Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, Dept Prod Anim, Fac Med Vet & Zootecnia, Dist Rubiao Jr S-N,Caixa Postal 560, BR-18618970 Botucatu, SP, Brazil FAPESP: 2009/00938-3 FAPESP: 2012/07510-1 CNPq: 486089/2013-14

The relationship between energy status and fertility in dairy cattle was retrospectively analyzed by comparing fertility with body condition score (BCS) near artificial insemination (AI; experiment 1), early postpartum changes in BCS (experiment 2), and postpartum changes in body weight (BW; experiment 3). To reduce the effect of cyclicity status, all cows were synchronized with Double-Ovsynch protocol before timed AI. In experiment 1, BCS of lactating dairy cows (n = 1,103) was evaluated near AI. Most cows (93%) were cycling at initiation of the breeding Ovsynch protocol (first GnRH injection). A lower percentage pregnant to AI (P/AI) was found in cows with lower (≤ 2.50) versus higher (≥ 2.75) BCS (40.4 vs. 49.2%). In experiment 2, lactating dairy cows on 2 commercial dairies (n = 1,887) were divided by BCS change from calving until the third week postpartum. Overall, P/AI at 70-d pregnancy diagnosis differed dramatically by BCS change and was least for cows that lost BCS, intermediate for cows that maintained BCS, and greatest for cows that gained BCS [22.8% (180/789), 36.0% (243/675), and 78.3% (331/423), respectively]. Surprisingly, a difference existed between farms with BCS change dramatically affecting P/AI on one farm and no effect on the other farm. In experiment 3, lactating dairy cows (n = 71) had BW measured weekly from the first to ninth week postpartum and then had superovulation induced using a modified Double-Ovsynch protocol. Cows were divided into quartiles (Q) by percentage of BW change (Q1 = least change; Q4 = most change) from calving until the third week postpartum. No effect was detected of quartile on number of ovulations, total embryos collected, or percentage of oocytes that were fertilized; however, the percentage of fertilized oocytes that were transferable embryos was greater for cows in Q1, Q2, and Q3 than Q4 (83.8, 75.2, 82.6, and 53.2%, respectively). In addition, percentage of degenerated embryos was least for cows in Q1, Q2, and Q3 and greatest for Q4 (9.6, 14.5, 12.6, and 35.2% respectively). In conclusion, for cows synchronized with a Double-Ovsynch protocol, an effect of low BCS (≤ 2.50) near AI on fertility was detected, but change in BCS during the first 3 wk postpartum had a more profound effect on P/AI to first timed AI. This effect could be partially explained by the reduction in embryo quality and increase in degenerate embryos byd 7 after AI in cows that lost more BW from the first to third week postpartum.

The objectives of the current study were to investigate the efficacy of PGF₂α as a therapy to reduce the prevalence of subclinical endometritis and improve pregnancy per artificial insemination (P/AI) in cows subjected to a timed artificial insemination (AI) program. A total of 1,342 lactating Holstein dairy cows were allocated randomly at 25 ± 3 d in milk (DIM) to remain as untreated controls (control, n=454) or to receive a single PGF₂α treatment at 39 ± 3 DIM (1PGF, n=474) or 2 treatments with PGF(α at 25 ± 3 and 39 ± 3 DIM (2PGF, n=414). All cows were enrolled in the double Ovsynch program at 48 ± 3 DIM and were inseminated at 75 ± 3 DIM. A subset of 357 cows had uterine samples collected for cytological examination at 25 ± 3, 32 ± 3, and 46 ± 3 DIM to determine the percentage of polymorphonuclear leukocytes (PMNL). Subclinical endometritis was defined by the presence of ≥ 5% PMNL. Vaginal discharge score was evaluated at 25 ± 3 DIM and used to define the prevalence of purulent vaginal discharge. Body condition score was assessed at 25 ± 3 DIM. Pregnancy was diagnosed 32 d after AI and reconfirmed 28 d later. At 32 ± 3 DIM, the prevalence of subclinical endometritis was reduced by treatment with PGF₂α at 25 ± 3 DIM in 2PGF (control=23.5% vs. 1PGF=28.3% vs. 2PGF=16.7%); however, this benefit disappeared at 46 ± 3 DIM, and 14% of the cows remained with subclinical endometritis. One or 2 treatments with PGF₂α did not influence P/AI on d 32 or 60 after timed AI, which averaged 39.9 and 35.2%. Similarly, treatment with PGF₂α had no effect on pregnancy loss between 32 and 60 d of gestation (11.9%). Cows diagnosed with both purulent vaginal discharge and subclinical endometritis had the lowest P/AI and the highest pregnancy loss compared with those diagnosed with only 1 of the 2 diseases or compared with cows having no diagnosis of uterine diseases. Interestingly, subclinical endometritis depressed P/AI and increased pregnancy loss only when it persisted until 46 DIM. On d 32 after AI, cows not diagnosed with subclinical endometritis and those that resolved subclinical endometritis by 46 DIM had greater P/AI than those that remained with subclinical endometritis at 46 DIM (45.4 and 40.0 vs. 25.0%, respectively). Similar to P/AI, cows not diagnosed with subclinical endometritis and those that resolved subclinical endometritis by 46 DIM had less pregnancy loss than those with subclinical endometritis at 46 DIM (9.6 and 13.5 vs. 43.9%, respectively). One or 2 treatments with PGF₂α before initiation of the timed AI program were unable to improve uterine health, P/AI, and maintenance of pregnancy in lactating dairy cows. Cows diagnosed with both purulent vaginal discharge and subclinical endometritis had the greatest depressions in measures of fertility at first AI, particularly when subclinical endometritis persisted in the early postpartum period.

Reproductive management programs that synchronize ovulation can ovulate a smaller than normal follicle, potentially resulting in inadequate progesterone (P4) concentrations after artificial insemination (AI). Ovulation of the dominant follicle of the first follicular wave with human chorionic gonadotropin (hCG) treatment can produce an accessory corpus luteum and increase circulating P4 concentrations. This manuscript reports the results of 2 separate analyses that evaluated the effect of hCG treatment post-AI on fertility in lactating dairy cows. The first study used meta-analysis to combine the results from 10 different published studies that used hCG treatment on d 4 to 9 post-AI in lactating dairy cows. Overall, pregnancies per artificial insemination (P/AI) were increased 3.0% by hCG treatment post-AI [34% (752/2,213) vs. 37% (808/2,184); Control vs. hCG-treated, respectively]. The second study was a field research trial in which lactating Holstein cows (n=2,979) from 6 commercial dairy herds were stratified by parity and breeding number and then randomly assigned to one of 2 groups: control (no further treatment, n=1,519) or hCG [Chorulon i.m.: 2,000 IU (in 3 of the herds) or 3,300 IU (in 3 herds); n=1,460] on d 5 after a timed AI (ovulation synchronized with Ovsynch, Presynch-Ovsynch, or Double-Ovsynch). In a subset of cows, the hCG profile and P4 changes were determined. Treatment with hCG increased P4 (4.3 vs. 5.3 ng/mL on d 12). Pregnancies per AI were greater in cows treated with hCG (40.8%; 596/1,460) than control (37.3%; 566/1,519) cows. Interestingly, an interaction among treatment and parity was observed; primiparous cows had greater P/AI after hCG (49.7%; 266/535) than controls (39.5%; 215/544). In contrast, older cows receiving hCG (35.7%; 330/925) had similar P/AI to controls (36.0%; 351/975).Thus, targeted use of hCG on d 5 after TAI enhances fertility about 3.0% (based on meta-analysis) to 3.5% (based on our field trial). Surprisingly, this fertility-enhancing effect of hCG was very large in first-lactation cows but not observed in older cows in the field study. Future research is needed to confirm these intriguing results and to determine why older cows did not have improved fertility after hCG treatment.

Previous studies reported increased fertility using Ovsynch for presynchronization before Ovsynch (Double-Ovsynch), as compared with presynchronization with two prostaglandin F(2α) (PGF(2α)) treatments before Ovsynch (Presynch-Ovsynch). This study compared ovarian follicular dynamics and hormone concentrations during Double-Ovsynch versus Presynch-Ovsynch. Lactating Holstein cows (N = 193) were assigned to one of two treatment groups: (1) Presynch (N = 93), two injections of PGF(2α) 14 days apart, followed by the Ovsynch-timed AI protocol 12 days later; and (2) Double-Ovsynch (N = 100), one injection of GnRH, PGF(2α) 7 days later, and GnRH 3 days later, followed by the Ovsynch-timed AI protocol 7 days later. All cows received the same Ovsynch-timed AI protocol: GnRH (G1) at 68 ± 3 days in milk (mean ± SEM), PGF(2α) 7 days later, and GnRH (G2) 56 hours after PGF(2α). Ultrasonographic evaluations of the ovaries and blood sampling were performed at G1, PGF(2α), G2, and 6 days after the G2 injection of the Ovsynch-timed AI protocol. Double-Ovsynch decreased the percentage of cows with low circulating progesterone (P4) concentrations (0.50 ng/mL) at G1 (12.0% vs. 30.1%; P = 0.003) and increased the percentage of cows with medium P4 concentrations (0.50P4 ≤ 3.0 ng/mL) at G1 (80.0% vs. 57.0%; P0.01), and with CL at G1 (94.0% vs. 67.8%; P0.01). Double-Ovsynch also increased the percentage of cows with high P4 (3.0 ng/mL) at PGF(2α) (88.0% vs. 76.3%; P = 0.04) and tended to increase average circulating P4 at PGF(2α) (3.52 ± 0.17 ng/mL vs. 3.09 ± 0.21 ng/mL; P = 0.11). Double-Ovsynch also tended to increase percentage of cows ovulating to G1 (80.0% vs. 69.9%; P = 0.11) and G2 (98.0% vs. 93.5%; P = 0.08). Thus, presynchronization of cows with Double-Ovsynch induced ovulation in noncycling cows and appeared to increase most aspects of synchronization during the Ovsynch protocol.

It was hypothesized the lower fertility of repeat-breeder (RB) Holstein cows is associated with oocyte quality and this negative effect is enhanced during summer heat stress (HS). During the summer and the winter, heifers (H; n = 36 and 34, respectively), peak-lactation (PL; n = 37 and 32, respectively), and RB (n = 36 and 31, respectively) Holstein cows were subjected to ovum retrieval to assess oocyte recovery, in vitro embryonic developmental rates, and blastocyst quality [terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and total cell number]. The environmental temperature and humidity, respiration rate, and cutaneous and rectal temperatures were recorded in both seasons. The summer HS increased the respiration rate and the rectal temperature of PL and RB cows, and increased the cutaneous temperature and lowered the in vitro embryo production of Holstein cows and heifers. Although cleavage rate was similar among groups [H = 51.7% ± 4.5 (n = 375), PL = 37.9% ± 5.1 (n = 390), RB = 41.9% ± 4.5 (n = 666)], blastocyst rate was compromised by HS, especially in RB cows [H = 30.3% ± 4.8 (n = 244) vs. 23.3% ± 6.4 (n = 150), PL = 22.0% ± 4.7 (n = 191) vs. 14.6% ± 7.6 (n = 103), RB = 22.5% ± 5.4 (n = 413) vs. 7.9% ± 4.3 (n = 177)]. Moreover, the fragmentation rate of RB blastocysts was enhanced during the summer, compared with winter [4.9% ± 0.7 (n = 14) vs. 2.2% ± 0.2 (n = 78)] and other groups [H = 2.5% ± 0.7 (n = 13), and PL = 2.7% ± 0.6 (n = 14)] suggesting that the association of RB fertility problems and summer HS may potentially impair oocyte quality. Our findings provide evidence of a greater sensitivity of RB oocytes to summer HS.

Made available in DSpace on 2013-09-27T14:54:12Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Made available in DSpace on 2013-09-30T18:36:09Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:42:31Z No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Made available in DSpace on 2014-05-20T13:42:31Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The objectives of this study were to determine the interval from ovulation to deviation and the diameter of the dominant (DF) and largest subordinate (SF) follicles at deviation in buffalo (Bubalus bubalis) heifers. Two methods of evaluation (observed vs. calculated) were used. FSH and LH profiles encompassing follicle deviation (Experiment 1), and the follicular diameter when the DF acquired ovulatory capacity (Experiment 2) were also determined. The time of deviation and the diameter of the of and the largest SF at deviation did not differ between observed and calculated methods. Overall, follicle deviation occurred 2.6 +/- 0.2 d (mean SEM) after ovulation, and the diameters of the DF and SF at deviation were 7.2 +/- 0.2 and 6.4 +/- 0.2 mm, respectively. No changes in plasma levels of FSH or LH were observed (P= 0.32 and P= 0.96, respectively). Experiment 2 was conducted in two phases according to the diameter of the of during the first wave of follicular development at the time of LH challenge (25 mg of pLH). In the first phase, follicles ranging from 5.0 to 6.0 mm (n = 7), 6.1 to 7.0 mm (n = 11), or 7.1 to 8.0 mm (n= 9) were used, and in the second phase, follicles ranging from 7.0 to 8.4 mm (n=10), 8.5 to 10.0 mm (n = 10), or 10.1 to 12.0 mm (n = 9) of diameter were used. After the pLH treatment, the of was monitored by ultrasonography every 12 h for 48 h. No ovulations occurred in heifers in the first phase. However, in the second phase, an effect of follicular diameter was observed on ovulation rate [7.0-8.4 mm (0.0%, 0/10), 8.5-10.0 mm (50.0%, 5/10), and 10.0-12.0 mm (55.6%, 5/9)]. In summary, follicle deviation occurred 2.6d after ovulation in buffalo (B. bubalis) heifers, when the diameters of the DF and SF were 7.2 and 6.4 mm, respectively. No significant changes in plasma concentrations of FSH or LH were detected. Finally, the acquisition of ovulatory capacity occurred when the of reached 8.5 mm in diameter. (C) 2010 Elsevier B.V. All rights reserved. FMVZ USP, Dept Anim Reprod, BR-05508270 São Paulo, Brazil APTA Registro, Registro, SP, Brazil Autonomous Veterinarian, Vicosa, MG, Brazil UFMA, Ctr Ciencias Agr & Ambientais, Chapadinha, MA, Brazil IB UNESP, Dept Bioestatist, Botucatu, SP, Brazil IB UNESP, Dept Farmacol, Botucatu, SP, Brazil Lab Endocrinol UNESP, Dept Apoio & Prod Saúde Anim, Aracatuba, SP, Brazil Univ Saskatchewan, Western Coll Vet Med, Saskatoon, SK S7N 0W0, Canada IB UNESP, Dept Bioestatist, Botucatu, SP, Brazil IB UNESP, Dept Farmacol, Botucatu, SP, Brazil Lab Endocrinol UNESP, Dept Apoio & Prod Saúde Anim, Aracatuba, SP, Brazil FAPESP: 03/10203-4

The mouse dermal assay has long been used to assess the dermal tumorigenicity of cigarette smoke condensate (CSC). This mouse skin model has been developed for use in carcinogenicity testing utilizing the SENCAR mouse as the standard strain. Though the model has limitations, it remains as the most relevant method available to study the dermal tumor promoting potential of mainstream cigarette smoke. In the typical SENCAR mouse CSC bioassay, CSC is applied for 29 weeks following the application of a tumor initiator such as 7,12-dimethylbenz[a]anthracene (DMBA). Several endpoints are considered for analysis including: the percentage of animals with at least one mass, latency, and number of masses per animal. In this paper, a relatively straightforward analytic model and procedure is presented for analyzing the time course of the incidence of masses. The procedure considered here takes advantage of Bayesian statistical techniques, which provide powerful methods for model fitting and simulation. Two datasets are analyzed to illustrate how the model fits the data, how well the model may perform in predicting data from such trials, and how the model may be used as a decision tool when comparing the dermal tumorigenicity of cigarette smoke condensate from multiple cigarette types. The analysis presented here was developed as a statistical decision tool for differentiating between two or more prototype products based on the dermal tumorigenicity.

A total of 177 Nelore heifers were examined by ultrasonography to determine the presence or absence of a corpus luteum (CL) and received a 3 mg norgestomet ear implant plus 2 mg of estradiol benzoate i.m. On Day 8, implants were removed and 150 μg of d -cloprostenol i.m. was administered. At the time of norgestomet implant removal, heifers with or without CL at the time of initiating treatment were assigned equally and by replicate to be treated with 0 IU ( n = 87) or 400 IU ( n = 90) eCG i.m. All heifers received 1 mg of EB i.m. on Day 9 and were submitted to fixed-time artificial insemination (FTAI) 30–34 h later. The addition of eCG increased the diameter of the largest follicle (LF) at FTAI (10.6 ± 0.2 mm vs. 9.5 ± 0.2 mm; P = 0.003; mean ± SEM), the final growth rate of the LF (1.14 ± 0.1 mm/day vs. 0.64 ± 0.1 mm/day; P = 0.0009), ovulation rate [94.4% (85/90) vs. 73.6% (64/87); P = 0.0006], the diameter of the CL at Day 15 (15.5 ± 0.3 mm vs. 13.8 ± 0.3 mm; P = 0.0002), serum concentrations of progesterone 5 days after FTAI (6.6 ± 1.0 ng/ml vs. 3.6 ± 0.7 ng/ml; P = 0.0009), and pregnancy per AI [P/AI; 50.0% (45/90) vs. 36.8% (32/87); P = 0.04]. The absence of a CL at the beginning of the treatment negatively influenced the P/AI [30.2% (16/53) vs. 49.2% (61/124); P = 0.01]. Therefore, the presence of a CL (and/or onset of puberty) must be considered in setting up FTAI programs in heifers. In addition, eCG may be an important tool for the enhancement of follicular growth, ovulation, size and function of the subsequent CL, and pregnancy rates in progestin-based FTAI protocols in Bos indicus heifers.

The aim of the present study was to compare a synchronization of time of ovulation protocol for fixed-timed embryo transfer (FTET) with the usual administration of a single dose of prostaglandin associated with detection of estrus. Also, the effect of the presence of CL at the beginning of FTET protocol was evaluated. Lactating Holstein cows (n=651) with three previous artificial inseminations were classified according to presence or absence of a corpus luteum (CL). Cows with a CL were randomly assigned to two additional treatments and submitted to embryo transfer after detection of estrus (PGF-Estrus) or FTET (FTET-CL). Cows without CL were allocated to the FTET-NoCL treatment. On a random day of the estrous cycle (Day 0), cows in the PGF-Estrus treatment (n=229) were treated with 150 microg d-cloprostenol (PGF) i.m. followed by detection of estrus from Day 1 through Day 5 after PGF. Embryos were transferred 6-8 days after estrus detection. Cows in the FTET-CL (n=208; presence of CL) and FTET-NoCL (n=214; absence of CL) treatments received a norgestomet ear implant plus 2mg estradiol benzoate (EB) and 50mg progesterone i.m. on Day 0. On Day 8, the implant was removed and 400 IUeCG, 150 microg d-cloprostenol and 1mg estradiol cypionate i.m. were administered. No detection of estrus was performed and Day 10 was arbitrarily considered as the estrus day. Ultrasonographic exams were performed in all recipients and only cows with a single CL> or =15 mm or multiple CL received a fresh or frozen-thawed embryo on Day 17. Pregnancy was diagnosed by ultrasonography at 30 and 60 days of pregnancy. When FTET and PGF-Estrus were compared, the proportion of cows receiving an embryo (recipients transferred-to-treated rate) was greater in the FTET-CL (75.0% (156/208) than in PGF-Estrus (34.5%, 79/229; P

A tiered testing strategy based on a comparative chemical and biological testing program has been developed to evaluate the potential of tobacco processes, ingredients, or other technological developments to change the biological activity that results from burning tobacco. Cast sheet tobacco is a specific type of reconstituted tobacco sheet that can be used in the manufacture of cigarettes. The comparative chemical and biological testing program was used to compare the mainstream smoke and cigarette smoke condensate (CSC) from a Reference cigarette that did not contain cast sheet to that collected from Test cigarettes containing cast sheet at a final blend level of either 10% or 15%. Testing included mainstream cigarette smoke chemistry studies, in vitro studies (Ames assay, sister chromatid exchange assay, and neutral red cytotoxicity assay), and in vivo toxicology studies (13-week rat nose-only inhalation assay and 30-week mouse dermal tumor promotion assay). Certain statistically significant differences were observed in the chemical and biological studies when the Reference cigarette was compared to each of the Test cigarettes. However, when viewed collectively, the chemical and biological studies demonstrated that inclusion of cast sheet up to 15% in the final blend did not increase the inherent biological activity of mainstream cigarette smoke or CSC.

Two experiments were conducted to investigate the effects of equine chorionic gonadotropin (eCG) at progestin removal and gonadotropin-releasing hormone (GnRH) at timed artificial insemination (TAI) on ovarian follicular dynamics (Experiment 1) and pregnancy rates (Experiment 2) in suckled Nelore (Bos indicus) cows. Both experiments were 2 × 2 factorials (eCG or No eCG, and GnRH or No GnRH), with identical treatments. In Experiment 1, 50 anestrous cows, 134.5 ± 2.3 d postpartum, received a 3 mg norgestomet ear implant sc, plus 3 mg norgestomet and 5 mg estradiol valerate im on Day 0. The implant was removed on Day 9, with TAI 54 h later. Cows received 400 IU eCG or no further treatment on Day 9 and GnRH (100 μg gonadorelin) or no further treatment at TAI. Treatment with eCG increased the growth rate of the largest follicle from Days 9 to 11 (means ± SEM, 1.53 ± 0.1 vs. 0.48 ± 0.1 mm/d; P < 0.0001), its diameter on Day 11 (11.4 ± 0.6 vs. 9.3 ± 0.7 mm; P = 0.03), as well as ovulation rate (80.8% vs. 50.0%, P = 0.02), whereas GnRH improved the synchrony of ovulation (72.0 ± 1.1 vs. 71.1 ± 2.0 h). In Experiment 2 (n = 599 cows, 40 to 120 d postpartum), pregnancy rates differed (P = 0.004) among groups (27.6%, 40.1%, 47.7%, and 55.7% for Control, GnRH, eCG, and eCG + GnRH groups). Both eCG and GnRH improved pregnancy rates (51.7% vs. 33.8%, P = 0.002; and 48.0% vs 37.6%, P = 0.02, respectively), although their effects were not additive (no significant interaction). In conclusion, eCG at norgestomet implant removal increased the growth rate of the largest follicle (LF) from implant removal to TAI, the diameter of the LF at TAI, and rates of ovulation and pregnancy rates. Furthermore, GnRH at TAI improved the synchrony of ovulations and pregnancy rates in postpartum Nelore cows treated with a norgestomet-based TAI protocol.

The aim of the present study was to determine the relationship among body weight (BW), body condition score (BCS) and rump fat thickness (RFAT) measured by ultrasonography, and validate the relationship between BCS and RFAT over the time. Two hundred sixty and six Nelore cows had their BW, BCS and RFAT evaluated at five different moments during the production cycle: M1) weaning; M2) parturition, M3) 42 days post-partum; M4) 82 days post-partum and M5) 112 days post-partum. A BCS value was attributed for each cow following a 1 to 5 points scale. Ultrasonographic images for RFAT measurement were obtained using a 3.5 MHz linear transducer. Images were immediately analyzed as soon as they were formed and frozen. Body condition scores and ultrasound measurements were collected on the same day by a single trained technician. The relationship between BCS and RFAT values was investigated by regression models. The analysis of similarity among the five obtained models was performed using the proc MIXED from SAS and the correlations among variables were analyzed with proc CORR from SAS. The BCS was able to predict RFAT in Nelore cows in all different moments evaluated. Also, it was shown that BCS presented high correlation (r = 0.82 to 0.93) and relationship (R2 = 0.73 to 0.92) with RFAT. However, both BCS and RFAT showed low correlation (r = 0.37 to 0.50) and relationship (R2 = 0.13 to 0.25) with BW. The BCS classification by visual method using a 1 to 5 point scale, was able to predict the RFAT in Nelore cows over the time.

This study evaluated whether the four gonadorelin products that are commercially available in the United States produce comparable ovulation responses in lactating cows. Dairy cows at 7 d after last gonadotropin-releasing hormone (GnRH) treatment of Ovsynch (Day 7), with a corpus luteum (CL) >= 15 mm and at least one follicle >= 10 mm, were evaluated for response to GnRH treatment. Selected cows were randomized to receive (100 mu g; im): (1) Cystorelin (n = 146): (2) Factrel (n = 132): (3) Fertagyl (n = 140); or (4) Ovacyst (n = 140). On Day 14, cows were examined for Ovulation by detection of an accessory CL. Circulating luteinizing hormone (LH) concentrations were also evaluated in some cows after treatment with 100 mu g (n = 10 per group) or 50 mu g (n = 5 per group) GnRH. Statistical analyses were performed with the procedures MIXED and GLIMMIX of the SAS program. Percentage of cows ovulating differed (P < 0.01) among groups, with that for Factrel being lower (55.3%) than that for Cystorelin (76.7%), Fertagyl (73.6%), or Ovacyst (85.0%), There was no effect of batch, parity, or follicle size on ovulation response. but increasing body condition score decreased Ovulation response. There was a much greater LH release in cows treated with 100 mu g than in those treated with 50 mu g, but there were no detectable differences among products in time to LH peak, peak LH concentration, or area under the LH curve and no treatment effects nor treatment by time interactions on circulating LH profile. Thus, ovulation response to Factrel on Day 7 of the cycle was lower than that for other commercial GnRH products, although a definitive mechanism for this difference between products was not demonstrated. IVX Pharmaceutical Company United States Department of Agriculture (USDA) (2001-52101-11252) Wisconsin State Experiment Station

The present study investigated how the timing of the administration of estradiol benzoate (EB) impacted the synchronization of ovulation in fixed-time artificial insemination protocols of cattle. To accomplish this, two experiments were conducted, with EB injection occurring at different times: at withdrawal of the progesterone-releasing (P4) intravaginal device or 24h later. The effectiveness of these times was compared by examining ovarian follicular dynamics (Experiment 1, n=30) and conception rates (Experiment 2, n=504). In Experiment 1, follicular dynamics was performed in 30 Nelore cows (Bos indicus) allocated into two groups. On a random day of the estrous cycle (Day 0), both groups received 2mg of EB i.m. and a P4-releasing intravaginal device, which was removed on Day 8, when 400 IU of eCG and 150 microg of PGF were administered. The control group (G-EB9; n=15) received 1mg of EB on Day 9, while Group EB8 (G-EB8; n=15) received the same dose a day earlier. Ovarian ultrasonographic evaluations were performed every 8h after device removal until ovulation. The timing of EB administration (Day 8 compared with Day 9) did affect the interval between P4 device removal to ovulation (59.4+/-2.0 h compared with 69.3+/-1.7h) and maximum diameter of dominant (1.54+/-0.06 acm compared with 1.71+/-0.05 bcm, P=0.03) and ovulatory (1.46+/-0.05 acm compared with 1.58+/-0.04 bcm, P0.01) follicles. In Experiment 2, 504 suckling cows received the same treatment described in Experiment 1, but insemination was performed as follows: Group EB8-AI48 h (G-EB8-AI48 h; n=119) and Group EB8-AI54 h (G-EB8-AI54 h; n=134) received 1mg of EB on Day 8 and FTAI was performed, respectively, 48 or 54 h after P4 device removal. Group EB9-AI48h (G-EB9-AI48 h; n=126) and Group EB9-AI54 h (G-EB9-AI54 h; n=125) received the same treatments and underwent the same FTAI protocols as G-EB8-AI48 h and G-EB8-AI54 h, respectively; however, EB was administered on Day 9. Conception rates were greater (P0.05) in G-EB9-AI54 h [63.2% (79/125) a], G-EB9-AI48 h [58.7% (74/126) a] and G-EB8-AI48 h [58.8% (70/119) a] than in G-EB8-AI54 h [34.3% (46/134) b]. We concluded that when EB administration occurred at device withdrawal (D8), the interval to ovulation shortened and dominant and ovulatory follicle diameters decreased. Furthermore, when EB treatment was performed 24h after device removal, FTAI conducted at either 48 or 54 h resulted in similar conception rates. However, EB treatment on the same day as device withdrawal resulted in a lesser conception rate when FTAI was conducted 54 h after device removal.

This study evaluated a novel presynchronization method, using Ovsynch prior to the Ovsynch-timed AI protocol (Double-Ovsynch) compared to Presynch-Ovsynch. Lactating Holstein (n=337) cows, were assigned to two treatment groups: (1) Presynch (n=180), two injections of PGF 14 d apart, followed by the Ovsynch-timed AI protocol 12 d later; (2) Double-Ovsynch (n=157), received GnRH, PGF 7 d later, and GnRH 3 d later, followed by the Ovsynch-timed AI protocol 7 d later. All cows received the same Ovsynch-timed AI protocol: GnRH (G1) at 68+/-3 DIM (mean+/-SEM), PGF 7 d later, GnRH (G2) 56h after PGF, and AI 16 to 20h later. Pregnancy was diagnosed 39-45 d after timed AI. Double-Ovsynch increased the pregnancies per AI (P/AI) compared to Presynch-Ovsynch (49.7% vs 41.7%, P=0.03). Surprisingly, Double-Ovsynch increased P/AI only in primiparous (65.2% vs 45.2%; P=0.02) and not multiparous (37.5% vs 39.3%) cows. In a subset of 87 cows, ovarian ultrasonography and progesterone (P4) measurements were performed at G1 and 7 d later. Double-Ovsynch decreased the percentage of cows with low P4 (1ng/mL) at G1 (9.4% vs 33.3%) and increased the percentage of cows with high P4 (or =3ng/mL) at PGF (78.1% vs 52.3%). Thus, presynchronization of cows with Double-Ovsynch increased fertility in primiparous cows compared to a standard Presynch protocol, perhaps due to induction of ovulation in non-cycling cows and improved synchronization of cycling cows. Future studies are needed, with a larger number of cows, to further test the hypothesis of higher fertility with Double-Ovsynch, and to elucidate the physiological mechanisms that underlie apparent changes in fertility with this protocol.

A tiered testing strategy has been employed to evaluate the potential of tobacco processes, ingredients, or technological developments to change the biological activity resulting from burning cigarette tobacco. The strategy is based on comparative chemical and biological testing. The introduction of banded cigarette papers in cigarettes to meet New York state "Fire Safety Standards for Cigarettes" constitutes an example of a technological development evaluated utilizing this tiered testing strategy that included a comparison of the chemical and biological effects of cigarettes with and without the banded cigarette paper technologies (BCPT) (representative of current marketed technologies). Specific testing included mainstream cigarette smoke chemistry studies; in vitro studies included genotoxicity (Ames and sister chromatid exchange) and cytotoxicity studies (neutral red); in vivo studies included a 13-week inhalation study in Sprague-Dawley rats and a 30-week dermal tumor promotion study in SENCAR mice. Collectively, data indicated that cigarettes with and without BCPT had a similar toxicological profile in this test battery.

SUMMARY This article describes cooperation between Bradford University Library and the Department of Computing that has resulted in nine research projects over a twenty-five year period on various aspects of bibliographic control. It recounts the origins of the Universal Standard Bibliographic Code (USBC) and its development for the identification of both books and non-book material. It then describes various aspects of the projects including simulating the merging necessary to set up a national database, the cleaning of a database, its use in inter-library lending, and its application together with expert systems for the quality control of databases. The final project is BOPAC that has used modern technology to create faster and better access to a number of library catalogues worldwide and has demonstrated that authority control in its present form is not effective.

A comprehensive 2-year oral chronic toxicity/carcinogenicity study was conducted with smokeless tobacco using modern toxicological test methods and well-accepted standards. The study included a 1-year interim subgroup to assess toxicity at that intermediate time point. Test groups consisted of a tobacco blend (B) used in snus, and an aqueous tobacco extract of that tobacco blend (E) administered at 0.2, 2, or 5 mg nicotine/kg body weight/day via dosed feed to male and female Wistar Han rats. The dosages were selected to simulate potential exposure in humans ingesting smokeless tobacco or an aqueous extract of smokeless tobacco (the latter intended to simulate a snus extract, to enable bridging these data to snus epidemiology data). The following endpoints were evaluated: clinical observations, body weights, feed consumption (FC), ophthalmic exams, toxicokinetics, clinical pathology, gross pathology, and histopathology. During the 2-year study, clear treatment-related, dose-responsive effects included: (1) increases in plasma nicotine and cotinine (indicating that animals were appropriately exposed to levels relevant to human exposure) and (2) decreases in body weights with some alterations in FC. At the 2-year time point, two tumor types (in the highest B doses) displayed statistically significantly increased incidence trends vs. controls: (1) uterine carcinoma in females and (2) epididymal mesothelioma in males. Three tumor types displayed statistically significantly decreased incidence trends: (1) mammary gland adenomas in females, (2) skin basal cell carcinomas in females, and (3) thyroid follicular cell adenomas in males. These increases (and decreases) in tumor trends were interpreted as not being treatment-related because: (1) there were no preneoplastic or related non-neoplastic histopathological findings in the treated rats at the 1-year or 2-year time points to suggest that any of these neoplastic findings were treatment-related and (2) the tumor morphologies and incidences were generally within the expected range of historical controls for Wistar Han rats. Findings from this study indicate that chronic exposure of male and female Wistar Han rats to either a tobacco blend used in snus, or a tobacco extract of that blend does not lead to increased toxicity or carcinogenicity, based on the specified outcomes measured.

A tiered testing strategy has been developed to evaluate the potential for new ingredients, tobacco processes, and technological developments to alter the biological activity that results from burning tobacco. A series of studies was initially conducted with cigarettes containing 3% high fructose corn syrup (HFCS) as an alternate tobacco casing material to corn syrup/invert sugar, including determination of selected mainstream cigarette smoke (MS) constituent yields, Ames assay, sister chromatid exchange (SCE) assay in Chinese hamster ovary (CHO) cells, a 30-week dermal tumor-promotion evaluation of cigarette smoke condensate (CSC) in SENCAR mice, and a 13-week subchronic inhalation study of MS in Sprague-Dawley rats. A second series of studies was conducted with cigarettes containing 3%, 4% and 5% HFCS including MS chemistry, Ames assay, SCE assay in CHO cells, and a neutral red cytotoxicity assays. Collectively, mainstream smoke chemistry, genotoxicity, dermal tumor-promotion, and inhalation toxicity studies demonstrated no differences between cigarettes with 3% HFCS and cigarettes with 3% corn syrup/invert sugar. Also, mainstream smoke chemistry and genotoxicity of cigarettes with 4% and 5% HFCS were not different from cigarettes with 3% HFCS. In conclusion, the addition of up to 5% HFCS to cigarette does not alter the mainstream smoke chemistry or biological activity of mainstream smoke or mainstream smoke condensate as compared to cigarettes with 3% corn syrup/invert sugar with regard to the parameters investigated and presented.

Numerous chemical and toxicological studies indicate that smoke from ECLIPSE, a cigarette that primarily heats rather than burns tobacco, is simplified and reduced in specific chemicals believed to be associated with smoking-related diseases, and demonstrates reduced smoke toxicity and biological activity in vitro when compared to conventional tobacco burning cigarettes. These data led to the hypothesis that cigarette smoke condensate (CSC) from ECLIPSE should have lower tumorigenicity than 1R4F condensate in the SENCAR mouse dermal tumor promotion assay. Female SENCAR mice were initiated with a single topical application of 7,12-dimethylbenz[a]anthracene (DMBA) followed by promotion with ECLIPSE or 1R4F CSC. Dermal application of 10, 20, or 40 mg ECLIPSE or 1R4F CSC three times/week for 29 weeks did not alter body weights, survival or other indicators of subchronic toxicity. In DMBA-initiated mice, there were significant increases in both the number of microscopically confirmed tumor-bearing animals and total number of microscopically confirmed dermal tumors at all 1R4F CSC doses and the high-dose ECLIPSE CSC. However, the number of ECLIPSE tumor-bearing animals were reduced 83%, 93% and 67% at the low-, mid- and high-doses, respectively, compared to the 1R4F. Similarly, the total number of dermal tumors was reduced 91%, 94% and 87% at the low-, mid- and high-dose, respectively, compared to the 1R4F CSC. ECLIPSE CSC demonstrated dramatic reductions in dermal tumor promotion potential compared to 1R4F CSC.

A tiered testing strategy has been developed to evaluate the potential of tobacco processes, ingredients, or technological developments to change the biological activity resulting from burning tobacco. The strategy is based on comparative chemical and biological testing. Expanded shredded tobacco stems (ESS) constitute an example of a common tobacco components expansion process currently used in the manufacture of cigarettes to increase the tobacco blend filling capacity. As part of the toxicological evaluation of ESS, test cigarettes containing 9.5%, 18.5%, and 25% ESS were compared to control cigarettes containing 0% ESS. Testing included mainstream cigarette smoke chemistry studies, genotoxicity studies (Ames and sister chromatid exchange), a 13-week inhalation study in Sprague-Dawley rats, and a 30-week dermal tumor promotion study in SENCAR mice. Collectively, data indicated that cigarettes with and without ESS had a similar toxicological profile in this test battery.

A subchronic, nose-only inhalation study compared the effects of mainstream smoke from a 1R4F research cigarette to that of a 2R4F research cigarette. Male and female rats were exposed for 1 h/day, 5 days/wk, for 13 wk to mainstream smoke at 0, 0.06, 0.20, or 0.80 mg wet total particulate matter per liter of air. Clinical signs, body and organ weights, clinical chemistry, hematology, carboxyhemoglobin, serum nicotine, pulmonary plethysmography, gross pathology, and histopathology were determined. When histological changes resulting from exposure to smoke from the two types of cigarettes were compared, no biologically significant differences were observed. At the end of the exposure period, subsets of rats from each group were maintained without smoke exposures for an additional 13 wk (recovery period). At the end of the recovery period, there were no statistically significant differences in histopathological findings observed between the 1R4F and the 2R4F cigarettes. The complete toxicological assessment in this comparative inhalation study of 1R4F and 2R4F cigarettes suggests no overall biologically significant differences between the rats exposed to the two cigarettes.

A tiered testing strategy has been developed to evaluate the potential for new ingredients, tobacco processes, and technological developments to increase or reduce the biological activity that results from burning tobacco. In the manufacture of cigarettes, honey is used as a casing ingredient to impart both aroma and taste. The primary objective of this document is to summarize and interpret chemical and toxicological studies that have been conducted to evaluate the potential impact of honey on the biological activity of either mainstream cigarette smoke or cigarette smoke condensate. As part of ongoing stewardship efforts, cigarettes produced with honey (5% wet weight) as an alternative to invert sugar in tobacco casing material were subjected to extensive evaluation. Principal components of this evaluation were a determination of selected mainstream smoke constituent yields, Ames assay, sister chromatid exchange assay in Chinese hamster ovary cells, a 30-wk dermal tumor promotion evaluation of cigarette smoke condensate in SENCAR mice, and a 13-wk inhalation study of cigarette smoke in Sprague-Dawley rats. Comparative analytical evaluations demonstrated that the substitution of honey for invert sugar as a casing material in cigarettes had no significant impact on mainstream smoke chemistry. In addition, in vitro and in vivo studies demonstrated that cigarettes containing tobacco cased with honey had comparable biological activity to cigarettes containing invert sugar. Collectively, these data demonstrate that the use of honey as an alternative casing material in the manufacture of cigarettes does not alter the potential toxicity of cigarette smoke condensate (CSC) or cigarette smoke; therefore the use of honey as an ingredient added to cigarette tobacco is acceptable from a toxicological perspective.

A subchronic, nose-only inhalation study compared the effects of mainstream smoke from a cigarette containing 100% flue-cured tobacco cured by a direct-fired process to that of a cigarette containing 100% flue-cured tobacco cured by a heat exchanger process. The tobaccos and mainstream smoke from tobaccos cured by the heat exchanger process have been shown to have significantly lower levels of tobacco-specific nitrosamines than tobaccos cured by a direct-fired process. Male and female rats were exposed for 1 h/day, 5 days/wk, for 13 wk to mainstream smoke at 0, 0.06, 0.20, or 0.80 mg wet total particulate matter per liter of air. Clinical signs, body and organ weights, clinical chemistry, hematology, carboxyhemoglobin, serum nicotine, plethysmography, gross pathology, and histopathology were determined. When histologic changes resulting from exposure to smoke from the two types of cigarettes were compared, the only significant difference was increased epithelial hyperplasia of the anterior nasal cavity in males in the high-exposure group for the heat-exchanger cigarette. At the end of the exposure period, subsets of rats from each group were maintained without smoke exposures for an additional 13 wk (recovery period). At the end of the recovery period, there were no statistically significant differences in histopathological findings observed between the heat-exchanger-cured tobacco cigarette when compared to the direct-fired cured tobacco cigarette. The complete toxicological assessment in this study of heat exchanger and direct-fired tobaccos suggests no overall biologically significant differences between the two cigarettes.

ECLIPSE cigarettes utilize a special form of continuous glass filament (CGF) as an insulator around the carbon heat source. The average numbers of CGFs on the external barrel and cigarette filter end were determined subsequent to manufacture, subsequent to real-world consumer handling and subsequent to simulated consumer handling. The following were not statistically significantly different: the average number of CGFs on the external barrel of cigarettes retrieved from the test market compared to the average external barrel counts from cigarettes subsequent to manufacture or when subjected to simulated consumer handling, and the average number of CGFs on the external barrel of cigarettes subsequent to manufacture compared to the average external barrel counts from cigarettes subjected to simulated consumer handling. The average number of CGFs on the filter end of cigarettes retrieved from the test market was statistically significantly higher than average cigarette filter end counts from cigarettes subsequent to manufacture. The average number of CGFs on the cigarette filter end of cigarettes retrieved from the test market was statistically significantly lower than average cigarette filter end counts from cigarettes subjected to simulated consumer handling. Overall, results from this study suggest that consumer handling does increase the average numbers of CGFs on the external surfaces of the cigarette. Further, the results of this study demonstrate that for the purpose of CGF quantification, the simulated consumer handling protocol used in this study (i.e., based on laboratory measurements of forces) is a reasonably good model for actual consumer handling of cigarettes. Based on the minimal number of CGFs that could be transferred to the smoker and the deposition pattern governed by their physical characteristics, the potential to deposit CGFs from these cigarettes to the lungs of smokers is extremely remote. Therefore, no convincing information exists to suggest that smokers would be exposed to CGFs from any ECLIPSE-related source at a biologically significant level.

Made available in DSpace on 2014-12-03T13:11:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-03-27Bitstream added on 2014-12-03T13:23:33Z : No. of bitstreams: 1 WOS000333677500103.pdf: 1779985 bytes, checksum: c931d4a7396ce7ff5ecaa8c1f0f6e5f9 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Oocytes from dairy cattle and buffaloes have severely compromised developmental competence during summer. While analysis of gene expression is a powerful technique for understanding the factors affecting developmental hindrance in oocytes, analysis by real-time reverse transcription PCR (RT-PCR) relies on the correct normalization by reference genes showing stable expression. Furthermore, several studies have found that genes commonly used as reference standards do not behave as expected depending on cell type and experimental design. Hence, it is recommended to evaluate expression stability of candidate reference genes for a specific experimental condition before employing them as internal controls. In acknowledgment of the importance of seasonal effects on oocyte gene expression, the aim of this study was to evaluate the stability of expression levels of ten well-known reference genes (ACTB, GAPDH, GUSB, HIST1H2AG, HPRT1, PPIA, RPL15, SDHA, TBP and YWHAZ) using oocytes collected from different categories of dairy cattle and buffaloes during winter and summer. A normalization factor was provided for cattle (RPL15, PPIA and GUSB) and buffaloes (YWHAZ, GUSB and GAPDH) based on the expression of the three most stable reference genes in each species. Normalization of non-reference target genes by these reference genes was shown to be considerably different from normalization by less stable reference genes, further highlighting the need for careful selection of internal controls. Therefore, due to the high variability of reference genes among experimental groups, we conclude that data normalized by internal controls can be misleading and should be compared to not normalized data or to data normalized by an external control in order to better interpret the biological relevance of gene expression analysis. Univ Fed Sao Carlos, Ctr Ciencias Biol & Saude, Dept Genet & Evolucao, BR-13560 Sao Carlos, SP, Brazil Univ Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP, Brazil Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Reprod Anim, Sao Paulo, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Reprod Anim, Jaboticabal, SP, Brazil APTA, Unidade Pesquisa Desenvolvimento Registro, Registro, SP, Brazil Vida Reprod Consultoria, Cravinhos, SP, Brazil MSD Saude Anim, Sao Paulo, Brazil LTDA, WTA Tecnol Aplicada, Cravinhos, SP, Brazil Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Cirurgia, Sao Paulo, Brazil Univ Montreal, Fac Med Vet, Ctr Rech Reprod Anim, St Hyacinthe, PQ J2S 7C6, Canada Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Reprod Anim, Jaboticabal, SP, Brazil FAPESP: 09/00938-3 FAPESP: 10/13384-3 FAPESP: 10/09561-7 FAPESP: 11/14207-0 FAPESP: 12/07510-1 CNPq: 476229/2011-1

Contents The uterus plays an essential role in mammalian reproduction and is a target of several hormonal protocols used to improve fertility in cattle. Many studies highlighted the importance of eCG treatment following fixed-time artificial insemination in improving follicular growth, ovulation and pregnancy rates in cattle. Moreover, eCG has been implicated in angiogenesis, leading to important changes in uterine blood flow and vascularisation. However, there is still a lack of information regarding the specific alterations induced by eCG upon glandular and vascular characteristics of bovine uterus. To investigate the influence of eCG on: uterine thickness and area; uterine artery diameter and area; uterine vascular and gland density; and the expression of the VEGFA-system, the uteri of crossbred beef cows were collected. All cows were submitted to follicular wave emergence synchronization. On day four of protocol, cows submitted to superovulation (n = 6) received 2000 IU eCG, on day eight, after expected follicular deviation, cows submitted to stimulatory treatment (n = 5) received 400 IU eCG. Control cows (n = 5) did not receive eCG. On day five po cows were subjected to ultrassonographic evaluation and slaughtered for uterine tissue sampling on day six po. Uterine vessels and glands were quantified by the counting point stereological method. The VEGFA-system was localized in different cellular types, showing no qualitative or quantitative differences in the site of expression or the intensity of the positive signal among the groups. Vascular density was decreased in the endometrium of stimulated and myometrium of superovulated cows compared with the control ones, which showed higher vascular density in the myometrium and endometrium of the ipsilateral uterine horn. The uterine gland density was higher in superovulated compared with stimulated and control cows. Thus, we can infer that stimulatory or superovulatory treatments with eCG influence the vascular density in the endometrium and myometrium in cattle.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific nitrosamine, induces lung adenomas in A/J mice following a single intraperitoneal (i.p.) injection. However, inhalation of mainstream cigarette smoke does not induce or promote NNK-induced lung tumors in this mouse strain purported to be sensitive to chemically-induced lung tumorigenesis. The critical events for NNK-induced lung tumorigenesis in A/J mice is thought to involve O(6)-methylguanine (O(6)MeG) adduct formation, GC--AT transitional mispairing, and activation of the K-ras proto-oncogene. The objective of this study was to test the hypothesis that a smoke-induced shift in NNK metabolism led to the observed decrease in O(6)MeG adducts in the lung and liver of A/J mice co-administered NNK with a concomitant 2-h exposure to cigarette smoke as observed in previous studies. Following 2 h nose-only exposure to mainstream cigarette smoke (600 mg total suspended particulates/m(3) of air), mice (n=12) were administered 7.5 micromol NNK (10 microCi [5-3H]NNK) by i.p. injection. A control group of 12 mice was sham-exposed to HEPA-filtered air for 2 h prior to i.p. administration of 7.5 micromol NNK (10 microCi [5-3H]NNK). Exposure to mainstream cigarette smoke had no effect on total excretion of NNK metabolites in 24 h urine; however, the metabolite pattern was significantly changed. Mice exposed to mainstream cigarette smoke excreted 25% more 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) than control mice, a statistically significant increase (P0.0001). Cigarette smoke exposure significantly reduced alpha-hydroxylation of NNK to potential methylating species; this is based on the 15% reduction in excretion of the 4-(3-pyridyl)-4-hydroxybutanoic acid and 42% reduction in excretion of 4-(3-pyridyl)-4-oxobutanoic acid versus control. Detoxication of NNK and NNAL by pyridine-N-oxidation, and glucuronidation of NNAL were not significantly different in the two groups of mice. The observed reduction in alpha-hydroxylation of NNK to potential methylating species in mainstream cigarette smoke-exposed A/J mice provides further mechanistic support for earlier studies demonstrating that concurrent inhalation of mainstream cigarette smoke results in a significant reduction of NNK-induced O(6)MeG adduct formation in lung and liver of A/J mice compared to mice treated only with NNK.

Demonstrates through case studies how authority control simply does not work. Shows how the case studies were carried out using BOPAC2 which provides facility for downloading large files and a greater range of displays than normal OPACs. Stresses that authority control is important not only to library catalogues but also to information on the Internet. Because it is so important and because it is so expensive priority action is needed to rectify the situation. Suggests a number of ways in which this might be done.

The efficiency of different doses of rBST in a MOET protocol was verified in buffaloes. The animals received an intravaginal progesterone device (DIB) plus 2.0 mg of estradiol benzoate (Ric-BE, i.m.) and 0 mg, 250 mg or 500 mg of rBST. Ovarian follicular growth was stimulated by treatment with 200 mg of FSH in decreasing amount (12/12hs). Buffaloes received injections of PGF2α on D6 and on D7. The DIB was removed on D7. On D8, 24 hours after DIB removal, ovulation was induced with GnRH. The follicular emergence and the response to FSH treatment were evaluated by ultrasound on D4 and D8. The artificial insemination (AI) was performed 12 and 24 hours after the GnRH administration. The embryonic structures were collected 5.5 days after the first AI. Data were analyzed by ANOVA. No statistical differences were found between groups. The results indicated that rBST, associated to a MOET protocol at the different dosages used, does not to improve the efficiency of the technique in buffalo.

This paper describes a Research Project, funded by the British Library Research and Innovation Centre, from September 1996 to January 1998.1 The Project developed and extensively tested and evaluated a World Wide Web front end called BOPAC2 that allows access to a number of library catalogues via Z39.50 either simultaneously or individually. BOPAC2 is designed to make access to large and complex retrievals simpler. Similar records are clustered together and retrievals may be sorted in a number of ways and by different criteria. The design, development and evaluation of the system are described along with suggestions for future work.

Three factors are likely to force dramatic changes in the libraries of the future. They are the increase in electronic publishing, the digitising of conventional library materials, and the Internet which is creating a second information revolution. New concepts are needed for cataloguing functions in the Internet environment. Cataloguing, now a precoordinate function should shift to a post-coordinate activity. Authority control should become an activity of the searching stage rather than the cataloguing stage. Uniform titles will be needed as one of the linking mechanisms. The value of ISBD (International Standard Bibliographic Control) needs to be justified. Issues connected with the Internet and search engines are explored. A scenario for the Catalogue 2000 is given that envisions the cataloguers' aims being the inclusion of library catalogue material that is relevant but not necessarily in the library.

This study was designed to determine if the Eclipse prototype 9-014 cigarettes, which use a special form of continuous glass filament (CGF) as an insulator around the carbon heat source, yield CGFs via mainstream smoke. A previously developed method (Higuchi et al., 2000) that employed electrostatic precipitation-with a greater than 99% collection efficiency of mass-was used to capture CGFs transferred to mainstream smoke. The cigarettes were smoked using an exaggerated puffing condition more than twice the Federal Trade Commission (FTC) standard. Prior to smoking, cigarettes were subjected to handling procedures that simulated commercial shipping conditions. Using a modified standard addition method, and utilizing a mixture of water and glycerol as a mock condensate, CGFs were intentionally added to a series of (mock condensate) samples to develop knowledge of CGF recovery efficiency. The linear regression model of the recovered CGFs demonstrated a recovery efficiency of 86%. This efficiency rate was applied to the number of CGFs recovered from samples of smoke condensate and associated background samples. The number of CGFs in smoke condensate collected from the Eclipse 9-014 prototype was approximately 0.32 +/- 0.17 CGFs per cigarette (+/- standard deviation), including the background counts of CGFs, and 0.16 CGFs per cigarette when corrected for background contributions. The number of CGFs found in the smoke condensates for this prototype was statistically (p =.00031) distinguishable from zero and background in these experiments. The low number of CGFs seen in the transfer data from this prototype studied, the unique physical characteristics of the filaments (e.g., controlled physical dimensions), and the absence of biological activity of similar glass filaments/fibers indicate that biologically significant exposure to the Eclipse smoker does not occur.

4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific nitrosamine, induces lung adenomas in A/J mice, following a single intraperitoneal (ip) injection. However, inhalation of tobacco smoke has not induced or promoted tumors in these mice. NNK-induced lung tumorigenesis is thought to involve O 6 -methylguanine (O 6 MeG) formation, leading to GC→AT transitional mispairing and an activation of the K-ras proto-oncogene in the A/J mouse. NNK can be metabolized by several different cytochromes P 450 , resulting in a number of metabolites. Formation of the promutagenic DNA adduct O 6 MeG is believed to require metabolic activation of NNK by cytochrome P 450 -mediated α-hydroxylation of the methylene group adjacent to the N-nitroso nitrogen to yield the unstable intermediate, methanediazohydroxide. Nicotine, cotinine (the major metabolite of nicotine), and aqueous cigarette tar extract (ACTE) have all been shown to effectively inhibit metabolic activation of NNK to its mutagenic form, most likely due to competitive inhibition of the cytochrome P 450 enzymes involved in α-hydroxylation of NNK. The objective of the current study was to monitor the effects of cotinine and cigarette smoke (CS) on the formation of O 6 MeG in target tissues of mice during the acute phase of NNK treatment. To test the effect of cotinine, mature female A/J mice received a single intraperitoneal injection of NNK (0, 2.5, 5, 7.5, or 10 μmole/mouse) with cotinine administered at a total dose of 50 μmole/mouse in 3 separate ip injections, administered 30 min before, immediately after, and 30 min after NNK treatment. To test the effect of whole smoke exposure on NNK-related O 6 MeG formation, mice were exposed to smoke generated from Kentucky 1R4F reference cigarettes at 0, 0.4, 0.6, or 0.8 mg wet total particulate matter/liter (WTPM/L) for 2 h, with a single ip injection of NNK (0, 3.75, or 7.5 μmole/mouse) midway through the exposure. Cigarette smoke alone failed to yield detectable levels of O 6 MeG. The number of O 6 MeG adducts following ip injection of NNK was significantly (p < 0.05) reduced in both lung and liver by cotinine and by cigarette smoke exposure. Our results demonstrate that NNK-induced O 6 MeG DNA adducts in A/J mice are significantly reduced when NNK is administered together with either cotinine, the major metabolite of nicotine, or the parental complex mixture, cigarette smoke.

The objectives of the present work were to determine follicle deviation assessed by ultrasonography and profile of plasmatic gonadotrophins, and ovulatory capacity in buffalo species, since these information are not available in the literature. The knowledge of follicular development and gonadotrophins profile during deviation and the follicular response to an exogenous ovulation inducer can be useful tools for follicular manipulation of estrous cycle.

Evaluation of the Genotoxic and Cytotoxic Potential of Mainstream Whole Smoke and Smoke Condensate from a Cigarette Containing a Novel Carbon Filter. Bombick, D. W., Bombick, B. Reed, Ayres, P. H., Putnam, K., Avalos, J., Borgerding, M. F., and Doolittle, D. J. (1997). Fundam. AppL Toxicol. 39, 11-17. A novel carbon filter has been developed which primarily reduces the amount of certain vapor phase constituents of tobacco smoke with greater efficiency than the charcoal filters of cigarettes currently in the market In vitro indicators of genotoxic and cytotoxic potential were used to compare the cigarette smoke condensate (particulate phase) or whole cigarette smoke (vapor phase and particulate phase) from cigarettes containing the novel carbon filter with smoke condensate or whole smoke from commercial or prototype cigarettes not containing the novel carbon filter. Ames bacterial mutagenicity, sister chromatid exchange (SCE) in Chinese hamster ovary (CHO) cells, and neutral red cytotoxicity assays in CHO cells were utilized to assess the genotoxic and cytotoxic potential of the cigarette smoke condensates. SCE and neutral red cytotoxicity assays were utilized to assess the genotoxic and cytotoxic potential of the whole smoke. As expected, the novel carbon filter did not significantly affect the genotoxic or cytotoxic activity of the smoke condensate, although we did observe that the use of low-nitrogen tobacco reduced the mutagenicity of the condensate in Salmonella typhimurium strain TA98. However, the whole smoke from cigarettes containing the novel carbon filter demonstrated significant reductions in genotoxic and cytotoxic potential compared to cigarettes without the novel carbon filter. The toxicity of the smoke was correlated (r = 0.7662 for cytotoxicity and r — 0.7562 for SCE induction) to the aggregate mass of several vapor phase components (acetone, acetaldehyde, acrolein, acrylonitrile, 1,3-butadiene, ammonia, NO,, HCN, benzene, isoprene, and formaldehyde) in the smoke of the cigarettes utilized in this study. In conclusion, this novel carbon filter, which significantly reduced the amount of carbonyls and other volatiles in mainstream cigarette smoke, resulted in significant reductions in the genotoxic and cytotoxic activity of the smoke as measured by