Sero-Monitoring of Horses Demonstrates the Equivac® HeV Hendra Virus Vaccine to Be Highly Effective in Inducing Neu-Tralising Antibody Titres

Hendra virus (HeV) is a high consequence zoonotic pathogen found in Australia. The HeV vaccine was developed for use in horses and provides a One Health solution to the prevention of human disease. By protecting horses from infection, the vaccine indirectly protects humans as well, as horses are the only known source of infection for humans. The sub-unit-based vaccine, containing recombinant HeV soluble G (sG) glycoprotein, was released by Pfizer Animal Health (now Zoetis) for use in Australia at the end of 2012. The purpose of this study was to collate post-vaccination serum neutralising antibody titres as a way of assessing how the vaccine has been performing in the field. Serum neutralization tests (SNTs) were performed on serum samples from vaccinated horses submitted to the laboratory by veterinarians. The SNT results have been analysed, together with age, dates of vaccinations, date of sampling and location. Results from 332 horses formed the data set. Provided horses received at least three vaccinations (consisting of two doses 3–6 weeks apart, and a third dose six months later), horses had high neutralising titres (median titre for three or more vaccinations was 2048), and none tested negative.

ANALISIS PERBANDINGAN BIAYA ANTARA SEBELUM DAN SETELAH PENGETATAN BIAYA LANDED COST PADA PT. PFIZER INDONESIA (PFIZER ANIMAL HEALTH)

Tujuan dalam penelitian adalah untuk mengetahui pengaruh kebijakan pengetatan biaya yang dilakukan oleh PT Pfizer Indonesia (Pfizer Animal Health) dengan membandingkan biaya sebelum dan setelah kebijakan pengetatan biaya landed cost pada tahun 2015 melalui uji komparasi dengan mengamati variabel landed cost yang terdiri dari tiga indikator yang digunakan yaitu: pembelian atau purchase, bea masuk (BM), biaya proses kepabeanan (BPK) dengan menggunakan analisis F atau uji analisis ragam atau ANOVA (analysis of variance) digunakan untuk menganalisis perbandingan perubahan kebijakan pengetatan biaya landed cost periode tahun fiskal 2013 sampai dengan 2015 dengan memusatkan pada faktor perubahan kebijakan pengetatan biaya landed cost dengan mengunakan data internal dengan tiga indikator utama yaitu: pembelian (purchase), bea masuk (BM) atau import duty dan biaya proses kepabeanan (BPK) atau Custom clearance. Berdasarkan analisis data menggunakan uji analisa ragam (ANOVA) satu arah menunjukkan bahwa terdapat perbedaan signifikan pada biaya landed cost sebelum dan setelah diterapkannya pengetatan biaya dimana hal tersebut dapat dilihat dari nilai Fhitung (28,27) lebih besar dari pada Ftabel (3,68). Ini berarti bahwa penerapan pengetatan biaya landed cost memberikan perbedaan yang signifikan terhadap biaya landed cost setelah penerapan pengetatan biaya. Atau artinya terdapat perbedaan yang signifikan pada biaya landed cost setelah penerapan kebijakan pengetatan biaya pada tahun 2015 dibandingkan dengan sebelum pengetatan biaya diterapkan. Hasil penelitian ini secara keseluruhan menjelaskan bahwa penerapan kebijakan pengetatan biaya memberikan kontribusi besar dalam efisiensi biaya yang akan meningkatkan pendapatan perusahaan dan menjaga kestabilan harga pokok penjualan. Kata Kunci: Pengetatan biaya, analisis landed cost

ANALISIS PERBANDINGAN BIAYA ANTARA SEBELUM DAN SETELAH KEBIJAKAN PENGETATAN BIAYA LANDED COST PADA PT. PFIZER INDONESIA (PFIZER ANIMAL HEALTH)

Tujuan dalam penelitian adalah untuk mengetahui pengaruh kebijakan pengetatan biaya yang dilakukan oleh PT Pfizer Indonesia (Pfizer Animal Health) dengan membandingkan biaya sebelum dan setelah kebijakan pengetatan biaya landed cost pada tahun 2015 melalui uji komparasi dengan mengamati variabel landed cost yang terdiri dari tiga indikator yang digunakan yaitu: pembelian atau purchase, bea masuk (BM), biaya proses kepabeanan (BPK) dengan menggunakan analisis F atau uji analisis ragam atau ANOVA (analysis of variance) digunakan untuk menganalisis perbandingan perubahan kebijakan pengetatan biaya landed cost periode tahun fiskal 2013 sampai dengan 2015 dengan memusatkan pada faktor perubahan kebijakan pengetatan biaya landed cost dengan mengunakan data internal dengan tiga indikator utama yaitu: pembelian (purchase), bea masuk (BM) atau import duty dan biaya proses kepabeanan (BPK) atau Custom clearance. Berdasarkan analisis data menggunakan uji analisa ragam (ANOVA) satu arah menunjukkan bahwa terdapat perbedaan signifikan pada biaya landed cost sebelum dan setelah diterapkannya pengetatan biaya dimana hal tersebut dapat dilihat dari nilai Fhitung (28,27) lebih besar dari pada Ftabel (3,68). Ini berarti bahwa penerapan pengetatan biaya landed cost memberikan perbedaan yang signifikan terhadap biaya landed cost setelah penerapan pengetatan biaya. Atau artinya terdapat perbedaan yang signifikan pada biaya landed cost setelah penerapan kebijakan pengetatan biaya pada tahun 2015 dibandingkan dengan sebelum pengetatan biaya diterapkan. Hasil penelitian ini secara keseluruhan menjelaskan bahwa penerapan kebijakan pengetatan biaya memberikan kontribusi besar dalam efisiensi biaya yang akan meningkatkan pendapatan perusahaan dan menjaga kestabilan harga pokok penjualan. Kata kunci: pengetatan biaya, analisis landed cost

PREVALENCE, DIAGNOSTICS AND TREATMENT OF OVARIAN FOLLICULAR CYSTS IN DAIRY COWS

Ovarian cysts are among the primary causes for poor reproduction efficiency and economic losses in dairy cattle farming. Material and metods: The studies were performed in 2015-2017 in three dairy cattle farms.In the experiment, 79 cows with follicular ovarian cysts were divided randomly into 3 groups. To the first group (n=33), a standard double Ovsynch was applied with 0.01 mg GnRH (Veterelin, Laboratorios Calier, Spain) on day 0, 150 µg PGF2α (D – Cloprosstenolsodium, Indupart, Vetpharma Animal Health, Spain) on day 7, 0.01 mg GnRH (Veterelin, Laboratorios Calier, Spain) on day 10, and after 7 days, Ovsynch (GnRH-PGF-GnRH). The cows from the second group (n=28) were submitted to double Ovsynch and additional progesterone treatment through placement of Controlled Internal Drug Release device (EAZI BREED CIDR Cattle insert, 1.38 g progesterone; Pfizer Animal Health, USA) for 7 days between the first GnRH at day 0 and prostaglandin on day 7. The control group comprised 18 untreated cows. Results: First-service conception rate in cows treated by double Ovsynch and vaginal inserts was higher 28.6% compared to that in standard double Ovsynch: 24.2%. Cumulative pregnancy percentage after applied treatments was 60.7% in double Ovsynch plus progesterone and 54.5% in standard double Ovsynch.

98 COMMERCIAL APPLICATION OF A NEW COW-SIDE LH ASSAY FOR DETERMINING OPTIMUM AI INTERVALS IN SUPEROVULATED BEEF DONORS IN THE USA: A PRELIMINARY STUDY

Successful superovulation and embryo collection require donors to be managed for precise insemination intervals. At times, determinations of these intervals can become difficult because of oestrus detection subjectivity. The objective of this study was to determine the feasibility of performing a rapid cow-side LH assay (Predi′Bov®, ReproPharm, Nouzilly, France) on superovulated donors with emphasis on determining how to use the results in a commercial program to time inseminations. This study was conducted at Ovagenix (Bryan, TX, USA) on 24 superovulated donors. On Day 0 cows received an intravaginal progesterone-releasing device (CIDR®, Pfizer Animal Health, New York, NY, USA) and were administered 25 mg of progesterone and 2.5 mg of oestradiol 17β intramuscularly (IM, 2 mL). Superstimulation was induced using 150–300 mg of Folltropin-V® (FSH, Bioniche Animal Health Inc., ON, Canada) administered with 7 injections at 12-h intervals in decreasing dosages (Day 4–7). On Day 6, two doses of Estrumate® (equaling 1 mg, Intervet Inc., Merck Animal Health, Summit, NJ, USA) were given IM, concurrently with FSH injections 5 and 6 with CIDR removal on Day 7 (a.m.). HeatWatch® transmitters (Cow Chips LLC, Denver, CO, USA) were applied to donors on Day 7 to determine onset of standing oestrus. Blood samples were collected in heparinized tubes via tail vein puncture starting at CIDR removal continuing every 6 h until a positive Predi′Bov® test was acquired or 36 h after CIDR removal. Whole blood (0.5 mL–1) was submitted to a proprietary 3-step procedure using a tube-stick applicator (40 min) to determine whether increased concentrations of LH were present. Stick applicators that turned blue were positive for increased concentrations of LH. Donors were artificially inseminated 12 and 24 h after a positive test, with specified sires, and embryos were nonsurgically collected 7 days after insemination. The Predi′Bov® test determined 91.6% (22/24) of the LH peaks occurred within 12 to 24 h after CIDR removal; 45.8% (11/24) occurred at 18 h; and 4% (1/24) at 30 h. A total of 203 viable embryos (average 8.5/donor) was collected with 174 embryos (average 9.2 ± 4.7/donor) from donors that exhibited oestrus and produced a positive test (19/24; 79.2%) and 29 embryos (average 5.8 ± 6.3/donor) from donors exhibiting silent oestrus that produced a positive test (5/24; 20.8%). One donor exhibited oestrus and never produced a positive test, but contributed 8 viable embryos to the total. In conclusion, Predi′Bov® cow-side usage could offer commercial utility when identifying oestrus is difficult or nonexistent. Increased detection of ovulation could help to identify optimal AI intervals, thereby increasing embryo production and limiting the use of expensive semen for desired donors.

162 FERTILITY OF BEEF RECIPIENTS FOLLOWING A FIXED-TIME EMBRYO TRANSFER PROTOCOL WHICH INCLUDED FOLLICLE STIMULATING HORMONE DILUTED IN HYALURONAN

This study was performed to test the viability of administering Folltropin-V® (FSH, Bioniche Animal Health) diluted in hyaluronan (MAP-5 50 mg, sodium hyaluronate, Bioniche Animal Health) to beef cows enrolled in a recipient synchronization protocol to evaluate its effect on recipient fertility. All recipients were administered an estradiol 17β (2.5 mg, IM) and progesterone (50 mg, IM) combination injection on Day 0, a CIDR® (progesterone 1.34 g, Pfizer Animal Health, Groton, CT, USA) was inserted for 7 days. Lutalyse® (dinoprost tromethamine, Pfizer Animal Health, 25 mg, IM) was administered at the time of CIDR removal on Day 7, and estradiol 17β (1 mg, IM) was administered on Day 8. On Day 16, the presence of at least one corpus luteum, detected via ultrasound, resulted in the recipient receiving an embryo (both fresh and frozen–thawed embryos were used). Embryos were not transferred into cows that did not show ultrasonic evidence of a CL. Dependent variables for which data were collected included circulating progesterone levels at the time of transfer and CL diameter, area, and circumference; measured in millimeters. The total study (n = 274) consisted of both wet (n = 85) and dry (n = 189) cows and included both Bos indicus (Brahman-influenced) crossbred (n = 93) and Bos taurus (Angus-based) cows (n = 181). The experiment consisted of cows being placed in either the treated or control groups, with treated cows receiving a single 40 mg (1 mL) IM injection of Folltropin-V in hyaluronan on Day 5 and control cows receiving no additional injections. Results are shown in Table 1. Transfer rate, conception rate, and pregnancy rate were tested for significance with chi-square analysis and remaining statistics were analyzed with a t-test: two-sample assuming equal variances. There were no significant differences found between the treated and control groups for transfer rate, conception rate, or pregnancy rate. Corpus luteum diameter was shown to be larger in control cows (P < 0.05); however, CL area and circumference did not differ significantly. Folltropin-V given with hyaluronan at a 40-mg dose on Day 5 did not improve fertility, induce a larger CL, or increase circulating progesterone levels in synchronized beef recipients as hypothesized. Further work is needed with Folltropin-V in hyaluronan to determine if an alternative dose or timing of administration would be more appropriate for improving fertility in recipients. Table 1.Fertility data in beef recipients following synchronization for fixed-time embryo transfer with a protocol that included (Treated) or did not include (Control) FSH in hyaluronan

317 EFFECT OF TIME OF ONSET OF FOLLICLE-STIMULATING HORMONE TREATMENT ON SUPEROVULATORY RESPONSE AND EMBRYO YIELD OF SANTA INES EWES DURING BREEDING SEASON

This study was designed to investigate if the time of onset of FSH treatment [near the emergence of first or last follicular wave on progesterone (P4) protocol] influenced the superovulatory response and embryo yield in Santa Ines ewes during breeding season. Days of emergence of the follicular waves were defined in a previous study that evaluated the follicular dynamic in oestrus synchronization treatments (Oliveira et al. 2011 Acta Sci. Vet. 40). We observed emergence of the first and last follicular wave on 5.69 ± 0.42 and 11.25 ± 0.39 days of protocol, respectively. Twenty Santa Ines ewes were submitted to 2 superovulatory protocols according to the time that FSH treatments were initiated (G-first wave, n = 10; G-last wave, n = 10). On Day 0, all ewes received a P4 device (CIDR®; Pfizer Animal Health, New York, NY, USA) and injection of 37.5 µg of d-cloprostenol, IM. The FSH treatments started on Day 6 and Day 11 of protocol for G-first and G-last, respectively. The superovulatory regimen consisted of 8 IM injections of pFSH administrated twice daily (40, 40, 30, 30, 20, 20, 10, and 10 mg of pFSH). The P4 device was removed on Day 8 and Day 13 for G-first and G-last, respectively. At these times, all ewes received another injection of 37.5 µg of d-cloprostenol and a dose of 200 IU of eCG. During 4 days after the P4 device removal, ewes were mated by a fertile ram. Embryo collections were accomplished 7 days after CIDR withdrawal. The ovaries were evaluated by ultrasonography (3 times daily, during the mating period) and laparotomy (concomitantly with embryo collection). The superovulatory response was observed by classified by score: 0 = 4 or fewer corpora lutea (CL); 1 = between 5 and 10 CL; and 2 = 11 or more CL. Data were analysed by GLIMMIX using SAS software (SAS Institute Inc., Cary, NC, USA). All donors from G-first had superovulatory response classified as score 2, whereas 60% of ewes from G-last were classified as score 2, 20% as score 1, and 20% as score 0 (P < 0.05). There were effects between treatments (P < 0.05) for ovulation rate (G-first: 97.9 ± 1.4% v. G-last: 88.5 ± 4.4%) and number of ovulations (G-first: 17.0 ± 2.3 v. G-last: 12.5 ± 2.6). The numbers of luteinized unovulated follicles were 0.7 ± 0.5 for G-first and 1.2 ± 0.4 for G-last (P > 0.05). There was no difference between G-first and G-last (P > 0.05) in the rate of ova/embryos recovered (54.9 ± 5.7% v. 49.3 ± 8.5%), mean number of ova/embryos recovered (9.0 ± 1.4 v. 6.3 ± 1.1), number of viable embryos (3.8 ± 1.5 v. 3.4 ± 0.8), or viability rate (40.3 ± 10.8 v. 53.4 ± 12.1). In conclusion, the FSH treatment started near the emergence of the first follicular wave of progesterone protocol in Santa Ines ewes during the breeding season resulted in a higher superovulatory response than that started near the last follicular wave; however, no improvements in embryo yield were observed. Financial support: FAPESP.

18 PREGNANCY RATES IN LACTATING DAIRY COWS TREATED WITH GONADOTROPIN-RELEASING HORMONE-BASED SYNCHRONIZATION PROGRAMS AND INSEMINATED AT A FIXED TIME

An experiment was designed to compare pregnancy rates in lactating dairy cows synchronized with a 7-day CIDR-Synch or a 5-day CIDR-Synch program and to determine if the addition of a second prostaglandin F2α (PGF) injection to the 7-day CIDR-Synch program would improve pregnancy rates following fixed-time AI (FTAI). The experiments were performed on 2 dairy farms in Argentina, with year-round calving and a mixed feeding system (35% grazing plus 65% corn silage and grain). Cows (n = 621) were 39.3 ± 6.5 days in milk (DIM, mean ± SD) when they were enrolled in the program, had 2.4 ± 1.5 lactations and a body condition score (BCS) of 3.1 ± 0.2 (range: 2.7 to 4.0). All cows received a pre-synchronization treatment with 2 doses of prostaglandin (PGF, 25 mg of dinoprost, Lutalyse, Pfizer Animal Health, Argentina) 14 days apart, and 11 days after the second PGF (Day 0) received 10 µg of Buserelin (GnRH, Receptal, MSD-Intervet, Argentina) and a CIDR device (1.9 g of progesterone, Pfizer Animal Health). Cows were randomly allocated to 1 of 3 groups. The CIDR devices were removed and PGF was administered to cows in Groups 1 and 2 on Day 7. A second GnRH was given 56 h later and cows experienced FTAI 16 h after gonadotropin-releasing hormone (GnRH) injection (i.e. 72 h after CIDR removal). Cows in Group 2 also received a second PGF injection on the afternoon of Day 7. Cows in Group 3 had the CIDR removed and received 2 PGF injections 12 h apart on Day 5. A second dose of GnRH was given and FTAI was performed at the same time, on Day 8 (i.e. 72 h after CIDR removal). All cows were examined by ultrasonography (Aloka 500V, Aloka, Tokyo, Japan) on the day of the first PGF injection and at CIDR removal to determine the presence and number of corpora lutea (CL), and 30 days after FTAI to determine pregnancy status. Data were analyzed by logistic regression to determine the effects of treatment, parity, days postpartum, milk production, BCS, presence of a CL at enrollment, and number of CL at the time of CIDR removal on pregnancy rates. Overall pregnancy rates did not differ among groups: 32.9% (68/207), 38.2% (78/204), and 38.3% (80/209) for Groups 1, 2, and 3, respectively (P = 0.2). Although the number of CL present at CIDR removal did not significantly affect pregnancy rates (P = 0.4), pregnancy rates in cows with 1 CL in Groups 1 and 2 tended to differ [29.0% (11/38) v. 48.9% (21/43); P < 0.07], but neither differed from that in Group 3 [37.2% (16/43)]. No differences were detected among groups in cows without a CL at CIDR removal [overall pregnancy rate: 29.4% (5/17)] and those with ≥2 CL [overall pregnancy rate: 36.1% (173/479)]. Among the other variables evaluated, first-parity cows had 1.96 (1.38–2.78) times more chance of getting pregnant than second-or-more-parity cows (P = 0.002) and cows with BCS >3 had 1.63 (1.16–2.28) times more chance of getting pregnant than those with BCS <3 (P = 0.003). Finally, herd, days postpartum, milk production, and presence of a CL at enrollment did not significantly affect pregnancy rates. We concluded that the 3 treatments resulted in similar pregnancy rates for lactating dairy cows and that the benefit of adding a second PGF injection to the 7-day protocol was only marginal in cows with 1 CL at CIDR removal.

318 EFFECT OF BREED AND FOLLICULAR STATUS ON RESPONSE TO SUPEROVULATION IN SOUTH AFRICAN GOATS

This study evaluated the effect of breed and follicular status on quantity and quality of embryos recovered following superovulation in South African goats. Mature, nonlactating Boer (n = 10) and unimproved indigenous (n = 10) goats were used in this trial. Oestrous cycles of does were synchronised with controlled internal drug release (CIDR®; Pfizer Animal Health, New York, NY, USA) dispensers inserted for 9 days and injected with prostaglandin at CIDR insertion. All does were superovulated with pFSH, administered in 7 dosages, at 12 h intervals, starting from 48 h before CIDR removal. Ultrasonographic scans were performed at initiation of the superovulation treatment. Does were cervically inseminated with fresh undiluted semen at 36 and 48 h following CIDR withdrawal. Embryos were surgically collected at Day 6 following AI. The total number of recovered structures (unfertilised ova and embryos) was evaluated microscopically for the stage of development and quality, using morphological criteria. The structures were classified as unfertilised ova (if there was no cleavage), degenerate embryos (embryos at 8-cell stage and earlier stage), or as transferable grade 1, 2, and 3 embryos (morphologically intact compacted morulae, early blastocysts, and expanded blastocysts). All data were analysed using the ANOVA procedures of SAS (SAS Institute Inc., Cary, NC, USA). The time from CIDR removal to onset of oestrus did not differ significantly between breeds. Breed had no effect on the total number of corpora lutea, structures (unfertilised ova and embryos), unfertilised ova, embryos, degenerated embryos, and transferable embryos. The total number of follicles at the onset of pFSH treatment was positively correlated with the number of structures (unfertilised ova plus embryos) recovered. Similarly, the total number of embryos recovered was positively correlated with the number of follicles 2 to 3 mm and 4 to 5 mm, and the total number of follicles. The number of follicles 4 to 5 mm and the total number of follicles at the onset of pFSH treatment were positively correlated with total number of the transferrable embryos at the beginning of a superovulation treatment. The total number of transferable embryos was negatively correlated with the presence of follicles with a diameter >6 mm at the beginning of the superovulation treatment. In conclusion, breed had no effect on response to superovulation. The total number of follicles and follicle diameters 2 to 3 mm and 4 to 5 mm were positively correlated with embryo yield. The presence of follicles >6 mm at the beginning of superovulation negatively affected the quality of embryos. The results of this study indicate that breed has no effect on response to superovulation. However, the number and size of follicles at the beginning of a superovulation treatment affect both embryo yield and quality.