Use of prostaglandin F2α as ovulatory stimulus for synchronizing dairy cattle.

The aim of this study was to evaluate if prostaglandin F2α (PGF) can be used to induce ovulation in a GnRH-progesterone based protocol. In Experiment 1 crossbred dairy cows (n=32) were synchronized with a progesterone-GnRH based protocol for seven days, where the luteolytic dose of 150µg PGF was given 24h prior progesterone device removal (CIDR). On Day 8 cows were separated into two groups to receive: 1) 2mL of Saline (Control Group, n=15) or 2) 150µg of PGF (PGF Group, n=17). Ovulation rate was higher in the PGF than Control group (100% vs 53.3%, P=0.001, Odds ratio=30.88). The percentage of cows that ovulated synchronously tended to be higher in the PGF than Control group (P=0.1, Odds ratio=9.6). Experiment 2 was performed in a cross-over (3×3) design. Crossbred dairy cows (n=25) received a CIDR for seven days and GnRH on Day 0. Seven days later 150µg of PGF was given and the progesterone device was removed, and 24h later cows were distributed into three groups to receive: 1) 2mL of Saline (Control Group, n=25), 2) 150µg of PGF (PGF Group, n=25) or 3) 1mg of ECP (ECP Group, n=23). Diameter of ovulatory follicle was larger in the PGF and Control than ECP Group (P=0.002, Effect size>4.0). Synchronized ovulation rate (between 72 and 96h after CIDR removal) tended to be higher in PGF group in Control group (P=0.1, Odds ratio=0.35). Results suggest that PGF is equally efficient to ECP to induce synchronized ovulation in dairy cows subjected to progesterone-GnRH based protocols.

Comparison between two estradiol-progesterone based protocols for timed artificial insemination in blocks in lactating Nelore cows.

The aim of this study was to compare the use of artificial insemination in time blocks (Artificial Insemination Blocks, AIB) using an 8 and 9 d estradiol-progesterone based protocol. In this experiment, lactating Nelore cows (n=253) were subjected to two estradiol-progesterone based TAI protocols. On the morning of Day 10 (8d group, n=124) or Day 11 (9d group, n=129), cows were examined by ultrasonography to evaluate the diameter of the preovulatory follicle and were inseminated once at one of the following time points, according to the diameter of the pre-ovulatory follicle (POF): Block 0 (POF≥15mm, TAI 0h after conventional TAI), Block 1 (POF 13.0-14.9mm, TAI 6h later), Block 2 (POF 10.1-12.9mm, TAI 24h later), and Block 3 (POF≤10.0mm, TAI 30h later). The pregnancy per AI (P/AI) did not differ between 8d and 9d groups (P>0.05). Considering only multiparous cows, however, P/AI tended to be greater in the 8d (64.1%) than in the 9d group (49.3%; P=0.08). Cows from the 9d group tended to have a larger POF than cows from the 8d group (P=0.07). In conclusion, these results provide evidence that there is no difference between 8d or 9d protocols when using the AIB technique. Use of the 8d estradiol-progesterone based protocol, however, tended to increase pregnancy in multiparous cows.

Timed artificial insemination in blocks: A new alternative to improve fertility in lactating beef cows.

The aim of this study was to evaluate whether changing the interval from CIDR removal to timed artificial insemination (TAI) according to the diameter of the preovulatory follicle (POF) would improve pregnancy per AI in cows. In Study 1, a retrospective analysis of TAI experiments (n=96 cows) was performed to characterize the time of ovulation according to the diameter of the dominant follicle. It was observed that cows with a larger POF had ovulations earlier than cows with smaller POF, according to the equation: y=0.72x(2)-26.74x+264.54 (R(2)=0.63; P<0.001). In Study 2, lactating Nelore cows (n=412) were subjected to an EB-CIDR based TAI protocol. On the morning of Day 10 (time of TAI), cows were randomized into Control (n=209) and Block (n=203) groups; (1) Cows in the Control Group were TAI 48 h after CIDR removal (08:00 am on Day 10), and; (2) Cows in the block group were inseminated once at one of the following time points, according to the diameter of the POF on Day 10: B0 (POF≥15mm, TAI 0 h after convetional TAI), B1 (POF 13-14.9 mm, TAI 6h later), B2 (POF 10.1-12.9 mm, TAI 24h later) and B3 (POF≤10mm, TAI 30 h later). The cows of the Block Group had greater pregnancy rates per AI than the Control Group (129/203, 63.5% when compared with 102/209, 48.8%, respectively; P<0.01). In conclusion, results of the present study demonstrate that adjusting the timing of TAI according to the diameter of the POF can be an effective practice for improving fertility of cows in TAI protocols.

The use of PGF2α as ovulatory stimulus for timed artificial insemination in cattle.

The objective of this study was to evaluate the effect of a PGF2α-analogue (PGF) on ovulation and pregnancy rates after timed artificial insemination (TAI) in cattle. In experiment 1, crossbred dual-purpose heifers, in a crossover design (3 × 3), were given an intravaginal progesterone-releasing insert (controlled internal drug release [CIDR]) plus 1 mg estradiol benzoate (EB) intramuscularly (im) and 250 µg of a PGF-analogue im on Day 0. The CIDR inserts were removed 5 days after follicular wave emergence, and the heifers were randomly divided into three treatment groups to receive the following treatments: (1) 1 mg of EB im (EB group, n = 13); (2) 500 µg of PGF im (PG group, n = 13); or (3) saline (control group, n = 13), 24 hours after CIDR removal. Ovulation occurred earlier in EB (69.81 ± 3.23 hours) and PG groups (73.09 ± 3.23 hours) compared with control (83.07 ± 4.6 hours; P = 0.01) after CIDR removal. In experiment 2, pubertal beef heifers (n = 444), 12 to 14 months of age were used. On Day 0, the heifers were given a CIDR insert plus 2 mg EB im. On Day 9, the CIDR was removed and the heifers were given 500 µg of PGF im. Heifers were randomly assigned into one of three treatment groups: (1) 1 mg of EB (EB group; n = 145); (2) 500 µg of PGF (PG group; n = 149), both 24 hours after CIDR removal; or (3) 600 µg of estradiol cypionate (ECP group; n = 150) at CIDR removal. Timed artificial insemination occurred 48 hours after CIDR removal in the ECP group and 54 hours in the PG and EB groups. The percentage of heifers ovulating was higher in the PG group compared with the other groups (P = 0.08). However, the pregnancy rates did not differ among groups (47.6%, 45%, and 46.6%, for EB, PG, and ECP, respectively; P = 0.9). In experiment 3, 224 lactating beef cows, 40 to 50 days postpartum with 2.5 to 3.5 of body condition score were treated similarly as described in experiment 2, except for the ECP group, which was excluded. The treatments were as follows: 1 mg EB (EB group; n = 117) or 500 µg PGF (PG group; n = 107), 24 hours after CIDR removal. The calves were temporarily separated from their dams from Days 9 to 11. No difference was detected on the pregnancy rate between the EB and PG groups (58.1% vs. 47.6%, respectively; P = 0.11). Taken together, the combined results suggested that PGF2α could be successfully used to induce and synchronize ovulation in cattle undergoing TAI, with similar pregnancy rates when compared with other ovulatory stimuli (ECP and EB).

Respiratory system of Gluconacetobacter diazotrophicus PAL5. Evidence for a cyanide-sensitive cytochrome bb and cyanide-resistant cytochrome ba quinol oxidases.

In highly aerobic environments, Gluconacetobacter diazotrophicus uses a respiratory protection mechanism to preserve nitrogenase activity from deleterious oxygen. Here, the respiratory system was examined in order to ascertain the nature of the respiratory components, mainly of the cyanide sensitive and resistant pathways. The membranes of G. diazotrophicus contain Q(10), Q(9) and PQQ in a 13:1:6.6 molar ratios. UV(360 nm) photoinactivation indicated that ubiquinone is the electron acceptor for the dehydrogenases of the outer and inner faces of the membrane. Strong inhibition by rotenone and capsaicin and resistance to flavone indicated that NADH-quinone oxidoreductase is a NDH-1 type enzyme. KCN-titration revealed the presence of at least two terminal oxidases that were highly sensitive and resistant to the inhibitor. Tetrachorohydroquinol was preferentially oxidized by the KCN-sensitive oxidase. Neither the quinoprotein alcohol dehydrogenase nor its associated cytochromes c were instrumental components of the cyanide resistant pathway. CO-difference spectrum and photodissociation of heme-CO compounds suggested the presence of cytochromes b-CO and a(1)-CO adducts. Air-oxidation of cytochrome b (432 nm) was arrested by concentrations of KCN lower than 25 microM while cytochrome a(1) (442 nm) was not affected. A KCN-sensitive (I(50)=5 microM) cytochrome bb and a KCN-resistant (I(50)=450 microM) cytochrome ba quinol oxidases were separated by ion exchange chromatography.