Effects of different doses of estradiol benzoate used in a cervical relaxation protocol on the success of non-surgical embryo recovery and luteal function in superovulated ewes.

This study investigated the effectiveness of different doses of estradiol benzoate (EB) to promote cervical relaxation and their effects on luteal function and outcomes of non-surgical embryo recovery (NSER) in sheep. Multiparous (MULT) and nulliparous (NULL) crossbred Lacaune X Santa Inês ewes were superovulated and naturally bred. Seven days after progesterone withdrawal, females were randomly assigned to one of three distinct cervical relaxation protocols, consisting of i.m. treatment with 37.5 µg d-cloprostenol and different doses of EB: 0.0 mg (0.0EB group; n = 3 NULL and 14 MULT); 0.5 mg (0.5EB group; n = 4 NULL and 12 MULT) or 1.0 mg (1.0EB group, n = 6 NULL and 11 MULT) 16 h before NSER. All ewes received 50 IU of oxytocin 20 min before NSER (D17). Blood samples were collected and ultrasound exams (B-mode and color Doppler) were performed at two timepoints: immediately before d-cloprostenol and EB treatments and prior to NSER. Estrous behavior, corpora lutea count and NSER success outcomes were not affected by EB treatments nor parity (P > 0.05). Embryo recovery rate was greater for ewes in the 0.5EB group and in the NULL ewes (P < 0.05). Ovarian biometrics differed between the two evaluation timepoints in all groups (P < 0.05). Plasma estradiol increased over time, reaching a significant greater level in 1.0EB ewes compared to controls on D17 (P < 0.05), whereas progesterone concentrations decreased over time in all groups (P > 0.05). In conclusion, treatments did not affect NSER success but they did affect luteal function by altering P4 and E2 concentrations. Therefore, the NSER technique can be successfully performed in ewes with or without prior treatment with EB.

Single dose of 300 IU hCG in the early luteal phase in superovulated ewes: Effects on corpora lutea, progesterone profile, and embryo recovery.

This study aimed to evaluate the effects of hCG treatment during the early luteal phase on ovarian function, progesterone profile, and embryo yield in superovulated ewes. Superovulated sheep were randomly assigned to receive 300 IU hCG i.m. (GhCG, n = 24) or not (GControl, n = 25) at 96 h after the removal of the progesterone (P4) device (D13). Non-surgical embryo recovery (NSER) was performed eight days after P4 withdrawal. Ultrasound evaluations were performed on D13, D14, D16, and D17. Blood samples were collected on D14, D16, and D17. Superovulation scores were recorded based on the number of corpora lutea (CL) as follows: 1 (≤ 2), 2 (3-5), 3 (6-8), and 4 (≥ 9). NSER efficiency, superovulation response, and luteal tissue area were similar in both groups (P > 0.05). Structural luteolysis tended to be higher in GControl (P = 0.07; 47.0 %) while functional luteolysis was similar in both groups (P > 0.05; 0.0 % and 5.9 %). The recovery rate was greater (P < 0.05) in GhCG (89.8 %) compared with GControl (71.0 %), with similar overall ova/embryo numbers observed for both groups (P > 0.05). GhCG showed a higher concentration of animals with a superovulatory response score of 4 (54.5 %; P < 0.05) compared with the lowest scores. Plasma progesterone on D16 was higher (P < 0.05) in GhCG ewes (11.1 ± 1.5 vs 6.9 ± 1.5 ng/mL). In conclusion, the hCG treatment improved circulating P4 and embryo recovery rate, tended to maintain luteal functionality, and thus constitutes an additional tool for improving embryo yield in superovulated ewes.

Hair as an alternative noninvasive matrix: sources of variation in testosterone levels.

Hair is one widely used alternative matrix for endocrine studies. Not only can it maintain hormone content during storage for long periods of time, but its collection also induces little to no stress. Noninvasive techniques have broadened the opportunities for endocrine research, particularly regarding wild animals. Despite its advantages, many sources of variation may affect the steroid concentration found in hair, such as body location harvested, fur color, reproductive status, and sex. Thus, domestic species, such as the dog, are an excellent and approachable model for understanding this variability. For such, we addressed diverse sources of variation in testosterone concentrations from 24 domestic dogs (Canis lupus familiaris) of the Poodle breed of various colors and neuter status, and from both sexes. The variation comprised the comparison between 2 different matrices (blood vs hair); 2 different extraction storage methods (refrigerator vs freezer); 3 body regions (head, torso, and limbs); 3 coat colors (black, brown, and white); different neuter status (intact vs castrated males) and, finally, sex. Our results showed no correlation between blood and hair testosterone concentrations. Additionally, we did not find differences related to the storage method, body region, or coat color. There were differences in concentration between males and females, but not between females and castrated males. We discuss hair testosterone levels exhibited reasonable stability, and we present practical applications for both domestic and wildlife animals.

Treatment with eCG and hCG to induce onset of estrous cycles in ewes during the non-breeding season: Effects on follicular development and fertility.

Although combining of eCG and hCG administrations is known to enhance LH-like actions, there have been few studies where there was comparison of the effects of treatment of anestrous ewes with eCG and hCG and eCG alone. In Experiment 1, 18 ewes in seasonal anestrus were administered an intravaginal device (IVD) containing medroxyprogesterone acetate for 12 days, and at the time of IVD removal (D0), were allocated into the following groups (n = 6/group): no further treatment (control); 400 IU eCG (eCG); or 400 IU eCG and 200 IU hCG (eCG + hCG). There was greater ovarian follicular growth in the groups treated with gonadotropins, compared to the control, and there were greater progesterone concentrations in the eCG + hCG group on D9 (P < 0.05). In Experiment 2, 66 ewe lambs were assigned to the same treatment groups described for Experiment 1, and subsequently there was natural mating with rams. There was a greater rate of behavioral estrous manifestation in the eCG (88.5 %; 23/26) and eCG+hCG (85.2 %; 23/27), than control (30.8 %; 4/13; P < 0.05) group. Pregnancy rate was also greater in the eCG (34.6 %; 9/26) and eCG+hCG (18.6 %; 5/27) than control (0 %; 0/13; P < 0.05) group, whereas conception rate, considering only ewe lambs that were mated, was only greater in the eCG group. Although there were greater progesterone concentrations 9 days after treatment in the eCG+hCG group, there was no difference in follicular growth in anestrous ewes, nor was there an effect on estrous behavior manifestation and pregnancy rates in ewe lambs, compared to treatment with only eCG.

Diversity in migratory patterns among Neotropical fishes in a highly regulated river basin.

Migratory behaviour of selected fish species is described in the Paraná River, Brazil-Argentina-Paraguay, to search for patterns relevant to tropical regulated river systems. In a 10 year mark-recapture study, spanning a 1425 km section of the river, 32 867 fishes composed of 18 species were released and 1083 fishes were recaptured. The fishes recaptured were at liberty an average 166 days (maximum 1548 days) and travelled an average 35 km (range 0-625 km). Cluster analysis applied to variables descriptive of movement behaviour identified four general movement patterns. Cluster 1 included species that moved long distances (mean 164 km) upstream (54%) and downstream (40%) the mainstem river and showed high incidence (27%) of passage through dams; cluster 2 also exhibited high rate of movement along the mainstem (49% upstream, 13% downstream), but moved small distances (mean 10 km); cluster 3 included the most fishes moving laterally into tributaries (45%) or not moving at all (25%), but little downstream movement (8%); fishes in cluster 4 exhibited little upstream movement (13%) and farthest downstream movements (mean 41 km). Whereas species could be numerically clustered with statistical models, a species ordination showed ample spread, suggesting that species exhibit diverse movement patterns that cannot be easily classified into just a few classes. The cluster and ordination procedures also showed that adults and juveniles of the same species exhibit similar movement patterns. Conventional concepts about Neotropical migratory fishes portray them as travelling long distances upstream. The present results broaden these concepts suggesting that migratory movements are more diverse, could be long, short or at times absent, upriver, downriver or lateral, and the diversity of movements can vary within and among species. The intense lateral migrations exhibited by a diversity of species, especially to and from large tributaries (above reservoirs) and reservoir tributaries, illustrate the importance of these habitats for the fish species life cycle. Considering that the Paraná River is highly impounded, special attention should be given to the few remaining low-impact habitats as they continue to be targets of hydropower development that will probably intensify the effects on migratory fish stocks.