Review: Non-surgical artificial insemination and embryo recovery as safe tools for genetic preservation in small ruminants.

Artificial insemination (AI) and in vivo embryo production (or multiple ovulation and embryo transfer, MOET) programs are both instrumental in accelerating the propagation of genetically and economically superior goats and sheep. The aim of this review was to present the current gestalt of non-surgical AI and embryo recovery (NSER) procedures in small ruminants. Small body size, precluding rectal palpation, and highly limited penetrability of the uterine cervix in ewes are the major reasons for the scarce use of non-surgical assisted reproduction techniques in this species. As a result, AI and embryo recovery techniques in sheep mainly involve laparoscopy or laparotomy (LAP). In does, however, the Embrapa method of AI allows for successful intrauterine deposition of semen, resulting in pregnancy rates from 50 to 80% under field conditions (>3 000 goats inseminated) when frozen-thawed semen is used. After the administration of prostaglandin F2α (PGF2α), non-surgical (transcervical) embryo recovery is also feasible in goats, with the cervical penetration rate approaching 100%. There is a paucity of information on the efficacy of non-surgical AI using frozen semen in sheep, but the results are satisfactory with fresh, cooled, or chilled ram semen. An application of the NSER technique in ewes has greatly improved over the last decade, and cervical penetration rates of ∼90% can be achieved when a hormonal cervical dilation protocol using PGF2α, oxytocin, and/or estradiol ester (e.g., estradiol benzoate) is applied. In some genotypes of sheep, sufficient cervical dilation can be induced without estradiol ester included in the protocol. Several studies indicated that recovery of transferable quality ovine embryos using NSER is comparable to that employing a ventral midline laparotomy, and NSER is evidently a method of choice when animal welfare is concerned. Considering both the number of retrievable embryos and animal well-being, the NSER is a viable alternative for surgical procedures. With further developments, it has the makings of a primary, if not exclusive, embryo recovery technique in small ruminants worldwide.

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.

Administration of a single dose of 300 IU of human chorionic gonadotropin seven days after the onset of estrus improves pregnancy rate in dairy goats by an unknown mechanism.

This study examined the effects of exogenous hCG administration on ovarian function and pregnancy rates in estrous-induced dairy goats during the transition into the breeding season. Eighty-six Toggenburg does received 60 mg of medroxyprogesterone acetate intravaginal sponge for 6 d plus 200 IU of equine chorionic gonadotropin and 30 µg of d-cloprostenol i.m. 24 h before sponge removal, and were then bred for 96 h. Seven days (D7) after first mating the does received either 1 mL of saline (the control group, n = 43) or 300 IU of hCG (the hCG-treated group, n = 43) i.m. Transrectal ovarian ultrasonography (B-mode and color Doppler) was performed on D7, D13, D17, and D21 and ultrasonographic pregnancy detection on D30. Pregnancy rate was higher (P < 0.05) in hCG-treated goats (90.7%; 39/43) than that in control animals (74.4%; 32/43). Accessory luteal structures (ALSs) were detected in 46.5% (20/43) of hCG-treated does. All hCG-treated does that had ALSs and 82.6% of goats without ALS post-treatment remained pregnant. The total luteal area increased (P < 0.05) from D7 to D13 in pregnant animals of both groups, whereas mean vascular area declined (P < 0.05) by D21 in all nonpregnant does. Serum progesterone concentrations increased (P < 0.05) on D21 in pregnant goats of both groups, but they were related to changes in luteal tissue content only in control does throughout the present study. Mean daily numbers of small- and medium-sized antral follicles decreased (P < 0.05) only in pregnant animals of both groups with a decline in medium follicle numbers occurring earlier in hCG-treated (D13) compared with control does (D17). To summarize, a single dose of hCG given on D7 after estrus was followed by a decrease in the number of medium-sized antral follicles in gestating hCG-treated does, induced the formation of ALSs in ~47% of all hCG-treated does, and significantly increased the pregnancy rate in estrous-induced Toggenburg goats in the transition to the breeding season.

Cervical penetration rates and efficiency of non-surgical embryo recovery in estrous-synchronized Santa Inês ewes after administration of estradiol ester (benzoate or cypionate) in combination with d-cloprostenol and oxytocin.

The effects of estradiol esters, d-cloprostenol and oxytocin on induction of cervical dilation prior to non-surgical embryo recovery in Santa Inês ewes (Days 6-7 estrous cycle) were assessed in this study. In Trial 1, transcervical embryo flushing was performed in estrous-induced ewes administered 37.5 µg of d-cloprostenol i.m. 10 h before and 50 IU of oxytocin i.v. 20 min before uterine flushing with (EB-PGF-OT; n = 13) or without (PGF-OT; n = 11) 1 mg of estradiol benzoate i.m. administered concurrently with d-cloprostenol injection. In Trial 2, the estrous-synchronized animals were treated with 1 mg of estradiol benzoate (EB-PGF-OT; n = 12) or estradiol cypionate (EC-PGF-OT; n = 12) i.m. along with 37.5 µg of d-cloprostenol i.m. 16 h before and 50 IU of oxytocin i.v. 20 min before uterine flushing. In Trial 1, uterine flushing could be accomplished in 38% of ewes in the EB-PGF-OT and 27% those in the PGF-OT (P>0.05) group. Flushing fluid recovery averaged 90% and there were 1.0 ± 1.1 embryos/ewe collected with mean duration of the flushing procedure being ˜36 min. In Trial 2, uterine flushing was accomplished in 78% of ewes in the EB-PGF-OT and 44% of those in the EC-PGF-OT group (P>0.05) with mean flushing fluid recovery rate being 88% and time elapsing to complete flushing being ˜33 min. Within the subsets of animals treated with EB, the percentages of successful transcervical penetrations were 38% compared with 78% in Trials 1 and 2, respectively (i.e., with EB administered 10 h compared with 16 h before uterine flushing: P<0.05). The interval from EB administration to the beginning of transcervical penetration can affect the efficacy of embryo recovery procedures utilizing a combined EB/d-cloprostenol/oxytocin pre-treatment.

Freezing goat embryos at different developmental stages and quality using ethylene glycol and a slow cooling rate / Congelação de embriões caprinos em diferentes estádios de desenvolvimento e qualidade utilizando etileno glicol e taxa de resfriamento lenta

The efficiency of an alternative freezing protocol for goat embryos of different morphology and quality was tested. Fifty-eight embryos on Day 6-7 stage were transferred as fresh or after freeze-thawing (n=29/group). For freezing, embryos were placed into 1.5M ethylene-glycol solution for 10min. During this time, they were loaded in the central part of 0.25mL straw, separated by air bubble from columns containing PBS/BSA 0.4% plus 20% BFS. Straws were then frozen using a freezing machine from 20ºC to -6ºC at a cooling rate of 3ºC/min, stabilization for 15min (seeding after 5min), from -6 C to -32ºC at 0.6 C/min,and plunged into liquid nitrogen. Frozen embryos were thawed for 30s at 37ºC in a water bath. Embryos subjected to fresh transfer were maintained in holding medium (37ºC). Fresh and frozen-thawed embryos were transferred at day 7 post-estrus to 30 recipients. Kidding and kid born rates were similar (P> 0.05), respectively, for recipients receiving fresh (66.7% or 10/15; 55.2% or 16/29) or frozen-thawed (60% or 9/15; 51.7% or 15/29) embryos. The cryopreservation of goat embryos using slow-freezing protocol and 1.5MEG resulted in similar efficiency rates of fresh embryos.(AU)

Este estudo testou a eficiência de protocolo alternativo de criopreservação de embriões caprinos de diferentes qualidades morfológicas. Foram utilizados 58 embriões, coletados entre o sexto e o sétimo dia do ciclo estral (n=29/grupo). Embriões congelados passaram por solução 1,5M etilenoglicol por 10min e foram aspirados durante esse tempo para parte central de palheta 0,25mL, separada por bolhas de ar de colunas contendo PBS 0,4% BSA e 20% SFB. As palhetas foram congeladas em máquina de congelação de 20ºC a -6ºC, com taxa de resfriamento de 3ºC/min, estabilização por 15min (seeding após 5min), -6ºC a -32ºC a 0,6ºC/min, e imersas em nitrogênio líquido. Os embriões foram descongelados por 30s a 37ºC, em água. Embriões frescos foram mantidos em solução de manutenção (37ºC). Embriões frescos e congelados/descongelados foram transferidos para 30 receptoras no sétimo dia do ciclo estral. A taxa de partos e a de crias nascidas (respectivamente) foram similares (P>0,05) para receptoras recebendo embriões frescos (66,7% ou 10/15; 55,2% ou 16/29) ou congelados/descongelados (60,0% ou 9/15; 51,7% ou 15/29). O protocolo de criopreservação de embriões utilizado no presente estudo resultou em índices de eficiência semelhantes aos de embriões frescos.(AU)

Relationship of antral follicular blood flow velocity to superovulatory responses in ewes.

The aim of this study was to examine the association between antral follicular blood flow velocity and the response of ewes to hormonal ovarian superstimulation. Ten Santa Inês ewes were subjected to a short- (7days; Group 1) or long-term (13days; Group 2) progesterone (CIDR®; InterAg, Hamilton, New Zealand) priming, and a superovulatory treatment with porcine follicle-stimulating hormone (pFSH; Folltropin®-V; Bioniche Animal Health, Belleville, ON, Canada), given twice daily for four consecutive days in decreasing doses and initiated four or ten days after CIDR insertion, respectively. Embryos were recovered surgically seven days after the last pFSH dose. From one day prior to until the end of the pFSH regimen (Days -1 to 3), all ewes underwent daily transrectal ultrasonography of ovaries. The number of high-velocity pixels (HVPs; 0.055-0.11m/s or upper 50% of recordable velocities) on Day 1 correlated directly with the number of corpora lutea (CL; r=0.92, P=0.0002) and of viable embryos (r=0.77, P=0.01). Correlations were also recorded between the number of HVPs on Day 3 and the recovery rate (r=-0.69, P=0.03), viability rate (r=-0.64, P=0.05), and percentage of degenerated embryos (r=0.65, P=0.04). The percentage of HVPs relative to the total area of ovarian cross section on Day 1 was correlated with the number of CL (r=0.95, P<0.001) and of viable embryos (r=0.85, P=0.002). This parameter on Day 3 was also correlated with the recovery rate (r=-0.69, P=0.03). The percentage of HVPs relative to the total Doppler area on Day 0 was correlated with the recovery rate (r=0.72, P=0.02). It can be concluded that sonographic assessment of high-velocity antral follicular blood flow has the makings of a useful non-invasive method to predict the outcome of the superovulatory treatment in ewes.

Effect of altering the intervals between consecutive superovulatory doses of porcine follicle-stimulating hormone on ovarian responses and embryo yields in anestrous ewes.

The effect of varying intervals between successive gonadotropin injections on the superovulatory outcomes in anestrous Rideau Arcott ewes superstimulated for ovarian follicular development with multiple doses of porcine FSH (pFSH) was evaluated in a single study. Twenty-five animals received six (1×2.5ml and 5×1.25ml) injections of Folltropin®-V given at 0800 and 1600h or at 0800 and 2000h in Group 1 (n=9) or Group 2 (n=16), respectively. An i.m. injection of 500 IU of equine chorionic gonadotropin (eCG; Folligon®) was given concurrently with the first pFSH dose. Time of estrus was synchronized among ewes with intravaginal sponges containing 60mg of medroxyprogesterone acetate (Veramix®) that were left in place for 14days; sponges were removed at the time of the 5th pFSH injection. Six days after insertion of MAP sponges, all ewes received an i.m. injection of estradiol-17ß dissolved in 1ml of sesame oil (350µg/ewe) to synchronize follicular wave emergence. Following the last pFSH dose, all animals were given a single i.m. injection of 50µg of gonadotropin-releasing hormone (GnRH; Cystorelin®) to induce ovulations before placing in a pen with four fertile rams for 36h. The ovarian responses were assessed and embryos recovered surgically 7days after GnRH injections. The mean number of corpora lutea was greater (P<0.05) in Group 1 compared with Group 2 ewes (21.0±2.9 compared with 10.4±1.6, respectively; mean±SEM) but there was no difference (P>0.05) in the number of transferable embryos (5.4±2.4 compared with 5.4±1.3/ewe, respectively), and Group 1 animals had significantly more degenerated embryos than Group 2 ewes (2.6±1.2 compared with 0.6±0.3/ewe, respectively). A superovulatory protocol wherein pFSH injections were given at 0800 and 1600h was more effective in terms of inducing multiple ovulations than the protocol with 12-h intervals between consecutive pFSH doses, but it was not associated with an increased production of transferable quality embryos by anestrous ewes.

Is progesterone the key regulatory factor behind ovulation rate in sheep?

Ovarian antral follicles in the ewe grow in an orderly succession, producing 3 to 4 waves per estrous cycle. In prolific sheep, some large antral follicles from the second-to-last wave of the estrous cycle are added to the ovulatory follicles emerging just before estrus to give a higher ovulation rate; it is feasible that regression of these follicles is prevented by an increase in serum concentrations of FSH or LH pulsatility at proestrus. Prolific sheep tend to have a shorter luteal phase than nonprolific ewes and there is a great deal of evidence that luteal progesterone (P4), in addition to regulating LH release, may govern the secretion of FSH heralding the emergence of follicular waves. The specific purpose of this study was to determine whether or not extending the duration of the luteal phase in prolific sheep to that typically seen in nonprolific breeds would alter the follicle wave dynamics and ovulation rate. In 2 separate experiments, exogenous P4 (7.5 mg per ewe intramuscularly) was administered on day 11 at PM and day 12 at AM (day 0 = first ovulation of the interovulatory interval studied) in moderately prolific Rideau Arcott × Polled Dorset ewes (experiment 1, n = 8) and highly prolific Olkuska ewes (experiment 2, n = 7; TRT), whereas the equinumerous groups of animals served as controls (CTR). Transrectal ovarian ultrasonography was performed daily, and jugular blood samples were drawn twice a day from day 9 until the next ovulation. Progesterone injections resulted in relatively uniform increments in serum P4 levels, but the mean duration of the interovulatory interval did not differ (P > 0.05) between TRT and CTR groups of ewes in either experiment. The mean ovulation rate post-treatment was 1.6 ± 0.2 vs 3.2 ± 0.4 (experiment 1, P < 0.001) and 3.2 ± 0.8 vs 4.0 ± 1.0 (experiment 2, P > 0.05) in TRT vs CTR, respectively. The number and percentage of ovulating follicles from the penultimate wave of the interovulatory interval studied was 0.25 ± 0.16 vs 1.75 ± 0.45 (P < 0.01) and 25.0 ± 16.4% vs 75.0 ± 16.4% (P < 0.05) in experiment 1, and 0.50 ± 0.30 vs 1.60 ± 0.40 (P < 0.05) and 13.8 ± 9.0% vs 53.4 ± 16.7% (P < 0.05) in experiment 2, for TRT vs CTR, respectively. In summary, administration of P4 at the end of diestrus decreased the incidence of ovulations from the penultimate wave of the estrous cycle in both the moderately and highly prolific strains of sheep, but it reduced the ovulation rate only in moderately prolific ewes.

Follicular wave emergence in Santa Inês ewes subjected to long-term, progesterone-based estrous synchronization protocols at different times of the year.

This study was conducted to document the pattern of antral follicular wave emergence throughout the 14-day, progesterone (P4)-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 P4 treatment on ensuing ovarian follicular wave development was examined. Sexually mature Santa Inês 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 F2α 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<0.05) during the ID period than during the TP and DD periods (4.0±0.1, 3.4±0.1 and 3.6±0.1, respectively). The presence of medium-sized antral follicles (4.0 to 5.75mm) in the absence of corpora lutea at the time of CIDR insertion tended to advance follicular wave emergence. Although the long-term P4 treatment was not originally designed to synchronize follicular waves, there was a distinctive pattern of follicular wave dynamics during the period of application of CIDRs that was affected mainly by the number of emerging follicular waves and ovarian status at CIDR insertion.

Effects of season and ovarian status on the outcome of long-term progesterone-based estrus synchronization protocols and ovulatory follicle development in Santa Inês ewes under subtropical conditions.

This study set out to investigate the efficiency of long-term estrus synchronization protocols and ovulatory follicle dynamics in ultrasonographically monitored Santa Inês ewes during lengthening (LD; September-October) and shortening photoperiods (SD; April-May), and the transitional period (TP; January). In addition, the influence of ovarian status (e.g., size of antral follicles and/or presence of corpora lutea) at the outset of the estrus synchronization protocols on the ensuing development of ovulatory follicles was examined. Seventy sexually mature Santa Inês ewes were subjected to one of the two estrus synchronization regimens; on Day 0 (random day of the estrous cycle or anovulatory period), the ewes were fitted with an intravaginal progesterone (P4)-releasing (controlled intrauterine drug release [CIDR]) device, which was left in place for 14 days (G-1CIDR, n = 35) or replaced on Day 7 (G-2CIDR, n = 35), and received an intramuscular injection of 10 mg of PGF2α. The ewes allocated to the G-1CIDR group had mean serum P4 concentrations less than 2 ng/mL during the last 4 days of the synchronization protocol. There were no differences (P > 0.05) in mean ovulation rates between the two protocols tested nor among the ewes varying in ovarian status or studied at different times of the year, but ovulations occurred ∼ 12 hours later in the TP compared with the SD period (P < 0.05). Ovulatory follicles emerged earlier (P < 0.05) in the G-1CIDR group than in the G-2CIDR group (Day 8.3 ± 0.5 vs. 9.2 ± 0.4) and during LD (Day 7.1 ± 0.6) compared with the TP (Day 9.1 ± 0.6) and SD (Day 9.9 ± 0.5 of the protocol). In conclusion, the replacement of CIDR devices prevented the occurrence of lower-than-normal luteal phase levels of P4 at the end of the 14-day estrus synchronization protocol. However, although this procedure and seasonal influences altered certain growth characteristics of ovulatory follicles, there were no effects of these factors on the mean ovulation rate.

Conventional and Doppler ultrasonography on a goat with gangrenous mastitis / Ultrassonografia convencional e Doppler em cabra com mastite gangrenosa

Objetivou-se descrever os achados clínicos e ultrassonográficos de uma cabra leiteira apresentando mastite gangrenosa. Com a ultrassonografia convencional, observaram-se alterações consistentes com o avanço do quadro clínico, como alterações na ecogenicidade do leite e parênquima mamário. A ultrassonografia Doppler revelou aumento da velocidade de pico sistólico nos três momentos estudados, passando de 15 para 17 e 29cm/s. Houve redução, também, dos valores do índice de resistência, de 0,71 para 0,61 e 0,43. O índice de pulsatividade sofreu redução entre o segundo e o terceiro dia de observação, passando de 0,98 para 0,71. Foi possível estabelecer relação direta entre os achados ultrassonográficos e a gravidade da doença. A ultrassonografia convencional e Doppler podem ser utilizadas para o estudo hemodinâmico do úbere de cabras leiteiras, favorecendo diagnóstico e prognóstico de alterações nesse órgão...

Profile of IL-2, IL-4, IL-10, IFN-gamma, TNF-alpha and KC-like cytokines in pregnant bitches / Perfil de citocinas IL-2, IL-4, IL-10, IFN-gamma, TNF-alpha e células KC-like em cadelas gestantes

The aim of this study was to determine the profile of IL-2, IL-4, IL-10, IFN-γ, and TNF-α cytokines and KC-like cells (natural killer) in pregnant bitches, unpublished values for the species. A total of 27 females of the Shi Tzu, Pug, English Bulldog and French breeds, weighing 4-20kg and aged 4-6 years were used. Blood samples were collected from bitches during the anestrous and on the 2nd, 5th, 6th, 7th and 8th week of pregnancy. Serum levels of cytokines were measured by panel MILLIPLEX MAP (CCYTO-90K, MILLIPORE, Billerica, Massachusetts, USA) validated for dogs. Twenty four females showed physiological pregnancy and three bitches showed pathological pregnancy. There was no difference between cytokine values during anestrous and gestational weeks of bitches (P>0.05). However, it was possible to verify the physiological behavior of serum levels during modulation of immune response in the gestational process of animals. In animals with gestational disorders, abnormal values for IL-2, IL-4 and INF-y were noted. It was concluded that serum levels of cytokines evaluated in pregnant bitches can help the better understanding of physiological and pathological gestational processes and correlated immunology in this species...

Objetivou-se por meio deste estudo determinar o perfil das citocinas IL-2, IL-4, IL-10, IFN-γ, TNF-α e células KC-like (natural killer) em cadelas gestantes, valores que são inéditos para a espécie. Foram utilizadas 27 fêmeas das raças Shi-tzu, Pug, Bulldog Inglês e Francês, pesando de 4 a 20 kg e com idade entre quatro e seis anos. Foram coletadas amostras sanguíneas das cadelas durante o anestro e na segunda, quinta, sexta, sétima e oitava semanas gestacionais. Os níveis séricos de citocinas foram dosados pelo painel MILLIPLEX MAP (CCYTO-90K, MILLIPORE, Billerica, Massachusetts, EUA) validado para cães. Vinte e quatro fêmeas apresentaram gestação fisiológica e três cadelas, gestação patológica. Não foi verificada diferença entre os valores das citocinas durante anestro e nas semanas gestacionais das cadelas (P>0,05). Entretanto, foi possível verificar o comportamento fisiológico dos níveis séricos durante a modulação da resposta imune no processo gestacional dos animais. Verificaram-se, nos animais com afecções gestacionais, valores alterados para IL-2, IL-4 e INF-y. Pôde-se concluir que os valores séricos das citocinas avaliadas, nas cadelas gestantes, podem auxiliar no maior conhecimento dos processos fisiológicos e patológicos da gestação, correlacionados à imunologia, nessa espécie...

Echobiometrics kidney and renal artery triplex doppler of canine fetuses / Ecobiometria renal e Doppler Triplex da artéria renal de fetos caninos

The aim of this study was to assess the sogographic parameters and biometry of canine fetal kidneys using the B mode, and to determinate the vascular index of the fetal renal arteries using the Doppler Triplex. Twenty four Shi-tzu and Pug, weighting between 4 and 10kg, aging between 4 and 6 years old were evaluated. The B mode, the fetal renal echobiometry and regularity of the renal surface, echotexture and cortex:medular ratio were evaluated during the 5th, 6th, 7th and 8th weeks of pregnancy. At the same time point of the B mode evaluation, the Doppler Triplex was carried out to assess the sistolic peak velocity (SPV), end diastolic velocity (EDV), vascular resistive (RI) and pulsatility index (PI). B mode revealed no fetal renal abnormalities and echobiometry showed important measurements during fetal development (P<0.0001). The values of the fetal renal arteries index (PSV and EDV) increased during the course of the pregancy (P<0.05) and remained constant for PI and RI (P>0.05). B mode and Doppler Triplex were important tools for the assessment of fetal renal development, using echobiometry and renal arterial index in canie fetuses...

O objetivo deste estudo foi avaliar, por meio da ultrassonografia convencional modo B, as características sonográficas e a biometria dos rins de fetos caninos, bem como determinar os índices vasculares da artéria renal dos conceptos ao Doppler Triplex. Foram utilizadas 24 fêmeas Shi-tzu e Pugs pesando de quatro a 10 kg e com idade entre quatro e seis anos. Ao modo B, a ecobiometria renal fetal, a regularidade da superfície renal, a ecotextura e a relação córtico-medular foram avaliadas durante a quinta, sexta, sétima e oitava semanas gestacionais. Ao Doppler Triplex, durante o mesmo período em que se realizou o exame convencional, foram determinados o pico de velocidade sistólica (PVS), a velocidade diastólica final (EDV) e o índice de resistência vascular (RI) e de pulsatividade (PI). Ao modo B, não foram detectadas alterações em rins fetais, e à ecobiometria renal dos fetos, foi possível determinar medidas renais importantes, verificando-se aumento das biometrias no decorrer do desenvolvimento fetal (P<0,0001). Ao Doppler Triplex, determinaram-se os índices vasculares da artéria renal fetal, sendo que os valores para PSV e EDV aumentaram no decorrer das semanas gestacionais (P<0,05) e permaneceram constantes para PI e RI (P>0,05). Concluiu-se que o modo B e o Doppler Triplex são ferramentas importantes para a avaliação do desenvolvimento renal fetal, com a utilização da ecobiometria renal e avaliação dos indices vasculares da artéria renal de fetos caninos...

Reproductive efficiency of adult and prepubertal goats subjected to repeated follicular aspiration / Eficiência reprodutiva em cabras adultas e pré-púberes após repetidas sessões de aspiração folicular

The objective of this study was to assess the reproductive response of adult and prepubertal goats subjected to repeated laparoscopic ovum pick-up (LOPU). The study animals were divided into two groups, specifically, adult nanny goats (GA, n=10) and prepubertal nanny goats (GP, n=10), which were subjected to estrous synchronization and ovarian stimulation for LOPU. Both groups underwent six LOPU procedures at seven-day intervals and were subsequently subjected to controlled mating and pregnancy diagnosis to evaluate their future fertility. The study showed a reduction in the number of follicles visualized and in the amount and quality of the oocytes that were recovered and exposed to in vitro maturation. As indicated by the fertility test, however, no complications were found during the laparoscopic procedures that would impair the reproductive future of the animals. Therefore, a viable number of oocytes were obtained even with the decreased reproductive efficiency, proving that repeated LOPUs do not interfere with the reproductive of adult and prepubertal nanny goats. These results indicate a positive aspect of this procedure, allowing for increasing reproductive performance of this kind, when used for the production in vitro.

O objetivo deste estudo foi avaliar a resposta reprodutiva de cabras adultas e pré-púberes submetidas a repetidas aspirações foliculares por videolaparoscopia (LOPU). Os animais do estudo foram divididos em dois grupos, especificamente cabras adultas (GA, n = 10) e pré-púberes (GP, n = 10), que foram submetidas a sincronização de estro e estimulação ovariana para LOPU. Ambos os grupos foram submetidos a seis procedimentos LOPU em intervalos de sete dias e foram posteriormente submetidos à monta controlada e posterior diagnóstico de gestação para avaliar sua fertilidade. O estudo mostrou uma diminuição do número de folículos visualizados, na quantidade e na qualidade dos oócitos que foram recuperados e expostos à maturação in vitro. Tal como indicado pelo teste de fertilidade, no entanto, não foram observadas grandes complicações durante os procedimentos laparoscópicos que pudessem prejudicar o futuro reprodutivo dos animais. Portanto, foi obtido um número de oócitos viáveis, mesmo com a diminuição da eficiência reprodutiva, provando que repetidas LOPU não interferem na reprodutividade de animais adultos e pré-púberes. Estes resultados indicam um aspecto positivo desse procedimento, possibilitando um aumento da eficiência reprodutiva dessa espécie quando utilizado para a produção in vitro.

The effects of parturition season and suckling mode on the puerperium of Santa Ines ewes and on the weight gain of lambs / Efeitos do período das parições e dos sistemas de amamentação sobre o puerpério de ovelhas Santa Inês e desempenho ponderal de cordeiros

Seventy-seven ewes were randomly divided into groups according to parturition season and suckling mode [Rainy season: continuous (n=14) and controlled (n=13); Dry season: continuous (n=25) and controlled (n=25)]. The controlled suckling mode, in both seasons, resulted in a decrease in variables, intervals between parturition and first estrus (reduction of 27.11 and 11.46 days for rainy and dry season, respectively; P<0.05) and between parturition and estrus of conception (reduction of 12.81 and 13.58 days, for rainy and dry season, respectively; P<0.05). As to the lambs, the weight gain was higher in animals subjected to controlled suckling in relation to continuous, especially when lambing occurred in the rainy season (17.83±0.56 vs. 13.95±0.52kg for Lambs' weight at 90 days old, respectively; P<0.05). Therefore, it was concluded that controlled suckling management is better indicated for the Amazonian region, since it allows the ewes to have a shorter puerperium period and the lambs exhibit higher weight gain.

Foram utilizadas 77 ovelhas, divididas aleatoriamente em grupos de acordo com o período de parição e o manejo de amamentação [chuvoso: contínua (n=14) e controlada (n=13); seco: contínua (n=25) e controlada (n=13)]. O sistema de amamentação controlado, em ambos os períodos, resultou em decréscimo para as variáveis intervalos entre parto e primeiro estro (redução de 27,11 e 11,46 dias para os períodos chuvoso e seco, respectivamente; P<0,05) e entre parto e estro da concepção (redução de 12,81 e 13,58 dia para os períodos chuvoso e seco, respectivamente; P<0,05). Quanto ao desempenho ponderal dos cordeiros, o ganho de peso foi maior para os animais submetidos ao regime de amamentação controlada em relação à contínua, especialmente quando os nascimentos ocorreram no período chuvoso (17,83±0,56 vs. 13,95±0,52kg para o peso dos cordeiros aos 90 dias de idade, respectivamente; P<0,05). Contudo, o manejo de amamentação controlada é indicado para a região amazônica, por permitir menor tempo de puerpério das ovelhas e maior ganho de peso dos cordeiros.

Laparoscopic ovum collection in sheep: gross and microscopic evaluation of the ovary and influence on ooctye production.

Gross and microscopic evaluations of the ovaries of sheep subjected to successive ovum collection were performed with the objective of identifying if there was interference in oocyte production and morphology of gonads. Gross evaluation of the internal genital tract was also performed. Eighteen ewes of the Santa Inês breed were randomly distributed into three experimental groups of six animals each; G0, G1 and G9 with no, one and nine repetitions, respectively. Estrous synchronization was achieved with a short protocol using MAP followed by single dose stimulation with 80mg of FSHp and 300 IU of eCG (IM). Laparoscopic ovum pick-up was performed 36h later, with 7-day intervals. The number of visualized follicles, aspirated follicles and oocytes recovery was recorded. After the last intervention, ovariectomy was performed for evaluation of gross and microscopical appearance and existence of lesions caused by follicular puncture, which were classified as absent (0), mild (1), moderate (2) and severe (3). The number of visualized follicles, aspirated follicles and oocytes recovery were 13.2±2.0, 11.3±3.0 and 5.8±2.3, respectively, with a recovery rate of 51.7%. No statistical difference was found between the nine sessions (P>0.05). Nine sessions of superovulation and ovum collection procedures did not cause ovarian lesions and did not interfere with the production of follicles in ewes of the Santa Inês breed.