Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

Abstract The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.

Relationships between follicle and corpus luteum size and vascularization with ovulation, progesterone production, and pregnancy in Nellore beef cattle

ABSTRACT The objective of this study was to evaluate the relationships between preovulatory follicle (POF) and corpus luteum (CL) diameters, and POF and CL vascular perfusion with progesterone production, ovulation, and pregnancy in Nellore cows subjected to timed artificial insemination (TAI). Nellore cows (n = 201) were subjected to ovulation synchronization and later to ultrasound evaluation of POF and CL at the time of insemination (D0) and seven days later (D7), respectively. Females were divided into three categories according to the POF diameter assessed at the time of insemination: small (SF), medium (MF), and large (LF) follicles. The LF group had a greater number and intensity of pixels in the POF ultrasound exam compared with the SF group. The CL flow intensity and progesterone concentration were also higher in the LF group. The SF group showed lower flow intensity and lower ovulation rate compared with the others. When non-pregnant females were compared to pregnant ones, no difference was observed in any of the analyzed variables. The results show for the first time in Nellore cattle the relationship between the size of ovarian structures and blood flow (quantity and intensity) as well as the ability of the CL to produce progesterone. The intensity of the POF pixels proved to be relevant, demonstrating correlations with the size and flow of the CL, which were not found when evaluating only the number of pixels, thus revealing the importance of evaluating complementary characteristics of the flow.

Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.(AU)

Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Uso da ultrassonografia Doppler colorido na reprodução de pequenos ruminantes / Use of color Doppler ultrasonography in the reproduction of small ruminants

A ultrassonografia (US) Doppler colorido fornece uma ferramenta não invasiva valiosa para detectar e monitorar mudanças dinâmicas na rede vascular e fluxo sanguíneo em vários órgãos e tecidos reprodutivos. Em apoio as biotécnicas da reprodução animal assistida, a US Doppler colorido tem mostrada alta eficiência no monitoramento funcional das estruturas ovarianas. A previsão de respostas ovarianas e produções embrionária em ovinos pela identificação de sinais Doppler na parede folicular já se mostrou eficiente. A aplicação da US Doppler colorido para a identificação da funcionalidade do tecido luteal é ainda maior, desde acompanhamentos fisiológicos e diagnósticos de disfunções luteais até ampla aplicação em conjunto as diferentes biotécnicas reprodutivas. Destaca-se em ovelhas e cabras, a aplicação comercial da US Doppler colorido para o diagnóstico de gestação precoce, de disfunções luteais, de determinação de respostas ovarianas em fêmeas doadoras e receptoras de embriões, para identificar efeitos luteotrófico de estratégias hormonais, e ainda para amparar as estratégias de ressincronização de estro.(AU)

Color Doppler ultrasonography (US) provides a valuable non-invasive tool for detecting and monitoring dynamic changes in the vascular network and blood flow in various reproductive organs and tissues. In support of assisted animal reproduction biotechniques, color Doppler US has shown high efficiency in the functional monitoring of ovarian structures. The prediction of ovarian responses and embryonic production in sheep by identifying Doppler signals in the follicular wall has already proved to be efficient. The application of color Doppler US for identifying the functionality of the luteal tissue is even greater, from physiological monitoring and diagnosis of luteal dysfunctions to wide application together with different reproductive biotechniques. It stands out in sheep and goats, the commercial application of color Doppler US for the diagnosis of early pregnancy, luteal dysfunctions, determination of ovarian responses in embryo donor and recipient females, to identify luteotrophic effects of hormonal strategies, and even to support estrus resynchronization strategies.(AU)

Importance of preovulatory estradiol on uterine receptivity and luteal function

Animals that exhibited estrus had greater pregnancy success compared to animals that did not exhibit estrus before fixed-time AI (FTAI). Estradiol is synthesized in bovine ovarian follicles under gonadotropin regulation and can directly and indirectly regulate the uterine receptivity and luteal function. Estradiol concentrations at FTAI impacted oviductal gene expression and has been reported to play an important role in establishing the timing of uterine receptivity. These changes have been reported to impact uterine pH and sperm transport to the site of fertilization. After fertilization, preovulatory estradiol has been reported to improve embryo survival likely by mediating changes in uterine blood flow, endometrial thickness and changes in histotroph. Cows with greater estradiol concentrations at the time of GnRH-induced ovulation also had a larger dominant follicle size and greater circulating progesterone concentrations on day 7. Therefore, it is impossible to accurately determine the individual benefit of greater estradiol concentrations prior to ovulation and greater progesterone concentrations following ovulation to pregnancy establishment, as these two measurements are confounded. Research has indicated an importance in the occurrence and timing of increasing preovulatory concentrations of estradiol, but increasing estradiol concentrations by supplementation may not be sufficient to increase fertility. Increased production of estradiol by the preovulatory follicle may be required to enhance fertility through the regulation of sperm transport, fertilization, oviductal secretions, the uterine environment, and embryo survival.(AU)

Dr Association of bone morphogenetic protein 15 and growth differentiation factor 9 with litter size in livestock: a review study

Litter size is one of the crucial factors in livestock production and is of high economic value, which is affected by ovulation rate, hormones, and growth factors.Growth factors play a multifaceted role in reproductive physiology. This review aims to investigate the association of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9)with litter size in livestock.The transforming growth factor ß(TGF-ß) superfamily includes more than 34 members; GDF9 and BMP15 are among the most significantfactors for regulating fertility and litter size in most livestock species. Ovarian follicles release BMP15 and GDF9 that are involved in the maturation of primary follicles into the basal form, proliferation of granulosa and theca cells, steroidogenesis, ovulation, and formation of the corpus luteum. Besides, these factors are highly expressed in oocytes and are necessary for female fertility and multiple ovulation in several livestock species. Animals with two inactive copies of these factors are sterile, while those with one inactive copy are fertile. Thus, the present reviewprovides valuable information on the association of BMP15 and GDF9with litter size in livestock that can be used as biological markers of multiple ovulation or for improving fertility in livestock.(AU)

Effect of ovarian status on reproductive performance in dairy heifers inseminated with sexed semen

Background: The intensive reproductive management in the dairy farms requires inclusion of a large group of replacement heifers in the breeding program for a shorter period. In this aspect, a creation of effective estrus synchronization protocols with timed artificial insemination (TAI) by sexed semen and optimization of the current ones have a crucial role for obtaining high pregnancy rate. These protocols are beneficial, because they led to reduced interval to first AI, lack of need for estrus detection, and allow obtaining a large group of female calves for on time. Because of limited fertilizing potential of sexed spermatozoa this type of semen is applied mainly for heifers, as fertility is higher compared to lactating cows. The main objective of the present study was to investigate the effect of ovarian status on the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Materials, Methods & Results: Forty-eight Holstein healthy heifers separated in 2 groups were subjected to PGF2α-GnRH pre-treatment and Ovsynch or PRID-5-day estrus synchronization protocol and timed artificial insemination (TAI) with sex-sorted semen, staring 6 days after end of the hormonal pre-treatment. The ovarian status (presence of follicles with or not a corpus luteum) of the heifers at the begin of the treatment, on day of TAI and pregnancy rate in different groups were determined and compared. Additionally, the ovarian status at the begin of the treatment, ovarian status and size of preovulatory follicle (PF) on day of TAI and total values for both groups according to reproductive performance (pregnant or non-pregnant) were also analyzed. On day of TAI the animals with PF and a lack of corpus luteum (CL) in both groups were more that those with PF and CL (39.3% and 30% vs. 60.7 % and 70%), with significant (P < 0.05) difference in PRID-5-day group. The pregnancy rate tended to be higher in PRID-5 day than Ovsynch treatment (65% vs. 35.7%). A higher percentage (100% and 67.9%) of the pregnant animals in both treatments had not CL on day of TAI, and the size of the PF (1.58 ± 0.12 cm and 1.64 ± 0.13 cm) was increased (P < 0.05). Similar effects of the ovarian status on reproductive performance were obtained after a comparison of the total values between pregnant and non-pregnant animals. The percentage of heifers with observed PF without CL on day of TAI was significantly greater (P < 0.05) in pregnant compared to non-pregnant group (91.3% vs. 40%). The opposite dependence was determined for the parameter presence of follicles and corpus luteum (8.7% vs. 60%; P < 0.05). Furthermore, the size of the PF measured immediately before TAI was increased in animals became pregnant (1.60 ± 0.12 cm vs. 1.34 ± 0.17 cm; P < 0.05). Discussion: The analysis of the obtained results showed that the ovarian status on day of TAI affects the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Hormonal pre-treatment with onset of PRID-5-day protocol 6 days later and TAI with sex-sorted semen ensure acceptable pregnancy rate. The greater preovulatory follicle and a lack of corpus luteum before insemination provide significantly (P < 0.05) more pregnant animals, compared to the cases when CL is presented. Ultrasound determination of the ovarian function before insemination can be used in selection of heifers for TAI with sex-sorted semen.

Effect of the presence and location of corpus luteum on competence of bovine cumulus-oocyte complexes

This study aimed to determine the effect of presence of the corpus luteum (CL) and its influence on cumulus­oocyte complexes (COCs) obtained from the ipsilateral or contralateral ovary in bovine on the recovery and capacity of the oocytes to sustain mono-spermic fertilization, undergo preimplantation development, and develop to the blastocyst stage. Ovaries were collected at a local slaughterhouse and kept in pairs corresponding to the same animal. In the first experiment the variables evaluated were compared between cows with (CCL+) and without (CCL- ) CL, and for the second experiment, comparisons were made between ovaries with an ipsilateral (CL+), contralateral (CL−), and no (NCL). The recovery rate of COCs was higher in ovaries from CCL− cows, and a higher proportion of grade 1 COCs were recovered from this group. A higher proportion of metaphase I oocytes at 7 h of maturation, and a higher rate of cleavage were observed in the CCL+ group; however, a higher proportion of embryos were obtained from the CCL− group. Besides, COCs from the CL+ group had a lower proportion of grades 1 and 2 morphological qualities, lower rate of metaphase II oocytes at 22 h of maturation, and lower rate of formation of two pronuclei, whereas a higher proportion of unfertilized oocytes after in vitro fertilization. On the other hand, the COCs from the CL− group displayed a lower proportion of oocytes with more than two pronuclei, higher cleavage rate, and higher final blastocyst production were obtained when compared to CL+. Thus, the effects of CL on the competence of bovine COCs are different depending on the anatomical proximity of their location in the animal, negatively affecting the quality of COCs located in the same ovary, but not having negative effects on the competence of COCs in the ovaries contralateral to their location.(AU)

Hiperplasia endometrial cística em uma coelha-doméstica (Oryctolagus cuniculus domesticus) / Cystic endometrial hyperplasia in a domestic rabbit (Oryctolagus cuniculus domesticus)

Background: Cystic endometrial hyperplasia is a hormone-dependent disease induced by systemic increase in progesteronethat can occur in several domestic species, such as the rabbit. This disease may be associated with sex steroid hormones,especially progesterone, and may be asymptomatic, and it is diagnosed using complementary imaging tests such as totalabdominal ultrasound. However, surgical excisional biopsy with histopathological tissue analysis is the gold standard. Thisstudy reports a case of asymptomatic cystic endometrial hyperplasia in a female Miniature Lion Lop rabbit (Oryctolaguscuniculus domesticus) treated with therapeutic ovariohysterectomy.Case: A domestic, adult, female Miniature Lion Lop rabbit (Oryctolagus cuniculus domesticus), aged approximately 5 yearsand weighing 3.2 kg, was referred to specialized care to undergo ovariohysterectomy, an elective procedure. The tutor onlyreported the occurrence of a single episode of vulvar secretion more than 2 years ago, treated with antibiotics, with remissionof clinical signs. In the intraoperative period after celiotomy, the uterine horn and uterine body showed a significant increasein volume, with abnormal color changes and tissue consistency; however, both changes were clinically asymptomatic. Subsequently, biopsy was performed during the ovariohysterectomy procedure. The excised uterus and ovaries were placed in 10%formalin and histopathologically analyzed. The macroscopic histopathological examination of the sectioned tissue revealed aslight amount of brownish fluid inside the uterine horns, in addition to multiple cystic areas in the uterine mucosa. Microscopicexamination revealed marked hyperplasia of well-differentiated endometrial epithelial cells, occasionally forming cystic structuresof different sizes. Moderate congestion, mild multifocal hemorrhage, and mild multifocal inflammatory infiltrate in the lamina...

Pathomorphological lesions in the ovaries of water buffaloes

Background: Buffalo breeding is common in many countries. Buffalo's milk is used in the production of mozzarella, yoghurt, ice cream, and various dairy desserts; meat is preferred in sausage production. The female buffaloes are bred to benefit from their milk and to obtain offspring. These animals, which are not suitable for feeding in barns, generally live in pastures, especially on wet land, and are very difficult to follow. Therefore, diseases occur randomly in slaughterhouses. Studies on genital system problems are very limited. Water buffaloes (Bubalus bubalis) are animals with economic value as many buffalo products are provided. In this respect, inspection and control of female genital disorders is fundamental to ensure good reproductive performance of female buffaloes. The aim of the study was to investigate pathomorphological lesions occurring in the ovaries of water buffaloes which were sent to slaughterhouses. Materials, Methods & Results: A total of 198 ovaries of water buffaloes were collected from various slaughterhouses located in Adapazari, Istanbul, Kocaeli, Samsun and Trabzon cities of Turkey. After macroscopic examination; tissue samples were fixed in 10 % buffered formalin, processed routinely and were stained with hematoxylin-eosin (HE). As a histochemical staining, Masson's trichrome staining was applied to characterize the lesions. Immunohistochemistry (IHC) was performed on 10 % neutral formalin fixed, paraffin-embedded, 4-6-µm-thick sections from ovaries using progesterone receptor (PR) with streptavidin-biotin complex peroxidase (StrepABC-P) method. Histopathologically, follicular cysts (n:147) and luteal cysts (n:22) were seen. While the cyst lumens were sometimes surrounded by granulosa and/or luteal cells, most of them were limited by the connective tissue capsule structure. This capsule structure was shown in blue by Masson's trichrome staining. Hemorrhage was observed in some cystic corpus luteum, diagnosed as corpus hemorrhagicum. In addition, cysts giving papillary extension into the lumen and inflammation of some ovaries were observed. Immunohistochemically, the staining with PR antibody in ovarian cysts showed no immunolabelling around the follicular cyst, while the nuclei of some of the luteal cells forming the luteal cyst had strongly nuclear positivity and slightly cytoplasmic positivity. In the biochemical examination of the fluids obtained from cystic ovaries (n: 37), the average of estradiol was 2.84 ng/mL (min: 0.01 ng/mL, max: 4.30 ng/mL) and progesterone average is 49.09 ng/mL (min: 1.88 ng/mL, max: 254.2 ng/mL). Discussion: Ovarian cysts in buffaloes seem to be among the serious fertility problems as in cattle. Although the exact cause of ovarian cyts has not been determined yet, it is known that the hypothalamic-pituitary-gonadal (HPG) axis is disrupted in cyst formation in general. In this study, the mean estradiol value was within the standard range but close to the lower limit; progesterone value was above the limit. The increase in progesterone level was compatible with the pathogenesis of cyst genesis. Beside this result, staining with PR was positive in the luteal cells that formed the luteal cyst immunohistochemically. In addition, although the hemorrhages observed in the corpus luteum are considered physiological, it should not be forgotten that they can be vital if they rupture. All these results show us the animals sent for slaughtering mostly have serious genital problems threatening their fertility.

Relationships between antral follicle counts and ovarian morphology of Bos indicus of different ages / Relações entre a contagem de folículos antrais e a morfologia ovariana de Bos indicus de diferentes idades

This study aimed to evaluate the relationship between the antral follicle count (AFC) and the ovarian morphology of Bos indicus in different age groups: 12-23 months (G1), 24-35 months (G2), 36-47 months (G3), 48-59 months (G4), and >60 months (G5). Ovaries were collected individually and sent to the laboratory, where we measured AFC, the diameter and weight of the ovaries, dominant follicle (DF, >8mm), corpus luteum (CL), and small follicles (SF, <8mm). AFC were classified as high, intermediate-high, intermediate-low, and low. A group of ovaries was subjected to follicular aspiration to evaluate the morphological quality of the recovered cumulus oocyte complexes (COCs). Mean AFC did not vary between ages. The morphological characteristic that was most closely correlated with AFC was the weight of the small follicles. There was no relationship between AFC and the weight and diameter of the DF and CL. Quality of the COCs was superior in ovaries in which the CL was present, but it did not vary between the AFC classes. We conclude that AFC can be performed on a single ovary, regardless of the presence, diameter, and weight of the CL and DF in zebu between 12 and 60 months old.

O presente estudo teve como objetivo avaliar a relação entre a contagem de folículos antrais (CFA) e a morfologia ovariana de Bos indicus em diferentes faixas etárias: 12-23 meses (G1), 24-35 meses (G2), 36-47 meses (G3), 48-59 meses (G4) e >60 meses (G5). Os ovários foram coletados individualmente e enviados ao laboratório, onde suas características morfológicas foram medidas: CFA, diâmetro e peso dos ovários, do folículo dominante (FD, ≥8 mm), do corpo lúteo (CL) e dos folículos pequenos (SF, <8 mm). A CFA foi classificada como alta, intermediária-alta, intermediária-baixa e baixa. Um grupo de ovários foi submetido à aspiração folicular para avaliar a qualidade morfológica dos complexos cumulus oócitos (COCs) recuperados. A CFA média não variou entre as idades. A característica morfológica que se correlacionou mais intimamente com a CFA foi o peso dos folículos pequenos. Não houve relação entre a CFA e o peso e o diâmetro do FD e do CL. A qualidade dos COCs foi superior em ovários em que o CL estava presente, mas não variou entre as classes de CFA. Concluiu-se que a CFA pode ser realizada em um único ovário, independentemente da presença, do diâmetro e do peso do CL e do FD em zebuínos entre 12 e 60 meses de vida.

Função luteal e hipoluteoidismo em cadelas / Función lútea y hipoluteismo en perras / Luteal function and hypoluteoidism in dogs

O controle da função luteal é realizado por um balanço de fatores luteotróficos e luteolíticos, e nenhuma luteolisina de origem uterina está envolvida na regulação do corpo lúteo cíclico canino. Todavia, ao final da gestação, um pico de PGFM reflete a síntese e secreção de PGF2α envolvida na luteólise pré-parto. Em cadelas, a prolactina e o LH, mas também a PGE2, P4 e E2, e possivelmente citocinas e fatores de crescimento agem como fatores autócrinos e parácrinos para regular a função luteal. O hipoluteoidismo é definido como uma diminuição precoce da secreção de progesterona, com concentração insuficiente para a manutenção da gestação, causando perdas gestacionais. Entretanto, o valor de corte da concentração de progesterona para o diagnóstico ainda é controverso. A suplementação exógena com progestágenos é eficaz em manter a gestação, mas nas fases iniciais da gestação pode causar efeitos teratogênicos nos fetos.(AU)

El control de la función lútea se lleva a cabo por un equilibrio de factores luteotróficos y luteolíticos, y ninguna luteolisina de origen uterino está involucrada en la regulación del cuerpo lúteo cíclico canino. Sin embargo, al final del embarazo, un pico de PGFM refleja la síntesis y secreción de PGF2α involucrada en la luteólisis anteparto. En las perras, la prolactina y la LH, pero también la relaxina, PGE2, la P4 y la E2, y posiblemente citoquinas y factores de crecimiento, actúan como factores autocrinos y paracrinos para regular la función lútea. El hipoluteismo se define como una disminución precoz de la secreción de progesterona, con una concentración insuficiente para el mantenimiento del embarazo, provocando pérdidas gestacionales. Sin embargo, el valor de corte de la concentración de progesterona para el diagnóstico sigue siendo controvertido. La suplementación exógena con progestágenos es eficaz para mantener el embarazo, pero puede causar efectos teratogénicos en los fetos.(AU)

Strategic re-use of intravaginal progesterone devices increases conception rate in primiparous Nellore cows / Reutilização estratégica de dispositivo intravaginal de progesterona aumenta a taxa de concepção em vacas Nelore primíparas

In the present study, we investigate the effect of the presence or absence of corpus luteum (CL) at the beginning of a fixed-time artificial insemination (FTAI) protocol and to evaluate the impact of one-time use of intravaginal progesterone device (P4 device) in cows with or without CL. A total of 776 primiparous Nellore cows were subjected to FTAI approximately 45 days postpartum. In Experiment 1, 476 cows were divided into two experimental groups: with (CL-present, n=113) or without (CL-absent, n=363) CL, after ultrasound evaluation. On day 0 (D0), all cows received a new P4 device (1.0 g) and 2.0 mg estradiol benzoate (EB). Eight days later (D8), the P4 devices were withdrawn, and prostaglandin (15 mg), estradiol cypionate (0.5 mg), and eCG (300 IU) were administered i.m. All cows were inseminated 48 h after P4 device withdrawal (D10). In Experiment 2, the cows (n= 300) received (at D0) P4 devices that were previously used once in other cows with (n=109) or without CL (n=191) and 2 mg of EB. The same protocol as that used in Experiment 1 was performed from D8 onwards. In Experiment 1, the overall conception rate after FTAI was 55% (262/476). No difference was found in the conception rate between CL-present and CL-absent cows (52.2 vs. 55.5%). In Experiment 2, the conception rate obtained with P4 devices previously used in cows with CL (58.7%) was greater (P<0.05) than that obtained with P4 devices previously used in cows without CL (42.9%). Thus, this strategy resulted in a 15.8% increase in conception rate. In conclusion, the presence or absence of CL at the beginning of the FTAI protocol did not affect the conception rate in cows synchronized with the new P4 device, but the insertion of P4 devices previously used in cows with CL enhanced the conception rates in cows without CL.

No presente estudo, investigamos o efeito da presença ou ausência de corpo lúteo (CL) no início de um protocolo de inseminação artificial em tempo fixo (IATF) e avaliamos o impacto do uso único de dispositivo intravaginal de progesterona (dispositivo P4) em vacas com ou sem CL. Um total de 776 vacas Nelore primíparas, aproximadamente 45 dias pós-parto foram submetidas à IATF. No Experimento 1, após avaliação ultrassonográfica, 476 vacas foram divididas em dois grupos experimentais: com (CL-presente, n=113) ou sem (CL-ausente, n=363) CL. No dia 0 (D0), todas as vacas receberam um novo dispositivo de P4 (1,0 g) e 2.0 mg de benzoato de estradiol (BE). Após 8 dias (D8), os dispositivos P4 foram retirados e prostaglandina (15 mg), cipionato de estradiol (0,5 mg) e eCG (300 UI) foram administrados i.m. Todas as vacas foram inseminadas 48 horas após a retirada do dispositivo de P4 (D10). No Experimento 2, as vacas (n= 300) receberam (no D0) um dispositivo de P4 previamente utilizado uma única vez em outras vacas com (n=109) ou sem CL (n=191) e 2 mg de BE. O mesmo protocolo utilizado no Experimento 1 foi realizado a partir do D8. No experimento 1, a taxa geral de concepção após IATF foi de 55% (262/476). Não foi encontrada diferença na taxa de concepção entre as vacas com CL presente e CL ausente (52,2 vs. 55,5%). No Experimento 2, a taxa de concepção obtida com dispositivos P4 previamente utilizados em vacas com CL-presente (58,7%) foi maior (P<0,05) quando comparada aos dispositivos P4 previamente utilizados em vacas com CL-ausente (42,9%). Essa estratégia resultou em aumento de 15,8% na taxa de concepção. Em conclusão, a presença ou ausência de CL no início do protocolo de IATF não afetou a taxa de concepção em vacas sincronizadas com dispositivo novo de P4; e a eficácia dos dispositivos de P4 previamente utilizados em vacas com CL é maior durante seu segundo uso em vacas sem CL.

Níveis de progesterona e os parâmetros reprodutivos no período periovulatório de vacas Nelore submetidas ao protocolo de IATF / Progesterone levels and reproductive parameters in the periovulatory period of Nellore cows under FTAI protocol

Background: Fixed Time Artificial Insemination (FTAI) has achieved a significant evolution in the last 18 years, however, despite the progress achieved by modern FTAI programs, the conception rates obtained are still low. Therefore, this study aimed to evaluate the interrelation between progesterone levels in the periovulatory period and reproductive parameters of Nellore cows submitted to an FTAI protocol. Materials, Methods & Results: On a random day, called day 0 (D0), 57 cows received a P4 device associated with the intramuscular (IM) application of 2.0 mg of estradiol benzoate. On D9, the P4 devices were removed and then were administered 500 µg of cloprostenol sodium IM; 0.6 mg of estradiol cypionate IM and 300 IUI of Equine Chorionic Gonadotrophin IM. Blood samples were collected for the determination of serum P4 concentrations on D9 and D11 of the protocol. The evaluations of follicular diameter (DFOL), follicular wall area (AFOL) and the vascularization area of the follicle wall (VFOL) were carried out on D11 using B-mode ultrasonography examination and colour Doppler, and then the artificial inseminations were performed. The evaluation of the corpus luteum diameter (CLD), of the total corpus luteum area (CLA), of the area of corpus luteum vascularization (CLV) and blood sampling for determination of postovulatory P4 levels (Post-P4) were performed on D24. For the analysis of the P4 concentration the chemiluminescence method was used, with a sensitivity of 0.1 ng/mL. According to the P4 concentrations on D11, cows were divided into 2 groups, LOW LEVELS OF P4 and HIGH LEVELS OF P4. The diagnosis of pregnancy was performed using transrectal ultrasonography on D45, at this point the cows were divided into 2 groups, PREGNANT and NON-PREGNANT. The correlation between DFOL and P4 dosage on D11 was moderate, negative and significant and between the AFOL and the serum P4 levels on D9, was moderate, negative and significant. As for the other correlations between follicular and luteal parameters and serum P4 levels, these were low to moderate, negative and not significant. Cows in the LOW LEVELS OF P4 group had significantly larger diameter and follicular areas than the cows in the HIGH LEVELS OF P4 group, the other follicular and luteal parameters showed no statistical difference. Of the total 57 cows that were inseminated, 30 cows became pregnant. Cows in the PREGNANT group had serum P4 levels on D9 equivalent to that obtained by the NON-PREGNANT group. However, at D11 the cows that became pregnant presented significantly lower serum P4 levels than cows that did not become pregnant. Discussion: The results of the interrelation between follicular parameters and P4 levels obtained in the present study, pointed out that the lower the levels of P4, the higher the follicular parameters, corroborating with other authors. Thus, larger preovulatory follicles provided high ovulation rates. Periovulatory serum P4 levels did not significantly affect the morphofunctional parameters of the CL. Such findings may be justified by high periovulatory P4 levels resulting from less efficient luteolysis, exert a negative effect on the results of FTAI protocols, because progesterone inhibits the release of LH pulses. It is concluded that lower periovulatory P4 levels established a favourable condition for follicular development and fertility, however, morphofunctional parameters of the corpus luteum were not affected.

Indicações de ovariohisterectomia terapêutica em gatas / Indications of therapeutic ovariohisterectomy in female cats

Ovariohisterectomia (OH) é o procedimento cirúrgico mais realizado na rotina veterinária. Tal cirurgia pode ser realizada como tratamento de enfermidades, sendo denominada de OH terapêutica. Assim, o objetivo desta revisão é determinar quais as indicações de OH terapêutica em gatas. Para isso, foi realizada uma busca em sites de artigos científicos utilizando-se termos relacionados ao tema, abrangendo o período de 2012 a 2022 (10 anos). Os artigos sobre indicação de OH terapêutica em gatas foram analisados e os dados obtidos foram dispostos em tabela e gráfico. Foram analisadas 27 publicações, perfazendo 1.264 casos de OH terapêutica em gatas. Piometra foi a enfermidade mais frequente, totalizando 1.010 casos (79,90%). Cisto ovariano foi a segunda causa mais frequente, compreendendo 95 casos (7,51%). A terceira causa mais frequente de OH terapêutica foi a distocia, representando 81 casos (6,40%). A hiperplasia mamária foi indicação de OH terapêutica em 30 casos (2,37%). Prolapso uterino e torção uterina obtiveram a mesma frequência (11 casos; 0,87%). Maceração fetal foi uma indicação terapêutica em três relatos (0,23%) nos artigos analisados. Ocorreram dez outras indicações menos comuns de OH terapêutica em gatas. Desta forma, os resultados indicam a piometra como a doença mais frequente, entretanto, foram encontrados poucos artigos sobre o tema, indicando a necessidade de mais estudos envolvendo a espécie felina.

Ovariohysterectomy (OH) is the most performed surgical procedure in the veterinary routine. Such surgery can be carried out as a treatment for diseases, being called therapeutic OH. Thus, the present review aims to determine the indications for therapeutic OH in cats. For this purpose, a search was carried out on scientific article websites using terms related to the topic, covering the period from 2012 to 2022 (10 years). The articles on the indication of therapeutic OH in cats were analyzed and the data obtained were arranged in a table and graph. Twenty-seven publications were analyzed, totaling 1,264 cases of therapeutic OH in cats. Pyometra was the most frequent disease, totaling 1,010 cases (79.90%). Ovarian cyst was the second most frequent cause, comprising 95 cases (7.51%). The third most frequent cause of therapeutic HO was dystocia, representing 81 cases (6.40%). Breast hyperplasia was an indication for therapeutic OH in 30 cases (2.37%). Uterine prolapse and uterine torsion had the same frequency (11 cases; 0.87%). Fetal maceration was a therapeutic indication in three reports (0.23%) in the analyzed articles. There were ten other less common indications of therapeutic OH in cats. Thus, the results indicate pyometra as the most frequent disease; however, few articles were found on the subject, indicating the need for more studies involving the feline species.

Reproductive performance of fixed-time artificial insemination in heifers supplemented with a homeopathic complex / Desempenho reprodutivo à inseminação artificial em tempo fixo em novilhas suplementadas com complexo homeopático

For this study, ½ blood Nelore × Aberdeen Angus heifers (n = 40; 14 months) were divided into two groups: control (n=20; 310 kg) and Pró-Cio (n=20; 304 kg). For Pró-Cio group, 20g of homeopathic product (Pró-cio®, Real H, Campo Grande, Brazil) was supplied together with corn eighteen days prior (D-18) to the beginning of the fixed-time artificial insemination protocol (FTAI). On D0, all females received an intravaginal P4 device and 2 mg BE. On D5, the AFC was assessed by ultrasound and blood was collected for AMH dosage. On D8, P4 device was removed; 300 IU eCG, 0.530 mg of cloprostenol sodium and 0.5 mg of EC were administered; an estrus-identifying adhesive was fixed at the tail insertion and the diameter of the dominant follicle (DF) was measured. On D10, all heifers were inseminated, estrus manifestation was assessed and the preovulatory follicle (POF) was measured. On D20, the CL area was measured and blood flow was assessed using Doppler ultrasound. Data were analyzed by Student, Mann-Whitney or Friedman test. The rates of heat expression, ovulation, and blood flow score with Doppler were analyzed using Fisher's exact test and pregnancy rate by binary logistic regression model (P ≤ 0.05). There was no difference in AFC or AMH dosage (P > 0.05). Pró-Cio group had greater diameters of the DF (9.94 ± 0.42 mm) and POF (11.61 ± 0.56 mm) than the controls (DF: 7.72 ± 0.34 mm and POF: 9.91 ± 0.37 mm), as well a larger CL area (3.26 ± 0.26 versus 2.35 ± 0.16 cm2) and a higher mean CL vascularization score (3.06 versus 2.26; P < 0.05). There was no significant difference (P > 0.05) between estrus manifestation or conception rate for control and Pró-Cio groups. Heifers supplemented with the homeopathic product showed larger follicular diameters at the end of the FTAI protocol and CL with better blood flow scores compared to the control group.

Para esse estudo, novilhas ½ sangue Nelore x Aberdeen Angus (n = 40; 14 meses) foram divididas em dois grupos: Controle (n = 20; 310 kg) e Pró-cio (n = 20; 304 kg). Para o grupo Pró-Cio, 20 g/animal de produto homeopático (Pró-cio®, Real H, Campo Grande, Brazil) foi fornecido juntamente com o milho dezoito dias antes (D-18) de inicar o protocolo de inseminaçãoa artificial em tempo-fixo (IATF). No D0, todas as fêmeas receberam um dispositivo intravaginal de P4 e 2 mg BE. No D5, avaliou-se a contagem de folículos antrais (CFA) por ultrassonografia e realizou-se coleta de sangue para dosagem de AMH. No D8, procedeu-se a retirada do dispositivo de P4; aplicação de 300 UI de eCG, 0,530 mg de cloprostenol sódico e 0,5 mg de CE; um adesivo identificador de estro foi fixado na inserção da cauda e foi feita a mensuração do diâmetro do folículo dominante (FD). No D10, todas as novilhas foram inseminadas, foi realizada a avaliação da manifestação de estro e mensuração do folículo pré-ovulatório (FPO). No D20, realizou-se mensuração da área e avaliação do fluxo sanguíneo do CL por ultrassonografia Doppler. Os dados foram analisados pelos testes de Student, Mann-Whitney ou Friedman. As taxas de expressão de estro, ovulação e escore de fluxo sanguíneo com Doppler foram analisadas usando o teste exato de Fisher e taxa de prenhez pelo modelo de regressão logística binária (p ≤ 0,05). Não houve diferença na CFA nem na dosagem de AMH (p > 0,05). O grupo PróCio apresentou maior diâmetro do FD (9,94 ± 0,42 mm) e FPO (11,61 ± 0,56 mm) em relação ao controle (FD: 7,72 ± 0,34 mm e FPO: 9,91 ± 0,37 mm), além de maior área do CL (3,26 ± 0,26 versus 2,35 ± 0,16 cm2; p < 0,05) e maior escore de vascularização médio do CL (3,06 versus 2,26; p < 0,05). Não houve diferença (p > 0,05) na taxa de manifestação de estro e nem na taxa de concepção entre os grupos Controle e Pró-Cio, respectivamente. Novilhas suplementadas com o produto homeopático apresentaram maiores diâmetros foliculares ao final do protocolo de IATF e CL com melhores escores de fluxo sanguíneo em relação ao grupo controle.

Influence of eCG and reproductive management in the resynchronization of ovulation in dairy goats

Resynchronization protocols have been proposed as a way of shortening females' unproductive time in the flock, with good results in cattle and sheep. In goats, initial studies have shown that a second progestogen device inserted before luteolysis and pregnancy diagnosis does not interfere with the corpus luteum lifespan or functionality. This study aimed to evaluate the follicular growth, ovulation pattern and pregnancy rate after insertion of a second and new progestogen device for resynchronizing, with or without equine Chorionic Gonadotrophin (eCG), submitted to natural mating (NM) or artificial insemination (AI) to propose a viable resynchronization protocol for dairy goats. A total of 38 multiparous Saanen goats underwent a short-term progesterone protocol [six days exposed to medroxyprogesterone acetate (MAP) intravaginal sponges + 200 IU eCG and 0.12 mg of cloprostenol sodium on the 5th day + 0.025 mg of lecirelin 34 hours after sponge withdrawal] and, on day 16th after the ovulation, received a new MAP device which was retained until day 21. At this moment females were split into four groups: GeCG+NM ­ 100 IU eCG with NM; GSal+NM ­ saline solution with NM; GeCG+AI ­ 100 IU eCG with AI; and GSal+AI ­ saline solution with AI. Ultrasound scans were performed every 12 h from sponge withdrawal (day 21) until 108 h after sponge withdrawal (day 25) for follicular dynamics evaluation, at 240 h (day 31) for assessing the presence of active corpus luteum, and on day 60 for pregnancy diagnosis. No differences were found regarding ovulation time, synchronization and follicle size. However, GeCG+NM presented a greater estrus manifestation rate (100%) and pregnancy rate (62.5%) when compared to GSal+AI. In conclusion, resynchronization protocols in dairy goats may present satisfactory results.(AU)

Impacts of trace element supplementation on productive/reproductive postpartum performances of grazing dairy heifers from volcanic soils

This study has been designed to evaluate the effect of trace elements (TE) on the reproductive and productive performances of postpartum Holstein Friesian heifers, fed on pasture with origin on volcanic soils, poor in TE. Twenty-six heifers pregnant were divided into two groups: experimental (EG) and control (CG) groups (n=13, each group). For animals belonging to the EG, two intra-ruminal capsules with TE were administrated 60 days before partum, while in the CG, no boluses were administered. All animals' blood was collected weekly to assess progesterone levels by the ELFA technique for 11 weeks after partum. TE were evaluated on blood by AAS when the experience started, on the day of delivery, and after 60 days. Before calving, no statistical differences were observed between groups for the trace elements, although it has been noticed that heifers had deficiencies in Selenium, Copper, and Iodine. On the calving day and 60 days after, a statistical increase (p<0.05) in serum Copper and Selenium was observed in the animals belonging to the EG. For the other TE, no statistical differences were observed. Concerning reproductive characteristics, the EG, at five weeks postpartum, 70% of the cows were cyclic, while in the CG in the same period, 33% of animals showed signs of ovarian activity. At the level of productive parameters, no differences were observed between groups. The results obtained by the present study allow concluding that, under our experimental conditions, the administration of trace elements, in addition to reducing postpartum anestrus, increases the quality of the corpus luteum in the postpartum period of heifers.

Antral follicle count, productive and reproductive parameters in Bos indicus and Bos indicus-taurus prepubertal heifers with early puberty induction / Contagem de folículos antrais, parâmetros produtivos e reprodutivos em novilhas pré-púberes Bos indicus e Bos indicus-taurus com indução precoce de puberdade

Prepubertal Nelore (G-N = 15) and crossbred Nelore x Aberdeen Angus heifers (G-NA = 15) were used for this study. AFC, live weight, body condition score (BCS), ovary and dominant follicle (DF) diameters were determined in each animal. Puberty induction was performed by insertion of a 4th use progesterone device (D0) which was removed on D12. Also, 1 mg estradiol benzoate was administered, and estrus intensity was classified (D12). At D21, the presence and diameter of the corpus luteum (CL) were registered. AFC was highly repeatable, regardless of hormone induction in both G-N (r=0.79) and G-NA (r=0.90). The mean AFC was greater in G-N compared to G-NA (24.2±8.5 vs. 17.7±9.0 follicles). A variation in BCS throughout the study occurred in G-NA, but not in G-N. The average weight gain (AWG) was greater in G-NA compared to G-N (0.69±0.33 vs. 0.40±0.29kg/day). The G-NA resulted in a larger diameter of DF at D12 than G-N (11.6±2.7 vs. 9.3±1.5mm). In conclusion, AFC was greater in Nelore heifers, although in both breeds this count was highly repeatable during puberty induction. Crossbred heifers had greater BCS and AWG with greater diameter of DF, indicating higher precocity when compared to Nelore heifers.

Novilhas pré-púberes Nelore (G-N=15) e mestiças Nelore x Aberdeen Angus (G-NA=15) foram utilizadas neste estudo. CFA, peso vivo, escore de condição corporal (ECC), diâmetros do ovário e folículo dominante (FD) foram determinados em cada animal. A indução da puberdade foi realizada pela inserção de um dispositivo de progesterona de quarto uso (D0), que foi retirado no D12. Além disso, 1mg de benzoato de estradiol foi administrado e a intensidade do estro foi classificada (D12). No D21, foram registrados a presença e o diâmetro do corpo lúteo (CL). A CFA foi altamente repetível, independentemente da indução hormonal em G-N (r=0,79) e G-NA (r=0,90). A CFA média foi maior em G-N em comparação com G-NA (24,2±8,5 vs. 17,7±9,0 folículos). Uma variação no ECC ao longo do estudo ocorreu em G-NA, mas não em G-N. O ganho de peso médio (GPM) foi maior em G-NA em comparação com G-N (0,69±0,33 vs. 0,40±0,29kg/dia). O G-NA resultou em um diâmetro maior de FD em D12 do que o G-N (11,6±2,7 vs. 9,3±1,5mm). Em conclusão, a CFA foi maior em novilhas Nelore, embora em ambas as raças essa contagem tenha sido altamente repetível durante a indução da puberdade. Novilhas mestiças apresentaram maior ECC e GPM com maior diâmetro de FD, indicando maior precocidade quando comparadas às novilhas Nelore.