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This study aimed to evaluate the effects of Protium heptaphyllum fruit essential oil (PHEO) on the physiology of silver catfish (Rhamdia quelen) during anesthesia and recovery, through studying echocardiograms, oxidative status, and metabolic parameters. Three experiments were performed: (1) 50 silver catfish juveniles were submitted to anesthesia and recovery tests with 300, 400, 500, 600, and 700 mg L-1 of PHEO. (2) Echocardiogram analysis was performed in anesthetized and non-anesthetized fish. (3) Biochemical parameters were evaluated at 0, 30, 60, and 120 min of recovery after being anesthetized for 3 min with 600 mg L-1 PHEO. Times to sedation and deep anesthesia were reduced with PHEO increasing concentrations. The echocardiogram showed a higher cardiac rate in anesthetized fish. Plasma glucose levels increased in control fish through recovery time, but anesthetized fish showed lower levels than controls at 120 min of recovery. Metabolic parameters such as plasma and hepatic glucose did not show changes considering the recovery time of up to 120 min. Hepatic glycogen, lactate, and triglycerides reduced their levels over recovery times. Fish anesthetized enhanced superoxide dismutase activity and thiobarbituric acid reactive substances levels but decreased reduced glutathione (GSH) levels at 30 min compared to controls. After 60 min, GSH values were significantly higher in anesthetized fish than in controls. These results suggest that PHEO at 600 mg L-1 is an effective anesthetic for the rapid handling of silver catfish, providing stable metabolic parameters and enhanced antioxidant responses during recovery. Echocardiogram analysis confirms the anesthetic effect, supporting PHEO as a viable and efficient option for fish anesthesia in aquaculture. The use of PHEO in aquaculture can enhance fish welfare by reducing stress during handling and transportation, potentially leading to improved growth, health, and survival rates.

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Embryonic mortality in cattle is high, reaching 10-40 % in vivo and 60-70 % in vitro. Death of embryos involves reduced expression of genes related to embryonic viability, inhibition of DNA repair and increased DNA damage. In follicular granulosa cells, FGF18 from the theca layer increases apoptosis and DNA damage, so we hypothesized that FGF18 may also affect the oocyte and contribute to early embryonic death. The aims of this study were to identify the effects of FGF18 on cumulus expansion, oocyte maturation and embryo development from cleavage to blastocyst stage using a conventional bovine in vitro embryo production system using ovaries of abattoir origin. Addition of FGF18 during in-vitro maturation did not affect FSH-induced cumulus expansion or rates of nuclear maturation. When FGF18 was present in the culture system, rates of cleavage were not affected however, blastocyst and expanded blastocyst development was substantially inhibited (P < 0.05), indicating a delay of blastulation. The number of phosphorylated histone H2AFX foci per nucleus, a marker of DNA damage, was higher in cleavage-stage embryos cultured with FGF18 than in those from control group (P < 0.05). Furthermore, FGF18 decreased accumulation of PTGS2 and IFNT2 mRNA in blastocysts. In conclusion, these novel findings suggest that FGF18 plays a role in the regulation of embryonic death during the early stages of development by impairing DNA double-strand break repair and expression of genes associated with embryo viability and maternal recognition of pregnancy during the progression from oocyte to expanded blastocysts.

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We hypothesized that the hCG modulates the expression of IFNT-pathway and ISGs in bovine endometrium during early pregnancy. The aim of the current study is to evaluate the effect of hCG on IFNT-pathway signals and ISGs expression in endometrial cells. For this, 29 non-lactating cross-bread cows were used in the study and submitted to a 9-day fixed-time artificial insemination (FTAI) protocol. The day of the AI was considered Day 0 (D0), and five days (D5) after the FTAI, the cows were allocated into two groups: Control and hCG group, when a hCG group received a single dose of 2.500UI of hCG. On day 18 after FTAI (D18) cows were slaughtered and endometrial tissue samples were collected. There was no difference between the embryo recovery rate of the cows in C compared to the hCG. The hCG group increased the accessory corpus luteum formation rate. The hCG resulted in greater serum progesterone concentration in the hCG group compared to the C on Day 14. Only the expression of IFNAR2 and STAT1 were upregulated on pregnant cows of the hCG group compared to the C group. The pathway genes (JAK1, STAT2, and IRF9) were not regulated. The mRNA abundance of ISG15, MX1, MX2, and OAS1 was upregulated in pregnant cows for hCG group, compared to C group. The results show that the administration of hCG, 5 days after AI, in addition to increasing the serum progesterone, modulates the expression of IFNT-pathway and ISGs on bovine endometrium on Day 18 of pregnancy.

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Complex regional pain syndrome type I (CRPS-I) is a disabling pain condition without adequate treatment. Chronic post-ischemia pain injury (CPIP) is a model of CRPS-I that causes allodynia, spontaneous pain, inflammation, vascular injury, and oxidative stress formation. Antioxidants, such as alpha lipoic acid (ALA), have shown a therapeutic potential for CRPS-I pain control. Thus, we aim to evaluate if ALA repeated treatment modulates neuroinflammation in a model of CRPS-I in mice. We used male C57BL/6 mice to induce the CPIP model (O-ring torniquet for 2 h in the hindlimb). For the treatment with ALA or vehicle (Veh) mice were randomly separated in four groups and received 100 mg/kg orally once daily for 15 days (CPIP-ALA, CPIP-Veh, Control-ALA, and Control-Veh). We evaluated different behavioral tests including von Frey (mechanical stimulus), acetone (cold thermal stimulus), rotarod, open field, hind paw edema determination, and nest-building (spontaneous pain behavior). Also, hydrogen peroxide (H2O2) levels, NADPH oxidase and superoxide dismutase (SOD) activity in the sciatic nerve and spinal cord, and Iba1, Nrf2, and Gfap in spinal cord were evaluated at 16 days after CPIP or sham induction. Repeated ALA treatment reduced CPIP-induced mechanical and cold allodynia and restored nest-building capacity without causing locomotor or body weight alteration. ALA treatment reduced SOD and NADPH oxidase activity, and H2O2 production in the spinal cord and sciatic nerve. CPIP-induced neuroinflammation in the spinal cord was associated with astrocyte activation and elevated Nfr2, which were reduced by ALA. ALA repeated treatment prevents nociception by reducing oxidative stress and neuroinflammation in a model of CRPS-I in mice.

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Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high incidence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24 h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P < 0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P ≥ 0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6 h and BRCA1 at 3, 6, 12 and 24 h (P < 0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium.

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CONTEXT: The establishment of pregnancy in cows requires uterine activity regulation of the main Hippo signalling effector yes-associated protein 1 (YAP). It remains unknown (1) how YAP activity at the corpus luteum (CL) correlates with early pregnancy-related events in ruminants; and (2) if YAP activity in the uterus and CL can be affected by metabolic disorders that may lead to pregnancy failure in ruminants. AIMS AND METHODS: To determine the effect of early pregnancy on total and phospho-YAP expression and its transcriptional activity in the CL, we compared non-pregnant vs pregnant ewes. To understand the YAP activity dysregulation with disorders that may result in pregnancy loss, we induced negative energy balance in pregnant ewes. KEY RESULTS AND CONCLUSIONS: Our main results indicate that early pregnancy alters the expression and activity patterns of YAP in the ovine CL but not in the endometrium. In addition, while our NEB-induced model fails to alter YAP activity at the endometrium level, we found that fasting during the first but not second week of pregnancy affects YAP activity in the CL of pregnant ewes. IMPLICATIONS: The data presented herein provide considerable insight into the activity of a signalling pathway that may be a key player in pregnancy recognition and establishment in ewes.

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The Hippo pathway is involved in the proliferation of intrafollicular cells and in early embryonic development, mainly because effectors of this pathway are key transcription regulators of genes such as CTGF and CYR61, which are involved in cell proliferation. Recent studies by our group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development, leading to the hypothesis that FGF18 may have a role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the expression of Hippo pathway target genes, CTGF and CYR61, during oocyte maturation and early embryonic development. Three experiments were carried out, with in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) and embryo culture. In experiment one, FGF18 (100 ng/ml) induced an increase (P < 0.05) in CTGF gene expression at 12 h post-exposure. In experiment two, FGF18 (100 ng/ml) induced a reduction (P < 0.05) in CTGF expression at 3 h post-exposure. In the third experiment, day 7 embryos exposed to FGF18 during oocyte IVM expressed greater CTGF mRNA abundance, whereas FGF18 exposure during embryo in vitro embryo culture did not alter CTGF expression in comparison with untreated controls. The preliminary data presented here show that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.

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One of the major causes of early pregnancy loss is heat stress. In ruminants, interferon tau (IFNT) is the embryo signal to the mother. Once the interferon signaling pathway is activated, it drives gene expression for interferon-stimulated genes (ISGs) and alters neutrophils responses. The aim of the present study was to evaluate interferon (IFN) pathway, ISGs and gene expression in polymorphonuclear leukocytes (PMN) and oxidative stress in dairy cows under heat stress. Pregnant cows had their estrous cycle synchronized and randomly assigned to a comfort or heat stress group. Blood samples were collected at artificial insemination (AI) and on Days 10, 14 and 18 following AI. Pregnant cows were pregnancy checked by ultrasound on Day 30 and confirmed on Day 60 post-AI. Results are presented as mean ± SEM. The corpus luteum (CL) diameter was not different between groups of pregnant cows; concentration of progesterone of pregnant cows on Day 18 following AI was greater in comfort group compared to heat stressed group. Comfort pregnant cows had higher expression of all analyzed genes from interferon pathway, except for IFNAR1, on both Days 14 and 18. Conversely, heat stressed cows did not show altered expression of IFNT pathway genes and ISGs between Days 10, 14, and 18 after AI. The oxidative stress, determined as malondialdehyde (MDA) levels, was greater in heat stress group on Days 10, 14 and 18, independent of pregnancy status. Heat stress negatively influences expression of ISGs, IFN pathway gene expression in neutrophils, and oxidative stress. Our data suggest that lower conception rates in cows under heat stress are multifactorial, with the association of interferon pathway activation and the unbalanced oxidative stress being main contributing factors.

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Background: The Flemish Red is one of the oldest breeds of French cattle and, in France, the crosses with other red breeds and replacing them with more productive breeds caused a severe reduction in the number of purebred Flemish Red animals. Due to this drastic decrease in the Flemish Red population, this breed is at risk of extinction. The objective of the present study is to describe follicular development and serum progesterone (P4) profiles in Flemish Red cows raised in Southern Brazil for the improvement of reproductive management of this breed, considering that these animals may exhibit traits which disappeared in European animals due to genetic dilution.Materials, Methods & Results: The estrus cycles of two groups of post-pubertal non-lactating Flemish Red (FR, n = 7) and Holstein (HOL, n = 7) cows were synchronized with a Prostaglandin F2α analog. From the day of estrus, the ovaries were evaluated by transrectal ultrasound every 24 h for 21 days or until the detection of the second ovulation. The diameters of the dominant follicle (DF) and subordinate follicles (SF) were recorded according to the day of the estrus cycles and analyzed based on the diameter of both DF and SF on the day of follicular emergence, the day and diameter of DF on the day of divergence, daily growth rates of DF and FS, and the maximum diameter of both the ovulatory follicle and the dominant non-ovulatory follicle (DNOF). Samples of blood were collected every five days for P4 measurement (RIA). During emergence of the first follicle wave, the DF measured 3.97 ± 0.19 mm for FR and 4.00 ± 0.35 mm for HOL, while the SF reached 3.40 ± 0.22 mm in FR and 3, 07 ± 0.26 mm in HOL. The daily follicle growth rates was 1.10 ± 0.04 mm for DF and 0.67 ± 0.06 mm for the SF in FR, and were 1.01 ± 0.05 mm for DF and 0.72 ± 0.09 mm for the SF in HOL.[...](AU)

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Background: The Flemish Red is one of the oldest breeds of French cattle and, in France, the crosses with other red breeds and replacing them with more productive breeds caused a severe reduction in the number of purebred Flemish Red animals. Due to this drastic decrease in the Flemish Red population, this breed is at risk of extinction. The objective of the present study is to describe follicular development and serum progesterone (P4) profiles in Flemish Red cows raised in Southern Brazil for the improvement of reproductive management of this breed, considering that these animals may exhibit traits which disappeared in European animals due to genetic dilution.Materials, Methods & Results: The estrus cycles of two groups of post-pubertal non-lactating Flemish Red (FR, n = 7) and Holstein (HOL, n = 7) cows were synchronized with a Prostaglandin F2α analog. From the day of estrus, the ovaries were evaluated by transrectal ultrasound every 24 h for 21 days or until the detection of the second ovulation. The diameters of the dominant follicle (DF) and subordinate follicles (SF) were recorded according to the day of the estrus cycles and analyzed based on the diameter of both DF and SF on the day of follicular emergence, the day and diameter of DF on the day of divergence, daily growth rates of DF and FS, and the maximum diameter of both the ovulatory follicle and the dominant non-ovulatory follicle (DNOF). Samples of blood were collected every five days for P4 measurement (RIA). During emergence of the first follicle wave, the DF measured 3.97 ± 0.19 mm for FR and 4.00 ± 0.35 mm for HOL, while the SF reached 3.40 ± 0.22 mm in FR and 3, 07 ± 0.26 mm in HOL. The daily follicle growth rates was 1.10 ± 0.04 mm for DF and 0.67 ± 0.06 mm for the SF in FR, and were 1.01 ± 0.05 mm for DF and 0.72 ± 0.09 mm for the SF in HOL.[...]

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Alto grau de insucesso reprodutivo resulta de falhas no funcionamento ovariano. Tal insucesso implica em prejuízo financeiro para produtores de animais domésticos e limita a propagação de espécies selvagens. Assim, a compreensão dos mecanismos endócrinos, celulares e moleculares envolvidos no controle dos processos ovarianos é fundamental para embasar procedimentos, estratégias e tecnologias que visem a manipulação de tais processos para aumentar sua eficiência e conseqüentemente o sucesso reprodutivo. Especificamente, serão tratados na presente revisão os seguintes tópicos: a dinâmica da formação e depleção da população de folículos ovarianos, a dinâmica da ativação de folículos e a dinâmica do desenvolvimento e funcionamento folicular pós-ativação e sua relação à ovulação, fertilização e gestação inicial.

A significant proportion of reproductive failure results from poor ovarian function. Such failure results in financial losses to domestic animal commercial operations and limits wildlife species propagation. Therefore, the understanding of endocrine, cellular and molecular mechanisms that control ovarian processes is critical to provide the basis for procedures, strategies and technologies that aim the manipulation of such processes to increase their efficiency and, thereby, reproductive success. Specifically, in the present review the following topics will be addressed: the dynamics of formation and depletion of ovarian follicle population, the dynamics of activation of follicles and the dynamics of follicle development and function after activation and their relationship to ovulation, fertilization and early gestation.

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Alto grau de insucesso reprodutivo resulta de falhas no funcionamento ovariano. Tal insucesso implica em prejuízo financeiro para produtores de animais domésticos e limita a propagação de espécies selvagens. Assim, a compreensão dos mecanismos endócrinos, celulares e moleculares envolvidos no controle dos processos ovarianos é fundamental para embasar procedimentos, estratégias e tecnologias que visem a manipulação de tais processos para aumentar sua eficiência e conseqüentemente o sucesso reprodutivo. Especificamente, serão tratados na presente revisão os seguintes tópicos: a dinâmica da formação e depleção da população de folículos ovarianos, a dinâmica da ativação de folículos e a dinâmica do desenvolvimento e funcionamento folicular pós-ativação e sua relação à ovulação, fertilização e gestação inicial. (AU)

A significant proportion of reproductive failure results from poor ovarian function. Such failure results in financial losses to domestic animal commercial operations and limits wildlife species propagation. Therefore, the understanding of endocrine, cellular and molecular mechanisms that control ovarian processes is critical to provide the basis for procedures, strategies and technologies that aim the manipulation of such processes to increase their efficiency and, thereby, reproductive success. Specifically, in the present review the following topics will be addressed: the dynamics of formation and depletion of ovarian follicle population, the dynamics of activation of follicles and the dynamics of follicle development and function after activation and their relationship to ovulation, fertilization and early gestation. (AU)

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