Resumo em Inglês:

Abstract The aim of the study was to evaluate the use of Acustic Radiation Force Impulse (ARFI) elastography in mammary parenchyma and supramammary lymph nodes, for detection of active mastitis in sheep with naturally infected chronic fibrous lesions. 27 female sheep were included and B-mode ultrasound and ARFI elastography images were obtained, acquiring qualitative (echogenicity and echotexture) and quantitative (shear rate, depth and short/long axis ratio) variables of 48 mammary glands. The glands were divided into three experimental groups: control group (CG) - healthy animals; LSCC- animals that presented fibrous lesions and SCC (somatic cell count) less than 500 x 103 cls/mL; HSCC: animals that presented fibrous lesions and SCC (somatic cell count) more than 500 x 103 cls/mL; The qualitative variables using B-mode ultrasonography, including echotexture and echogenicity, showed no significant differences between the evaluated groups and tissues (p = 0.9336 and p = 0.233, respectively) .In healthy areas of the gland, it was an increasing in shear wave velocity (SWV) in LSCC than in HSCC (p=0.04). When comparing the fibrosis in the LSCC and HSCC groups with their respective normal areas, the velocity increased in both groups: LSCC (p= 0,0007) and HSCC (p= 0,0001). When comparing the areas of fibrosis in LSCC and HSCC with the CG parenchyma, there was an increase in LSCC (p=0.001) and HSCC (p=0.0001). B-mode ultrasound indicate predominance of hypoechoic echogenicity in lymph nodes and reduced short/long axis ratio in cases of active subclinical mastitis. The supramammary lymph node showed increased SWV when comparing the CG with HSCC groups (p=0.02) and GC with LSCC (p=0.04). B-mode ultrasonography is useful for evaluating the mammary parenchyma, however, its application as a standalone diagnostic technique is not recommended. ARFI elastography indicates potential cutoff points for differentiating subclinical mastitis from healed mastitis, highlighting its importance as a tool for distinguishing normal areas from fibrous parenchymal areas. While this study did not establish specific cutoff points due to sample size limitations, further research with larger sample sizes could explore and define these critical thresholds.

Resumo em Inglês:

Abstract This study evaluated two surgical sterilization techniques in free-ranging female capybaras (n = 21). The first group underwent uterine horn ligature (HL; n = 11), while the second was subjected to partial salpingectomy (S; n = 10). We assessed total operative time, incision length, the ease of identifying reproductive structures, the adequacy of exposure for surgical performance through flank or midline approaches, and the extent of abdominal viscera manipulation for each method. The HL method emerged as faster, with an average operative time difference of 16 minutes. In the S group, a flank mini-laparotomy over the ovarian topography facilitated easy exposure of the ipsilateral ovary and uterine tube, enabling ligature and partial resection of the uterine tube but not the uterine horn exposure. However, accessing the contralateral uterine tube without a bilateral incision was impractical, thus prolonging the total operative time due to the need for patient repositioning and new antisepsis procedures. Conversely, a post-umbilical approach for the HL method necessitated only one mini-laparotomy incision, offering ample uterine exposure for hysterotomy in pregnant females. Both methods involved minimal abdominal viscera manipulation and resulted in no fatalities or postoperative complications. Although direct comparison is limited by the distinct sterilization techniques and surgical approaches, this study underscores the challenges and surgical access of each method. Our findings endorse the HL technique as an effective contraception method for female capybaras to prevent the birth of seronegative offspring that could amplify Rickettsia sp., the causative agent of Brazilian spotted fever.

Resumo em Inglês:

Abstract The objective of the present study was to evaluate the effects of a novel Inhibin (INH) DNA vaccine (C500/pVAX-asd-IS) on the immune response, reproductive hormone levels, and spermatogenesis of rats. Forty healthy male rats were divided into four groups, and respectively immunized (thrice, 14 d apart) with 1×108, 1×109, and 1×1010 CFU of the recombinant inhibin vaccine (group C500/pVAX-asd-IS-L, C500/pVAX-asd-IS-M, and C500/pVAX-asd-IS-H) or 1×1010 CFU C500. P/N values increased after vaccination and differed (p <0.05) at 7 d, and sharply increased at 14 d following the booster vaccination (p <0.01); The weight and volume of testes in C500/pVAX-asd-IS groups were increased (p < 0.05) at decapitation, respectively; Histological evaluation showed that the number of spermatogenic cells in the lumen was increased, and the cytoplasmic remnants of sperms were allergy increased significantly compared with the control group. Oral vaccination with INH DNA reduced (P < 0.05) serum concentrations of INH B, enhanced serum concentrations of testosterone (T) and FSH. Furthermore, mRNA expressions of VIM and SMAD4 in the testes were increased in C500/pVAX-asd-IS-M and C500/pVAX-asd-IS-H groups (p < 0.05 or p < 0.01). The mRNA amount of INHβ-B in C500/pVAX-asd-IS-M group was greater than control group (p < 0.05).These results suggested that neutralization of endogenous INH through oral vaccination with INH DNA delivered by C500 strain successfully elicited a humoral immune response. INH gene immunization may have a positive effect on spermatogenesis and reproductive efficiency in male rats.

Resumo em Inglês:

Abstract The current investigation aimed to explore the effects of Myrciaria dubia liquid extract (MDLE) as the primary component of an extender for breeder rooster semen over different periods at room temperature. Fifteen breeder roosters (40 weeks of age, average body weight of 2.05±0.12) with confirmed fertility were used. Employing a factorial design (3x4), the treatments consisted of semen in natura and two semen extenders (an experimental based on MDLE and a commercial) subjected to four periods at room temperature post-collection (5, 10, 15 and 20 minutes) with four replicates (tubes) each. All variables evaluated in this study yielding significant results (p<0.05). Analyzed individually, the experimental extender based on MDLE exhibited a linear reduction (p<0.05) in motility and vigor results, while it caused an increase in pH values and percentages of sperm defects evaluated. When compared with semen in natura and commercial extender, the efficiency of MDLE as a semen extender was inferior to that observed with the commercial extender and similar to the results observed with semen in natura. Nonetheless, the experimental extender based on MDLE yielded satisfactory results for up to 15 minutes of storage time. In conclusion, MDLE can be considered as an alternative for composing a roosters’ semen extender, maintaining sperm characteristics within acceptable limits for up to 15 minutes at room temperature. However, this experimental extender demonstrated lower efficiency than the commercial extender in maintaining the sperm quality at room temperature across all periods tested.

Resumo em Inglês:

Abstract PICK1 plays a crucial role in mammalian spermatogenesis. Here, we integrated single-molecule long-read and short-read sequencing to comprehensively examine PICK1 expression patterns in adult Baoshan pig (BS) testes. We identified the most important transcript ENSSSCT00000000120 of PICK1, obtaining its full-length coding sequence (CDS) spanning 1254 bp. Gene structure analysis located PICK1 on pig chromosome 5 with 14 exons. Protein structure analysis reflected that PICK1 consisted of 417 amino acids containing two conserved domains, PDZ and BAR_PICK1. Phylogenetic analysis underscored the evolutionary conservation and homology of PICK1 across different mammalian species. Evaluation of protein interaction network, KEGG, and GO pathways implied that interacted with 50 proteins, predominantly involved in glutamatergic synapses, amphetamine addiction, neuroactive ligand-receptor interactions, dopaminergic synapses, and synaptic vesicle recycling, and PICK1 exhibited significant correlation with DLG4 and TBC1D20. Functional annotation identified that PICK1 was involved in 9 GOs, including seven cellular components and two molecular functions. ceRNA network analysis suggested BS PICK1 was regulated by seven miRNA targets. Moreover, qPCR expression analysis across 15 tissues highlighted that PICK1 was highly expressed in the bulbourethral gland and testis. Subcellular localization analysis in ST (Swine Tesits) cells demonstrated that PICK1 significantly localized within the cytoplasm. Overall, our findings shed new light on PICK1’s role in BS reproduction, providing a foundation for further functional studies of PICK1.

Resumo em Inglês:

Abstract The study aimed to evaluate the effectiveness of combining two injectable progesterone (iP4) formulas for estrous synchronization in ewes and to compare it with traditional intravaginal progesterone devices. Additionally, the study assessed whether the inclusion of GnRH enhances the reproductive outcomes of the iP4 treatment. Two experiments were conducted. In the first experiment, 20 Santa Inês ewes were divided into two groups: one group received intravaginal progesterone devices, and the other received combined long-acting and short-acting iP4. In the second experiment, 30 Corriedale ewes were divided into two groups: one received the combined iP4 with GnRH, and the other without GnRH. Estrous, ovulation, follicular populations, and progesterone concentrations were monitored. The combined iP4 treatment induced an artificial luteal phase and produced reproductive responses similar to those obtained with intravaginal devices. In the first experiment, the iP4 treatment tended to result in more synchronized ovulation compared to the control (P=0.095). In the second experiment, adding GnRH enhanced the quality of the corpus luteum, as indicated by increased diameter and vascularization on Day 23 (P=0.047 and P=0.02, respectively). The combined administration of long-acting and short-acting iP4 effectively synchronized estrous in ewes and showed similar efficacy to traditional intravaginal devices. The inclusion of GnRH improved luteal quality, suggesting potential benefits for reproductive management in ewes. Further studies are needed to evaluate the fertility outcomes of these protocols under field conditions.

Resumo em Inglês:

Abstract Anticancer therapy often leads to premature ovarian insufficiency (POI) and infertility due to the extreme sensitivity of the ovarian follicle reserve to the effects of chemotherapy. Withanolides are known for their cytotoxic effect on cancer cells and low cytotoxicity on non-malignant or healthy cells. Therefore, this study aimed to investigate the in vivo effects of three withanolides derivatives: 27-dehydroxy-24,25-epoxywithaferin A (WT1), 27-dehydroxywithaferin A (WT2), and withaferin A (WTA) on fertility, and the ovarian preantral follicles of young female mice. To achieve this, mice received 7 intraperitoneal doses of WT1, WT2, or WTA at a concentration of 2 mg/kg (Experiment I) and 5 or 10 mg/kg (Experiment II) over 15 alternate days. In experiment I, two days after administration of the last dose, half of the mice were mated to evaluate the effects of withanolides on fertility. The other half of the mice, as well as all mice from experiment II, were sacrificed for histological, inflammation, senescence, and immunohistochemical analyses of the follicles present in the ovary. Regardless of the administered withanolide, the concentration of 2 mg/kg did not show toxicity on the follicular morphology, ovarian function, or fertility of the mice. However, at concentrations of 5 and 10 mg/kg, the three derivatives (WT1, WT2, and WTA) increased follicular activation, cell proliferation, and ovarian senescence without affecting inflammatory cells. Furthermore, at a concentration of 10 mg/kg, the three withanolides showed intensified toxic effects, leading to DNA damage as evidenced by the labeling of γH2AX, activated Caspase 3, and TUNEL. We conclude that the cytotoxic effect of the tested withanolide derivatives (WT1, WT2, and WTA) in the concentration of 2 mg/kg did not show toxicity on the ovary. However, in higher concentrations, such as 10 mg/kg, toxic effects are potentiated, causing DNA damage.

Resumo em Inglês:

Abstract The cooling of Litopenaeus vannamei shrimp spermatophores for assisted insemination can enable the transfer of gametes between reproduction laboratories. This study aimed to assess three extenders for cooling L. vannamei spermatophores for assisted insemination. Spermatophores were chilled at 15 °C for 24 or 48 hours using powdered coconut water ACP® (PCW), mineral oil (MO), and sterilized seawater (SSW) as extenders. All treatments demonstrated consistent responses over time. Apparent viability and morphological integrity percentages remained above 60% and 70%, respectively, across treatments and storage durations. Focusing on diluents, normal cell percentages for MO, SSW, and PCW treatments were 74.9±9.20%, 77.3±9.40%, and 78.1±6.35%, respectively, irrespective of storage time. The highest hatching rate was observed in the SSW treatment (80.67±12.01%), which was significantly superior to the PCW treatment (50.15±20.75%). The hatching rates observed in the MO treatment (71.47±18.83%) did not statistically differ from either PCW or SSW treatments. The cooling protocol successfully preserved the spermatophores' ability to maintain favorable levels of apparent viability, normal morphology, and hatching rates after 48 hours of storage at 15 °C using mineral oil, seawater, or ACP® as extenders. Sterilized seawater emerged as the most efficient diluent, delivering superior hatching rates following artificial insemination.

Resumo em Inglês:

Abstract Polyethylenimine (PEI) has been explored as an efficient non-viral system for delivering genes to cells; however, there were no protocols for its use in porcine fetal fibroblasts (PFF). Therefore, we compared different concentrations of FITC-PEI (0.625, 1.25, 2.5, 5, 10, 20, 40, or 80 µg/mL) and incubation times (30 min, 1 h, or 2 h). It was observed that the incubation time did not affect the internalization of the PEI-FITC and that 30 min was sufficient to capture the complex. The concentrations higher than 10 µg/mL could reach many marked PFF (>90%). Then, two PEI concentrations were tested, 10 or 40 µg/mL, combined with an N/P of 2 with the pmhyGENIE-5 for 30 min. The percentage of PFF-GFP positive was similar between the PEI concentrations in the evaluation time points (24 h, 48 h, and 72 h). However, 40 µg/mL caused higher membrane damage rates. Thus, it can be concluded that concentrations between 10 – 80 µg/ml of PEI promote high incorporation rates, even in periods as short as 30 minutes. Furthermore, it can be stated that the transfection condition used in Polyplexes 1 (10 µg/mL of PEI and 37.5 µg/mL of pmhyGENIE-5 for 30 min) efficiently produces genetically edited porcine fetal fibroblasts with low cell damage.

Resumo em Inglês:

Abstract This study was conducted to evaluate manila duck’s (Cairina moschata) frozen semen quality after cryopreservation in lactated ringer's egg yolk-astaxanthin (LREY-A) with 5 different concentrations of dimethyl sulfoxide (DMSO). Methodology: Semen was collected from 3 manila ducks (Cairina moschata) using the cloaca massage technique twice a week. Fresh semen was evaluated macro and microscopically then polled and divided into 5 tubes of treatments. Each tube was diluted in DMSO4, DMSO6, DMSO8, DMSO10, and DMSO12. The semen of each treatment was loaded into a 0.25 mL straw and equilibrated at 5 °C for 2 h. Freeze above nitrogen vapor and stored a container of liquid nitrogen at -196 °C, then semen thawed in a water bath at 37 °C for 30 sec. Data were analyzed using One-Way ANOVA Analysis. Results of this showed that post-equilibration sperm motility and sperm viability have differed significantly (P<0.05) for each treatment, with the highest % sperm motility DMSO8 and DMSO6, this is also shown in post-thawing sperm motility and viability which have differed significantly (P<0.05) and the highest % sperm viability were DMSO8 and DMSO6. In conclusion, Frozen semen extender formulation of DMSO8 and DMSO6 which are used in manila duck semen cryopreservation was the best to other treatments to maintain % sperm motility and % sperm viability in post-equilibration and post-thawing. The highest sperm motility recovery rate was in DMSO8. The lowest sperm live and dead abnormality was in DMSO8. It is concluded that the combination of DMSO8 was the best in maintaining the quality of manila duck frozen semen.

Resumo em Inglês:

Abstract Deslorelin is a GnRH agonist used in veterinary medicine to temporarily inhibit reproduction in domestic animals and is sometimes tested in captive species in zoo to control population or tame aggressive behaviours in males. However, some studies have revealed the inefficacy of deslorelin specifically in males, contrary to females that follow a classic long-term inhibition of the reproductive hypothalamic-pituitary axis through sexual steroid negative feedback. We implanted 5 males and 6 females grey mouse lemurs (Microcebus murinus), long-day breeders that display a complete inhibition of the reproductive system during winter, at the end of the short-day period, a few weeks before the breeding season. Contrary to females, which exhibited a classic inhibitory response to deslorelin, males’ testosterone levels increased as well as their testis size, which suggests a sex-specific sensitivity to the negative feedback of sexual steroids before the mating period. We propose that this sex-imbalance is related to the different life-history of males as opposed to females concerning reproductive tasks and behaviour.

Resumo em Inglês:

Abstract The relationships between members of the groups include behaviors related to affiliation, dispute for dominant positions, parental care, and facing disputes for food and territory. All these activities are under hormone modulation and those of a steroidal nature are heavily involved. Despite this, only few data are available on steroid hormones in free-ranging marmosets of the Callithrix genus, which limits the understanding of the physiological functioning and modulation of the socio-sexual behavior by steroid hormones of this taxon. In this study, we characterized fecal concentrations of progesterone, estrogens, and glucocorticoids of six breeding and non-breeding females from two groups of free-ranging Callithrix penicillata (É. Geoffroy, 1812). The concentration of progesterone was significantly higher in females which gave birth, compared to non-breeding females. The levels of fecal estrogens and glucocorticoids did not differ between breeding and non-breeding females. The data are in agreement with the few studies on steroid values of wild and captive marmosets. This study shows the concentrations of progesterone and glucocorticoids in free-ranging C. penicillata for the first time, and it is the only study reporting the concentration of fecal estrogens in wild marmosets. Overall, the high levels of progesterone associated with pregnancy in free-ranging C. penicillata as well as levels of estrogens and glucocorticoids close to those reported for other species, suggest a conserved pattern of hormonal secretion between Callithrix species that have been studied in captivity.

Resumo em Inglês:

Abstract Due to high annual culling rates, pig farms require a constant income of replacement gilts. Gilts typically reach puberty at nearly six months of age. Puberty may be induced through early boar exposure, therapy with steroid hormones and chorionic gonadotropins, and optimized by identifying biological predictors and risk factors. Old age at the time of the first mating is associated with an increased risk of premature culling, often attributed to reproductive failures and locomotor problems. While female prolifacy has increased substantially during the last few decades, selecting for litter size to optimize lifetime productivity would be more efficient after two parities. Additionally, uterine capacity and the number of functional teats should be considered in selecting future dams. For each female, the cost-effective number of parities at removal is determined by the cumulative number of pigs born and weaned during the total herd days.

Resumo em Inglês:

Abstract Intracytoplasmic Sperm Injection (ICSI) has increased usage in cases of stallion fertility issues, particularly for older stallions, those with reduced sperm numbers or quality, or stallions that have passed away, and only a limited amount of frozen semen is available. By manipulating the frozen semen through thawing, diluting, and refreezing or by cutting the straw under liquid nitrogen, the supply of semen for ICSI can be extended. While ICSI requires a minimal number of spermatozoa per procedure, it is important to consider sperm quality as a crucial factor affecting fertilization and embryo development. Although it is possible to produce healthy embryos and offspring from low quality sperm samples, it is preferable to process and select morphologically and functionally superior sperm to maximize the chances of successful fertilization and embryo development. Several techniques are available for selecting the spermatozoa for ICSI, such as swim-up, washing, density gradient centrifugation, microfluidic sorting, and some combinations. In this review, we will focus on semen type, handling, recent breakthroughs, stallion effects on ICSI efficiency and the prospects of this technology within the equine industry.

Resumo em Inglês:

Abstract Oviduct fluid extracellular vesicles (oEV) are essential for periconceptional events. The presence of EV has already been identified in the oviduct fluid (OF) from mammalian species, except in caprine. Therefore, this study aimed to isolate and characterize the caprine oEV (coEV). Initially, in Experiment 1, coEV were isolated from the OF of either each animal individually or from a pool of three animals. In experiment 2, coEV were isolated during the follicular or luteal phases of the estrous cycle. The coEV were characterized by size distribution, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS) analysis, as well as, by transmission electron microscopy (TEM) and dot blotting (DB). Our results indicated that the physicochemical characteristics of the coEV were similar (P > 0.05), regardless of the isolation method (individual or pool). However, coEV collected during the luteal phase were larger (P < 0.05) than those during the follicular phase. The TEM showed spherical and cup-shaped particles, characteristic of exosomes. The DB revealed the presence of exosomal proteins involved in the biogenesis of coEV. In conclusion, it is possible to isolate and characterize coEV from a single caprine female and the estrous cycle phase influences the vesicles average size and PDI.