Maricy Apparicio
@fmvz.unesp.br
Department of Veterinary Surgery and Animal Reproduction
UNESP
RESEARCH, TEACHING, or OTHER INTERESTS
Veterinary, Animal Science and Zoology, Obstetrics and Gynecology
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Marina Vilela Estevam, Gilson Helio Toniollo, and Maricy Apparicio
Elsevier BV
Renatha A. Araújo, Nathali A. A. Sales, Roberta C. Basile, Walter H. Feringer-Junior, Maricy Apparício, Guilherme C. Ferraz, and Antonio Queiroz-Neto
MDPI AG
Firocoxib is a non-steroidal anti-inflammatory drug specifically formulated for veterinary medicine and selectively acts on inhibiting the cyclooxygenase 2 enzyme (COX-2). This study evaluated the possible adverse effects of administering oral therapeutic firocoxib on gastric mucosa, hematological parameters, coagulation cascade, and hepatic and renal biochemistry in healthy horses. Nine clinically healthy Arabian horses, approximately 9 years old, received 0.1 mg/kg of oral firocoxib for 14 days. The gastroscopic examination was conducted 1 day before starting treatment (D0) and two days after the last blood collection (D23). Venous blood samples were obtained for laboratory tests on day 1, immediately prior to the initiation of treatment (D1), after 7 and 14 days of treatment (D7 and D14), and 7 days after the conclusion of treatment (D21. No changes were found in the gastroscopic and hematological tests. Coagulation and serum biochemistry levels remain between these species’ average values. However, the increased activated partial thromboplastin time (aPTT) and prothrombin time (PT) indicate reduced blood coagulation capacity, which contradicts the expected effect of treatment with selective COX-2 inhibitors, as these drugs theoretically promote coagulation. Administering firocoxib to horses is safe as it does not cause significant adverse reactions. Therefore, it is a suitable option for managing inflammatory conditions in these animals with attention to an unexpected adverse anti-coagulopathy effect, and further study is warranted.
Samara Beretta, Maricy Apparicio, Gilson Hélio Toniollo, and Marita Vedovelli Cardozo
FapUNIFESP (SciELO)
Abstract The neonatal period represents a critical stage for the establishment and development of the gut microbiota, which profoundly influences the future health trajectory of individuals. This review examines the importance of intestinal microbiota in humans and dogs, aiming to elucidate the distinct characteristics and variations in the composition between these two species. In humans, the intestinal microbiota contributes to several crucial physiological processes, including digestion, nutrient absorption, immune system development, and modulation of host metabolism. Dysbiosis, an imbalance or disruption of the gut microbial community, has been linked to various disorders, such as inflammatory bowel disease, obesity, and even neurological conditions. Furthermore, recent research has unveiled the profound influence of the gut-brain axis, emphasizing the bidirectional communication between the gut microbiota and the central nervous system, impacting cognitive function and mental health. Similarly, alterations in the canine intestinal microbiota have been associated with gastrointestinal disorders, including chronic enteropathy, such as inflammatory bowel disease, food allergies, and ulcerative histiocytic colitis. However, our understanding of the intricacies and functional significance of the intestinal microbiota in dogs remains limited. Understanding the complex dynamics of the intestinal microbiota in both humans and dogs is crucial for devising effective strategies to promote health and manage disease. Moreover, exploring the similarities and differences in the gut microbial composition between these two species can facilitate translational research, potentially leading to innovative therapeutic interventions and strategies to enhance the well-being of both humans and dogs.
Aracelle Elisane Alves, Tathiana Ferguson Motheo, Maricy Ferreira Apparicio, Giuliano Queiroz Mostachio, Ricarda Maria dos Santos, Wilter Ricardo Russiano Vicente, and Gaia Cecilia Luvoni
FapUNIFESP (SciELO)
Abstract Logistic and economical limitations are often the causes of dog owners not accurately monitoring the estrous cycle and the optimal insemination time. The aim of this study was to evaluate in vivo early embryonic development in bitches, after the analysis of sequential vaginal cytologies associated to single progesterone measurement and single laparoscopic insemination with high quality semen (fresh and with high spermatozoa concentration) or low-quality semen (frozen/thawed and with low spermatozoa concentration) at 48 h post- ovulation time predicted on a single progesterone measurement. Ten bitches were inseminated with 250 x 106 fresh spermatozoa (80% motility), and ten with 80 x 106 frozen/thawed spermatozoa (60% motility) in the cranial part of each uterine horn. Seven days later, ovariohysterectomy was performed and the oviducts and uterine horns and body were flushed to recover embryos and unfertilized oocytes. In 80% of the bitches inseminated with fresh and 50% of bitches inseminated with frozen/thawed semen, embryos at 2 to 8 cells stage were recovered mostly from the, oviducts. This study indicates that pregnancies can be obtained with a single laparoscopic intrauterine insemination after single serum progesterone measurement, although with a low number of embryos. This result should be taken into account in case economic or logistic restrictions that affect the possibility of owners to plan an accurate monitoring of the optimal breeding time using fresh and frozen semen.
Marina Vilela Estevam, Samara Beretta, Nathalia F. Smargiassi, Maricy Apparício, Gilson Helio Toniollo, and Gener T. Pereira
Frontiers Media SA
The popularity of brachycephalic dogs has increased in recent years due to their docile temperament and peculiar features. The historical inbreeding and consequent lack of genetic diversity involved in the development of these breeds led to an increase in the manifestation of deleterious genes that may lead to malformations. In addition, there are serious health issues intrinsic to the conformation, mainly attributed to these extreme characteristics. Therefore, this retrospective study aimed to observe the frequency of malformations in brachycephalic dogs compared to the pure and mixed breeds (MB). The medical records of pregnant bitches admitted at the Service of Obstetrics and Animal Reproduction (SORA) from January 2017 to December 2021 were retrieved from the hospital's computer system and analyzed one by one. Seven hundred sixty-eight neonates born from 168 litters were included in this study. Of these litters, 72.6% (122/168) were brachycephalic. Malformations were found in 52 puppies, with an incidence of 6.77% (52/768). Of the 32 litters that produced malformed puppies, 28 were brachycephalic (87.5%). In total, 23 types of malformations were registered, the most common being cleft palate (1.30%) and anasarca (1.17%). Ten of the puppies (10/52; 19.23%) presented two or more associated malformations. Bitches above 7 years were more prone to present malformed puppies in their litters. Brachycephalic breeds were 3.03 times more likely to present malformed neonates when compared to other breeds; the odds ratio increased to 5.07 when modern brachycephalic was compared to ancestral brachycephalic. Regarding the mode of delivery, elective cesarean sections accounted for 66.6% of births while 19.64% were eutocic vaginal deliveries, and 13.69% were dystocic. The presence of malformed puppies in a litter causes suffering for the owner, the bitch and for the puppy itself, therefore, the veterinarian plays a key role in this scenario. Knowledge about congenital abnormalities, their causes, diagnosis, and approach is essential to reduce the incidence of malformations and improve the quality of life of these animals.
Beatrice Ingrid Macente, Carlos Eduardo Fonseca-Alves, Georgia Modé Magalhães, Mariana Riboli Tavares, Cleber Fernando Menegasso Mansano, Lara Mouttham, Maricy Apparício, Gilson Hélio Toniollo, and Pierre Comizzoli
Mary Ann Liebert Inc
The objective of the study was to evaluate the integrity of cat testicular tissues after vitrification with different devices followed by different warming conditions. The influence of vitro culture for 24 hours after warming also was examined. Testicular tissues from adult domestic cats were dissected in small fragments that were vitrified using Cryotop® or threaded on fine needles, warmed (directly at 37°C or with a preliminary 10 seconds exposure to 50°C), and/or cultured in vitro for an additional 24 hours. For each treatment group, tissues were assessed based on histology, apoptosis, and sperm DNA integrity. Results showed that fragments of testicular tissues were efficiently cryopreserved (maintaining the quality of all cell types) with vitrification with Cryotop followed by direct warming at 37°C, and additional culture of 24 hours at 38.5°C. These encouraging results are paving the road to optimize preservation protocols and use them for systematic banking of tissues from genetically valuable felids.
Martina Colombo, Maria Giorgia Morselli, Maricy Apparicio, and Gaia Cecilia Luvoni
Wiley
AbstractFollicle‐like structures are three‐dimensional matrices joint with living cells that allow the in vitro development of female gametes in more physiological conditions. They have been shown to be beneficial to fresh oocytes in different species, and in this study, domestic cat (Felis silvestris catus) granulosa cells were used to create a functional follicle‐like structure aimed at supporting the in vitro maturation of conspecific vitrified oocytes, key players of fertility preservation programmes that usually struggle to acquire their full developmental competence after warming. Cat granulosa cells were cultured for up to 6 days in three‐dimensional barium alginate microcapsules (i.e. follicle‐like structures) or in two‐dimensional monolayers, and their steroidogenic ability (estradiol and progesterone secretion) was assessed to confirm their functionality. The same systems were used (on day 2 or 6 of granulosa cells culture) for the in vitro maturation (IVM) of Cryotop® vitrified immature cat oocytes and compared with microdrops of IVM medium without cells (control). Granulosa cells were able to maintain their functionality in vitro in both the conditions, even if with a different extent of hormonal secretion along culture (p = .02). Vitrified oocytes resumed meiosis at higher rates when cultured with 2 days old granulosa cells (p = .03), but full maturation rates slightly raised when granulosa cells were cultured longer, albeit without differences with the control group. This study paved the road for the creation of enriched culture systems in the domestic cat, but innovations are strongly needed for vitrified oocytes that deserve better chances to develop in vitro.
B.I. Macente, M. Apparicio, C.F.M. Mansano, M.R. Tavares, C.E. Fonseca-Alves, B.P. Sousa, P.H.L. Bertolo, R.O. Vasconcelos, E.S. Teixeira, and G.H. Toniollo
Elsevier BV
Martina Colombo, Maria Giorgia Morselli, Mariana Riboli Tavares, Maricy Apparicio, and Gaia Cecilia Luvoni
MDPI AG
Cryoinjuries severely affect the competence of vitrified oocytes (VOs) to develop into embryos after warming. The use of culture conditions that provide physical and chemical support and resemble the in vivo microenvironment in which oocytes develop, such as 3D scaffolds and coculture systems, might be useful to improve VOs outcomes. In this study, an enriched culture system of 3D barium alginate microcapsules was employed for the in vitro embryo production of domestic cat VOs. Cryotop vitrified-warmed oocytes were in vitro matured for 24 h in the 3D system with or without fresh cumulus-oocyte complexes (COCs) in coculture, whereas a control group of VOs was cultured in traditional 2D microdrops of medium. After in vitro fertilization, presumptive embryos were cultured in 3D or 2D systems according to the maturation conditions. Vitrified oocytes were able to mature and develop into embryos in 3D microcapsules (17.42 ± 11.83%) as well as in 2D microdrops (14.96 ± 8.80%), but the coculture with companion COCs in 3D resulted in similar proportions of VOs embryo development (18.39 ± 16.67%; p = 1.00), although COCs presence allowed for blastocyst formation (0.95 ± 2.52%). In conclusion, embryos until late developmental stages were obtained from cat VOs, and 3D microcapsules were comparable to 2D microdrops, but improvements in post-warming conditions are still needed.
Caio de Faria Tiosso, Fabiana Azevedo Voorwald, Maurício Veloso Brun, Luciana Cristina Padilha Nakaghi, Roberta Martins Crivelaro, Maricy Apparicio, and Wilter Ricardo Russiano Vicente
FapUNIFESP (SciELO)
ABSTRACT: SILSTM via an umbilical hernia opening was used in the correction of an abdominal cryptorchidism in a dog. The SILSTMport was inserted through a 2.5 cm skin incision on the umbilical hernia and pneumoperitoneum was established with CO2. A 10mm telescope, 5mm laparoscopic Babcock clamp, and a clamp with combined cutting and coagulation features were used for the dissection and removal of the testicle through the SILSTM port. The treatment proved to be effective, easy, and quick to perform. However, the lack of triangulation between clamps and telescope limited the movement of the instruments, making it more laborious than a multiple-trocar approach. It can be concluded that the use of a SILSTM port through a pre-existing opening of the abdominal wall enables a reduction of the risks associated with multiple incisions; such as injury and manipulation of internal organs, pain, bleeding, and development of new hernias.
Camila M. Gorricho, Mariana R. Tavares, Maricy Apparício, Carlos E. Fonseca‐Alves, Beatrice I. Macente, Cleber F.M. Mansano, and Gilson H. Toniollo
Wiley
ContentsThe aim of the present study was to investigate whether or not the size of the ovarian fragment influences its resistance to cryostorage. For that purpose, ovaries were collected from 34 queens (various breeds, age 1–5 year) by routine ovariectomy, transported to the laboratory and then sectioned in different sizes (3 mm × 3 mm × 3 mm, 5 mm × 3 mm × 3 mm and 7 mm × 3 mm × 3 mm) and randomly assigned to a control (GC3, GC5 and GC7, respectively) or vitrified (GV3, GV5 and GV7, respectively) groups. Vitrified‐warmed fragments were evaluated by histomorphology and immunohistochemistry (for apoptotic rates by using cleaved caspase‐3). Histological examination reveals that 72.97% of the follicles in GV3 and 72.58% in GV5 were normal while only 42.86% of the follicles in GV7. The main morphological alteration presented in all groups was a detachment of the epithelial cells. Similarly, immunohistochemistry evaluation using caspase 3 revealed a small proportion of apoptotic cells in GV3 (8.43%) while in GV7 30.43% of the cells expressed cleaved caspase‐3. These findings indicate that fragments sectioned in 3 mm × 3 mm × 3 mm (27 mm3) seem more adequate for perfusion of the cryoprotectant, causing less damage to the cell after vitrification‐warming.
Beatrice Ingrid Macente, Raquel Ribeiro Gutierrez, Maricy Apparício, Cristiano de Carvalho Balieiro, Cleber Fernando Menegasso Mansano, Marcelo Maia Pereira, Juliana Corrêa Borges-Silva, Eliandra Antonia Pires-Buttler, André Luis Batista Galvão, Gilson Hélio Toniollo,et al.
Colegio Brasileiro de Reproducao Animal - CBRA
The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) – epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 – the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the α-tocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.
TF Motheo, DR Arnold, LC Padilha‐Nakaghi, EA Pires‐Buttler, AE Alves, M Apparicio, WRR Vicente, and FL Lopes
Wiley
ContentsPost‐translational modifications of histones, such as acetylation, are involved in regulating chromatin remodelling and gene expression. Proper in vitro maturation (IVM) of canine oocytes, for many reasons, is up to now inefficient. This study aimed to evaluate the post‐translational histone H4 acetylation at lysine 5 (H4K5) in immature and post‐IVM canine oocytes. Oocyte nuclear stage was assessed using Hoechst 33342 staining. Acetylation patterns were determined by indirect immunofluorescence staining of immature and post‐IVM oocytes, using an antibody against the acetylated lysine 5 residue on histone 4 (H4K5ac). The experiment was repeated four times, with a total of 7–17 oocytes evaluated per stage. Immunofluorescence signal was quantified using the NIHimagej software. Data were expressed as a percentage of the average fluorescence intensity of the specific antibody over the intensity of DNA, as determined by Hoescht staining. H4K5ac displayed a significantly higher acetylated pattern in immature oocytes (0.97 ± 0.08) when compared to post‐IVM oocytes at different nuclear stages. There was a decrease in the fluorescence level of the matured oocytes with the progression of meiosis (GVBD: 0.47 ± 0.06 and MI/MII: 0.35 ± 0.04). Similarly to other domestic species, we hypothesized that post‐translational modification of histone acetylation takes place during meiosis of in vitro matured canine oocytes. However, it remains to be investigated whether these changes occur during in vivo maturation.
BI Macente, GH Toniollo, M Apparicio, CFM Mansano, HE Thomé, CL Canella, MEG Tozato, and RR Gutierrez
Wiley
ContentsThis study aimed to evaluate tissue damage of feline testicles sectioned in two different sizes (0.3 or 0.5 cm3) and submitted to different cryoprotectants (propanediol or glycerol). Testicles obtained from 12 domestic cats were sectioned in 0.3 and 0.5 cm3 sized pieces and immediately evaluated by TBARS and semi‐quantitatively by histomorphology. The remaining fragments were placed in cryotubes with 1 ml Egg yolk Tris Equex STM extender containing 3% glycerol or 3% propanediol and cryopreserved by fast‐freezing technique. Frozen‐thawed fragments were also evaluated by TBARS and histomorphology. Statistical analysis was performed using one‐way ANOVA with Student–Newmann–Keuls post hoc test, with p < .05. Fresh and cryopreserved tissues generally exhibited a similar morphology concerning detachment of cells from the basement membrane and observation of nucleoli, with a great proportion scored as 0 (no alteration). When present, alterations were slight and the morphology was considered to be good (most classified in scores 1). Pyknosis was the main anomaly observed as score 2 in 54.6% and 58.4% of 0.3‐cm3 fragments cryopreserved in propanediol and glycerol, respectively (16.7% scored 2 in fresh tissue). In TBARS evaluation, 0.5‐cm3 fragments cryopreserved in glycerol produced less free radical compared to the 0.3 cm3 cryopreserved in glycerol or propanediol. Our results showed that glycerol was more efficient than propanediol to cryopreserve 0.5‐cm3 fragments; this might be attributed to the fact that glycerol molecular weight is larger than propanediol and so its perfusion in the testicular tissue is slower.
AE Alves, LC Padilha‐Nakaghi, EA Pires‐Butler, M Apparicio, NAM Silva, TF Motheo, WRR Vicente, and GC Luvoni
Wiley
ContentsIn vitro culture of ovarian preantral follicles has emerged as a reproductive technology aimed at obtaining large amount of oocytes for in vitro embryo production. The addition of growth factors (GF) in the in vitro culture of preantral follicles of different species has provided superior results of follicular development, antrum formation and proliferation of granulosa cells. However, there are only few reports regarding the use of these factors on feline preantral follicle in vitro culture. Thus, the aim of this study was to investigate the effect of a combination of IGF‐1 and EGF on in vitro viability and growth of preantral follicles and enclosed oocytes collected from domestic cats. A total of 64 follicles characterized by multilayer granulosa cells were isolated and individually cultured for 6 days (T6) in minimum essential medium supplemented with IGF‐1+ EGF (100 ng/ml each) or without (control). A higher percentage of follicles were viable after culture with GF than without, and an increase in size when IGF‐1+ EGF were added to the medium (170 ± 32.4 μm (T0) vs. 201 ± 22.3 μm (T6); p < .05) was observed. An increase in the diameter was also observed in follicles cultured without GF, but this increase was only 8.3% compared to 15.4% of those cultured with GF (p < .05). No differences were found in the diameter of oocytes contained in follicles cultured in the non‐supplemented or supplemented media (107.9 ± 11.8 μm (T0) vs. 113.2 ± 15.6 μm (T6); p > .05). These data suggest that the addition of IGF‐1 and EGF to the culture medium promotes the in vitro development of preantral follicles of cats.
M Apparicio, VG Santos, DFO Rocha, CR Ferreira, BI Macente, GM Magalhães, AE Alves, TF Motheo, LC Padilha‐Nakaghi, EA Pires‐Buttler,et al.
Wiley
ContentsWith the purpose of identifying factors involved in early stages of embryo development in the domestic cat, matrix‐assisted laser desorption/ionization imaging mass spectrometry (MALDI‐IMS) was used for the first time to describe the spatial localization of proteins in the oviducts of queens. Oviducts were obtained from two 2 and 4 years old cross‐bred queens, divided into three segments, snap‐frozen in liquid nitrogen and then stored at −80°C until use. Next, they were sectioned in a cryostat, fixed on ITO (indium tin oxide) conductive glass slides for MALDI‐IMS and serial sections were collected on microscope slides for histology. As confirmed by histology, MALDI‐IMS was able to show contrasting protein distributions in the oviductal infundibulum, ampulla and isthmus. Mass spectra were characterized by abundant ions of m/z 1,259, 4,939, 4,960 and 10,626, which have been tentatively attributed to keratin, thymosin β10, thymosin β4 and S100, respectively. Keratin and thymosins are involved in the biological response to tissue damage. S100 proteins are calcium‐modulated proteins implicated in a variety of cellular activities, including cell differentiation and regulation of cell motility. These results suggest that protein composition differs between segments of the cat oviduct, which corresponds to morphological changes within these sections. Further functional studies could elucidate the effects of these proteins on feline reproductive physiology.
Giuliano Queiroz Mostachio, Maricy Apparício, Tathiana Ferguson Motheo, Aracélle Elisane Alves, and Wilter Ricardo Russiano Vicente
FapUNIFESP (SciELO)
<p>A fisiopatologia da hiperplasia prostática benigna (HPB) não está totalmente compreendida, no entanto, a diidrotestosterona é o principal hormônio envolvido. Recentemente, o efeito da toxina botulínica A (TB-A) foi investigado, mostrando que esta induz atrofia do parênquima e redução do volume prostático. Com base nisso, este estudo teve como objetivos comparar os efeitos da administração da TB-A com a orquiectomia no tratamento da HPB, além de avaliar os efeitos da TB-A sobre a libido e qualidade do sêmen. Para tanto, 16 cães adultos foram submetidos à castração ou administração de 500U de TB-A, e avaliados durante 16 semanas. A orquiectomia mostrou-se um excelente tratamento, promovendo redução de 80% do volume prostático. Aplicação da TB-A não ocasionou alterações na libido, ereção ou qualidade e características seminais. Por outro lado, reduziu significamente o volume da próstata. Os resultados sugerem que a administração intraprostática de TB-A é um tratamento alternativo efetivo e sem efeitos colaterais para cães destinados a programas de reprodução. No entanto, esta terapia apresenta reduções inferiores do volume prostático, quando comparada com a orquiectomia.</p>
Maricy Apparicio, Giuliano Q. Mostachio, Tathiana F. Motheo, Aracelle E. Alves, Luciana Padilha, Eliandra A. Pires-Butler, Paula A. P. Savi, Ricardo A. R. Uscategui, Gaia C. Luvoni, and Wilter R. R. Vicente
CSIRO Publishing
The aim of this study was to evaluate the influence of different bi-phasic systems with gonadotrophins and steroids on in vitro maturation rates of oocytes obtained from bitches at different reproductive stages (follicular, luteal, anoestrous). In System A (control) oocytes were matured for 72 h in base medium (BM) with 10 IU mL–1 human chorionic gonadotrophin (hCG), 1 μg mL–1 progesterone (P4) and 1 μg mL–1 oestradiol (E2); in bi-phasic System B oocytes were matured for 48 h in BM with hCG and for 24 h in BM with P4; in bi-phasic System C oocytes were matured for 48 h in BM with hCG, P4 and E2, and for 24 h in BM with P4; in System D, oocytes were cultured in BM without hormonal supplementation. Data were analysed by ANOVA. There was a positive effect of the bi-phasic systems on germinal vesicle breakdown, metaphase I and metaphase II rates, irrespective of reproductive status (P < 0.05). Bi-phasic systems were also beneficial for cortical granule distribution (an indication of cytoplasmic maturation) and its relationship to nuclear status: 74.5% of the oocytes cultured in System B and 85.4% of those cultured in System C presented both nuclear and cytoplasmic maturation (P < 0.001). The stage of the oestrous cycle did not influence maturation rates.
LC Padilha, PPM Teixeira, EA Pires‐Buttler, M Apparício, TF Motheo, PAP Savi, EYO Nakaghi, AE Alves, and WRR Vicente
Wiley
ContentsThe success of embryo production in vitro depends upon the use of an efficient oocyte retrieval technique, and the best results have been obtained by laparoscopic aspiration. The aim of this study was to evaluate the effect of consecutive sessions of follicular aspiration on the quantity, quality and in vitro maturation competence of oocytes obtained from ewes subjected to hormonal stimulation. Six Santa Ines ewes underwent nine sessions of follicular aspiration by laparoscopy with a 7‐day interval between sessions, totalling 56 aspirations. After 24 h of culture, oocytes were stained and classified according to the stage of nuclear and cytoplasmic maturation. Oocyte retrieval rate was 61.4 ± 2%, resulting in a total of 249 oocytes. No significant variation was observed between sessions (p > 0.05). The average number of oocytes retrieved from each ewe was 6.4 ± 2 per session and 42 ± 4 in total. No significant difference was observed between the frequencies of the different stages of nuclear maturation: 32.72% mature, 40.74% immature and 26.54% degenerated/indeterminate oocytes; however, a significant difference was observed between the frequencies of the different stages of cytoplasmic maturation: 10.7% mature, 73.25% immature and 16.05% degenerated/indeterminate oocytes. No significant difference was observed in nuclear or cytoplasmic maturation between the weeks of procedure. We conclude that after nine consecutive sessions of follicular aspiration, the quantity and quality of retrieved oocytes remained unchanged as well as the levels of nuclear and cytoplasmic maturation obtained, demonstrating the viability of this technique for repetitive follicular aspirations on the same donor.
Tathiana Ferguson Motheo, Aracélle Elisane Alves, Giuliano Queiroz Mostachio, Maricy Apparício, Alexandre Pinto Ribeiro, Fabiana Ferreira de Souza, Maria Denise Lopes, and Wilter Ricardo Russiano Vicente
FapUNIFESP (SciELO)
This study aimed to determine the effects of different concentrations of botulinum toxin type A (BT-A) on semen parameters, and seminal plasma biochemical and protein profiles of dogs with benign prostatic hyperplasia (BPH). Eighteen sexually intact male dogs with BPH were randomly divided in three groups, and received an intraprostatic injection of saline solution (control group - CG), 250UI (GI) or 500UI (GII) of BT-A under transabdominal ultrasound guidance. Semen was collected at baseline, 2, 4 and 8 weeks after treatment. Semen parameters were determined and seminal plasma pH, total protein (TP), total chlorides (TC), calcium (Ca), potassium (K), and sodium (Na) concentrations were assessed. One-dimensional sodium dodecyl sulfatepolyacrilamide gel eletrophoresis (SDS- PAGE) was performed to determine seminal plasma protein profile. Sperm parameters and seminal plasma pH, TP, TC, Ca and K mean values did not change significantly at any time point and among treated groups (P>0.05). The SDS-PAGE analysis of the pooled fractions identified 31 protein bands with molecular weights ranging from 3.9 to 106.2kDA in all treatment groups during the entire evaluation period. Regardless the used dose, intraprostatic BT-A injection do not alter semen parameters and seminal plasma biochemical and protein profiles of dogs with BPH.
M Apparicio, E Ruggeri, and GC Luvoni
Wiley
ContentsThe aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n = 72) using a vitrification kit for bovine embryo or slow frozen (n = 69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48 h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n = 92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p < 0.01). Morphological integrity after culture was similar in vitrified (73.6%) and slow frozen oocytes (76.8%); however, only 37.5% of the morphologically normal oocytes resumed meiosis after vitrification compared to 60.9% of those submitted to slow freezing procedure (p < 0.01). Fresh oocytes showed higher morphological integrity (91.3%) and meiosis resumption rates (82.6%, p < 0.002) than cryopreserved oocytes, irrespective of the procedure used. These results suggest that immature cat oocytes vitrified with a kit for bovine embryos retain their capacity to resume meiosis after warming and culture, albeit at lower rates than slow frozen oocytes. Vitrification and slow freezing methods show similar proportions of oocytes with normal morphology after culture, which demonstrate that thawed and warmed oocytes that resist to cryodamage have the same chances to maintain their integrity after 48 h of culture.
CRF Gadelha, WRR Vicente, APC Ribeiro, M Apparicio, GJ Covizzi, and ACN Campos
SciELO Agencia Nacional de Investigacion y Desarrollo (ANID)
The incidence of prostatic malignancy has increased the use of tissue markers to detect cancer. Tissue specific antigens or differentiation antigens are foun...
EA Alves, L Padilha, PA Savi, MF Apparicio, GQ Mostachio, TF Motheo, EA Pires‐Buttler, WRR Vicente, and GC Luvoni
Wiley
ContentsOptimal conditions for in vitro culture of feline ovarian follicles have not yet been defined. Follicular development is regulated by intraovarian growth factors, as insulin‐like growth factor (IGF‐1), and during the different stages of the oestrous cycle, follicles are exposed to specific hormonal environments. The aim of this study was to investigate the effect of IGF‐1 on in vitro growth and granulosa cell (GC) viability of preantral follicles collected from domestic cats at follicular and luteal phases of the oestrous cycle. Oestrus and ovulation were induced in 12 cats. A total of 39 and 32 follicles collected at the follicular and luteal phases, respectively, were individually cultured in vitro for 6 days in minimum essential medium media supplemented with or without IGF‐1 (100 ng/ml). Follicles collected during the follicular phase and cultured without IGF‐1 displayed a significant increase in size and higher GC viability (46.5 ± 22.1 μm, 66.7%, respectively) than that of follicles collected at the luteal phase and cultured without IGF‐1 (26.7 ± 14.4 μm, 50%, respectively; p < 0.05). In contrast, when IGF‐1 was added to the culture medium, no differences were observed in size or GC viability between follicles collected at the two phases of the cycle. Nonetheless, follicles collected at the luteal phase and cultured with IGF‐1 had a significant increase in their diameter and GC viability (31.9 ± 15.9 μm, 63.6%, respectively) than that cultured without IGF‐1 (26.7 ± 14.4 μm, 50%, respectively; p < 0.05). These data suggest that in vitro growth and GC survival of feline preantral follicles are affected by the oestrous cycle phase, and the IGF‐1 exerts a positive effect on follicles collected at the luteal phase.
Giuliano Queiroz Mostachio, Maricy Apparício, Tathiana Ferguson Motheo, Aracélle Elisane Alves, and Wilter Ricardo Russiano Vicente
Elsevier BV